Locus-specific analysis of human leukocyte antigen class I expression in melanoma cell lines (original) (raw)
Surface expression of human leukocyte antigen (HLA) class I antigens on melanoma lines was evaluated by locus-specific monoclonal antibodies (mAbs) with three different techniques: Fluorescence-activated cell sorting (FACS), immunohistochemistry with cytospin preparation (ICP), and complement-mediated cytotoxicity (CMC). Eleven HLA class I-expressing cell lines developed from metastases were used. Specific expression of HLA loci was examined under routine culture conditions and after 48-h incubation in interferon-γ (IFN-γ: 500 U/ml). Loss of allelic expression was seen in one line (586-MEL): Products of genes coding for HLA-A29 and-B44, in strong linkage disequilibrium, were not detectable. HLA-A antigens were consistently detected by all methodologies and minimally affected by pretreatment with IFN-γ. HLA-B antigens were detectable in 8 of 11 lines by ICP and 3 of 11 lines by CMC. By FACS the supratypic specificity HLA-Bw6 was expressed at low levels in most lines (mean fluorescence 47.2 ± 13.4 and rose to 259.8 ± 45.9 after incubation with IFN-γ; P < 0.001). HLA-Cw antigen detection by CMC correlated with HLA-B (p < 0.01), suggesting that down-regulation and sensitivity to IFN-γ are shared by the two loci. This low expression of the HLA-B antigens may play a role in the evasion of the host immune response and its up-regulation may be useful in allowing tumor antigen recognition. Keywords Melanoma; Human leukocyte antigen class I; Immunotherapy There is strong experimental evidence that major histocompatibility complex (MHC) class I antigen expression is important in tumor immunology and immunotherapy. In animal studies the expression of MHC class I antigens appears essential for tumor rejection (1,2). In vitro studies with either human or mouse tumor-infiltrating lymphocytes (TILs) as effectors have shown that target cell lysis is MHC class I restricted (3-5). Moreover, adoptive transfer of TILs to patients with advanced cancer has resulted in tumor regression (6), suggesting that TIL recognition of tumor antigen in the context of MHC class I may be responsible for the antitumor antigen response.
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