Alloreactive CD8+ T cells may persist in animals made tolerant of transplanted tissues; their function is controlled through continuous censorship by regulatory CD4+ T cells. We sought to establish the stage at which such censorship operates. We found that monospecific CD8+ T cells introduced into tolerant animals responded to the tolerated tissue antigen as if they had received CD4+ T cell “help”: they proliferated and accumulated normally. However, they did show compromised graft rejection, interferon-γ production and cell-mediated cytotoxicity. These findings suggest that tolerance mediated by regulatory T cells acts by censoring immune effector functions rather than by limiting the induction of T cell responses.
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We thank S. Humm for preparing many of the mAbs used in vivo and the PSB staff—especially M. Coates, K. Steventon and S. Laynes. Supported by a program grant from the Medical Research Council, UK and the Chang-Gung Memorial Hospital, Taiwan (to C. L.).
Tolerance induction to fully mismatched skin allografts. CBA mice were treated with three doses of 1 mg of the anti-CD4, anti-CD8 and anti-CD154 nondepleting mAbs over 1 week following B10 skin transplantation at day 0. These mice accepted the allografts indefinitely as well as new B10 skin transplants grafted at day 100 (filled squares, n = 6, MST > 200 days). The same mice remained competent to reject third-party BALB/c skin grafts, transplanted at day 145 (open squares, n = 6, MST = 16 days). Mice treated with the same mAbs without an initial tolerising skin allograft, rejected B10 skin grafts when transplanted at day 100 (filled triangles, n = 5, MST = 16 days) as well as BALB/c grafts transplanted at day 145 (open triangles, n = 5, MST = 15 days). Untreated controls readily rejected B10 skin allografts (filled circles, n = 10, MST = 10 days). (PDF 114 kb)
An agonist CD40 mAb can bypass help for CD8+ T cell expansion. Euthymic or thymectomized and CD4-depleted CBA mice were injected intravenously with 1 × 107 CFSE-labeled Des+ CD8+ T cells. The following day, mice were immunized intravenously with 1 × 106 (CBA × B10)F1 APCs or no APCs as a control. In addition, some mice received agonist CD40 mAb (FGK-45, 1 mg per mice) by i.p. injection. (a) The average number of divisions was calculated by the equation listed in the Methods. (b) Total number of Des+ CD8+ T cells in spleens of host mice. Six mice were included in each group. Data were pooled from three independent experiments. (PDF 129 kb)
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