Murine renal transplantation procedure.

Renal orthotopic transplantation in mice is a technically challenging procedure. Although the first kidney transplants in mice were performed by Russell et al over 30 years ago (1) and refined by Zhang et al years later (2), few people in the world have mastered this procedure. In our laboratory we have successfully performed 1200 orthotopic kidney transplantations with > 90% survival rate.

Spontaneous renal allograft acceptance associated with "regulatory" dendritic cells and IDO.

MHC-mismatched DBA/2 renal allografts are spontaneously accepted by C57BL/6 mice by poorly understood mechanisms, but both immune regulation and graft acceptance develop without exogenous immune modulation. Previous studies have shown that this model of spontaneous renal allograft acceptance is associated with TGF-beta-dependent immune regulation, suggesting a role for T regulatory cells. The current study shows that TGF-beta immune regulation develops 30 days posttransplant, but is lost by 150 days posttransplant.

Evaluation of immune regulation in transplant patients using the trans vivo delayed type hypersensitivity assay.

Allograft recipient IL-10 and/or transforming growth factor-beta (TGF-beta) dependent anti-inflammatory T-cell delayed type hypersensitivity (DTH) responses to donor derived antigens, or regulatory T-cell responses, have been demonstrated in rodents and transplant patients using a previously described trans vivo DTH assay. We used this assay to determine the incidence of recipient anti-inflammatory T-cell responses to donor antigens in a large cohort (n = 420) of primary kidney and simultaneous kidney-pancreas transplant patients tested a mean of 4.8 years after transplantation.

A flow cytometry-based method for detecting antibody responses to murine cytomegalovirus infection.

An assay based on target cells infected with green fluorescent protein labeled murine cytomegalovirus (GFP-MCMV) and dual color flow cytometry for detecting antibody to MCMV is described. After optimizing conditions for this technique, kinetics of anti-MCMV IgG antibody response was tested in susceptible (BALB/c) and resistant (C57BL/6) mouse strains following primary MCMV infection. Previously published antibody kinssetics were confirmed in susceptible mice, with peak IgG response seen approximately 8 weeks after primary infection, decreasing by 20 weeks after infection.

Oxygen sensing by primary cardiac fibroblasts: a key role of p21(Waf1/Cip1/Sdi1).

In mammalian organs under normoxic conditions, O2 concentration ranges from 12% to <0.5%, with O2 approximately 14% in arterial blood and <10% in the myocardium. During mild hypoxia, myocardial O2 drops to approximately 1% to 3% or lower.

Allograft acceptance despite differential strain-specific induction of TGF-beta/IL-10-mediated immunoregulation.

We examined the immune approaches that C57BI/6 and BALB/c mice take when treated to accept cardiac allografts. C57BI/6 mice accept DBA/2 cardiac allografts when treated with gallium nitrate (GN) or anti-CD40L mAb (MR1). These allograft acceptor mice fail to mount donor-reactive delayed type hypersensitivity (DTH) responses, and develop a donor-induced immunoregulatory mechanism that inhibits DTH responses.

Evidence that a similar range of alloimmune responses can develop in murine and human allograft recipients.

Based on discrepancies between various laboratory and clinical observations, skepticism has developed regarding the pre-clinical value of rodent immunologic studies. In this article, we review the progression of our murine and human studies over the last several years, which have demonstrated that humans and mice can make very similar types of immune responses in vivo to allografts. Early studies by ourselves and others, demonstrated that mice can make either pro-inflammatory (rejection) or anti-inflammatory (acceptance) immune responses to graft alloantigens.

The graft helps to define the character of the alloimmune response.

Introduction: In mice, kidney and liver allografts may be spontaneously accepted, whereas cardiac and skin allografts in the same strain combinations are rapidly rejected. The reasons for this dichotomy in murine response outcomes remains to be determined.

Methods And Results: When DBA/2 (H-2d) cardiac allografts were placed in C57BL/6 (H-2b) recipients, they were rejected within 10 days, unless the allograft recipients were transiently treated with gallium nitrate (GN), at which time the allografts were accepted for > 150 days.