"Pruning" of alloreactive CD4+ T cells using 5- (and 6-)carboxyfluorescein diacetate succinimidyl ester prolongs skin allograft survival.
Removal of alloreactive cells by either thymic deletion or deletion/anergy in the periphery is regarded as crucial to the development of tolerance. Dyes, such as CFSE, that allow monitoring of cell division suggest that in vitro proliferation could be a used as a way of "pruning" alloreactive cells while retaining a normal immune repertoire with retention of memory to previously encountered pathogens. This would overcome the problems occurring as a result of therapies that use massive depletion of T cells to allow acceptance of organ transplants or bone marrow grafts. We therefore used a skin graft model of CD4-mediated T cell rejection across a major H-2 mismatch (C57BL/6 (H-2(b)) to BALB/c (H-2(d)) mice) to evaluate whether nondividing CD4(+) T cells derived from a mixed lymphocyte culture would exhibit tolerance to a skin graft from the initial stimulator strain. We demonstrate that selective removal of dividing alloreactive CD4(+) T cells resulted in marked specific prolongation of allogeneic skin graft survival, and that the nondividing CD4(+) T cells retained a broad TCR repertoire and the ability to maintain memory. This novel way of depleting alloreactive T cells may serve as a useful strategy in combination with other mechanisms to achieve transplant tolerance.
Watson
Debbie
D
Center for Kidney Research, The Children's Hospital at Westmead, New South Wales, Australia.
Zhang
Geoff Yu
GY
Sartor
Mary
M
Alexander
Stephen I
SI
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism / Q000637:transplantation; D002455:Cell Division / Q000276:immunology; D002469:Cell Separation / Q000379:methods; D018984:Epitopes, T-Lymphocyte / Q000008:administration & dosage / Q000276:immunology; D005260:Female; D005452:Fluoresceins / Q000378:metabolism*; D006082:Graft Enhancement, Immunologic / Q000379:methods; D006084:Graft Rejection / Q000517:prevention & control*; D007156:Immunologic Memory; D008213:Lymphocyte Activation; D007959:Lymphocyte Culture Test, Mixed; D008212:Lymphocyte Depletion / Q000379:methods; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D011948:Receptors, Antigen, T-Cell / Q000096:biosynthesis; D016038:Skin Transplantation / Q000276:immunology* / Q000379:methods / Q000473:pathology; D013045:Species Specificity; D013388:Succinimides / Q000378:metabolism*; D016176:T-Lymphocyte Subsets / Q000166:cytology / Q000276:immunology / Q000378:metabolism / Q000637:transplantation; D014184:Transplantation, Homologous
C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D015415:Biomarkers; D018984:Epitopes, T-Lymphocyte; D005452:Fluoresceins; D011948:Receptors, Antigen, T-Cell; D013388:Succinimides