Characterization and expression of the HLA-DC antigens defined by anti-Leu 10 |
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Authors: | Yu-Xian Chen Robert L. Evans Marilyn S. Pollack Lewis L. Lanier Joseph H. Phillips Carmen Rousso Noel L. Warner Frances M. Brodsky |
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Affiliation: | a Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA b Becton Dickinson Monoclonal Center, Mountain View, CA 94043, USA c Visiting Investigator from the Department of Immunology, Cancer Sciences, Peking, China |
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Abstract: | The expression of HLA-DC antigens on peripheral blood mononuclear cells, in tonsil and lymph node tissue sections, on tumor cell lines, and on activated T cells was studied using monoclonal antibody, anti-Leu 10. Anti-Leu 10 reacts with HLA-DC molecules on homozygous B cell lines expressing HLA-DR 1,2,4,5,6,8, and 9. It reacts with heterozygous B lymphocytes expressing DR7 and DRw10, suggesting it also recognizes HLA-DC molecules linked to DRw10. The HLA-DC molecules detected by anti-Leu 10 are expressed on all Ig-positive and DR-positive peripheral B lymphocytes and an apparent subpopulation of DR-positive periperal blood monocytes. Two-color immunofluorescence experiments using phycoerythrin-anti-HLA-DR (L243) and FITC-anti-Leu 10 demonstrated a correlation of the amounts of HLA-DR and DC antigens expressed on B lymphocytes. Cells expressing relatively low or high amounts of one Class II molecule express respectively low or high amounts of the other Class II molecule. Anti-Leu 10 reacted with all B lymphocyte derived tumor cell lines not with lines of myeloid or erythroid origin, and with only one T cell derived line, HUT-78 which has an activated T cell phenotype. Consistent with this result, anti-Leu 10 binding suggested the presence of HLA-DC on activated T cells in lymphoid tissue, in addition to staining B cells. HLA-DC was also detected on mitogen and MLC activated T cells by anti-Leu 10 binding. Anti-Leu 10 is, therefore, a useful reagent for further studies of the role of HLA-DC in T cell activation and in normal B cell and monocyte functions. |
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Keywords: | PBS phosphate buffered saline 7.4 PE phycoerythrin FACS fluorescence activated cell sorter PHA phytohemagglutinin FITC fluorescein isothiocyanate FCS fetal calf serum PBM peripheral blood mononuclear MLC mixed lymphocyte culture HBSS Hank's balanced salt solution |
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