title: Linked suppression across an MHC-mismatched barrier in a miniature swine kidney transplantation model.
abstract: We have demonstrated previously that a 12-day course of FK506 permits the induction of tolerance to fully MHC-mismatched renal transplants in miniature swine. In the present study, we examined the mechanism of this tolerance by assessing the possibility that the survival of one-haplotype mismatched third-party kidneys might be prolonged via linked suppression. Ten SLA(d/d) miniature swine received fully MHC-mismatched renal allografts from SLA(c/c) donors with 12 days of FK506. Six animals received second SLA(c/c) kidneys without immunosuppression to confirm tolerance. Regulatory mechanisms were assessed by mixed lymphocyte reaction (MLR) and cell-mediated lympholysis coculture assays and ELISA for regulatory cytokines. Linked suppression was investigated by transplanting SLA(a/c) or SLA(a/d) allografts into long-term tolerant recipients without immunosuppression. All recipients showed donor-specific unresponsiveness in standard cell-mediated lympholysis and MLR assays. Tolerant cells prestimulated with donor Ag and then cocultured with naive recipient MHC-matched cells inhibited antidonor responses, confirming the presence of regulatory cells. ELISA and MLR assays showed that TGF-beta2 was involved in mediating the suppression in vitro. SLA(a/d) renal allografts transplanted into tolerant recipients were rejected by postoperative day 8 (median, 7 days; range, 6-8). In contrast, SLA(a/c) allografts showed markedly prolonged survival (median, 52 days; range, 28-78; p = 0.0246), suggesting linked suppression. Animals not challenged with a second donor-matched graft did not manifest linked suppression consistent with in vitro data showing that re-exposure to tolerated Ags is important for generation of regulatory cells. To our knowledge, these data represent the first evidence of linked suppression across fully MHC-mismatched barriers in a large animal model.
authors.affiliation: Transplantation Biology Research Center, Massachusetts General Hospital, Boston, MA 02129, USA.
authors.forename: Adam D
authors.initials: AD
authors.lastname: Griesemer
authors.forename: John C
authors.initials: JC
authors.lastname: Lamattina
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Okumi
authors.forename: Justin D
authors.initials: JD
authors.lastname: Etter
authors.forename: Akira
authors.initials: A
authors.lastname: Shimizu
authors.forename: David H
authors.initials: DH
authors.lastname: Sachs
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Yamada