Bardin, F Clerget, B Prum, F Corn is, for ECRAF de G ique Clinique, H ital Lariboisi e, Paris, France de Recherche Europ n pour la Polyarthrite Rhumato e, ECRAF-Universit d’Evry et de Paris VII, Evry-Genopole, France INSERM U, Le Kremlin Bic re, France Laboratoire de Biologie, Centre Hospitalier Sud Francilien, Corbeil Essonne, France Laboratoire Statistique et G ome, Genopole, Evry, France Arthritis Res Ther , (suppl):GenHotelLaboratoire UnitGene and cell therapy Effect of synovial fluid on adeno-associated virus mediated gene transfer to chondrocytesV Cottard, G Falgarone,, D Lutomski, MC Boissier,, N BessisUPRES EA – and Rheumatology Department, UniversitParis XIII, Paris, France Avicenne Hospital, Bobigny, France Arthritis Res Ther , (suppl):Background: HLA-DRB alleles coding for the shared epitope (SE), an amino acid motif in the third hypervariable region with the beta chain of HLA-DR, are connected with RA. The underlying biological mechanism is unknown. A model was not too long ago proposed (Reviron et al. Arthritis Rheum ; :), according to the electric charge (EC) of your fourth antigenic NAN-190 (hydrobromide) peptide-binding pocket (P) on the HLA-DR molecule. SE alleles carrying a sturdy positive EC at P, the model postulates that the negative EC of your P features a protective impact for RA that modulates the impact from the SE. Objective: To test that model, taking advantage of familial-based association evaluation. Strategies: The DNA of Caucasian French families with one particular RA patient and both parents had been genotyped for HLA-DRB. GenotypesBackground: Direct intra-articular gene tranfer with adeno-associated virus (AAV) vectors is a promising strategy to let effective therapeutic transgene expression within the joint. Even so, more than of humansSArthritis Research Therapy SupplAbstracts with the rd European Workshop for Rheumatology Researchdemonstrate antibodies against AAV which could alter AAV transduction efficiency in vivo. PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/23118721?dopt=Abstract Objective: Our objective was to determine the influence of synovial fluid (SF) and serum from arthritic sufferers on AAV-mediated gene transfer to chondrocytes in vitro. Solutions: SF and sera derived from individuals suffering from various (inflammatory or mechanical) origins have been collected. Neutralizing activity against AAVIL- was determined by assessing the capability of SF or serum to inhibit transduction of AAVIL- into the CA human chondrocyte cell line. Total IgGs have been purified from SF by salt-dependent chromatography. Results: In SF and sera, inhibition of AAV-mediated gene transfer to chondrocytes was observed in all arthritic individuals. Purified IgG from SF exhibited inhibition patterns equivalent to these observed with unfractionated SF. A correlation was observed involving levels of inhibition by the SF and the serum (P r .). Lastly, we’ve shown that the inhibition of AAVIL- infection on CA cells by SF and sera may be reversed by rising the number of AAVIL- particles, using a dose-dependent effect. Conclusion: SF and sera from all sufferers show a neutralizing activity against AAV infection on chondrocytes. Neutralizing factors had been IgG antibodies 
present ACP-196 biological activity inside the SF. This inhibition is often reversed by escalating AAV doses. In the future, these data may well be helpful to adapt intraarticular AAV gene therapy to every individual patient. Immunotherapy through injection of immature dendritic cells prevents collagen-induced arthritisP Plence, R Pfannes, D No , F Apparailly, J Sany, C Jorgensen Immuno Rheumatology, University Hospital Montpellier, France Arthritis.Bardin, F Clerget, B Prum, F Corn is, for ECRAF de G ique Clinique, 
H ital Lariboisi e, Paris, France de Recherche Europ n pour la Polyarthrite Rhumato e, ECRAF-Universit d’Evry et de Paris VII, Evry-Genopole, France INSERM U, Le Kremlin Bic re, France Laboratoire de Biologie, Centre Hospitalier Sud Francilien, Corbeil Essonne, France Laboratoire Statistique et G ome, Genopole, Evry, France Arthritis Res Ther , (suppl):GenHotelLaboratoire UnitGene and cell therapy Impact of synovial fluid on adeno-associated virus mediated gene transfer to chondrocytesV Cottard, G Falgarone,, D Lutomski, MC Boissier,, N BessisUPRES EA – and Rheumatology Department, UniversitParis XIII, Paris, France Avicenne Hospital, Bobigny, France Arthritis Res Ther , (suppl):Background: HLA-DRB alleles coding for the shared epitope (SE), an amino acid motif within the third hypervariable region on the beta chain of HLA-DR, are connected with RA. The underlying biological mechanism is unknown. A model was recently proposed (Reviron et al. Arthritis Rheum ; :), according to the electric charge (EC) of your fourth antigenic peptide-binding pocket (P) with the HLA-DR molecule. SE alleles carrying a robust constructive EC at P, the model postulates that the negative EC on the P includes a protective impact for RA that modulates the impact from the SE. Objective: To test that model, taking advantage of familial-based association evaluation. Procedures: The DNA of Caucasian French families with one particular RA patient and each parents were genotyped for HLA-DRB. GenotypesBackground: Direct intra-articular gene tranfer with adeno-associated virus (AAV) vectors is usually a promising method to enable effective therapeutic transgene expression inside the joint. On the other hand, over of humansSArthritis Study Therapy SupplAbstracts in the rd European Workshop for Rheumatology Researchdemonstrate antibodies against AAV which could alter AAV transduction efficiency in vivo. PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/23118721?dopt=Abstract Objective: Our objective was to determine the influence of synovial fluid (SF) and serum from arthritic patients on AAV-mediated gene transfer to chondrocytes in vitro. Techniques: SF and sera derived from individuals struggling with several (inflammatory or mechanical) origins had been collected. Neutralizing activity against AAVIL- was determined by assessing the ability of SF or serum to inhibit transduction of AAVIL- into the CA human chondrocyte cell line. Total IgGs were purified from SF by salt-dependent chromatography. Benefits: In SF and sera, inhibition of AAV-mediated gene transfer to chondrocytes was observed in all arthritic individuals. Purified IgG from SF exhibited inhibition patterns equivalent to these noticed with unfractionated SF. A correlation was observed amongst levels of inhibition by the SF plus the serum (P r .). Lastly, we’ve got shown that the inhibition of AAVIL- infection on CA cells by SF and sera could possibly be reversed by escalating the number of AAVIL- particles, with a dose-dependent impact. Conclusion: SF and sera from all sufferers show a neutralizing activity against AAV infection on chondrocytes. Neutralizing factors have been IgG antibodies present in the SF. This inhibition is often reversed by increasing AAV doses. Within the future, these information may well be beneficial to adapt intraarticular AAV gene therapy to each and every person patient. Immunotherapy by means of injection of immature dendritic cells prevents collagen-induced arthritisP Plence, R Pfannes, D No , F Apparailly, J Sany, C Jorgensen Immuno Rheumatology, University Hospital Montpellier, France Arthritis.