Conformation and surface expression of free HLA-CW1 heavy chains in the absence of β2-microglobulin

Aline Martayan, Michele Fiscella, Andrea Setini, Gina Ciccarelli, Roberto Gambari, Giordana Feriotto, Alberto Beretta, Antonio G. Siccardi, Ettore Appella, Patrizio Giacomini

Research output: Contribution to journalArticlepeer-review

Abstract

A β2-microglobulin (β2m)-deficient kidney carcinoma cell line and three monoclonal antibodies to the α1 (L31), α2 (W6/32), and α3 (Q1/28) domain of class 1 HLA molecules were selected to assess the role of β2m in regulating the conformation and surface expression of HLA-C molecules. HLA- A2, -B27, and -CW1 molecules synthesized by β2m-deficient cells were compared to heavy chains synthesized in transfectants expressing a large excess of β2m. As assessed by differential binding with monoclonal antibodies and partitioning studies in the detergent TX-114, no HLA-A2, - B27, or -CW1 molecules can be expressed, in a correct conformation, by β2m- deficient cells. These cells, however, do express low but significant amounts of free HLA-CW1 heavy chains at the cell surface. Transfection with β2m causes a coordinate change in the antibody reactivity of the three domains of HLA-CW1 molecules, thereby providing the first experimental demonstration that assembly with β2m affects the folding of not only the α1 and α2, but also of the α3 domain. HLA-CW1 heavy chains, when free of β2m, are less soluble in the detergent TX-114 than free HLA-B27 heavy chains, and when associated with β2m share an α3 domain epitope with free HLA-A2 and -B27 heavy chains. Moreover, their assembly with β2m is largely incomplete. These data additionally demonstrate an impaired ability of HLA-CW1 to properly fold and established a close similarity of HLA-CW1 to murine Db and L(d) molecules. Although the functional role, if any, of free HLA-CW1 heavy chains remains to be determined, the present study demonstrates that the absence of β2m does not completely ablate class I expression in neoplastic cells of human origin.

Original languageEnglish
Pages (from-to)23-33
Number of pages11
JournalHuman Immunology
Volume53
Issue number1
DOIs
Publication statusPublished - Mar 1997

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

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