5^th European Immunology Conference July 21-23, 2016 Berlin, Germany

Biography:

Abstract:

Immunology and metabolism have always been considered as distinct disciplines. However, recent advances in the understanding of immune functions under HIV-1 infection associate these branches with intricate networks. Today, we show that inflammation and kynurenines whose plasma levels are higher in HIV-1-infected subjects when compared to those of uninfected controls reduced the ability of memory CD4 T-cells to respond properly to IL-2. We found that in vitro treatment of memory CD4 T-cells with kynurenine results in oxygen reactive species (ROS) production that was responsible for inhibiting the phosphorylation of STAT5 factor following IL-2 stimulation. Interestingly, we found that memory CD4 T-cells from HIV-1-infected subjects, even those with less than 3 months of presumed infection, displayed reduced ability to protect themselves against Fas-mediated apoptosis in the presence of IL-2 help. The effect of kynurenines and subsequent oxidative stress was abrogated when HIV-1-infected subjects were treated with combined antiretroviral therapy (cART) early during the time course of their infection. Collectively, these data highlight the key role of inflammation and perturbed metabolism during HIV-1 infection, particularly in the survival of memory cells and the nature of HIV-1 reservoir.

Biography:

Mikhailova Valentina Anatolevna has graduated from Saint-Petersburg State University, Russia, Biological Faculty in 2009. She has completed her PhD and defended her thesis in 2015. She is a Research Fellow in the Laboratory of Intercellular Interactions of Department of Immunology. She is the author of more than 45 publications

Abstract:

Preeclampsia is a complication of pregnancy that affects health of woman and fetus. Pathogenesis of preeclampsia is closely connected with inflammation in fetal-maternal interface. The aim of the study was to assess leucocyte phenotype and functions in case of healthy pregnancy and preeclampsia. Ethical Committee of FSBSI (The Research Institute of Obstetrics, Gynecology and Reproductology named after D.O.Ott), Saint-Petersburg, Russia, approved the study. Peripheral blood was obtained from 422 women (healthy non pregnant women, healthy pregnant women (38-39 w.g.) and pregnant women with preeclampsia (38-39 w.g.). Such methods as flow cytometry (BD FacsCanto II, USA), atomic force microscopy (NT-MTD, Russia) and cell cultures (THP-1, Ea.Hy926) were used. We showed that expression of CD11c byNK-cells, CD18 by CD8+T-cells and CD119 by monocytes increased in case of preeclampsia comparing with healthy pregnancy. So does in vitro adhesion of lymphocytes and monocytes to intact and TNFα activated endothelium. CD107a expression by NK cells was lower and TRAIL expression was higher in case of preeclampsia comparing with healthy pregnancy. Plasma of peripheral blood of all studied patients contained cellular microparticles (average size: 330 nm). In case of preeclampsia the amount of microparticles with receptors CD45 and CD16 increased comparing with healthy pregnancy. Microparticles from plasma of women with preeclampsia in vitro influenced expression of CD181, CD18 and CD54 by monocytic cells (THP-1). Observed changes in adhesion receptors expression, adhesion function of T-cells, NK cells and monocytes, TRAIL expression of NK cells are a result of inflammation in decidua, which can be triggered by cellular microparticles present in plasma.

Biography:

Francisco Assis de Andrade has completed his MD from Barcelona University-Hospital Clínic (Systemic autoimmune diseases) and he is carrying out PhD in Medical Sciences from Univerisidade do Estado do Rio de Janeiro, School of Medicine. He is the Chief of Ophthalmology Department at Centro Oftalmológico de Botafogo, Rio de Janeiro, Brazil, Ophthalmologist Speaker of Bausch-Lomb and Janssen-Cilag. He has published 4 papers in reputed journals, chapter of a book and he has also been serving as an Editorial Board Member of repute.

Abstract:

The eye is divided anatomically in three distinguished layers: An outer or fibrous layer (cornea/sclera), middle or vascular layer (uvea, a continuous structure comprising iris, ciliary body and choroid) and an inner or sensorineural layer (retina). Due to its embryonic origin, its peculiar anatomy and the presence of physiological factors that modulate the immune response, the eye presents unique immunological characteristics. The eye is protected from invasive pathogens by two systems, an anatomical barrier and an immunological barrier. Under conditions of intraocular inflammation in experimental animals and humans, activated CD4+ T-cells infiltrate the eye and cause immune responses and inflammation, which damage vision related cells and tissues. However, the eye has a unique immune system to protect important cells and tissues from activated effector CD4+ T-cells. When Streilein and colleagues described ACAID, they provided the first evidence that ocular parenchymal cells participate in the so called ocular immune privilege. To fully understand the complex mechanism of ocular immune system and the immune response in general, the mechanisms by which the threat of autoimmune disease is kept in check, as well as those that play a role in the induction of autoimmunity, must be completely elucidated. It has been demonstrated that there are multiple anatomic and functional layers that are necessary for the preservation of the privileged immune protection of the eye.

Biography:

Andreia Ribeiro did her Biology Degree and Microbiology Master in Aveiro University in Portugal. Since 2010 she has been working as research assistant in flow cytometry and immunology groups, in Portugal and in Ireland. In 2013 she started her PhD in the DECIDE consoritum and completed in 2016 from National University of Ireland, Galway

Abstract:

Extracellular matrix (ECM) plays an important role in the tumour microenvironment and in biologic processes such as hematopoiesis. ECM contributes to regulation of cell survival, proliferation and cell differentiation. The aims of this project were i) to study the quality, quantity and biological role of ECM produced by a cloned mouse mesenchymal stromal cell line (MS5) on cell differentiation ii) to study the role of hypoxia on cell differentiation. To carry out these studies, we have used two methods of producing ECM in vitro. In both methods ECM is produced in normoxia and hypoxia. In method 1, cells are lysed by osmotic shock with a Tris/EDTA buffer, the standard way of preparing ECM in many studies. In method 2, MS5 were transduced with a caspase 9 vector, allowing induction of apoptosis in the cells following ECM production. Balb/c bone marrow mesenchymal stromal cells (MSC) were then seeded either in uncoated plastic dishes or in dishes covered with ECM and differentiation assays were performed, again either in normoxia or hypoxia. Results show that the two methods produce qualitatively different ECM and that hypoxia plays a role in ECM composition. Moreover, compared with hypoxia, normoxia is a better condition for adipogenic differentiation of fresh MSC. In contracts, osteogenic differentiation is better on ECM in hypoxia. In conclusion, different methods of preparing ECM in vitro lead to different protein composition and different outcomes in cell differentiation. Hypoxia also makes a difference in ECM composition and cell differentiation.

Biography:

Abstract:

Various parts of Malva sylvestris have different medicinal properties such as antimicrobial activity. In this study the effects of M. sylvestris extract on protein electrophoresis pattern and gamma interferon of C. albicans infected mice was studied. Sixty female mice were divided randomly in six groups (three treatment groups, Candida, placebo and control groups). Treatment groups received aquatic extract of M. sylvestrs (50, 100 and 200 mg/kg) for 20 days every other day via injection in peritoneum. C. albicans was injected once after sixth injection of extract. Results showed significant decrease in the amount of albumin in three treatment groups. The β-globulin amount of 50 and 100 mg/kg groups and gamma interferon amount of all three treatment groups were increased significantly in proportion to control group. Clearing the body from pathogen organisms depends on cellular responses. It seems that M. sylvestris is capable of stimulating cellular immune response and can be used in studies about C. albicans.

Biography:

Abstract:

Medicinal plants are believed to be an important source of new chemical substances with potential therapeutic effects. The aim of the present study was to evaluate the anti inflammatory effect of oral administration of Oliveria decumbens hyroalcholic extract by Carageenan tests in rats. In this research, 60 male Wistar rats weighing about 210±20 grams were divided into ten group (n=6) for evaluation of antinociceptive effects the formalin test induced pain. The nociceptive response develops two phase: First (0-5) min after formalin (first or acute phase) and (16-60) min after formalin injection (second or chronic phase). The animals pre-treated with oral dose of extracts (100, 200, 400 mg/kg), 60 min before administration formalin for anti-inflammatory effects, the carrageenan induced hind paw edema model in rats were used and the animals pre-treated with oral dose of extracts (100, 200, 400 mg/kg), 30 min before administration of carrageenan. The control group is without receiving any drug and the sham group receiving an equal volume from distilled water. Then the paw volume measured in the mercury from 0 to 2 hours and 30 min after carrageenan injection statistical analysis by ANOVA and T-Test used (p<0/05). Results shows there is decreased pain and inflammation in formalin and carrageenan tests in the group that received 400 mg/kg dose of extract in comparison with the control and sham groups (p<0.05). This results demonstrated extract dose-dependently following the receipt of formalin resulted in a decrease of nociceptive in acute and chronic phase. There is decreased nociception in chronic and acute phase of formalin test in the group that received only 400 (mg/kg) of extract. So, data shows there is decreased inflammation in Carrageenan test in the group that received 400 (mg/kg) dose of extract in comparison with the control and sham groups (P 0.05).

Biography:

Abstract:

Various parts of Malva sylvestris have different medicinal properties such as antimicrobial activity. In this study the effects of M. sylvestris extract on protein electrophoresis pattern and gamma interferon of C. albicans infected mice was studied. Sixty female mice were divided randomly in six groups (three treatment groups, Candida, placebo and control groups). Treatment groups received aquatic extract of M. sylvestrs (50, 100 and 200 mg/kg) for 20 days every other day via injection in peritoneum. C. albicans was injected once after sixth injection of extract. Results showed significant decrease in the amount of albumin in three treatment groups. The β-globulin amount of 50 and 100 mg/kg groups and gamma interferon amount of all three treatment groups were increased significantly in proportion to control group. Clearing the body from pathogen organisms depends on cellular responses. It seems that M. sylvestris is capable of stimulating cellular immune response and can be used in studies about C. albicans.

Biography:

Andreia Ribeiro did her Biology Degree and Microbiology Master in Aveiro University in Portugal. Since 2010 she has been working as research assistant in flow cytometry and immunology groups, in Portugal and in Ireland. In 2013 she started her PhD in the DECIDE consoritum and completed in 2016 from National University of Ireland, Galway.

Abstract:

Mesenchymal Stromal Cells (MSC) possess immunomodulatory and anti-inflammatory properties, having several effects on immune cells. For this reason MSC have been proposed as a potential therapeutic modality for osteoarthritis (OA) and rheumatoid arthritis (RA). However, there is a commercial need to have a reliable, rapid, quantifiable assay to assess the potency of allogeneic human MSC. The aims of this assay were to determine by flow cytometry the effects of MSC from bone marrow (BM MSC) and adipose tissue (ASC) on TNF-and IL-6 production by LPS stimulated monocytes of healthy and patient samples. A number of factors were considered prior to optimising the assay, including: Brefeldin A concentration, type of anticoagulant (heparin; K2EDTA or citrate), LPS concentration; blood dilution and incubation time. MSC numbers were then titrated and co-cultured with whole blood of healthy donors. All results are expressed as intracellular expression of TNF-α and IL-6, on gated monocytes identified as CD45+CD14+ cells using an Accuri four colour flow cytometer. Results show that BM MSC and ASC significantly reduced TNF-and IL-6 expression by monocytes from healthy donors and in OA and RA patients. Thus, we have established a rapid, reliable and quantifiable screening assay to determinate the effects of MSC on LPS activated monocytes. Such an assay could be used to screen the recipients’ monocytes for inhibition by the MSC preparation that will be injected, thereby contributing to personalized medicine. For this reason this assay is being used in the ADIPOA2 clinical trial.

Biography:

Germain J P Fernando has completed his PhD at the University of Arizona, USA and Postdoctoral studies at the Baylor College of Medicine in Texas, USA. He is currently working as a Senior Research Fellow at the Australian Institute for Bioengineering and Nanotechnology (AIBN), University of Queensland, Brisbane, Australia, developing needle free vaccines.

Abstract:

Vaccinations greatly reduce the burden of infectious diseases. The current vaccination methods using the needle and syringe have a number of drawbacks. Firstly, they require liquid vaccine for delivery as such there is a need for cold chain during transportation. Secondly, intradermal (ID) and intramascular (IM) vaccination rely on trained personnel to perform these techniques. Finally, using needle and syringe introduce risks of needle stick injury as well as the spread of blood borne diseases. The Nanopatch mitigates all these issues. Vaccine dry coated onto the Nanopatch retains its activity even after six months at 25° C. The Nanopatch is designed to be easily administrable using a hand held applicator with the potential for self administration. The Nanopatch delivers vaccine to the skin’s viable epidermis and dermis layers. This reduces the possibility of damaging the blood vessels due to the array microprojections measuring only 0.11 mm. We have previously shown that the Nanopatch is able to induce similar immune response with ID injection with 1/10th and IM injection with a 1/100th of the vaccine dose in mice. The Nanopatch has demonstrated to be a more advantageous way of vaccination than the conventional ID and IM needle and syringe injections in both immunogenicity and administration. However there is still a need for a broader understanding of the mechanisms that lead to the enhanced immune response induced by the Nanopatch. To approach this question, we used systems biology methods to investigate which genes are up/down regulated at the site of vaccination. Here, I will discuss the Nanopatch molecular profile compared to ID vaccination. This study will contribute towards the knowledge for new potent vaccine development and better understanding of the molecular mechanisms of the Nanopatch action. We envisage the outcome will allow Nanopatch technology to translate from murine to larger animal models and ultimately leading into human clinical trials.

Biography:

Sundeep Kumar Upadhyaya is a Senior Consultant at New Delhi’s Indraprastha Apollo Hospitals. He is involved in the treatment of autoimmune disorders like Arthritides, Vasculitides, Lupus and Spondyloarthritis since 16 years. He is an Associate Professor and Teaching Faculty at the Teaching Program at the AHERF and National Boards of the Apollo Group of Hospitals. He is involved with cutting edge clinical work on these disorders and has developed treatment algorithms for Lupus Nephritis, Early and Established Rheumatoid Arthritis and has been on various national bodies and involved in management protocols for Autoimmune Disorders.

Abstract:

Rheumatoid arthritis (RA) is conventionally treated with cDMARDs (like Methotrexate, Leflunomide etc) but more severe, cDMARD resistant RA, is treated with a combinations of cDMARDs and Biologics (example Anti-TNF Biologics). Despite close monitoring and follow-up only a fraction of treated patients achieve complete remission and attain a drug free state. A combination of cDMARDs and Biologics in the treatment of early RA has been able to achieve drug free remissions in only a small minority of patients. Complete remissions are rare when patients are treated with cDMARDs alone. Presented here is the clinical description of patients with RA who achieved long-term drug free clinical remissions (CR) with cDMARDs alone at a single rheumatology clinic in northern India and remained in CR over a long period (at least 6 months). The immune-biology of such states and the clinical factors leading to the prolonged CR will be discussed next.

Biography:

Dr. Hanan Al-Khalifa, obtained her Master's degree in Parasitological Diseases and Immunology at University of Manchester, UK. Then she completed her Ph-D in 2007 in the University of Reading (UK), investigating the effect of n-3 fatty acids on the immune response and general health status. Her interests include but are not limited to immunological techniques, parasitological diseases, effect of nutrition, espicially fatty acids, on the immune status in both humans and expermental animals. She executed many research projects that focused on the effect of nutrition on immunology. Dr. Al-Khalaifa attended many scientific events and published more than 60 papers in refereed journals and conference proceedings

Abstract:

The phagocytosis assay allows quantitative measurement of the percentage of phagocytes and the enzymatic activity of each phagocyte. Immunomodulation of fatty acids in flaxseed may alter phagocytosis activity. The objective of this work was to compare the effect of feeding normal broiler chickens 15% of dietary flaxseed on phagocytic activity of monocytes and heterophils in the peripheral blood. One-day-old broilers were used. Upon hatching, all chicks were given the same basal diet for 13 d. Following this, dietary supplementation of flaxseed started at 14 d of ages until the end of the cycle at 35 d of age. At slaughter, samples of blood were collected from each bird. The quantitative analysis of the phagocytic activity of peripheral blood mononuclear phagocytes in whole blood was performed using phagotestcommercial kits. Results were expressed as percentage of fluorescent cells (% phagocytosing cells) and mean fluorescence intensity (MFI). Feeding flaxseed at 15 % did not affect either the percentage of cells participating in phagocytosis, or the Mean fluorescence intensity (MFI). However, there was a trend towards a decrease in the % of monocytes involved in phagocytosis in birds fed diets containing 15% flaxseed.Also, there was a trend towards a decreased MFI (p= 0.056) for monocytes. In general, results of the current study showed no effect of flaxseed on phagocytosis of peripheral blood cells.

Biography:

Saeed El-Ashram is an Associate Professor of Parasitology, Faculty of Science, Kafr El-Sheikh University, Kafr El-Sheikh, Egypt. He is the holder of several issued and pending patents in the field of Parasitology. He is also the Inventor of “Compositions and methods of enhancing immune responses to Eimeria” (United States Patent 8956849).

Abstract:

The recognition and elimination of eukaryotic parasites appear to be a hardwired prerequisite for host survival. However, eukaryotic parasites such as Toxoplasma gondii, Leishmania sp., Trypanosoma cruzi, Giardia sp., and Schistosoma sp. employ the innate system of the host for their growth, replication and continuation of their life cycle. So far, however, there has been little discussion about the interaction of the eukaryotic parasites and the innate immune system. Driven by this need, this review provides an overview of the host oriented inherent measures and eukaryotic parasite countermeasures for evading host defenses. Taken together, this information could be exploited to discover new therapeutic and prophylactic intervention points for broad spectrum host oriented inherent measures and eukaryotic parasite countermeasures and to determine the infection consequence.

Biography:

Jessy S Deshane is a pulmonary Immunologist with expertise in immune regulation in asthma. She investigates myeloid-derived regulatory cell biology and free radical mechanisms that regulate their differentiation and function. She pioneered these investigations both in mouse models and human asthma. She has authored 46 peer-reviewed publications, including high impact journals like Journal of Experimental Medicine, Journal of Clinical Investigations, Journal of Allergy and Clinical Immunology, Immunity and Cancer Research. She serves on the Editorial Boards for the journals Allergy and American Journal of Respiratory Cell and Molecular Biology and serves on grant review committees.

Abstract:

Rationale: Myeloid-derived suppressor cells (MDSCs) have been well established as regulators of anti-tumor immunity. MDSCs modulate amino acid metabolism in the tumor microenvironment and suppress T-cell function. However, it is less clear whether MDSCs regulate B-cell responses during tumor progression. Methods: Using a syngeneic orthotopic model for lung cancer with murine Lewis Lung Carcinoma cells, we evaluated B-cell subsets in tumor bearing mice by multi parameter flow cytometry. The amount of serum IgG or IL-7 was determined by ELSIA. Phospho-STAT5 and total STAT5 were detected by immunoblotting. To investigate MDSC-mediated suppression of B cell lymphopoiesis, we adoptively transferred MDSCs derived from bone marrow of CD45.2+ tumor bearing mice intratibially into congenic CD45.1+ mice. B-cell subsets in recipient mice at day 7 post MDSC transfer were enumerated as above. In vitro B-cell inhibitory assay was performed by co-culturing CFSE-labeled pre-activated splenocytes with MDSCs purified from bone marrow of tumor-bearing mice at a ratio of 1:1 in the absence or presence of arginase inhibitor nor-NOHA (20 M), iNOS inhibitor 1400W (500 nM) or IDO inhibitor 1-D-MT (1 mM) for 48 hours. The percentage of CD19+CFSElow cells (proliferating cells) was determined by FACS analysis. Results: Percentages and absolute numbers of Pro-, Pre- and mature B-cells were reduced in bone marrow (BM) of tumor bearing mice. Moreover, percentage and absolute number of follicular B cells were reduced, while immature B-cells increased in the spleen of tumor bearing mice. Levels of serum IgG were reduced in tumor-bearing mice. Furthermore, IL-7 and downstream STAT-5 signaling were impaired in tumor bearing mice. Transfer of BM-derived MDSCs from tumor bearing mice into congenic recipients resulted in significant reduction in both percentages and absolute numbers of immature and mature B-cells in peripheral blood of recipient mice. Pre-B cells and immature B-cells also decreased in BM of MDSC transferred recipients. Additionally, MDSCs suppress B-cell proliferation and IgG production by B-cells in an arginase and iNOS dependent but IDO independent manner. Conclusions: In the present study, we demonstrate that B-cell differentiation in vivo is impaired in the BM and spleen of mice with lung cancer. Adoptive transfer studies with congenic mice demonstrate that MDSCs derived from Lewis Lung Carcinoma bearing mice may suppress B-cell differentiation in tumor naive mice. These results together suggest that tumor-related MDSCs may potentially regulate humoral immune responses to promote tumor survival.

Biography:

Ildiko Molnar has completed her PhD in the special field of Graves’ Ophthalmopathy at the Hungarian Academy of Science. Her work and research connected her to Kenezy County and Teaching Hospital from 1977 to 2008. Her research activities are on field of internal medicine, endocrinology, immunology and allergology. Currently she is the Chief of EndoMed, Immunoendocrinology and Osteoporosis Centre, Private Outpatient Clinic from 2008. She is an expert in laboratory methods (ELISA, blotting, allergy testing) and DXA measurement. She has published more than 53 papers in reputed journals, 16 chapters and 2 books.

Abstract:

Type II 5’-deiodinase enzyme (DII) activity is responsible for T4 conversion to T3 resulting in the majority of intracellular T3 concentration. DII is a membrane-anchored protein characterized by tissue-specificity; highly expressed in thyroid, pituitary, skeletal, eye and cardiac muscles, brain, adipose tissue and bone. Decreased DII activity leads to hypothyroidism in euthyroid sick syndrome. We demonstrated DII expression in thyroid, eye and skeletal muscle tissues by immunohistochemistry using immunized guinea pig and patients sera with Graves’ orbitopathy. Decreased DII activities were measured after adding proinflammatory cytokines and patients sera with hyperthyroid Graves’ orbitopathy and systemic sclerosis. Antibodies to DII inhibited the mitogen-activated protein kinase (MAPK) activation in thyroid tissue. Proinflammatory cytokines (IL-6, TNFα, IFNγ) inhibited thyroid DII activities in dose-dependent manner (Vmax: 4.1×10-3 pmol/mg/min for IL-6; 0.18 pmol/mg/min for TNFα; 0.23 pmol/mg/min for IFNγ). Hyperthyroid patient sera with Graves’ orbitopathy decreased better thyroid DII activities than eye muscle DII ones (3.99±5.79 vs. 7.66±10.49 pmol/mg/min, P<0.05, n=26). Patient sera with systemic sclerosis (SSc, n=19) decreased DII activities compared to those in controls (n=16) (4.99±1.04 vs. 2.88±0.61 pmol/mg/min, P<0.0001). Immunized guinea pig and Graves’ patient sera with anti-DII antibodies resulted in relevant inhibition of MAPK activation. In conclusion, DII protein can be a new autoantigen in thyroid autoimmunity, particularly in Graves’ orbitopathy. DII activity blocking cytokines could be responsible for low FT3 levels causing euthyroid sick syndrome in systemic sclerosis. The difference in tissue-specific DII activities could be implicated in the development of orbitopathy in hyperthyroid Graves’ disease.

Biography:

Rongtuan Lin is an Associate Professor in the Department of Medicine at McGill University and a Project Director of the Molecular Oncology Group at the Lady Davis Institute for Medical Research. He has received his PhD from Concordia University and completed Post-doctoral training at the Lady Davis Institute for Medical Research. He made important contributions in the fields of interferon signaling and innate antiviral immunity. He has a highly successful laboratory research program with 100 scientific publications, which have been cited more than 5,500 times. He was a recipient of a Chercheur-boursier Senior and Junior 2 from Fonds de la Recherche en Sante du Quebec. In 1996 and 1998, he has received the Milstein Young Investigator Award from the International Society for Interferon and Cytokine Research.

Abstract:

Oncolytic viruses (OVs) are novel anticancer agents that infect and effectively kill cancer cells but not normal cells. Although tumor growth is delayed or eliminated in numerous animal models following treatment with OVs, several cancer models remain partially or completely resistant to viral oncolysis. To overcome this resistance, experimental strategies are now combining OVs with different cytotoxic compounds to improve OV efficacy. Our laboratory has previously demonstrated that OV replication can be bolstered by co-administration of other chemical agents such as Triptolide, a natural molecule derived from the medicinal herb. In the current study, we investigated the capacity of sulforaphane (SFN); an anti-cancer compound naturally occurred in cruciferous vegetables with demonstrated potent antioxidant and possible anti-inflammatory actions to enhance vesicular stomatitis virus (VSV) oncolysis in OV-resistant cancer cells. We ultimately demonstrate that the resistant PC3 prostate cancer cell line can be sensitized to VSV by addition of SFN. Indeed, SFN dose-dependently enhances the replication of VSV. Neither VSV (MOI 0.1) nor SFN (20 uM) alone are toxic against PC3 cells in vitro; however, in combination they greatly increased the oncolytic capacity of VSV by reducing cancer cell viability and promoting apoptosis-mediated cell death. Furthermore, the potentiation of VSV oncolysis by SFN is dependent on the production of ROS and is associated with the induction of autophagy. SFN is known to induce phase II antioxidant genes via Nrf2 activation, which regulates ROS levels and stimulates autophagy in prostate cancer cells. Mechanistically, SFN inhibited the innate antiviral response by blocking the type-1 interferon (IFN) signaling pathway, through the activation of the Nrf2 transcription factor. Exogenous Nrf2 expression inhibits Interferon-Stimulated Response Element (ISRE) promoter activity in a dose dependent manner following virus infection or IFN treatment. Taken together, these results demonstrate for the first time the synergic effect of SFN and VSV and indicate that SFN treatment increases VSV replication and the subsequent apoptosis of tumor cells by inhibiting IFN signaling. We are currently investigating the molecular mechanism involved in VSV-induced oncolysis by Nrf2 activators and evaluating the therapeutic potential of the combination of OV and Nrf2 activators in a mouse model of prostate cancer.

Biography:

Meltem Elitas is a Faculty Member in Mechatronics Program at Sabanci University. Her background is in Electrical and Mechatronics Engineering. She has obtained her Doctorate from Bioengineering and Biotechnology Department at Ecole Polytechnique Federale de Lausanne. She has performed her Postdoctoral studies at Yale University Biomedical Engineering Department. She has published more than 25 papers in reputed journals. Her research interests are biomechatronics, cellular heterogeneity, cellular interactions, tumor microenvironment, live cell imaging and development of microfabricated tools for quantitative biology.

Abstract:

Understanding the interactions between tumor cells and immune cells in a quantitative manner will provide valuable information to reveal the mechanism of diseases, immune defense and development of new treatment reagents and strategies for the diseases. Today one of the biggest limitations relies on the traditional methods and tools that we use to investigate the rare cells and specific events in biology particularly in immunology. Since these techniques are not adequate enough to be selective, specific and quantitative, the rare cells such as the metastatic or drug resistant ones or the events such as onset symptoms of tumors and infections are being masked by majority of the cells or events in the population. Therefore, we cannot diagnosis on time or provide successful strategies. As a consequence, our approaches might not target the right cells at the right time in the right place. To overcome these limitations, we might profit from engineering approaches and tools. We can develop quantitative, accurate, reproducible and precise methods and use microfabricated tools to understand the nature and behavior of rare cells and events. The improvements from microfabricated tools in conjunction with microscopy might provide statistics from large numbers of single cells, short assay time, less sample consumption, less waste production, quantitative and reproducible data, single-cell resolution images, high-throughput, spatio-temporal tracking and real-time assays, etc. This talk will present recently developed microfabricated tools to understand the immune cell-tumor cell interactions. I will present our microfluidic applications and their preliminary data from my research group.

Biography:

Amina Dahmani is currently a PhD candidate in Microbiology-Immunology at Université de Montréal, Canada. She has completed her Master degree in Immunology at Univérsité Laval, Canada, in Cellular Therapy Lab directed by Dr Jacques P Tremblay, where she studied the development of immunological tolerance to allogeneic myoblasts transplantation as potential therapy for Duchenne Muscular Dystrophy. Later she has joined Dr Jean-Sébastien Delisle team's, dedicated to cancer and viral adoptive immunotherapy to complete her PhD. She is currently working to improve adoptive immunotherapy protocols.

Abstract:

Adoptive immunotherapy (AI) has emerged as a potentially curative therapy for advanced cancer and infections. Recent findings suggest that the transfer of T-cells with “early” memory features may improve the therapeutic potential of AI. TGF-β is a pleiotropic cytokine that controls a large spectrum of biological and pathological processes. In T-cell biology, TGF-β is mostly known for its immunoregulatory properties, but recent evidence has revealed a novel role of TGF-β in T-cell memory differentiation and maintenance. Thus, we investigated whether TGF-β could promote features of memory in ex vivo stimulated human T-cells to further improve the efficacy of clinical protocols for AI. Here we show that agonistic TGF-β stimulation leads to the expression of central memory markers without significantly altering T-cell expansion or polyfunctional cytokine secretion following stimulation. Furthermore, TGF-β exposure decreased expression of transcription factors responsible for effector differentiation (T-BET, GATA3 and BLIMP1) and increased those associated with memory differentiation, notably ID3. The knock-down of ID3 by specific siRNA revealed that TGF-β-driven T-cell memory differentiation largely depends on ID3. Moreover, TGF-β-exposed T-cells showed enhanced persistence, expansion and alloreactivity after adoptive transfer into NSG mice. Finally, using clinically relevant culture methods to generate T-cell lines against viral and tumor antigens, we found that TGF-β programmed the expression of early memory markers without significantly curtailing T-cell expansion or antigen-specificity. This finding provides a rationale for clinical use of TGF-β to optimize memory phenotype of ex vivo pathogen/antigen-specific T-cells expanded for AI.

Biography:

Omar El Bounkari worked in Klinikum der Universität München, Germany

Abstract:

Macrophage migration inhibitory factor (MIF) proteins (MIF and MIF-2) are chemokine-like inflammatory mediators with unique structural properties distinct from classical chemokines. MIF proteins play arole in the control of both physiological and pathophysiological immune responses. With MIF-2 only very recently identified, most evidence currently is available for MIF. In fact, MIF is a pivotal upstream mediator of innate immunity, while some contribution to the adaptive response has been reported. When dysregulated, MIF is an exacerbating promoter of several inflammatory diseases including atherosclerosis, a chronic inflammatory condition of large and medium-sized arteries and the major underlying cause of cardiovascular morbidity and mortality worldwide. MIF orchestrates the atherogenic recruitment of monocytes/macrophages and T lymphocytes through non-cognate interaction with the CXC chemokine receptors CXCR2 and CXCR4, respectively, and contributes to the inflammation and destabilization in atherosclerotic lesions. These processes have been considered as the effects of an innate chemokine on innate inflammatory cells in the atherosclerotic lesion area.Here we show that MIF also controls adaptive immune cells in atherosclerotic pathogenesis. We present data that MIF is a novel B cell chemokine that promotes B cell migration and proliferationvia the chemokine receptors CXCR4 and CXCR7 as well as CD74, the surface form of MHC class II invariant chain. MIF-driven B cell responses are mediated through the ZAP-70 and ERK1/2 signaling pathways and encompass the activation of calcium transients. We also studied the impact of Mif gene deletion in the proatherogenicApoE-/- genetic background in mice and have unraveled a surprising atherogenic phenotype with a previously unrecognized link between MIF and B cell pathobiology. This suggeststhat MIF could be a potential therapeutic target to induce protective B cell responses in such diseases.

Biography:

Denis Soulet has completed his PhD in Neuroimmunology from Laval University, Canada and Postdoctoral studies from Ycee Claude Bernard, Sweden. He is an Associate Professor at the Medicine Faculty of Laval University, Canada. He has published more than 40 papers in reputed journals and has been serving as an Editorial Board Member of SM Journal of Gastroenterology and Hepathology. He is leading a research team dedicated to study the role of peripheral inflammation in the enteric nervous system and its contribution to Parkinson’s disease. The ultimate goal of his research program is to design immunomodulatory based disease modifying drugs for PD.

Abstract:

Parkinson’s disease (PD) is a neurological disorder characterized by motor symptoms which are often preceded by non-motor symptoms, including gastrointestinal dysfunctions. Common treatments are only symptomatic; there is still no disease modifying drug available to cure patients. Since numerous pro-inflammatory markers have been measured in the central and peripheral nervous system, this deleterious immune response seems to be a potential target to develop new therapeutic strategies. Therefore, a better understanding of the role of the immune response in the etiology and progression of PD is essential. During my talk, I will present original data about the impact of the innate immune response on enteric neuronal damage in PD models. At first, we characterized the immune response induced by the neurotoxin MPTP in the enteric nervous system of partially immunodeficient mice. We demonstrated the timeline of inflammatory events occurring prior to the neuronal demise and the critical role of monocytes and macrophages in the gut. Thereafter, we tested various estrogenic compounds for their immunomodulatory and neuroprotective properties in PD models both in vivo and in vitro, delineating the major contribution of various estrogenic receptors, mainly the G Protein-coupled Estrogen Receptor 1 (GPER1). More recently, we successfully explored the therapeutic potential of a clinically approved selective estrogen receptor modulator, Raloxifene, for drug repurposing in PD. In conclusion, our data highlight the critical role of the immune response at early stages of PD and the immunomodulatory and neuroprotective potential of estrogen-based hormonotherapy at the pre-clinical level.

Biography:

Ganapathi Bhat Mugulthimoole is a Senior Consultant Medical Oncologist & Stem Cell Transplant Physician at Jaslok Hospital & Research Centre. He has completed his graduation in Medicine in 1993 and Post graduation in General Medicine in 2000. He has further trained in the field of oncology in various institutions in India and later gained expertise in stem cell transplant while working in Kuwait Cancer Control Centre (2002-2006). He has gained specialized training in stem cell transplantation as part of the ESH-EBMT (2007), 2011 (Labaule, France) and ICAS training program (2009) from Ulm University, Germany. He has numerous academic articles published in Indian, international journals and textbooks to his credit.

Abstract:

The tumor microenvironment is a principal feature of cancer biology that supports the initiation and progression of the tumor and response to therapy. Cells and molecules of the immune system are the essential elements of the tumor microenvironment. Therapeutic stratagems can harness the immune system to precisely target tumor cells by possibly stimulating tumor-specific immunological memory, which may lead to long term regression. Understanding the complexity of immunomodulation by tumors is important for the expansion and promotion of immunotherapy. Several approaches are being carried out to augment the anti-tumor immune responses. Among them are immunotherapeutic vaccines, adoptive cell transfer therapies and checkpoint blocking drugs. However, cancer cells try to evade the immune watchdogs by reducing surface tumor antigen or inducing cells that express certain proteins that affect immune cell inactivation or by promoting tumor proliferation and survival. Gaining a deeper understanding of the tumor immunogenicity by employing advanced techniques such as sequencing of the tumor DNA, will help to better address the challenges and gain an appreciation of the delicate association between cancer and our immune systems.

Biography:

Moshe Giladi has joined Novocure in 2005 and served as the Head of the NovoBiotic project until 2008. He was then promoted to Head of Novocure's Preclinical Research leading a team of experts of various fields: Cancer, immunology, cell biology and also responsible for research collaboration with academic institutes. He leads research activities studying tumor treating fields mechanism of action. He has received his PhD in Life Sciences from the Department of Molecular Microbiology and Biotechnology, Faculty of Life Sciences and his MBA from the Leon Recanati Graduate School of Business Administration both at the Tel Aviv University, Israel.

Abstract:

Tumor treating fields (TTFields) are an effective anti-neoplastic treatment modality delivered via non-invasive application of low intensity, intermediate frequency, alternating electric fields. This therapy is approved for the treatment of patients with glioblastoma. Previous investigations have shown that TTFields disrupt microtubules and septin filaments, both of which govern key roles in mitosis. Some of the outcomes of mitosis under TTFields application include abnormal chromosome segregation and ER stress which trigger different forms of cell death. The goal of this study was to evaluate whether TTFields induced cancer cell death can be perceived as immunogenic by the immune system and to explore the possibility of combining TTFields with immune-modulating drugs. Murine Lewis lung carcinoma (LLC) and ovarian surface epithelial (MOSE) cells were treated with TTFields using the inovitro system. The exposure of calreticulin (CRT) on the surface of treated cells was evaluated using flow cytometry. High-mobility group box 1 protein (HMGB1) secretion was measured using ELISA assay. For in vivo experiments, immunocompetent C57BL/6 mice were orthotopically implanted with LLC cells and treated with TTFields, anti-PD-1 or combination of the two modalities. Changes in tumor volume were monitored and flow cytometry analysis was performed for phenotypic characterization of tumor infiltrating immune cells. We demonstrate that application of TTFields leads to the exposure of CRT on the cell surface and also promotes release of HMGB1 from cancer cells in vitro. In vivo, the combined treatment of TTFields and anti-PD-1 led to a significant decrease in tumor volume compared to control group and to animals treated with anti-PD-1 alone. An increase in CD45+ tumor infiltrating cells was observed in both anti-PD-1 and TTFields+anti-PD-1 groups although statistical significance was reached only in the combination treatment group. Interestingly, CD45+ cells from the combination treatment group also demonstrated a significant upregulation of PD-L1 expression on the cell surface. Specifically, this upregulation in the PD-L1 expression was observed in both F4/80+CD11+ cells (macrophages) and CD11c+ cells (dendritic cells) whereas no significant effect on the infiltration pattern of these immune cell populations was noted. Taken together, our results demonstrate that TTFields application potentiates immunogenic cell death in cancer cells and that combining TTFields with specific immunotherapies such as anti-PD-1 might achieve tumor control by further enhancing antitumor immunity.

Biography:

Tali Feferman has completed her PhD in Molecular Biology at the Macquarie University and Postdoctoral Training at The Heart Research Institute in Sydney. She has then joined the Weizmann Institute of Science in 2000. Her initial interest involved exploring the mechanism of action and immunomodulation of Myasthenia Gravis (MG) and its model experimental autoimmune MG (EAMG). In recent years her interest is mainly focused on addressing questions important for optimizing cancer immunotherapy using cutting-edge imaging microscopic techniques in live mice.

Abstract:

Poor tumor vascularization is an obstacle to immunotherapy by CTLs. It impairs tumor infiltration but also introduces hypoxia, known to interfere with T-cell migration. It is yet unknown how suboptimal vascularization affects CTL migration and function within tumors. To study this question, we combined immunohistochemistry of human melanoma samples with two-photon imaging in live mice. Orthotopically implanted B16-OVA tumors were studied after adoptive transfer of in vitro matured antigen-specific OT-I CTLs. In patients, CD8 T-cells concentrated around peripheral vessels in the tumor and sparsely infiltrated avascular areas. In mice, CTLs crawled rapidly in oxygenated areas within 50 µm of flowing blood vessels. Occluding intratumoral blood vessels triggered immediate arrest of CTL motility, which was quickly reversed when flow was resumed. Immunohistology indicated that CTLs avoided hypoxic tumor areas. Live CTL imaging in vitro showed deceleration under hypoxic conditions and when oxidative phosphorylation was blocked. To circumvent intratumoral CTL dysfunction we attempted to increase vascular density by implanting tumors in matrices containing bFGF. bFGF-laced tumors were more easily rejected after transfer of CTLs and displayed delayed growth in untreated mice but were not affected in mice deficient in CD8 T-cells. CTLs infiltrated such tumors in normal numbers but displayed enhanced motility in highly vascular tumors, suggesting that enhanced rejection resulted from improved intratumoral CTL migration. Taken together, the results suggest that hypoxia limits CTL function away from blood vessels and that alleviating it may synergize with immunotherapy.

Biography:

Hisham Abd El Dayem is presently working in Ain Shams University, Egypt.

Abstract:

Purpose: To compare expression of multidrug-resistant protein 1/P-glycoprotein (MDR1/Pgp) in retinoblastoma in eyes treated by primary enucleation due to advanced tumor at initial presentation and those enucleated after being resistant to chemotherapy. Methods: This study was a prospective study. Twenty retinoblastoma patients presented to Retinoblastoma Clinic at Ophthalmology Department, Ain Shams University Hospitals. All patients had enucleation and were divided into 2 groups. Patients in group-1 underwent primary enucleation due to advanced tumor at presentation. Patients in group-2 underwent secondary enucleation after failure of conservative treatment. Immunohistochemical studies were performed searching for expression of multidrug-resistant protein 1/P-glycoprotein (MDR1/Pgp) in the two groups. Results: Analysis of the primary enucleation group showed high positive, low positive and negative expression in 1 (10%), 2 (20%) and 6 cases (70%) respectively. In secondary enucleation group: 5 cases (50%), 3 cases (30%) and 2 cases (20%) showed high positive, low positive and negative expression respectively. Conclusions: This pilot study though, not being able to demonstrate statistical significance in MDR1 expression in primary enucleated vs. secondary enucleated resistant cases, demonstrated p-value low enough to indicate a trend for more MDR1 expression in resistant cases (P=0.068). Further study with a larger sample size is warranted.

Biography:

Nan-Shan Chang is currently the Professor and Director of the Molecular Medicine Institute, National Cheng Kung University (NCKU) in Taiwan, and the Adjunct Professor with the SUNY Upstate Medical University and the NYS Institute for Basic Research in Developmental Disabilities, New York. Dr. Chang is most noted for his discovery of tumor suppressor WWOX in 2000. Recent Awards: Breast cancer and neurofibromatosis research awards from the Department of Defense, USA, in 2008 and 2010; NCKU Distinguished Professor Award 2010, 2013 and 2016; Distinguished Scientist Award 2011 from the Society of Experimental Biology & Medicine, USA.

Abstract:

Whether tumor suppressor WWOX stimulates immune cell maturation is largely unknown. Here, we determined that Tyr33-phosphorylated WWOX physically binds non-phosphorylated ERK and IκBα in immature acute lymphoblastic leukemia MOLT-4 T cell and in the naïve mouse spleen. The IκBα/ERK/WWOX complex was shown to localize, in part, in the mitochondria. WWOX prevents IκBα from proteosomal degradation. Upon stimulating MOLT-4 with ionophore A23187/phorbol myristate acetate (IoP), endogenous IκBα and ERK undergo rapid phosphorylation in less than 5 min, and subsequently WWOX is Tyr33 and Tyr287 de-phosphorylated and Ser14 phosphorylated. Three hr later, IκBα starts to degrade and ERK returns to basal or non-phosphorylation, and this lasts in the next 12 hr. Finally, expression of CD3 and CD8 occurs in MOLT-4, along with re-appearance of the IκBα/ERK/WWOX complex near 24 hr. Inhibition of ERK phosphorylation by U0126 or IκBα degradation by MG132 prevents MOLT-4 maturation. By time-lapse FRET microscopy, IκBα/ERK/WWOX complex exhibits an increased binding strength by 1-2 fold after exposure to IoP for 15-24 hrs. Meanwhile, a portion of ERK and WWOX relocate to the nucleus, suggesting their role in the induction of CD3 and CD8 expression in MOLT-4. (Supported by MOST and NHRI, Taiwan, and DoD, USA)

Biography:

Miao Dong is currently pursuing PhD in City University of Hong Kong. She has obtained her Bachelor degree from Liaoning University, China, majoring in Ecology and completed her Master degree from Shandong Agricultural University. During three years study, she investigated antioxidant defense system in zebrafish and has 6 publications (3 of 6 are as first author). During her PhD study, she has done research on interaction between innate immune proteins in fish blood and bacteria, also the collection of natural occurring antimicrobial peptides from medaka fish blood. The related manuscript has been submitted to JBC.

Abstract:

The excessive use of antibiotics in aquaculture contributes to the uprising of antibiotic resistance that threatens human health. We explore the innate immunity of fish for naturally occurring antimicrobial factors that can be developed into potential antibiotic agents. Antimicrobial peptides (AMPs) have been studied in many organisms but efforts on the systematic identification of AMPs in fish have been lacking. In this study, we systematically identified naturally occurring peptides in medaka plasma. Blood collected from medaka of different gender, age and infection status were combined and thereby providing a resource of plasma macromolecules under various possible physiological conditions. Peptides under the molecular weight of 3 kDa were fractionated and purified followed by mass spectrometry analysis. In total, 6483 unique peptides were identified against the medaka genome, constituting a database of circulating peptides in this organism. After evaluation with a combination of web based prediction tool and conserved physicochemical properties of AMPs, 83 potential antimicrobial peptides were predicted. One of them, a 13-residue peptide named VPS13D3241-3253, showed broad spectrum toxicity on fish and human pathogenic bacteria (Gram positive or Gram negative) without significant cytotoxicity on mammalian cell lines. Scanning electron microscopy indicated that VPS13D3241-3253 disrupted the cell wall of both Gram positive and negative bacteria. SOS response assay showed that this peptide efficiently induced DNA damage in bacteria. The identification of VPS13D3241-3253 illustrates the feasibility of the proteomic approach in the discovery of potentially novel AMPs from fish. These AMPs will form an important basis for the development of new antibacterial agents in the fishery.

Biography:

Anne Stinn is currently a PhD student at the Max Planck Institute for Infection Biology in Berlin, Germany. From 2008 to 2013 she studied Biology at the Justus Liebig University in Giessen, Germany. After completing her study she went to London as an Intern in the Research Group of Cell Death, Cancer and Inflammation (CCI) at the UCL Cancer Institute. She has started her PhD in the year 2014.

Abstract:

Acute myeloid leukemia (AML) is a highly malignant cancer of the myeloid cell lineage that is characterized by the rapid growth of abnormal white blood cells. Although AML is a relatively rare disease accounting for about 1% of all cancer cases (US), it is the type of leukemia showing the lowest survival rate. In the majority of all AML cases mutations in the kinase domain of the FMS-like tyrosine kinase III receptor (FLT3; CD135) are reported. Besides treatment based on chemo and radiation therapy as well as bone marrow transplantation, bispecific antibodies are studied for the use in immunotherapy against AML. These antibodies recognize tumor-associated antigens (TAAs) as well as the agonistic T-cell receptor/CD3 complex (TCR/CD3) and should thereby lead to a tumor cell-restricted activation of immune cells and specific lysis of cancer cells. In this study a structural analysis of the bispecific antibody NF-CU, the first known to date was performed. In addition to the structure of the NF-CU itself, the crystallographic structure of the antibody bound to FLT3 and CD3 was investigated. Deciphering the crystal structure of the antibody-antigens complex should give an inside into epitope recognition as well as the molecular mechanism leading to T-cell activation and tumor cell death.

Biography:

Maryam Golshani was graduated from the Pasteur Institute with a degree in Medical Bacteriology. Presently, she is a Post doctorate fellow and Junior Research Group Leader at IPI working on new Brucella vaccine candidates. Her research mainly focuses on in silico investing the immunogenicity of new vaccine targets and in vivo evaluating their protective efficacy against Brucella infection. She has involved in more than 11 projects and published 11 papers in reputed journals.

Abstract:

Objectives: Brucellosis is the most common bacterial zoonosis worldwide and no safe and effective vaccine is available for the prevention of human brucellosis. In humans, brucellosis is mostly caused by Brucella melitensis and Brucella abortus. According to our in silico studies, Omp2b is predicted to be potentially immunogenic antigen conserved in main Brucella pathogens. The aim of this study was to design truncated form of Omp2b and to evaluate the immunogenicity and protective efficacy of a recombinant protein vaccine encoding tOmp2b. Methods: Bioinformatics tools were used to design the truncated protein based on conserved domains and regions of epitopes with strong affinity for MHC molecules. The humoral/cellular immune response and protection levels against challenge with wild B. melitensis and B. abortus infections were evaluated in rtOmp2b+ adjuvants immunized mice and control groups. Results: Vaccination of BALB/c mice rtOmp2b provided the significant protection level against both B. melitenisis and B. abortus. Moreover, rtOmp2b elicited a strong specific IgG response (higher IgG2a titers) and significant IFN-γ/IL2 production. Conclusion: According to the results, rtOmp2b is able to induce cross-protection against B. melitensis and B. abortus infections. Therefore, it could be a new potential candidate for the development of Brucella subunit vaccines.

Biography:

Jalil Mehrzad has completed his PhD at the age of 32 years from Ghent University, Faculty of Veterinary Medicine, and postdoctoral studies from McGill University. He is an assciate professor of Immunology in Ferdowsi University of Mashhad (next year will move to Tehran University as full-time scientific member of departmet of Microbiology and Immunology). With H-index and citations of 16 and 1315, respectively, he has published more than 45 papers in reputed journals and has been serving as regular reviewer for many journals in the area of immunobiology, molecular biotechnology and medicine.

Abstract:

Caspases-mediated apoptosis/cell death activation is key regulatory response in many physiopathological conditions. Application of bioluminescence and the reaction of luciferase would provide a powerfully novel in vitro/vivo assay for apoptosis detecion. As key brain immune cells, astrocytes and microglials, are vital part of the central nervous system (CNS); they are the main responder to inflammation in CNS; any disruption on their function would lead to CNS damage. Aflatoxin B1 (AFB1) is commonly found in foodstuffs, and can be the cause of many diseases including cancer. AFB1 and its metabolites cause oxidative stress in especially the CNS-derived cells, adversely affecting their normal activities, thus leading to the neurodegenerative diseases including multiple sclerosis (MS), Alzheimer’s and Huntington’s diseases. Considering the importance of astrocytes and the inevitable existence of AFB1 in the feed/foods, worldwide, the study of astrocytes-AFB1 interactions is valuable. We therefore investigated the impact of AFB1 on the apoptosis of one of the key accessory supportive CNS, astrocytes, using several biochemical experimentations including intracellular ATP and caspases 3/7 measured by bioluminescence and luciferase reactions. The release of cytochrome c and apoptosis/necrosis of AFB1-treated astrocytes with various concentration of AFB1 and exposure time was also tested using Western blotting and flow cytometry techniques, respectively. Bioluminescence results revealed decreased intracellular ATP, increased caspases 3/7 activities, cytochrome-c release and apoptotic/necrotic of astrocytes particularly at higher timepoints and doses of AFB1. Considering the broad roles of astrocytes in CNS, this finding deepens our understanding of the molecular mechanisms and functional consequences of the neural cells damage neurotoxicity triggered by AFB1 exposure in mammals.

Biography:

Thomas Boldicke has completed his PhD at the Max Planck Institute for Molecular Genetics in Berlin. He has been working for 25 years in the field of recombinant antibodies particularly intrabodies. He has published more than 20 papers in reputed journals.

Abstract:

Intracellular antibodies (intrabodies) are targeted into a cell expressing the corresponding antigen, binding of the intrabody to the antigen results in inhibition of protein function. The advantages of high specificity, no off target effects and targeting of post translational modifications are the reasons that such molecules are very valuable in functional genomics. Two developments will boost the intrabody technology in the future: Cytoplasmic intrabodies can be stable expressed as single domain antibodies, mostly from camels. Alternatively, the construction of human VL and VH domains is ongoing. The single domain antibody approach is an effective alternative to other approaches for selection of stable cytoplasmic intrabodies such as the Intracellular Antibody Capture Technology (IACT) based on the yeast two hybrid system and Complementarity Determining Region (CDR) grafting or introduction of synthetic CDRs in stable frameworks. ER intrabodies: Selection of recombinant antibody fragments by in vitro display systems mainly phage and yeast display. One cloning step is sufficient to express scFv fragments as ER intrabodies. Most promising are these intrabodies retaining proteins passing the ER. Recently we demonstrated in mice a delay of metastasis of rhabdomyosarcoma tumor cells mediated by two specific intrabodies retaining two polysialyltransferases inside the ER. Finally transgenic ER intrabody mice have been generated. An intrabody mouse expressing an anti-VCAM intrabody is not lethal in comparison to the genetic knockout counterpart. 30% of genetic knockouts are lethal; therefore intrabody knockdown mice will be very useful in case the genetic knockdown is embryonically lethal.

Biography:

Hanan Al-Khalifa has obtained her Master’s degree in Parasitological Diseases and Immunology at University of Manchester and completed her PhD in 2007 in the University of Reading, UK, investigating the effect of n-3 fatty acids on the immune response and general health status. Her interests include but are not limited to immunological techniques, parasitological diseases, effect of nutrition, espicially fatty acids, on the immune status in both humans and expermental animals. She executed many research projects that focused on the effect of nutrition on immunology. She has attended many scientific events and published more than 60 papers in refereed journals and conference proceedings.

Abstract:

The phagocytosis assay allows quantitative measurement of the percentage of phagocytes and the enzymatic activity of each phagocyte. Immunomodulation of fatty acids in flaxseed may alter phagocytosis activity. The objective of this work was to compare the effect of feeding normal broiler chickens 15% of dietary flaxseed on phagocytic activity of monocytes and heterophils in the peripheral blood. One day old broilers were used. Upon hatching, all chicks were given the same basal diet for 13 days. Following this, dietary supplementation of flaxseed started at 14 days of ages until the end of the cycle at 35 days of age. At slaughter, samples of blood were collected from each bird. The quantitative analysis of the phagocytic activity of peripheral blood mononuclear phagocytes in whole blood was performed using PHAGOTEST commercial kits. Results were expressed as percentage of fluorescent cells (% phagocytosing cells) and mean fluorescence intensity (MFI). Feeding flaxseed at 15% did not affect either the percentage of cells participating in phagocytosis or the Mean Fluorescence Intensity (MFI). However, there was a trend towards a decrease in the percentage of monocytes involved in phagocytosis in birds fed diets containing 15% flaxseed. Also, there was a trend towards a decreased MFI (p=0.056) for monocytes. In general, results of the current study showed no effect of flaxseed on phagocytosis of peripheral blood cells.

Biography:

Laurence Macia is a Group Leader at the Charles Perkins Centre, School of Medicine of the University of Sydney. She has published over 30 articles in journals such as Nature Communication and Nature Reviews Immunology. Her research interest is the impact of diet on gut microbiota and development of inflammatory diseases. She has obtained her PhD in 2006 at the Pasteur Institute of Lille in France where she studied the inter-relation between metabolism and immunity. She has then worked at the Garvan Institute and at Monash University in Professor Mackay’s Lab to investigate the impact of diet on the development of Western diseases.

Abstract:

Incidence of food allergy has increased dramatically in recent decades particularly in Western countries. The diet hypothesis states that western diet enriched in fat and sugars while deprived in fibre contributes to development of western diseases such as allergy. Dietary fibre is potent prebiotic, reshaping beneficially gut microbiota. It is also fermented in the colon by anaerobic bacteria into short chain fatty acids (SCFA) that bind specific G-protein coupled receptors widely expressed in the host. The aim of this study was to determine the impact of diet enriched in dietary fibre and SCFA on the development of food allergy in mice. Mice were fed on diets either enriched or deprived in fibre in models of oral tolerance to peanut and of peanut allergy. In both models, dietary fibre was beneficial as enhanced oral tolerance and protection from food allergy were observed under high fibre feeding conditions. SCFA were behind these benefits as both acetate and butyrate protected from development of peanut allergy while propionate had no effects. Accordingly, mice knockout for GPR43 or for GPR109A, respectively preferential receptor for acetate and butyrate were not protected from food allergy development under high fiber feeding conditions. To determine the role of gut microbiota, germ free mice were reconstituted with microbiota isolated from high fibre vs. zero fibre fed mice and we found that the first was protective in food allergy. In conclusion, high fibre feeding protects from food allergy development by reshaping of gut microbiota and through the SCFA acetate and butyrate.

Biography:

Sundeep Kumar Upadhyaya is a Senior Consultant at New Delhi’s Indraprastha Apollo Hospitals. He is involved in the treatment of autoimmune disorders like Arthritides, Vasculitides, Lupus and Spondyloarthritis since 16 years. He is an Associate Professor and Teaching Faculty at the Teaching Program at the AHERF and National Boards of the Apollo Group of Hospitals. He is involved with cutting edge clinical work on these disorders and has developed treatment algorithms for lupus nephritis, early and established rheumatoid arthritis and has been on various national bodies and involved in management protocols for autoimmune disorders.

Abstract:

Rheumatoid arthritis (RA) is conventionally treated with cDMARDs (like Methotrexate, Leflunomide etc) but more severe, cDMARD resistant RA, is treated with a combinations of cDMARDs and Biologics (example Anti-TNF Biologics). Despite close monitoring and follow-up only a fraction of treated patients achieve complete remission and attain a drug free state. A combination of cDMARDs and Biologics in the treatment of early RA has been able to achieve drug free remissions in only a small minority of patients. Complete remissions are rare when patients are treated with cDMARDs alone. Presented here is the clinical description of patients with RA who achieved long-term drug free clinical remissions (CR) with cDMARDs alone at a single rheumatology clinic in northern India and remained in CR over a long period (at least 6 months). The immune-biology of such states and the clinical factors leading to the prolonged CR will be discussed next.

Biography:

Li Yu-Jung is currently a Lecturer at St. Mary’s Medicine, Nursing and Management College. He has completed his training program of Oral and Maxillofacial Surgery in Veterans General Hospital-Taipei, Taiwan during 2002-2006. He is also a Doctoral candidate majored in Mechanical and Electrical Engineering from National Taipei University of Technology. He has received his MS degree of Clinical Dental Science from Institute of Clinical Dentistry, National Yang-Ming University. He has also received MS degrees of Chemistry and Biophysics from Graduate Institute of Biophysics, National Central University and Institute of Chemistry, Tamkang University during 2006-2010 and Bachelor’s degree of Dentistry from Department of Dentistry, Chung Shan Medical University in 2002.

Abstract:

Due to lack of pulp structure and periodontal ligament (PDL), which are regarded as the mainly pain origins of tooth painful sensation, dental implant have the opportunity for painless molecular releasing and blood monitoring with long-term steady and continuous properties. The new pathway may allow functional peptides and even the bigger molecular releasing and monitoring, which is impossible to absorb throughout the gastrointestinal (GI) tract. Therefore we design the replaceable drug delivery and bio-sensing modules above the titanium dental implant fixture which is immobile inside the maxillary bone marrow. The drug delivery module contains the piezoelectric micro-pump, the drug container and the power supply inside, while the bio-sensing module is constructed by the integrated circuit (IC), the Bluetooth module and the power supply. The total loading volume of the drug delivery module is around 0.5-1 ml and the drug is polymerized due to safety concern. Therefore the drug releasing type throughout this module is slowly and continuously diffused into the surrounding blood pool inside the bone marrow. The released drug type also needs to be carefully selected to avoid surrounding bony destructions. In contrast, the biosensor may provide various molecular types of continuous blood monitoring within five minute intervals and lasting for about 1 month within current technology. With standard dental and medical protocol establishments, the device may provide more useful applications in clinical practice.

Biography:

Andreia Ribeiro has completed her Biology Degree and Microbiology Master in Aveiro University in Portugal. Since 2010 she has been working as Research Assistant in Flow Cytometry and Immunology Groups in Portugal and in Ireland. In 2013, she has started her PhD in the DECIDE consortium and completed in 2016 from National University of Ireland, Galway.

Abstract:

Extracellular matrix (ECM) plays an important role in the tumor microenvironment and in biologic processes such as hematopoiesis. ECM contributes to regulation of cell survival, proliferation and cell differentiation. The aims of this project were to study the quality, quantity and biological role of ECM produced by a cloned mouse mesenchymal stromal cell line (MS5) on cell differentiation and to study the role of hypoxia on cell differentiation. To carry out these studies, we have used two methods of producing ECM in vitro. In both methods ECM is produced in normoxia and hypoxia. In method 1, cells are lysed by osmotic shock with a Tris/EDTA buffer, the standard way of preparing ECM in many studies. In method 2, MS5 were transduced with a caspase 9 vector, allowing induction of apoptosis in the cells following ECM production. Balb/c bone marrow mesenchymal stromal cells (MSC) were then seeded either in uncoated plastic dishes or in dishes covered with ECM and differentiation assays were performed, again either in normoxia or hypoxia. Results show that the two methods produce qualitatively different ECM and that hypoxia plays a role in ECM composition. Moreover, compared with hypoxia, normoxia is a better condition for adipogenic differentiation of fresh MSC. In contracts, osteogenic differentiation is better on ECM in hypoxia. In conclusion, different methods of preparing ECM in vitro lead to different protein composition and different outcomes in cell differentiation. Hypoxia also makes a difference in ECM composition and cell differentiation.

Biography:

Roghaye Arezumand has completed her PhD from Pasteur Institute of Iran in 2015. She has published about 10 papers in scientific journals.

Abstract:

Nanobody is smallest antigen binding domain derived from camelids family. The small size and other evolutionary property could introduce it as novel drug candidate especially against cancer. Over expression of angiogenesis is a highlighted character of tumor tissues. Many angiogenic factors like VEGF family involved in new vessels formation in cancerous tissues. Placenta growth factor (PLGF) is highly expressed in pathologic condition of angiogenesis. Targeting of PLGF could have inhibitory effect on angiogenesis in cell and animal model. The aim of this study is targeting of PLGF by developed novel nanobody. We constructed a PLGF nanobody library in pHEN-4 phagemid vector. This library specify by biopanning on immobilized recombinant PLGF. After screening of individual colonies, we selected different nanobody for soluble expression. Affinity of this nanobody was done by ELISA based method and the effect of this nanobody on angiogenesis were assessment by proliferation, migration, invasion, 3D capillary formation and chorioallantoic membrane assay (CAM). This nanobody could inhibit the proliferation, migration and tube formation of HUVEC cells and invasion of MDA-MB231 breast cancer cells. In addition of in vitro assays, this nanobody could inhibit the neo-vascular formation of fertilized eggs. A novel and high affinity specific-Nb against PLGF with anti-angiogenesis effects on endothelial, breast cancer cells and on in vivo model was developed in this investigation.

Biography:

University of Baghdad, Iraq

Abstract:

Background: Hepatitis C virus (HCV) is a serious infectious disease that can cause lifelong infection. Infection with chronic hepatitis C virus (HCV) can lead to autoimmune hepatitis (AIH) in a minority of patients. Viral infection induces tumor necrosis factor (TNF-alpha) production in hepatocytes. On the other hand prolactin which is an endocrinal hormone acts as a cytokine and is also involved in immune responses. These findings suggest that both parameters may have an important role in the patho-physiology of human liver diseases induced by viruses. Aim: The aim of the presents study was evaluate the role of the immunoendocrine system in the pathogenesis of the disease, by measuring serum prolactin and tumor necrosis factor-alpha. Subject and Methods: Sixty- one chronic hepatitis C patients were consequently selected from the Medical city, Gastrointestinal Hospital in Baghdad, Iraq, during the period from July 2014 to September 2014, their median age was 34.8 year, 29 of them were males and 32 were females. All patients were diagnosed having positive for HCV RNA by means of polymerase chain reaction. The study also included twenty apparently healthy adult age and sex matched considered as controls, which were negatively screened with hepatitis C virus. Peripheral blood sample of 2 ml was aspirated using disposal syringes. Samples were collected between (9.00a.m-12.00p.m). The blood was allowed to clot in plain tube for 30-45 minutes at room temperature. Sera were obtained by centrifugation of the collected blood and then stored in plain tubes at -20 c. ELISA method was used to measure (TNF and Prolactin) Results: The results of this study showed an increase in mean value of both TNF and prolactin hormone in chronic hepatitis C patients. However no significant correlations were found between both parameters studied. Conclusions: Chronic hepatitis C is associated with an immunological abnormality mainly represented by tumor necrosis factor-alpha and prolactine. This might shed a light of the type of therapy and drug of choice when managing the disease.

Biography:

Roman Mehling has studied Biology at the Karls Eberhard University of Tubingen (Tubingen, Germany), focusing on Microbiology. He has started his research as a PhD student in the field of Preclinical Imaging at the Werner Siemens Imaging Center, University of Tubingen in 2015. His research focuses on impact of reactive oxygen species and NF-kB signaling during hapten-induced skin inflammation.

Abstract:

Canonical NF-κB activation (p50) is involved in opposing immune functions (Cell 1995; 80:321-30). As the role of canonical NF-κB activation in adaptive immune responses is still ambiguous we analyzed acute and chronic contact hypersensitivity reactions in NF-κB-p50-/- mice. We sensitized wild-type and NF-κB-p50-/- mice at the abdomen (TNCB) and challenged one ear seven days later to elicit acute CHRS. Chronic CHSR was induced by repetitive TNCB-challenges for up to five times. CHSR was analyzed by measuring the ear thickness, histopathology and examination of ROS-stress non invasively in vivo with L-012-optical imaging (OI). In addition, we analyzed the leucocyte subsets in lymph nodes/spleens and the expression of NF-κB driven genes in inflamed ears. We determined a strongly enhanced acute and chronic CHRS in NF-κB-p50-/- mice when compared to WT mice. NF-κB-p50-/- mice exhibited an even up to two-fold increased ear thickness. H&E histology of NF-κBp50-/- mice revealed an impressively enhanced edema and a strongly pronounced leucocytic infiltrate dominated by polymorphonuclear leukocytes. Determination of ROS-stress exhibited significantly increased levels in ears from NF-κBp50-/- mice during acute and chronic CHSR. DHR-flow cytometry analysis of spleens of NF-κBp50-/- mice uncovered enhanced ROS expression by leucocytes and of CD4+CD69+ cells. Finally we uncovered an impressively enhanced IL-1β, IL-6 and TNF-alpha mRNA expression in ears of NF-κBp50-/- mice with acute and chronic CHSR. Our data suggest an anti-inflammatory role of canonical NF-κB activation in delayed-type hypersensitivity reactions. Thus, specific modulation of NF-κB signaling may be therapeutic tool for treatment of psoriasis or rheumatoid arthritis.

Biography:

Kerstin Fuchs has completed her PhD at the age of 31 years at the University of Tübingen and is actual involved in several postdoctoral studies from the Werner Siemens Imaging Center in Tübingen in the Department of Preclinical Imaging and Radiopharmacy. She is a group leader for imaging inflammation/immunology and has published 12 papers so far. She acts as reviewer of differnt journals and conferences mainly in the field of preclinal imaging.

Abstract:

Monocytic MDSCs (M-MDSC) and G-MDSCs are immature myeloid cells (CD11b+ Gr1+) and important regulators of basic immune responses. To date the role of MDSCs during RA is quite unknown; hence the aim of this study was to analyze the occurrence and homing of MDSCs during the effector phase in the GPI-serum RA mouse model. Further, we aimed to track 64Cu-PTSM or 64Cu-1A8 labeled G-MDSC after i.v. injection in RA mice by PET/MRI in vivo. BALB/c mice were injected with GPI-Ab to induce RA. We isolated cells from lavage of RA and healthy ankles at days 1, 3 and 6 and performed FACS analyis to identify G-MDSCs (Ly6G+) and M-MDSCs (Ly6C+). Ly6G antibody (Ab) or isotype Ab were used to deplete G-MDSCs. We labeled G-MDSCs intracellular with 64Cu-PTSM or an Ly6G-Ab (1A8) 64Cu-1A8. Labeled cells were i.v. injected in RA mice on day 6 and tracked their homing patterns by PET/MRI. On day 1 less than 10%, day 3 up to 55% and day 6 up to 70% of infiltrating CD11b+ cells were identified as G-MDSCs. Contrary the expression of M-MDSCs in arthritic ankle lavage was not affected. G-MDSC depletion with Ly6G Ab on day 6 after RA induction reduced the number of GMDSC to less than 10%. In vivo 64Cu-PTSM-G-MDSCs from RA ankles homed to arthritic ankles 1h post injection (5.5±0.6 %ID/cc) and 3.9±0.5%ID/cc after 24h. In contrast 64Cu 1A8-G-MDSCs showed enhanced homing into GPI-arthritic ankles (1h: 7.2%±3.0%ID/cc; 24h: 7.7±2.6%ID/cc) compared to 64Cu-PTSM-G-MDSCs. Thus, our data indicate that most of the infiltrating cells in inflamed ankles are G-MDSCs and depletion of Ly6G+ cells, lead to a massive decrease of G-MDSCs and arthritic joint inflammation, indicating a new therapeutic approach. 64Cu-1A8G cell labeling showed promising results for tracking G-MDSCs in vivo.

Biography:

Mohammad Sayyed Bakheet has completed his MD from Alazhar University and Postdoctoral studies from Alazhar University School of Medicine. He is the Assistant Professor of Biochemistry. He has published more than 25 papers in reputed journals and has been serving as an Editorial Board Member of repute.

Abstract:

TNF-α directly damages the vascular endothelial cells, reduces regional blood flow, causes occlusion of vessels and increases endothelial permeability. Endothelial cell injury after TNF-α mediated activation of immune system may result in secretion of vasoactive substances and increase in vascular permeability and intravascular coagulation. TNF-α may be involved in the pathogenesis of preeclampsia and may identify the patients who are at high risk of PE and can be a potential marker of the severity of the preeclampsia syndrome. Women with preeclampsia had deranged lipid profile due to abnormal lipid metabolism; this alteration of lipid metabolism may play a key role in the development of symptoms of Pre-eclampsia. Furthermore, changes to lipid metabolism may contribute towards the endothelial lesions observed in pre-eclampsia.

Biography:

Stephen D Miller is the Judy Gugenheim Research Professor of Microbiology, Immunology at Northwestern University Feinberg School of Medicine and Director of the Northwestern Interdepartmental Immunobiology Center. He is internationally recognized for his research on pathogenesis and regulation of autoimmune diseases. He has published over 370 journal articles, reviews and book chapters and has trained multiple generations of scientists. His work has significantly enhanced understanding of immune inflammatory processes underlying chronic autoimmune diseases focusing on translation of tolerance therapies induced by antigen linked biodegradable PLG nanoparticles for the treatment of autoimmunity, allergy and tissue/organ transplantation.

Abstract:

Ag specific tolerance is the desired therapy for immune mediated diseases. Our recent phase-1 clinical trial showed that infusion of myelin peptide coupled autologous apoptotic PBMCs induces dose dependent regulation of myelin specific T cell responses in MS patients. Antigen coupled apoptotic leukocytes accumulate in the splenic marginal zone (MZ) and are engulfed by F4/80+ MZ macrophages and CD8+ DCs inducing up regulation of PD-L1 in an IL-10 dependent manner. Tolerance results from the combined effects of PD-L1/PD-1 dependent T cell anergy and activation of Tregs recapitulating how tolerance is normally maintained in the hematopoietic compartment in response to uptake of senescing blood cells. To further advance clinical translation of tolerogenic therapies, we have shown that long lasting tolerance is inducible by I.V. administration of (auto) antigens covalently linked to or encapsulated within 500 nm carboxylated poly(lactide-co-glycolide) (PLG) nanoparticles (Ag-NP) abrogating development of Th1/Th17 mediated autoimmune diseases (EAE, T1D and celiac disease) and Th2 mediated allergic airway disease when used prophylactically and ameliorating progression of established disease when administered therapeutically. Ag-NP induced tolerance is mediated by the combined effects of cell intrinsic anergy and Treg activation and is dependent on route of administration, particle size and charge, uptake by MZ and live APCs via the MARCO scavenger receptor. As with tolerance induced by Ag-coupled apoptotic PBMCs, Ag-NP tolerance is induced and maintained by the combined effects of PD-L1/PD-1 dependent T cell anergy and activation of both Foxp3+ iTregs and Tr1 regulatory cells. These findings demonstrate the utility of Ag-NP as a novel, safe and cost effective means for inducing antigen specific tolerance for (auto) immune mediated diseases using an FDA approved biomaterial easily manufactured under GMP conditions.

Biography:

Johannes Schwenck studied Medicine at the Eberhard Karls University Tubingen. His MD thesis was recently awarded by the “Promotionspreis” of the University of Tubingen. Currently he is working as a Resident in the Department of Nuclear Medicine (University Hospital of Tubingen) and as a PhD Student in the Werner Siemens Imaging Center (University Hospital of Tubingen).

Abstract:

NF-B and ROS are crucial regulators of inflammation with considerable crosstalk between each other. Using non invasive in vivo optical imaging (OI) we uncovered the dynamics of ROS and NF-B-activation in a model of acute and chronic DTHR. We assessed NF-B-activation in NF-B luciferase reporter mice (n=10) and ROS expression with a ROS detecting chemiluminescence probe (L-012, n=8) using in vivo OI. Mice were sensitized with TNCB at the abdomen and challenged at the right ear 7 days later (acute DTHR). To induce chronic DTHR we challenged mice every 48 hours up to 5 times. Ear-thickness (ET) and in vivo optical imaging (OI) was measured 0-24 hours after the 1st, 3rd and 5th challenge. We analysed ear tissue by histology and RT-PCR. In acute CHSR ET increased steadily until 24 hours, whereas ROS and NF-B activity was first detectable 12 hours after the 1st challenge and peaked at 24 hour. After the 3rd challenge we measured at 4 hours enhanced ROS and NF-B activity, which culminated at 12 hours. After the 5th challenge ROS peaked at 4 hours, whereas NF-B activity increased till 12 hours. Histology revealed a strong edema in acute CHSR and massive tissue remodelling and a dense leukocyte infiltrate during chronic CHSR. ROS induced HO-1-mRNA was enhanced after the 1st and 3rd challenge. NF-B driven IL-1β and IL-6-mRNA was impressively elevated after the 3rd, but not after the 1st and 5th challenge. In vivo OI uncovered the differential temporal dynamics of ROS and NF-B expression during acute and chronic CHSR. This opens new windows of opportunity for innovative treatments of DTHR like rheumatoid arthritis.

Biography:

Kerstin Fuchs has completed her PhD at the University of Tubingen and is involved in several Postdoctoral studies from the Werner Siemens Imaging Center in Tubingen in the Department of Preclinical Imaging and Radiopharmacy. She is a Group Leader for Imaging Inflammation/Immunology and has published 12 papers so far. She acts as Reviewer of different journals and conferences mainly in the field of preclinical imaging.

Abstract:

Monocytic MDSCs (M-MDSC) and G-MDSCs are immature myeloid cells (CD11b+ Gr1+) and important regulators of basic immune responses. To date the role of MDSCs during RA is quite unknown; hence the aim of this study was to analyze the occurrence and homing of MDSCs during the effector phase in the GPI-serum RA mouse model. Further, we aimed to track 64Cu-PTSM or 64Cu-1A8 labeled G-MDSC after I.V. injection in RA mice by PET/MRI in vivo. BALB/c mice were injected with GPI-Ab to induce RA. We isolated cells from lavage of RA and healthy ankles at days 1, 3 and 6 and performed FACS analysis to identify G-MDSCs (Ly6G+) and M-MDSCs (Ly6C+). Ly6G antibody (Ab) or isotype Ab were used to deplete G-MDSCs. We labeled G-MDSCs intracellular with 64Cu-PTSM or Ly6G-Ab (1A8) 64Cu-1A8. Labeled cells were I.V. injected in RA mice on day 6 and tracked their homing patterns by PET/MRI. On day 1 less than 10%, day 3 up to 55% and day 6 up to 70% of infiltrating CD11b+ cells were identified as G-MDSCs. Contrary the expression of M-MDSCs in arthritic ankle lavage was not affected. G-MDSC depletion with Ly6G Ab on day 6 after RA induction reduced the number of GMDSC to less than 10%. In vivo 64Cu-PTSM-G-MDSCs from RA ankles homed to arthritic ankles 1 hour post injection (5.5±0.6 % ID/cc) and 3.9±0.5% ID/cc after 24 hours. In contrast 64Cu 1A8-G-MDSCs showed enhanced homing into GPI-arthritic ankles (1 hour: 7.2%±3.0% ID/cc; 24 hours: 7.7±2.6% ID/cc) compared to 64Cu-PTSM-G-MDSCs. Thus, our data indicate that most of the infiltrating cells in inflamed ankles are G-MDSCs and depletion of Ly6G+ cells lead to a massive decrease of G-MDSCs and arthritic joint inflammation, indicating a new therapeutic approach. 64Cu-1A8G cell labeling showed promising results for tracking G-MDSCs in vivo

Biography:

Abstract:

Objectives: To describe the clinical outcome and safety of rituximab (RTX) treatment in systemic lupus erythematosus (SLE) patients with severe manifestations or refractory to standard immunosuppressive therapy treated at a single center. Methods: Retrospective analysis of all patients with SLE treated with RTX at one center between June 2000 and December 2013. Clinical outcome was assessed by determining BILAG scores, anti dsDNA and C3 levels before and six months after RTX treatment. For safety analysis, adverse events and deaths were recorded. Results: Of a total of 115 patients, 93.9% were female; mean age at diagnosis was 26.39±11.90 years: Mean disease duration at first RTX treatment was 91.96±84.80 months. A BILAG score variation of -11.26±11.38 (p<0.001) was recorded six months after first RTX treatment; 40% of patients had a complete response and 27% had a partial response; in 36.5% of patients, C3 levels increased over 25% and in 33.5% dsDNA levels decreased over 50%. Depletion of CD19+ cells was achieved in 94.0% of patients. Hypogammaglobulinemia was detected in 14.9% of patients with significant reduction for IgM (p<0.001) and IgG (p=0.001) levels. Severe infections, infusion related and hypersensitivity reactions occurred in 7%, 3.5% and 2.6% of patients. Of the 115 patients, 62 patients had repeated RTX treatments with an average number of 1.95±1.17 cycles per patient and a mean interval between infusions of 21.44±20.11 months. At the end of follow up, 11 patients were deceased; 6 had cardiovascular events. Conclusion: RTX treatment was effective in decreasing disease activity with low incidence of adverse effects.

Biography:

Sankar Bhattacharyya has completed his PhD from Bose Institute, India and Postdoctoral studies from NIAID-NIH, USA and Department of Molecular Pathology, University of Wuerzburg, Germany. He is presently a Faculty Member of the Department of Zoology, Sidho Kanho Birsha University. He has published more than 20 papers in reputed journals such as Immunity, Journal of Experimental Biology, cancer research etc., and received INSPIRE Faculty Award from Department of Science and Technology (DST), Government of India.

Abstract:

Established tumor cause suppression of host immune system and recent studies indicate that a chronic pro inflammatory situation develops along with neoplastic progression. MDSCs are a group of heterogeneous immature myeloid lineage cells with potent T-cell suppressor activity that accumulate during inflammation and cancer. Although a good deal is known about suppressive effect of established tumor on host immune system, very little reports are available on effect of growing tumor on host immune system during very initial days. In order to study role of tumor immune interaction and influence of pro inflammatory milieu on tumor establishment we used mice model and a liquid transplantable murine tumor, EAC and carried out immunophenotyping studies. So far our study indicates that during very initial stage of tumor establishment a large number of T cells, majority of which is of activated/memory status infiltrate the tumor site. As tumor start to establish itself through direct and indirect intervention it depletes the tumor site T cell population and/or make them functionally inactive. Secondary lymphoid organ T cell reserve is not significantly affected though. We also observed significant infiltration of MDSC at tumor sit and increased proportion of these cells in bone marrow of tumor bearing animals, spleen remained relatively unaffected. An increased presence of macrophage was noticed in the ascites during initial stage but replaced by immature cells latter on. Together our data provides preliminary indication that tumor initially progress like pathogenic infection and immune system reacts similarly to this initial tumor cell population as it does against classic infection, causing increased infiltration of T cells and macrophages at tumor site, but as tumor starts to establish it manipulates microenvironment to deplete infiltrating T cells or render them ineffective.

Biography:

Abstract:

It has been shown that two types of cells (Th1/Th2 cells) play critical roles in defensive immune responses and in immunopathological disorders such as allergic reactions and autoimmune diseases and the methods of detecting Th1 and Th2 cells have become more important. The purpose of this research is to compare the level of Th1 cells (IL-2, IFN-γ), Th2 cells (IL-6, IL-10) and serum cortisol hormone in young athlete and non athlete men. This is an applied research that its data is gathered by free method. So in order to carry out the research among healthy and volunteer people, 20 of them (weight: 78± 4.1 Kg, height: 179 ± 3.59 ms, age: 26±4.38 BMI: 24±1.51) are sorted in athletes group (scientific group) and 12 of them (weight: 79±4.62 Kg, height: 181 ± 2.51 ms, age: 25±4.32 BMI: 24±1.63) in non athletes group (control group). The scientific group includes that athlete who has at least 6 months of regular aerobic exercise and the control group is the peoples who do not have any sport experience. Blood samples for evaluating the above factors in ELISA method are taken from both groups. Considering that the data were normal, with a T test, the independent student in the meaningful level of (P≤0/005) is examined. Comparing the average amount of IL-2, IL-6, IL-10, IFN-γ and serum Cortisol hormone in both groups, the amount of IL-6 and cortisol hormones in the scientific group has reduced considerably (p<0/05). The results of this research showed that aerobic exercise changed the balance of Th1/Th2 to TH1; this change is an effect of the reduction of TH1 anti-inflammatory cytokines and can be helpful in cure of rheumatic and allergic sickness.

Biography:

Abstract:

The ability to mount a strong immune response against pathogens is crucial for mammalian survival. However, excessive and uncontrolled immune reactions can lead to autoimmunity. Unraveling how the reactive versus tolerogenic state is controlled might point toward novel therapeutic strategies to treat autoimmune diseases. The surface receptor Toso/Faim3 has been linked to apoptosis, IgM binding and innate immune responses. In this study, we used Toso deficient mice to investigate the importance of Toso in tolerance and autoimmunity. We found that Toso(-/-) mice do not develop severe experimental autoimmune encephalomyelitis (EAE), a mouse model for the human disease multiple sclerosis. Toso(-/-) dendritic cells were less sensitive to Toll-like receptor stimulation and induced significantly lower levels of disease associated inflammatory T-cell responses. Consistent with this observation, the transfer of Toso(-/-) dendritic cells did not induce autoimmune diabetes indicating their tolerogenic potential. In Toso(-/-) mice subjected to EAE induction, we found increased numbers of regulatory T cells and decreased encephalitogenic cellular infiltrates in the brain. Finally, inhibition of Toso activity in vivo at either an early or late stage of EAE induction prevented further disease progression. Taken together, our data identify Toso as a unique regulator of inflammatory autoimmune responses and an attractive target for therapeutic intervention.

Biography:

Abstract:

Background: Leukocyte Fc receptors for immunoglobulin G (FcRs) play a major role in the handling of immune complexes and pathogens in systemic lupus erythematosus (SLE) and periodontitis. Both diseases have been shown to be partly influenced by genetic components including FcR genotype. The aim of the present study was therefore to evaluate whether FcR gene polymorphisms are associated with periodontitis risk in Egyptian SLE patients. Methods: The study subjects consisted of 100 SLE patients with periodontitis (SLE+P), 100 SLE patients without periodontitis (SLE), 100 healthy subjects with periodontitis (P) and 100 healthy subjects without periodontitis (H), who were all unrelated Egyptian non smokers. Genomic DNA was isolated from peripheral blood and FcγR genotypes for 3 biallelic polymorphisms (FcγRIIa-R131/H131, FcγRIIIa-158V/158F and FcγRIIIb-NA1/NA2) were determined by PCR-RFLP and allele specific polymerase chain reactions. Results: The SLE+P group was found to have more levels of periodontal destruction than the P group (P<0.001). A significant over representation of the FcγRIIa-R131 allele was found in the SLE+P group compared to the H group (SLE+P versus H: P<0.001, odds ratio, OR=10.553, 95% confidence interval, 95% CI=6.33-17.594). The FcγRIIa-R131 allele was also found to be overrepresented in the SLE+P group compared to the SLE group (SLE+P versus SLE: P-value <0.001, OR=4.317, 95% CI=2.818-6.613). The FcγRIIa-R131 allele was also found to be overrepresented in the SLE+P group compared to the P group (SLE+P versus P, P-value=0.001, OR =1.987, 95% CI=1.335-2.958). The frequencies of FcyRIIIa alleles among the subject groups showed SLE+Periodontitis group to have statistically significantly higher percentage of (F) allele than control group (SLE+Periodontitis vs. Control, VxF, P-value=0.001, OR=2.154, 95% CI=1.370-3.385). Same results were obtained when SLE+Periodontitis group was compared to periodontitis group (SLE+Periodontitis vs. periodontitis, VVxFF, P value<0.001, VxF, P value<0.001, OR=2.667, 95% CI=1.705-4.171). Same results were obtained when SLE+Periodontitis group was compared to SLE group (SLE+Periodontitis vs. SLE, VVxFF, P value=0.002). Same results were also obtained when SLE group was compared to control group (VVxFF, P value=0.030, VxF, P value=0.029, OR=1.615, 95% CI=1.048-2.489). Same results were also obtained when periodontitis group was compared to control group (VVxFF, P value<0.001). Conclusion: These results show the FcγRIIA-R131, FcγRIIIA-158F alleles to be associated with periodontitis risk in Egyptian SLE patients.

Biography:

Mohammad Afzal Khan has completed his PhD from Aligarh Muslim University, India and Postdoctoral studies from Stanford University School of Medicine, USA. He is a Scientist at Organ Transplant Centre in King Faisal Specialist Hospital and Research Centre, KSA. He has published more than 40 papers in reputed journals and has been serving as an Editorial Board Member of reputed Journal Cell & Tissue Transplantation & Therapy

Abstract:

Microvascular loss may be a root cause for chronic rejection in all solid organ transplants, which leads to the bronchiolitis obliterans syndrome (BOS), a fibrotic remodeling resulting in progressive narrowing of small airways. Previous research implicates T regulatory cell (Treg) as a potential mediator of microvascular repair. However, Treg has never been examined as an actual cause of graft hypoxia and ischemia during allograft rejection. We have reported the importance of functional microvasculature in the prevention of epithelial loss and fibrosis due to CD4+ T cells mediated rejection. The orthotopic tracheal transplant (OTT) model is ideal for studying the role of Treg mediated immune suppression in the rejection-associated airway hypoxia and ischemia implicated in chronic lung transplant rejection. In this study, we investigated that Treg mediated immune suppression (CD4+ T cells) promotes microvascular reestablishment and thus affects the progress of chronic rejection. Balb/C→C57/Bl6 allografts were adoptively transfer with Tregs (1x10^6) I.V. at d0 and allografts were monitored from day-2 to da-28 for tissue pO2, blood perfusion and functional microvasculature during acute rejection. Our data demonstrate that targeted immune suppression by Tregs significantly improves tissue pO2, microvascular flow at day 10 post transplantation, followed by sharp rise in IL-10 and IL-5 gene expression compared to untreated WT controls. However, Tregs treatment in WT is able to significantly (p<0.05) delay acute (from d10 to d14) rejection alone but not able to prevent acute rejection. These findings conclude that protecting airway microvasculature with Treg therapy facilitates microvascular re-establishment and shortens the phase of hypoxia in allograft. These findings can be translated to adjunct therapy in combination with existing transplant therapies especially with low-dose rapamycin (promotes Treg expansion) to improve the efficacy of Treg therapy.

Biography:

Mohammad Afzal Khan has completed his PhD from Aligarh Muslim University, India and Postdoctoral studies from Stanford University School of Medicine, USA. He is a Scientist at Organ Transplant Centre in King Faisal Specialist Hospital and Research Centre, KSA. He has published more than 40 papers in reputed journals and has been serving as an Editorial Board Member of reputed Journal Cell & Tissue Transplantation & Therapy

Abstract:

Microvascular loss may be a root cause for chronic rejection in all solid organ transplants, which leads to the bronchiolitis obliterans syndrome (BOS), a fibrotic remodeling resulting in progressive narrowing of small airways. Previous research implicates T regulatory cell (Treg) as a potential mediator of microvascular repair. However, Treg has never been examined as an actual cause of graft hypoxia and ischemia during allograft rejection. We have reported the importance of functional microvasculature in the prevention of epithelial loss and fibrosis due to CD4+ T cells mediated rejection. The orthotopic tracheal transplant (OTT) model is ideal for studying the role of Treg mediated immune suppression in the rejection-associated airway hypoxia and ischemia implicated in chronic lung transplant rejection. In this study, we investigated that Treg mediated immune suppression (CD4+ T cells) promotes microvascular reestablishment and thus affects the progress of chronic rejection. Balb/C→C57/Bl6 allografts were adoptively transfer with Tregs (1x10^6) I.V. at d0 and allografts were monitored from day-2 to da-28 for tissue pO2, blood perfusion and functional microvasculature during acute rejection. Our data demonstrate that targeted immune suppression by Tregs significantly improves tissue pO2, microvascular flow at day 10 post transplantation, followed by sharp rise in IL-10 and IL-5 gene expression compared to untreated WT controls. However, Tregs treatment in WT is able to significantly (p<0.05) delay acute (from d10 to d14) rejection alone but not able to prevent acute rejection. These findings conclude that protecting airway microvasculature with Treg therapy facilitates microvascular re-establishment and shortens the phase of hypoxia in allograft. These findings can be translated to adjunct therapy in combination with existing transplant therapies especially with low-dose rapamycin (promotes Treg expansion) to improve the efficacy of Treg therapy.

Biography:

Barbara Franziska Schorg has studied at the University of Hohenheim (Stuttgart, Germany), focusing on molecular biology and virology. After her studies, she started her research as a PhD student in the field of Preclinical Imaging at the Werner Siemens Imaging Center, University of Tuebingen. Her research focuses on T-cell based immunotherapies and tumor microenvironment in a progressive pancreatic cancer mouse model. Moreover, she puts considerable effort in the supervision and execution of several important PET/MRI cooperation studies.

Abstract:

Tumor cells and immune cells express inhibitory immune-checkpoint (ICP) ligands that paralyze tumor infiltrating T-cells. ICP-specific antibodies can restore T-cell-functions. Moreover, tumor-antigen (TA)-specific IFN-y secreting CD4+ T-cells (Th1) mediate strong anti-tumoral effects and can induce senescence in cancer cells. Thus, we establish a highly efficient TA-Th1-cell and checkpoint inhibitor based combined immunotherapy (CIT) in RIP1-Tag2 (RT2) mice with advanced endogenous pancreatic insular cell carcinomas where ICP-blockade and TA-Th1-cells alone are not sufficient. We started our CIT in 10-11 weeks old RT2-mice which usually die with 14 weeks. At this stage BGL are reduced due to enhanced insulin secretion. Mice underwent a single 2Gy whole body radiation followed by repeated injections of TA-Th1-cells (1x weekly) and of LAG-3+PD-L1-mAbs (1-2x weekly) or isotype-Abs. During CIT-treatment, the median BGL of RT2-mice increased from 80 mg/dl to nearly normal values of 91 mg/dl (week 14, n=30, p<0.01) while the BGL of RT2-mice treated with TA-Th1-cells+isotype mAbs already dropped to 66 mg/dl (n=30). Treated with LAG-3+PD-L1-mAbs alone was inefficient (week 14: 40 mg/dl, n=23). CIT-treatment extended the lifespan of the CIT-treated RT2-mice from 14 to 21 weeks. CIT-treated RT2-mice revealed very small tumors and a strong lymphocytic infiltrate. In contrast, LAG-3+PD-L1-mAbs treated RT2-mice exhibited large, vascularized endocrine tumors with a slight lymphcytic infiltrate. Insular cell carcinomas of CIT-treated RT2-mice revealed high p16 and low Ki67 expression but opposite results in the control groups. Thus, our CIT is applicable to reinforce Th1-cell based immunotherapies and to induce carcinoma regression even in mice with progressed carcinomas.

Biography:

Abstract:

Background: Induction of a strong hepatitis C virus (HCV) specific T-helper 1 (Th1) T-cell response plays a pivotal role in control and clearance of the virus. A multi-epitope vaccine containing T-cell epitopes could be a promising vaccination strategy against HCV, but further computational evaluations are important before initiating the experimental study. Method: In the present study, we have employed various approaches to design an efficient multi-epitope vaccine. First, CD8+ cytolytic T-lymphocytes (CTLs) epitopes, helper epitopes and adjuvant which are three essential components of peptide vaccine were determined. CTL epitopes were selected from HCV genotype 1a/1b, consensus regions of non structural protein 3 (NS3) and non structural protein 4A (NS4A) by various servers. NS3 derived sequences by various servers were used to induce CD4+helper T-lymphocytes (HTLs) responses. Heparin-Binding Hemagglutinin (HBHA), a novel TLR4 agonist, was applied as an adjuvant to polarize CD4+T cells toward T-helper 1 to induce strong CTL responses. Then obtained epitopes were linked together by appropriate linkers to enhance epitope presentation. 3D model of protein was generated and physicochemical properties, stability and allergenicity of the protein were predicted using bioinformatics tools and servers. Results: Our results indicated that more than 90% residues locate in favorite or additional allowed region of Ramachandran Plot. Also, based on Ramachandran plot analysis this protein could be classified as a stable fusion protein. In addition, this multi-epitope protein had strong potential to induce specific T-cell response against HCV. Conclusion: Our results supported that this multi-antigenic vaccine could be effectively considered as an efficient vaccine for prophylactic or therapeutic usages.

Biography:

Philip R Hardwidge is an Associate Professor at Kansas State University. His laboratory is interested in understanding, treating and preventing diarrheal disease caused by bacterial pathogens. His research team has discovered several mechanisms by which bacterial proteins subvert the host innate immune system to promote bacterial colonization and transmission. He is directing his knowledge of these proteins and their mammalian targets to innovative studies of metabolic syndromes, autoimmune disorders and cancer. He is also developing proteomic techniques to identify vaccine targets in other organisms.

Abstract:

Many bacterial pathogens utilize a type-III secretion system (T3SS) to inject virulence proteins (effectors) into host cells to subvert various biological functions. Effector subversion of pro-inflammatory host responses is well studied, but less attention has been given to the potential inhibition of host interferon (IFN) signaling. Type-I IFNs are important both to maintaining intestinal homeostasis and to responding to pathogen infection. Pathogens have evolved strategies to interfere with host type-I IFN production. A recent study found both that IFN-β is induced by enteropathogenic E. coli (EPEC) infection and that the EPEC T3SS effector NleD inhibits IFN-β induction. IFN expression is known to be important to limiting Citrobacter rodentium infection but whether C. rodentium T3SS effectors inhibit host IFN-β induction is unclear. We screened C. rodentium strains bearing deletions in individual T3SS effectors to determine the extent to which this pathogen might inhibit the host IFN-β response. To determine if C. rodentium T3SS effectors inhibit the host type-I IFN response, we monitored the survival of a recombinant vesicular stomatitis virus (VSV). Since TRAF3 is critical to IFN signaling, we also monitored effector mediated inhibition of the TNF receptor (TNFR) associated factor 3 (TRAF3) ubiquitination in RAW264.7 cells. Supernatants from cells infected with C. rodentium escN inhibited VSV to levels similar to those induced by LPS treatment. By contrast, supernatants from cells infected with WT C. rodentium did not inhibit VSV-GFP growth. These data suggested that a T3SS-effector inhibits the production of a host factor involved in virus inhibition. We then infected HeLa cells with C. rodentium strains lacking individual T3SS effector genes and screened the cell supernatants for anti-viral activity. nleB inhibited virus replication most significantly. By monitoring TRAF3 activity in C. rodentium infected cells, we also revealed the selective impact of NleB on K63 linked TRAF3 ubiquitination. We conclude that the T3SS effector NleB inhibits host IFN-β production by reducing the extent of the activation associated K63 linked TRAF3 ubiquitination.

Biography:

Bozena Futoma-Koloch was graduated from University of Wrocław in 2004. She has received her PhD in Microbiology from the University of Wrocław in 2008. After that, she has held an academic position of Assistant Professor in the Department of Microbiology in the Institute of Genetics and Microbiology at University of Wroclaw. She is a Member of Polish Society of Microbiologists, European Society of Clinical Microbiology and Infectious Diseases and International Complement Society. She has published more than 50 papers in national and international journals, 30 presentations at congresses with several awards. Her research program focuses on bacterial surface antigens as molecular targets of the protective immune response and their role in bacterial resistance to disinfectants.

Abstract:

The mechanisms used by bacteria to avoid host immunological defenses are not entirely understood. Microorganisms getting into contact with human blood or plasma have developed a variety of strategies to evade complement attack. One strategy is the incorporation of the sialic acid into the bacterial surface glycoconjugates that usually results in an increase in serum resistance. Nothing is known about the influence of sialylated bacterial surface structures on C3 fixation in serum. The role of the outer membrane proteins (OMP) in Salmonella susceptibility to serum has not also been investigated thoroughly. Therefore, C3 deposition on the O48 group of Salmonella bacteria has been studied. The tested microorganisms Salmonella O48 are characterized by sialylated lipopolysaccharide (LPS) and different patterns of OMP. Our investigations showed that bacteria were sensitive to human serum (HS) although they possessed sialylated LPS. We found that the greatest C3 deposition occurred on Salmonella sv. Isaszeg cells with low content of sialic acid in LPS. A weaker C3 deposition ratio was noted to Salmonella vs. Ngozi and Salmonella subsp. arizonae with the high contents of sialic acid in LPS. Immunoblotting revealed that C3 complement protein bound to OMP common to three tested strains. We suggest that the differential sensitivity of tested bacteria to HS may be due to a weaker C3 activation on strongly sialylated LPS and a binding of C3 components to the OMP.

Biography:

Jessy S Deshane is a pulmonary Immunologist with expertise in immune regulation in asthma. She investigates myeloid-derived regulatory cell biology and free radical mechanisms that regulate their differentiation and function. She pioneered these investigations both in mouse models and human asthma. She has authored 46 peer-reviewed publications, including high impact journals like Journal of Experimental Medicine, Journal of Clinical Investigations, Journal of Allergy and Clinical Immunology, Immunity and Cancer Research. She serves on the Editorial Boards for the journals Allergy and American Journal of Respiratory Cell and Molecular Biology and serves on grant review committees.

Abstract:

Rationale: Bronchial thermoplasty (BT) is a therapeutic option for a subset of asthmatics who continue to be symptomatic despite high dose glucocorticoid therapy. Mechanisms underlying the impact of BT on airway inflammation are largely unknown. Myeloid-derived regulatory cells (MDRCs) are important regulators of chronic inflammation in human asthma. There is also increasing appreciation for a relationship between airway microbiome and respiratory inflammation. We hypothesized that the longitudinal changes in diversity and or abundance of the airway microbiome may modulate immune regulation by MDRCs following BT and contribute to the beneficial outcome of BT. Methods: Bronchial washings (BW) and peripheral blood samples were collected at each of the three bronchoscopic procedures for BT from 5 patients with severe asthma. Multi-parameter flow cytometry was performed to enumerate MDRC subsets and regulatory T-cells (Treg). Microbial genomic DNA was isolated, 16S rDNA genes were amplified using V4 primers and PCR products were sequenced using the Illumina MiSeq platform. Sequences were processed, integrated, analyzed and reported using QIIME and in-house software. Statistical significance of longitudinal changes in microbial signatures and cell proportions were determined by linear mixed model regression analysis. Correlation between microbiome composition and time points was determined by PERMANOVA. Microbial phyla and cell MDRC subsets were correlated by linear mixed model regressing one variable to the other adjusting time points as a fixed effect and sample ID as a random effect. All analyses were conducted in the lme4 R package. Results: Significant longitudinal changes were noted at the level of phyla in comparisons of operational taxonomic unit and time points of bronchoscopies. Significant reduction in both proteobacteria (P=0.04) and verrucomicrobia (P=0.020) was observed in the BW. Correlation analyses revealed significant correlations between microbial alterations and proportions of MDRC subsets in the BW following BT. Alterations in phyla tenericutes and verrucomicrobia significantly correlated (P=0.039,P=0.0131) with the reduction in proportions of immunosuppressive CD14+CD16-HLA-DR- MDRCs in BW. Changes in phyla proteobacteria and fusobacteria correlated (P=0.002, P=0.017) with the enhancement of CD14+CD16+HLA-DR- MDRCs in the BW. Importantly, changes in kingdom Archaea; phylum euryarchaeota correlated with the progressive reduction of pro-inflammatory CD163+HLA-DR+CD11b+CD33+MDRCs (P=0.00003) and enhancement of the CD4+CD25+CD127loFoxp3+Tregs (P=0.0135) and CD14+CD16+HLA-DR- MDRCs (P=0.044) in the BW. Conclusions: Correlation of progressive modulation of airway microbiome and myeloid cell dynamics suggest a functional relationship/interaction between these components and an important contribution to immune regulation in the airways that can account for some of the benefits observed in patients following BT.

Biography:

Hanan Al-Khalifa has obtained her Master’s degree in Parasitological Diseases and Immunology at University of Manchester and completed her PhD in 2007 in the University of Reading, UK, investigating the effect of n-3 fatty acids on the immune response and general health status. Her interests include but are not limited to immunological techniques, parasitological diseases, effect of nutrition, espicially fatty acids, on the immune status in both humans and expermental animals. She executed many research projects that focused on the effect of nutrition on immunology. She has attended many scientific events and published more than 60 papers in refereed journals and conference proceedings.

Abstract:

Abnormal numbers of specific types of leukocytes may indicate immuno-suppression or immunocompetence in response to an immunomodulator. The objective of this study is to investigate the effect of feeding broilers on diet containing flaxseed on splenocyte T and B-cells. Upon hatching, all chicks were given the same basal diet for 13 days. Dietary supplementation of flaxseed started at 14 days of ages until the end of the cycle at 35 days of age. At slaughter, samples of spleen were collected. Spleen cells were harvested in cell suspensions. Immune cells were then enumerated using the flow haemocytometer in the Physiology Laboratory. The overall differences between the dietary treatments were analyzed using one-way analysis of variance (ANOVA) and the general linear model procedure of Minitab. Statistically, there was no significant effect of flaxseed on the percentage positive or mean fluorescence intensity of the leukocyte subsets under investigation. However, there was a trend towards an increase in the proportion of B-cells in the spleen after feeding 15% of flaxseed, which approached significance (P=0.058). There was also a trend towards a decrease in the mean fluorescence intensity of CD8+ subsets in the spleen, which was close to statistical significance (P=0.054). This trend is particularly interesting, given the fact that the bursa in these chickens were significantly observed to be smaller and thinner. It suggests that flaxseed may either prevent the homing of B-lymphocytes to the bursa or encourage the release of B-lymphocytes from the bursa into the circulation and then in the spleen.

Biography:

Laboratory for Molecular Infection Medicine Sweden (MIMS), Sweden

Abstract:

Key to the activation of the innate immune system are the pattern-recognition receptors (PRRs) including Toll-like receptors (TLRs), RIG-I-like receptors (RLRs), and cytoplasmic DNA receptors (CDRs). While essential for protection against infections, activation of PRRs require tight control to avert inflammatory diseases. The mechanisms underlying this strict regulation are unclear. The reversible attachment to and removal of ubiquitins from proteins by ubiquitin ligases and deubiquitinases respectively is a versatile system that regulates diverse aspects of biology including the immune system. The H2A deubiquitinase MYSM1 (H2A-DUB) MYSM1 is a nuclear metalloprotease previously described as a key component of epigenetic signaling machinery. We now show that MYSM1 is a master negative regulator of PRR pathways. In response to infections or inflammation, we found that MYSM1 rapidly accumulates in the cytoplasm. In the cytoplasm MYSM1 interacts with and inactivates key signaling complexes for the TLR, RLR and CDR pathways via the removal of K63 linked polyubiquitin chains. Hence mice deficient in MYSM1 are hyper-responsive to various innate immune stimuli, exhibit resistance to viral infections but are more susceptible to inflammatory disease such as sepsis. These results highlight MYSM1 as a key negative regulator of the innate immune system that protects against an overzealous self-destructive immune response.

Biography:

Un-Hwan Ha has completed his PhD in Microbiology from the University of Florida in 2002 and Postdoctoral studies in Cellular Microbiology from House Ear Institute and University of Rochester Medical Center. In 2008, he has begun to serve as an Assistant Professor at the Department of Biotechnology and Bioinformatics, Korea University and is currently positioned as a Professor. He has published more than 20 papers in reputed journals since 2008.

Abstract:

Pseudomonas aeruginosa is a Gram negative opportunistic bacterial pathogen that has a notorious reputation about drug resistance against commonly used antibiotics as well as an infectious agent to the respiratory tract of immunocompromised patients along with other microbial invaders. P. aeruginosa possesses diverse secretory systems, which play critical parts in releasing a number of virulence factors that are involved in causing acute and chronic infections. The potential effects of these factors on the modulation of host defense responses have been proposed. However, the resulting modulation effects against competitive bacteria, such as Staphylococcus aureus, are unknown since the clinical impact of polymicrobial diseases caused by combinations of pathogens has received much attention from the medical community. Here, we report that components secreted from P. aeruginosa enhance the expression of bradykinin receptors which act as important host defense responses against invading microbes by interacting with a ligand, bradykinin. In addition to this, LPS as a well-known membrane associated molecule pattern of P. aeruginosa induces the expression of TLR2, which plays a dominant role in sensing PAMPs typically expressed by Gram positive bacteria. Up-regulation of TLR2 influences the magnitude of proinflammatory responses to the secondary S. aureus infection. Moreover, P. aeruginosa Ndk with the aid of flagellin, increases the expression of interleukin-1 which is an important pro-inflammatory cytokine via NF-κB/inflammasome pathways. Taken together, the results of this study demonstrate that P. aeruginosa is capable of modulating host defense responses through the actions of associated or released virulence factors and this may have impacts on against a secondary microbial infection.

Biography:

Eva E Avila is a Professor at the Universidad de Guanajuato, Mexico. Her research interest is in innate immunity defense, especially antimicrobial peptides and some virulence mechanisms of the parasites Trichomonas vaginalis and Entamoeba histolytica. She also enjoys teaching in bachelor and postgraduate levels.

Abstract:

Trichomonas vaginalis is a flagellated parasite that causes human trichomoniasis; this is the non-viral most common sexually transmitted disease worldwide. Trichomoniasis is associated with the premature birth of newborns, with infertility and with increased susceptibility to human immunodeficiency virus and papillomavirus infections. This infection is characterized by a heavy inflammatory response with abundant number of neutrophils. Neutrophils, the most abundant cells in the bloodstream, have a main function to eliminate the pathogenic microorganisms through phagocytosis, degranulation and the formation of neutrophil extracellular traps (NETs). NETs are DNA fibers associated with histones and antimicrobial peptides that trap and prevent the spread of pathogens. The purpose of this research was to characterize the interaction between Trichomonas vaginalis and human neutrophils in vitro. The formation of NETs was activated by trophozoites and by its surface lipophosphoglycan (LPG), which was reduced in the presence of an antibody to TLR-4, suggesting the participation of this receptor in NETs formation induced by T. vaginalis. NETs trapped trophozoites, as observed by confocal microscopy and after a 3-hour interaction, the viability of T. vaginalis decreased significantly. These results suggest that neutrophil extracellular traps are effective against T. vaginalis; however, the presence of excessive number of neutrophils during infection may also contribute to the damage of epithelial mucosa.

Biography:

Tewodros Firdissa Duressa has obtained his PhD at KU Leuven. His research focuses Locust cellular innate immunity.

Abstract:

Within the order Insecta, knowledge about the innate immune response is mainly based on studies regarding holometabolous model organisms: In both fruit fly and moths the cellular immune response of phagocytosis, nodule formation and encapsulation is supported by a strong humoral pro-inflammatory response including coagulation, prophenoloxidase activation and antimicrobial peptide synthesis. In hemimetabolous locusts, genes encoding the classical antimicrobial peptide precursors are missing and they mainly depend upon their efficient cellular immune response. This elaborated cellular immune response has its trade off as it results in a steep decrease in the number of circulating hemocytes. Immune challenge makes a GBP hemocyte spreading peptide activating the hemocytes. Most tissues except gut and hemocytes display a high basal transcription of this cytokine. Spreading and increased adhesive character of activated hemocytes partly explains their rapid disappearance and reappearance following infection. It turned out that in adult locusts the persisting “hematopoietic organ” is not involved in replenishment of lost hemocytes but rather has a prophylactic function as main phagocytotic organ at later age. The instant hemocyte replenishment by circulating prohemocyte stem cells supports this hypothesis, an immune challenge by either Gram positive or Gram negative bacteria but not by a challenge with fungi results in a significant selective increase in expression of insect angiotensin converting enzyme in Locusta migratoria hemocytes. Knockdown of Locusta ACE by both RNAi and captopril inhibitor elucidated the involvement of ACE, either direct or indirect, in the appearance of LPS induced hemolymph peptides of which most have a so far unidentified function.

Biography:

Tzung-Yan Lee has completed his PhD degree from National Yang-Ming University and Postdoctoral studies from Institute of Biological Chemistry, Academia Sinica. He has published more than 50 papers in reputed journals and has been serving as an Editorial Board Member of repute.

Abstract:

An important initiator of the inflammatory response to obesity is adipose tissue, which is involved in obesity induced insulin resistance and chronic inflammation. Electroacupuncture (EA) shows anti-inflammation and several pleiotropic effects that interact with metabolic pathways. Numerous studies have demonstrated the clinical efficacy of acupuncture in weight loss. However, the precise mechanism of its potential effect related to adipose tissue remains poorly understood. Obese animals treated with EA showed significantly reduced body weight. EA decreased the number of F4/80 and CD11b positive macrophages in epididymal adipose tissue. We found that EA at Zusanli (ST36) acupoints significantly alleviated macrophage recruitment and then improved the obesity associated factors of sterol regulatory element binding protein (SREBP)-1 and target genes expression in obese animals. Adipose tissue tumor necrosis factor-α (TNF-α), interleukin-1 (IL-6), monocyte chemotactic protein-1 (MCP-1) and CD68 mRNA expression were significantly reduced by EA treatment in obese animals. On the other hand, EA significantly down-regulated HIF-1α level in a time course dependent manner in ob/ob mice. The expression level of hypoxia related genes (VEGFA, Slc2al, GPX1) and inflammation related genes (TNF-α, IL-6, MCP-1) were also poorly expressed in adipose tissue after EA treatment. This phenomenon was paralleled by the levels of inflammatory cytokines, such as TNF-α, IL-6 and IL-1β in obese mice. We conclude that EA offers a beneficial effect on adipose tissue mass in obese animals, at least partly, via attenuation of lipogenesis signaling, thus resulting in improved inflammatory response. Therefore, EA prevents weight gain through modulation of HIF-1α-dependent pathways and inflammatory response in obese adipose tissues.

Biography:

Mira Barda-Saad is a returning Scientist from the National Cancer Institute at NIH in Maryland, Senior Lecturer at the Mina and Everard Goodman Faculty of Life Sciences. She is currently examining the molecular signaling mechanisms controlling immune cell response with the primary goal of relating this knowledge to pathophysiological conditions of the immune system. She believes that understanding the dynamic behavior of signaling and cytoskeletal molecules that control immune cell activation is essential for identification of targets relevant for the treatment of cancer, autoimmune diseases and immunodeficiencies.

Abstract:

Natural killer (NK) cells represent a powerful weapon of immune defense against viral infections and tumor growth via the cytotoxicity of target cells and the production of cytokines. NK cell function is regulated by a balance between activating and inhibitory signals. Cancer cells or viruses often perturb this balance by expressing ligands for activating NK cell receptors and by down-regulating ligands for the inhibitory receptors, i.e., MHC class I molecules, resulting in target cell killing. Engagement of inhibitory receptors, including the killer cell immunoglobulin-like receptor (KIR), antagonizes activating pathways through the recruitment and activation of the SH2-containing protein tyrosine phosphatase-1 (SHP-1) to the NK immunological synapse (NKIS). To date, only the signaling molecule VAV1 was clearly demonstrated as a direct substrate of SHP-1 in human NK cells. Since SHP-1 activity is the major mechanism that prevents NK cell autoimmune response, it is of great importance to determine whether additional substrates of SHP-1 exist and whether additional molecular mechanisms down-regulate NK cell activation. Moreover, the mechanisms that control SHP-1 activity remain to be unraveled. In the present study, we demonstrate that in response to KIR receptor engagement, SHP-1 and the E3 ubiquitin ligases Cbls negatively regulate the linker for the activation of T cells (LAT) and phospholipase Cγ (PLCγ) 1/2. LAT dephosphorylation by SHP-1 abrogated PLCγ 1/2 recruitment to NKIS and decreased calcium flux and degranulation, thus abolishing NK cell cytotoxicity. Furthermore, LAT ubiquitylation via c-Cbl and Cbl-b following NK cell inhibition leads to its degradation and to the down-regulation of NK cell activation. Using a cutting-edge microscope system, we follow this cellular signaling cascade from the moment of encounter through target-cell killing. Our data suggest that LAT phosphorylation triggers its ubiquitylation, implying a collateral inhibitory mechanism in which a pool of phosphorylated LAT that escapes SHP-1 dephosphorylation is targeted to proteasomal degradation. These mechanisms serve as a key checkpoint in tuning NK cell activation threshold and the immune response.

Biography:

Nayef Jarrous is currently working at “The Hebrew University of Jerusalem, Israel”. His research interest is based on “Human nuclear RNase P ribonucleoprotein in tRNA processing”. He has published many articles in reputed journals.

Abstract:

RNA polymerase III (Pol III) is a key player in innate immunity, as it serves as a sensor of viral and bacterial DNA of infected cells. This sensing asset is based on promoter independent recognition of foreign DNA templates in the cytoplasm and transcription via nonspecific initiation mechanism. The resulting 5’-triphosphate RNA transcripts activate the retinoic acid induced gene I (RIG-I), thus leading to induction of type-I interferon. We have previously shown that the human catalytic ribonucleoprotein RNase P is implicated in formation of proficient initiation complexes of nuclear Pol III on 5S rRNA and tRNA genes. However, it was unknown if this ribonucleoprotein is also implicated in nonspecific initiation of gene transcription by cytoplasmic Pol III. We will present preliminary results that show that the H1 RNA subunit of human RNase P is implicated in promoter independent initiation of transcription of synthetic circular DNA templates (COLIGOs) by cytoplasmic Pol III. The regulatory role of H1 RNA in this transcription system may explain the existence of RNase P like RNA genes in DNA viruses and the possible roles of their transcripts in evading antiviral innate immune responses.

Biography:

Sumru Savas is an Internal Medicine Specialist since 1999, Graduate of European Academy for Medicine of Ageing (2015). She is currently a PhD student in Elderly Health-Gerontology (From 2011) at Ege University Health Sciences Institute. She is an Internist and Lecturer at Geriatrics Section of Internal Medicine Department.

Abstract:

Lipoprotein associated phospholipase A2 (Lp-PLA2) is a reported risk factor for dementia. However, the relationship between Alzheimer’s disease (AD) and Lp-PLA2 is still debatable and to the best of our knowledge, no study has evaluated the associations between levels of Lp-PLA2, proinflammatory cytokines and neopterin in AD. In total, 59 patients with AD and 38 non-demented individuals were included in the case control study. Fasting serum concentrations of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), neopterin and Lp-PLA2 were determined using ELISA. The associations between AD and each of the variables were analyzed by logistic regression. The median Lp-PLA2 levels in AD and controls were similar (P=0.29, not significant). Median serum neopterin and IL-6 levels were significantly higher in patients with AD than in controls (P=0.0001 and P=0.03, respectively). In regression analyses, median neopterin levels, a lower level of education and female gender were significantly associated with AD when compared with controls (OR, 31.44, 95% CI 3.59-275.28, P=0.002; OR, 4.35, 95% CI 1.13-16.61, P=0.032; OR, 7.25, 95% CI 1.88-28.00, P=0.004, respectively). In contrast to previous evidence suggesting its role in dementia and AD, Lp-PLA2 enzyme levels were higher in the controls and no relationship between Lp-PLA2 and either proinflammatory cytokines or neopterin was identified in AD. Elevated neopterin levels may be considered inflammatory markers of AD.

Biography:

Andreia Marques Ribeiro has completed her graduate degree in Biology and Master’s degree in Microbiology from Aveiro University in Portugal. Since 2010, she has been working as a Research Assistant in Flow Cytometry and Immunology groups in Portugal and in Ireland. In 2013, she started her PhD in the DECIDE consortium and completed in 2016 from National University of Ireland, Galway.

Abstract:

Mesenchymal Stromal Cells (MSC) possesses immunomodulatory and anti-inflammatory properties, having several effects on immune cells. For this reason MSC have been proposed as a potential therapeutic modality for osteoarthritis (OA) and rheumatoid arthritis (RA). However, there is a commercial need to have a reliable, rapid, quantifiable assay to assess the potency of allogeneic human MSC. The aims of this assay were to determine by flow cytometry the effects of MSC from bone marrow (BM MSC) and adipose tissue (ASC) on TNF-α and IL-6 production by LPS stimulated monocytes of healthy and patient samples. A number of factors were considered prior to optimizing the assay, including: Brefeldin A concentration, type of anticoagulant (heparin; K2EDTA or citrate), LPS concentration; blood dilution and incubation time. MSC numbers were then titrated and co-cultured with whole blood of healthy donors. All results are expressed as intracellular expression of TNF-α and IL-6, on gated monocytes identified as CD45+CD14+ cells using an Accuri four color flow cytometer. Result show that BM MSC and ASC significantly reduced TNF-α and IL-6 expression by monocytes from healthy donors and in OA and RA patients. Thus, we have established a rapid, reliable and quantifiable screening assay to determinate the effects of MSC on LPS activated monocytes. Such an assay could be used to screen the recipients’ monocytes for inhibition by the MSC preparation that will be injected, thereby contributing to personalized medicine. For this reason this assay is being used in the ADIPOA2 clinical trial.

Biography:

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Abstract:

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