TY - JOUR
T1 - Regulatory mechanism of peripheral tolerance
T2 - In vitro evidence for dominant suppression of host responses during the maintenance phase of tolerance to renal allografts in miniature swine
AU - Wu, Anette
AU - Yamada, Kazuhiko
AU - Ierino, Francesco L.
AU - Vagefi, Parsia A.
AU - Sachs, David H.
N1 - Funding Information:
The authors would like to thank Drs Megan Sykes and Zachary L. Gleit for critical review of the manuscript, Jessica Sachs for editorial assistance and Lisa A. Bernardo for help in manuscript preparation. The authors would also like to thank Novartis Pharmaceutical, Co. for generously providing Cyclosporine. This work was supported in part by National Institutes of Health grants #2RO1 AI31046 and 2PO1 HL18646.
PY - 2003
Y1 - 2003
N2 - Background: Studies from our laboratory have demonstrated that a short course of cyclosporine leads to indefinite survival of renal allografts across an MHC class-I barrier in miniature swine. We have recently reported that a peripheral regulatory mechanism appears to be involved in the maintenance of this tolerance since peripheral blood lymphocytes (PBL), exposed to donor antigen in vitro specifically suppressed the generation of anti-donor cytotoxic activity by recipient-matched naive PBL. We have now further investigated the mechanism of this phenomenon to determine the level at which such regulation occurs, and investigated the phenotypes of the cells involved in maintaining dominant suppression. Materials and methods: PBL from long-term tolerant animals (> 6 months after renal transplantation) were pre-stimulated in vitro with donor-type PBL. These cells were then incubated with recipient-matched naive responders and donor-type PBL stimulators in MLR assays. The proliferative activity of the cells was measured by [3H]-thymidine incorporation. Suppression was measured by inhibition of proliferation of naive cells in response to donor PBL when co-cultured with tolerant cells. Flow cytometry was used to study the phenotypes of cells that were present in cell cultures. Results: Primed PEL from tolerant animals markedly suppressed the proliferative response of recipient-matched naive cells to donor-matched stimulators in vitro. No suppression of proliferation was observed in response to third party stimulators, indicating that the suppression was donor-specific. Primed PBL from naive animals stimulated with donor antigen and co-cultured with unprimed recipient-matched naive cells also demonstrated reduced proliferative responses. However, this decrease in proliferation appeared to be due to a 'burn-out' phenomenon, as assessed by kinetic studies, rather than due to true suppression. Expression of CD25 increased on a sub-population of T cells from tolerant animals following priming with donor antigen. These cell then markedly inhibited further activation of CD25 positive cells in co-cultures with naive responder cells, suggesting a possible mechanism of suppression. Conclusion: These results suggest that the mechanism of tolerance to class-I-mismatched renal allografts, involves a population of regulatory cells that are capable of suppressing proliferative anti-donor responses.
AB - Background: Studies from our laboratory have demonstrated that a short course of cyclosporine leads to indefinite survival of renal allografts across an MHC class-I barrier in miniature swine. We have recently reported that a peripheral regulatory mechanism appears to be involved in the maintenance of this tolerance since peripheral blood lymphocytes (PBL), exposed to donor antigen in vitro specifically suppressed the generation of anti-donor cytotoxic activity by recipient-matched naive PBL. We have now further investigated the mechanism of this phenomenon to determine the level at which such regulation occurs, and investigated the phenotypes of the cells involved in maintaining dominant suppression. Materials and methods: PBL from long-term tolerant animals (> 6 months after renal transplantation) were pre-stimulated in vitro with donor-type PBL. These cells were then incubated with recipient-matched naive responders and donor-type PBL stimulators in MLR assays. The proliferative activity of the cells was measured by [3H]-thymidine incorporation. Suppression was measured by inhibition of proliferation of naive cells in response to donor PBL when co-cultured with tolerant cells. Flow cytometry was used to study the phenotypes of cells that were present in cell cultures. Results: Primed PEL from tolerant animals markedly suppressed the proliferative response of recipient-matched naive cells to donor-matched stimulators in vitro. No suppression of proliferation was observed in response to third party stimulators, indicating that the suppression was donor-specific. Primed PBL from naive animals stimulated with donor antigen and co-cultured with unprimed recipient-matched naive cells also demonstrated reduced proliferative responses. However, this decrease in proliferation appeared to be due to a 'burn-out' phenomenon, as assessed by kinetic studies, rather than due to true suppression. Expression of CD25 increased on a sub-population of T cells from tolerant animals following priming with donor antigen. These cell then markedly inhibited further activation of CD25 positive cells in co-cultures with naive responder cells, suggesting a possible mechanism of suppression. Conclusion: These results suggest that the mechanism of tolerance to class-I-mismatched renal allografts, involves a population of regulatory cells that are capable of suppressing proliferative anti-donor responses.
KW - Regulatory cells
KW - Suppression
KW - Tolerance
KW - Transplantation
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U2 - 10.1016/S0966-3274(03)00006-6
DO - 10.1016/S0966-3274(03)00006-6
M3 - Article
C2 - 12967789
AN - SCOPUS:0141629820
SN - 0966-3274
VL - 11
SP - 367
EP - 374
JO - Transplant Immunology
JF - Transplant Immunology
IS - 3-4
ER -