Injection of Donor-Derived OX62+ Splenic Dendritic Cells With Anti-CD4 Monoclonal Antibody Generates CD4+CD25+FOXP3+ Regulatory T Cells That Prolong Allograft Skin Survival Indefinitely and Abrogate Production of Donor-Specific Antibodies in a Rat Model

• Published in: Transplantation proceedings 2009-10, Vol.41 (8), p.3363-3366
• Main Authors: Alawieh, M, Rifle, G, Bouchot, O, Malapert, G, Mousson, C, Martin, L
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - therapeutic use; Biological and medical sciences; CD4 Antigens - immunology; Dendritic Cells - transplantation; Forkhead Transcription Factors - immunology; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Graft Survival - immunology; Interleukin-2 Receptor alpha Subunit - immunology; Isoantibodies - immunology; Medical sciences; Models, Animal; Rats; Rats, Inbred BN; Rats, Inbred F344; Rats, Inbred Lew; Skin Transplantation - immunology; Surgery; Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases; T-Lymphocytes, Regulatory - immunology; Tissue, organ and graft immunology; Transplantation, Homologous - immunology
• ISSN: 0041-1345; 1873-2623
• DOI: 10.1016/j.transproceed.2009.08.038

Abstract Objective To examine in a rat model the ability of donor dendritic cells and anti-CD4 monoclonal antibody (mAb) to generate donor-specific CD4+CD25+ regulatory T cells (Tregs) and to evaluate the capacity of these Tregs to prolong skin allograft survival and abrogate the production of donor-specific antibodies after skin grafting. Materials and Methods OX62+ (nonplasmacytoid) splenic dendritic cells were isolated from Fischer rats using magnetic beads and injected (2 × 106 ) into Lewis rat recipients with or without treatment with a nondepleting anti-CD4 (W3/25) mAb. After 4 weeks, splenic CD4+CD25+FOXP3+ T cells were harvested using magnetic beads from conditioned animals and injected (1 × 106 ) into naïve Lewis recipients (day 1) before they received a skin graft from a Fischer (n = 4) or a third-party (Norway; n = 4) donor rat. Donor-specific antibodies were detected in recipient blood using flow cytometric cross-matches with donor lymphocytes from day 0 to day 30 after grafting. Results After injection of conditioned CD4+CD25+FOXP3+ T cells, Lewis recipients accepted skin grafts from Fischer donors indefinitely (>100 days) but rejected third-party skin grafts. Donor-specific antibodies were detected at low levels in only 1 recipient receiving conditioned Tregs before grafting. Naive Tregs did not prolong skin graft survival. Conclusion These preliminary data suggest that splenic dendritic cells in combination with an anti-CD4 mAb induce donor-specific Tregs that indefinitely prolong allogeneic skin graft survival and inhibit donor-specific antibody production. Experiments are under way to determine whether this protocol can inhibit chronic lesions after heart transplantation in this model.