Locus-Specific Analysis of Human Leukocyte Antigen Class I Expression in Melanoma Cell Lines

Francesco M. Marincola, Peter Shamamian, Toni B. Simonis, Andrea Abati, Julia Hackett, Teresita O'Dea, Patricia Fetsch, John Yannelli, Nicholas P. Restifo, James J. Mule, Steven A. Rosenberg

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Abstract

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.

Original languageEnglish (US)
Pages (from-to)13-23
Number of pages11
JournalJournal of Immunotherapy
Volume16
Issue number1
DOIs
StatePublished - Jul 1994
Externally publishedYes

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Keywords

  • Human leukocyte antigen class I
  • Melanoma
  • lmmunotherapy
  • ¯

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Pharmacology
  • Cancer Research

Cite this

Marincola, F. M., Shamamian, P., Simonis, T. B., Abati, A., Hackett, J., O'Dea, T., Fetsch, P., Yannelli, J., Restifo, N. P., Mule, J. J., & Rosenberg, S. A. (1994). Locus-Specific Analysis of Human Leukocyte Antigen Class I Expression in Melanoma Cell Lines. Journal of Immunotherapy, 16(1), 13-23. https://doi.org/10.1097/00002371-199407000-00002