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Frequency Analysis of Simian Virus 40-specific Cytotoxic T Lymphocyte Precursors in the High Responder C57BL/6 Mouse Strain

Stephen R. Jennings

Kerin L. Fresa

The Department of Microbiology, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, U.S.A.

Studies in this laboratory have shown that long term simian virus 40 (SV40)-specific cytolytic T lymphocyte (CTL) cultures established from the spleens of high responder C57BL/6 (B6; H-2^b) mice exhibit a preference for the selection of H-2D^b-restricted CTL clones. In this study, we have investigated the basis for this selection. Limiting dilution cultures were established using responder cells from the popliteal lymph nodes and the spleens of B6 mice immunized subcutaneously in the hind footpads or via the intraperitoneal route, respectively, with syngeneic SV40-transformed cells expressing a full length (1 to 708 amino acid residues) SV40 large T antigen. The relative frequency of CTL precursors (CTLp) able to expand in vitro in the presence of SV40-transformed stimulator cells and interleukin 2 and exhibit lytic activity against H-2^b cells expressing full length T antigen ranged from 1/1900 to 1/15000 in the popliteal lymph node and from 1/8000 to 1/55000 in the spleen. In these two experimental systems, CTLp restricted to H-2K^b were apparently present at higher frequency than H-2D^b-restricted CTLp. Furthermore, CTLp recognizing determinants within the amino-terminal or carboxy-terminal halves of T antigen were generated in approximately equal numbers. The relative affinity of SV40-specific CTL, assessed by inhibition with anti-Lyt 2 monoclonal antibody, indicated that CTL restricted to H-2D^b interacted with their target with greater affinity than CTL restricted to H-2K^b. These data suggest that the predominance of isolation of H-2D^b-restricted CTL clones from long term in vitro cultures may be a function of the relative affinity of this population as a whole, rather than due to the immunodominance of this subpopulation during the in vivo response to SV40 T antigen.

Keywords: SV40, CTL recognition, frequency

Present address: Department of Microbiology and Immunology, Louisiana State University Medical Center, School of Medicine in Shreveport, Shreveport, Louisiana 71130, U.S.A.

Present address: Department of Pathology, Hahnemann University School of Medicine, Philadelphia, Pennsylvania 19102, U.S.A.

Received 12 May 1988; accepted 6 July 1988.

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