Establishment of a Lymphocyte Clone Expressing Granzyme B and Perforin mRNAs Immortalized with Herpesvirus saimiri

概要

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Minor histocompatibility antigens play a crucial role in inducing graft-versus-host disease (GVHD) after human leukocyte antigen (HLA)-identical allogeneic bone marrow transplantation. To identify unknown antigens, we need cytotoxic T lymphocyte (CTL) recognized by specific antigens. In this study Herpesvirus saimiri (HVS) subgroup C488 was used to establish a stable CTL clone. First, normal mononuclear cells were infected with HVS in owl monkey kidney (OMK) supernatant and cultured in RPMI 1640 medium supplemented with 10% FBS and 20% T cell growth factor (TCGF). The most proliferative cells were then subjected to five limiting dilutions. After this, the presence of the CD3^+CD4^-CD8^+ T lymphocyte clone was observed. Interestingly, CD4^+ T cells soon disappeared in the presence of TCGF after being infected with HVS. To confirm whether our lymphocyte clone was cytotoxic, the presence of mRNAs for cytotoxic granule granzyme B and perform was examined using RT-PCR, and both mRNAs were identified. Our results thus demonstrate that the established clone might be a CD8^+ cytotoxic T lymphocyte clone and that HVS was a useful tool for the immortalization of CTL. The disappearance of CD4^+ T lymphocytes in our system suggests that some inhibitory factor may be present in TCGF.
神戸大学の論文
2000-12-27