Day 3 :
- Allergy and Therapathies Technological Innovations in Immunology Rheumatology Innate Immune Responses
Location: Berlin, Germany
Chair
Thomas Boldicke
Helmholtz Centre for Infection Research, Germany
Co-Chair
Hanan Al-Khalifa
Kuwait Institute for Scientific Research, Kuwait
Session Introduction
Thomas Boldicke
Helmholtz Centre for Infection Research, Germany
Title: Recent highlights of in vivo knockdown by intrabodies
Time : 10:00-10:20
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.
Hanan Al-Khalifa
Kuwait Institute for Scientific Research, Kuwait
Title: Effect of flaxseed supplementation on the innate immune response
Time : 10:20-10:40
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.
Laurence Macia
Charles Perkins Centre, School of medical sciences, University of Sydney, Australia
Title: Gut microbiota, bacterial metabolites and metabolite sensing GPCRs determine mucosal tolerance and protection against food allergy
Time : 10:40-11:00
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.
Sundeep Kumar Upadhyaya
Indraprastha Apollo Hospitals, India
Title: Long-term drug free clinical-remissions (CR) in patients with rheumatoid arthritis (RA) on cDMARDs followed prospectively at a single rheumatology clinic in northern India over two years
Time : 11:15-11:35
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.
Li Yu-Jung
St. Mary’s Junior College of Medicine, Taiwan
Title: Intra-maxillary molecular delivery and blood monitoring via dental implant
Time : 11:35-11:55
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.
Andreia Ribeiro
National University of Ireland, Ireland
Title: Effect of extracellular matrix and hypoxia on mesenchymal stromal cell differentiation
Time : 11:55-12:15
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.
Roghaye Arezumand
North Khorasan University of Medical Sciences, Iran
Title: Novel phage displayed derived nanobody against placenta growth factor inhibited in vitro and in vivo angiogenesis model
Time : 12:15-12:35
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.
Hedef Dhafir El-Yassin
University of Baghdad, Iraq
Title: The immune response of prolactin and the induction of tumor necrosis factor (TNF) in Iraqi patients infected with hepatitis C virus
Time : 12:35-12:55
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.