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Flow-Microfluorometric Monitoring of Oligoclonal CD8⁺ T Cell Responses to an Immunodominant Moloney Leukemia Virus-Encoded Epitope In Vivo¹

Pierre Brawand^*, Giovanni Biasi, Clotilde Horvath^*, Jean-Charles Cerottini^* and H. Robson MacDonald^2,*

^* Ludwig Institute for Cancer Research, Lausanne Branch, University of Lausanne, Epalinges, Switzerland; and Institute of Experimental Pathology, University of Ancona, Faculty of Medicine, Ancona, Italy

The TCR repertoire of CD8⁺ T cells specific for Moloney murine leukemia virus (M-MuLV)-associated Ags has been investigated in vitro and in vivo. Analysis of a large panel of established CD8⁺CTL clones specific for M-MuLV indicated an overwhelming bias for Vß4 in BALB/c mice and for Vß5.2 in C57BL/6 mice. These Vß biases were already detectable in mixed lymphocyte:tumor cell cultures established from virus-immune spleen cells. Furthermore, direct ex vivo analysis of PBL from BALB/c or C57BL/6 mice immunized with syngeneic M-MuLV-infected tumor cells revealed a dramatic increase in CD8⁺ cells expressing Vß4 or Vß5.2, respectively. M-MuLV-specific CD8⁺ cells with an activated (CD62L^-) phenotype persisted in blood of immunized mice for at least 2 mo, and exhibited decreased TCR and CD8 levels compared with their naive counterparts. In C57BL/6 mice, most M-MuLV-specific CD8⁺CTL clones and immune PBL coexpressed V3.2 in association with Vß5.2. Moreover, these Vß5.2⁺V3.2⁺ cells were shown to recognize the recently described H-2D^b-restricted epitope (CCLCLTVFL) encoded in the leader sequence of the M-MuLV gag polyprotein. Collectively, our data demonstrate a highly restricted TCR repertoire in the CD8⁺ T cell response to M-MuLV-associated Ags in vivo, and suggest the potential utility of flow-microfluorometric analysis of Vß and V expression in the diagnosis and monitoring of viral infections.

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