Analysis of the HLA-restricted influenza-specific cytotoxic T lymphocyte response in transgenic mice carrying a chimeric human-mouse class I major histocompatibility complex.

Abstract

SummaryTransgenic murine lines have been constructed that express a chimeric class I molecule composed of the cx1 and u2 domains of HLAA2.1 and the 0, transmembrane, and cytoplasmic domains of H-2Kb. Upon immunization with influenza virus, transgenic mice developed a strong A2.1Kb-restricted cytotoxic T lymphocyte (CTL) response specific for the same matrix protein epitope that serves as the dominant A2.1-restricted determinant in the equivalent human response. Fine specificity analysis of CTL clones using truncated peptides revealed strong similarity between the response repertoire of transgenic mice and that previously reported using influenza-specific A2.1-restricted CTL clones from humans. This suggests that even when considering T cell responses by different species, the a1 and ot2 domains of the restriction element play a dominant role in determining the CTL specific repertoire. Thus, substituting the a3 domain of A2.1 with a murine counterpart has permitted development of a transgenic strain that should serve as an excellent model system in studies of HLArestricted responses .

Responses of human T cells to peptides flanking the tandem repeat and overlapping the signal sequence of MUC1

“…Using mice transgenic for the chimeric human HLA-A*0201/murine H-2K b molecule, 2 peptides from the TR domain were identified as sequences recognised by murine CTLs. 17 They did not contain the anchor motif for HLA-A*0201 and bound to MHC with low affinity. When they were used to immunise mice, only modest CTL activity was observed.…”

Protein delivery by subviral particles of human cytomegalovirus

“…Since for both the neomycin transferase and the green fluorescent protein used as markers in our study, no epitopes for the cytolytic T-lymphocyte response were known, we used a well-defined CTL-target epitope from the pp65 protein of HCMV for testing. Stably growing CTL clones were established in CD8/HLA-A2 double transgenic mice, following established procedures, 10,11 against the peptide epitope pp65 495-503 , known to be presented by HLA-A2. 12,13 These clones were subsequently used to analyze the presentation of a defined A2-presented peptide of pp65 after transfer of this tegument protein by DB into both fibroblasts and DC.…”

“…CTLL were generated by immunizing HLA-A2/CD8 double-transgenic mice (a kind gift of L. Sherman, The Scripps Institute, LaJolla, USA) with DB resuspended in incomplete Freunds adjuvant, followed by spleenectonomy 10 days later, similar to procedures described previously. 5,10,11 Splenic cells were restimulated weekly with irradiated, pp65 495-503 -loaded feeder cells and TCGF according to established protocols. 10,11 IFN-g-based ELISPOT analyses were carried out as detailed by others.…”

Recognition of chlamydial antigen by HLA‐B27‐restricted cytotoxic T cells in HLA‐B*2705 transgenic CBA (H‐2k) mice

“…Furthermore, CTL from mice transgenic for a particular human HLA molecule recognized the same HLA-restricted peptide(s) as did CTL of human origin (40,41). It is therefore likely that species-specific differences in antigen processing by mouse and human cells have little effect (42)(43)(44)(45). This system has not previously been applied to bacterial epitopes.…”