Technical Report No. 6-91
Flow-cytometric measurement of CD4–8– T cells bearing T-cell receptor alpha beta chains: 1. Results for a normal population including two cases with unusually high frequencies
Kusunoki Y, Hirai Y, Kyoizumi S, Akiyama MEditor’s note: A publication based on this report was published in Blood 79(11):2965-72, 1992.
Summary
In this study we detected rare, possibly abnormal, T cells bearing CD3 surface antigen and T-cell receptor (TCR) alpha-beta chains but lacking both CD4 and CD8 antigens (viz., TCR-alpha-beta+CD4–8– cells, as determined by flow cytometry). The TCR-alpha-beta+CD4–8– T cells were detected at a mean frequency of 0.63 plus or minus 0.35% (mean plus or minus standard deviation) in peripheral blood TCR-alpha-beta+ cells of 119 normal persons. Two unusual cases besides the 119 normal persons showed extremely elevated frequencies of TCR-alpha-beta+CD4–8– T cells, viz., approximately 5% to 10% and 14% to 19% in whole TCR alpha-beta+ cells. Both individuals were males who were otherwise physiologically quite normal with no history of severe illness, and these high frequencies were also observed in blood samples collected 2 or 8 years prior to the current measurements. The TCR-alpha-beta+CD4–8– T cells of the two individuals were found to express mature T-cell markers such as CD2, 3, and 5 antigens, as well as natural killer (NK) cell markers, viz, CD11b, 16, 56, and 57 antigens, when peripheral blood lymphocytes were subjected to three-color flow cytometry. Lectin-dependent or redirected antibody-dependent cell-mediated cytotoxicities were observed for both freshly sorted TCR-alpha-beta+CD4–8– cells and in vitro established clones. Nevertheless, NK-like activity was not detected. Further, Southern blot analysis of TCR beta and gamma genes revealed identical rearrangement patterns for all the TCR-alpha-beta+CD4–8– clones established in vitro. These results suggest that the TCR-alpha-beta+CD4–8– T cells from these two men exhibit unique characteristics and proliferate clonally in vivo.
In this study we detected rare, possibly abnormal, T cells bearing CD3 surface antigen and T-cell receptor (TCR) alpha-beta chains but lacking both CD4 and CD8 antigens (viz., TCR-alpha-beta+CD4–8– cells, as determined by flow cytometry). The TCR-alpha-beta+CD4–8– T cells were detected at a mean frequency of 0.63 plus or minus 0.35% (mean plus or minus standard deviation) in peripheral blood TCR-alpha-beta+ cells of 119 normal persons. Two unusual cases besides the 119 normal persons showed extremely elevated frequencies of TCR-alpha-beta+CD4–8– T cells, viz., approximately 5% to 10% and 14% to 19% in whole TCR alpha-beta+ cells. Both individuals were males who were otherwise physiologically quite normal with no history of severe illness, and these high frequencies were also observed in blood samples collected 2 or 8 years prior to the current measurements. The TCR-alpha-beta+CD4–8– T cells of the two individuals were found to express mature T-cell markers such as CD2, 3, and 5 antigens, as well as natural killer (NK) cell markers, viz, CD11b, 16, 56, and 57 antigens, when peripheral blood lymphocytes were subjected to three-color flow cytometry. Lectin-dependent or redirected antibody-dependent cell-mediated cytotoxicities were observed for both freshly sorted TCR-alpha-beta+CD4–8– cells and in vitro established clones. Nevertheless, NK-like activity was not detected. Further, Southern blot analysis of TCR beta and gamma genes revealed identical rearrangement patterns for all the TCR-alpha-beta+CD4–8– clones established in vitro. These results suggest that the TCR-alpha-beta+CD4–8– T cells from these two men exhibit unique characteristics and proliferate clonally in vivo.