Selective increase in CD4-positive graft-infiltrating mononuclear cells among the infiltrates in class I disparate kidney grafts undergoing rejection

Long-term tolerance to kidney allografts across a two-haplotype class I disparity is uniformly induced in miniature swine with a short course of cyclosporine (CsA). In the absence of CsA, all recipients acutely reject kidney allografts within 2 weeks. Previous experiments have shown that graft-infiltrating mononuclear cells (GIC) migrate to the allograft in both CsA-treated and untreated animals. To evaluate the correlation between GIC phenotype and the clinical status, infiltrating cells were examined by flow cytometry, using selective gating to distinguish them from other renal cells. GIC from tolerant and rejector animals were mostly mature T cells, with 84% CD8+ cells, which consisted of 68% CD8+/CD4- and 16% CD8+/ CD4+ cells. This cellular phenotype was, however, markedly different from that of peripheral blood lymphocytes, suggesting a selective migration of cells into the graft. This selective process counterselected the CD3+/CD2- subset of GIC, which was never found in the graft. The distribution of GIC subsets was initially comparable in tolerated and rejected kidneys, but the CD4 single-positive subset then increased specifically in the allograft destined to rejection. The absence of CD4 single-positive cells in tolerated grafts was unlikely to be due to a direct effect of the CsA, because long-term tolerant animals, which received a second kidney without further immunosuppression, also showed no increase in CD4 single-positive cells. The fact that CD4 single-positive cells appeared only within the rejected kidneys, strongly suggests that this cell subset may be important in mediating immune rejection and supports the hypothesis that the development of tolerance in this model depends on a relative deficit of T-cell help.