Pathogenic antigens involved in the induction of Heymann nephritis (HN), an experimental rat model of human membranous nephritis, have been identified in megalin (gp330) and the receptor-associated protein (RAP) [1,2]. A pathogenic epitope has been identified in RAP (amino acid 1-86) that plays a significant role in the formation of immune deposits in glomeruli in HN. A synthetic peptide (P31-53) derived from RAP1-86 contains a pathogenic epitope recognized by antibodies eluted from glomerular immune deposits and includes two putative RT-1B1 MHC class II-binding motifs. We have investigated whether RAP P31-53 can be recognized by T cells. Five peptide-specific T cell lines were generated from regional lymph node (LN) T cells from Lewis rats immunized with P31-53. The T cell lines were characterized by using a T cell proliferation assay for their specificity, FACS and MHC restriction assay for the phenotype, reverse transcription-polymerase chain reaction for TCR Vbeta repertoire and cytokine expression, and cloning and sequencing for the analysis of the CDR3 sequence of TCR. The helper function of the T cell line was confirmed by autoantibody production in vitro. In this study, we clearly identify that the synthetic pathogenic peptide P31-53 contains a T cell epitope recognized by CD4+ Th2 cells in Lewis rats. This recognition was restricted by MHC class II RT1.B1. These CD4+ Th2 cells were able to promote B cells to produce specific antibodies and used a restricted set of TCR Vbeta genes with preferential usage of Vbeta18. A charged amino acid motif at the CDR3 region of predominant TCR Vbeta subfamilies may contribute to the specific ability of these cells to recognize the immunogenic T cell epitope within RAP peptide P31-53.

SummaryRegulatory T cells (T regs ) have been recognized as central mediators for maintaining peripheral tolerance and limiting autoimmune diseases. The loss of T regs or their function has been associated with exacerbation of autoimmune disease. However, the temporary loss of T regs in the chronic spontaneous disease model has not been investigated. In this study, we evaluated the role of T regs in a novel chronic spontaneous glomerulonephritis model of B cell lymphoma 2-interacting mediator (Bim) knock-out mice by transient depleting T regs . Bim is a pro-apoptotic member of the B cell lymphoma 2 (Bcl-2) family. Bim knock-out (Bim -/-

Islet cell transplantation as a therapy for type 1 diabetes has been limited by progressive graft loss. Significant immunosuppression including T-cell ablation has been used in an attempt to limit islet rejection. Here, we show that CD3+ lymphocytes depleted of alloreactive T cells selected from a mixed lymphocyte reaction (MLR), where responder BALB/c splenocytes stained with carboxyfluorescein succinimidyl ester (CFSE) were stimulated with irradiated C57BL/6 splenocytes for 5 days, infused into diabetic immunodeficient mice are capable of restoring a broad T-cell repertoire and specifically do not reject islet transplants from the strain (C57BL/6) used in the original depletion. These mice demonstrate reconstitution with CD4 + and CD8 + T cells, the capacity to reject third-party grafts (CBA), and restoration of interferon-g (IFN-g) responses to third-party alloantigens. Over time, both forkhead box P3-positive (Foxp3 + ) T regulatory cells (Tregs) and gd T cells expand, suggesting a role for peripheral tolerance, in addition to the initial depletion of alloreactive T cells, in long-term islet graft survival. Our results suggest that immune restoration with CD3 + lymphocytes where alloreactive T cells are removed can restore cognate immunity without islet allograft loss and recurrence of diabetes.