Transgenic mice expressing a soluble foreign H-2 class I antigen are tolerant to allogeneic fragments presented by self class I but not to the whole membrane-bound alloantigen.

Abstract

The properties of transmembrane and soluble transplantation antigens were compared with respect to the induction of tolerance and the selection of the T-cell repertoire.For this purpose, transgenic (H-2b X H-2d)F1 mice were constructed that carry integrated copies of a modified H-2Kk gene resulting in the secretion from various cell types including thymocytes of soluble H-2Kk molecules. Despite the presence of H-2Kk antigen, these mice were still able to generate an H-2Kk-specific T-cell response. This response was comparable to that produced by normal littermates when stimulated with cells expressing membrane H-2Kk in a mixed lymphocyte reaction. In contrast, only transgenic mice failed to generate a cytolytic T-cell response to soluble H-2Kk antigen expressed by recombinant vaccinia virus and presented by the H-2Db molecule. These data imply the presence of two populations of alloreactive cytolytic T cells. A small fraction of T cells recognizes alloantigen as antigenic peptide(s) presented by other major histocompatibility complex class I molecules and tolerance can be induced in this population by soluble alloantigen. The majority of T cells, however, require the whole cell membrane-expressed class I molecule for recognition. This population is not affected by tolerance induction to the soluble major histocompatibility complex class I molecule.

Defining the Requirements for Peptide Recognition in Gene Therapy-Induced T Cell Tolerance

“…Previous in vitro studies using T cells from transgenic mice engineered to express a secreted allogeneic MHC class I molecule have shown that alloreactive CTL precursors are able to kill targets through the direct pathway if exogenous IL-2 is present in the cultures even though MHC class I peptide-reactive T cells appear to be absent (22). However, in this study, the effect of expressing a soluble allogeneic MHC class I transgene on allograft survival was not examined.…”

Cross-Presentation of Listeria-Derived CD8 T Cell Epitopes Requires Unstable Bacterial Translation Products

“…I. Gentschev, Biozentrum, University Würzburg, Wurzburg, Germany) as a template and was subcloned into pUC19. An XbaI site in front of the ATG start codon was converted to a BamHI site by insertion of a self-complementary oligonucleotide (5Ј-cta gcg gca gga tcc tgc gc-3Ј), then the p60 open reading frame was transferred as a 1.5-kbp BamHI fragment into the vaccinia recombination vector pCS43 (13). Construction of the recombinant vaccinia virus was performed using the vaccinia virus strain Copenhagen and its temperature-sensitive mutant ts7 according to standard procedures as described previously (13).…”

Expression of the murine cytomegalovirus glycoprotein H by recombinant vaccinia virus

“…The generation of a fragment of the correct size was confirmed by gel electrophoresis. The fragment was isolated, cut with BamHI and inserted into the vaccinia virus recombination vector pCS43 (Arnold et al, 1990) downstream of the vaccinia virus early-late promoter p7.5. The correct position of the start codon was confirmed by sequencing of the resulting plasmid pCS43-gH.…”