Following organ transplantation soluble MHC class I is released from the graft and may contribute to alloimmunity. We determined in a well-established rat model whether DC are able to internalise soluble MHC class I alloantigen and then re-present intact alloantigen to B cells and T cells for generation of an alloantibody or CD8 T cell response. PVG.RT1 u BM-derived DC internalised (via an active process) and retained intact a recombinant soluble form of RT1-A a (sRT1-A a ). When PVG.RT1 u rats were immunised with sRT1-A a -pulsed syngeneic DC, they developed a strong anti-sRT1-A a alloantibody response and showed accelerated rejection of RT1-A a -disparate PVG.R8 heart grafts. Alloantibody production and accelerated heart graft rejection were both dependent on immunisation with viable sRT1-A a -pulsed DC. The alloantibody response to sRT1-A apulsed DC was directed exclusively against conformational epitopes expressed by sRT1-A a and not epitopes expressed, for example, by non-conformational sRT1-A a heavy chain. Immunisation with sRT1-A a -pulsed syngeneic DC did not stimulate a CD8 T cell response. Our findings suggest a novel alloantigen recognition pathway whereby soluble MHC class I alloantigen released from an allograft may be taken up by recipient DC and presented in an intact unprocessed form to B cells for the generation of an alloantibody response.
SUMMARYPristane-induced arthritis (PIA) is a murine disease resembling rheumatoid arthritis (RA) which is characterized by autoimmune responses to joint tissues. To identify the range of potential antigens targeted in PIA, proteins from arthritic or normal joint extracts were fractionated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and systematically screened for the ability to react with either serum IgG, or cultured splenic T cells, obtained from healthy or arthritic mice. Extracts from both normal and arthritic animals contained multiple proteins that were capable of reacting with murine serum IgG in immunoblotting experiments. In healthy controls, more bands were identified in extracts prepared from 30-week-old mice than from 8-week-old animals, but the widest range of proteins bound were derived from arthritic joints. Furthermore, the sera from PIA-positive mice reacted with more bands from each of the extracts than did normal sera. Fractionated extracts prepared from healthy joints failed to stimulate the in vitro proliferation of splenic T cells from either normal or arthritic animals. When arthritic joint components were screened, T cells from healthy mice responded weakly to some fractions, but multiple fractions elicited strong proliferation by T cells from mice with PIA. A band of apparent molecular mass 60 000 was the protein most commonly bound by serum IgG from arthritic mice, and the corresponding fraction stimulated the highest responses by T cells from PIA-positive animals. These results are consistent with the notion that the 60 000 MW mammalian heatshock protein is an important antigen in PIA, but that the autoimmune response diversifies with the development of arthritis to target multiple joint components.
The rat major histocompatibility complex loci RT1-B and RT1-D are equivalent to the human leucocyte antigens HLA-DQ and HLA-DR respectively. Here we describe the complementary DNA (cDNA) sequence encoding the alpha and beta chains of both the RT1-B and RT1-D locus genes of the rat RT1u haplotype. We have found entire sequence identity between five different inbred rat strains of the RT1u haplotype, which differs from previously published, incomplete sequences. This information is of considerable value for experimental studies of transplantation immunity and autoimmune disease.
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