Impaired Intracellular Transport and Cell Surface Expression of Nonpolymorphic HLA-E

Ulbrecht, Matthias; Kellermann, Josef; Johnson, Judith P. und Weiss, Elisabeth H. (1992): Impaired Intracellular Transport and Cell Surface Expression of Nonpolymorphic HLA-E. Evidence for Inefficient Peptide Binding. In: Journal of Experimental Medicine, Bd. 176: S. 1083-1090

Abstract

The assembly of the classical, polymorphic major histocompatibility complex class I molecules in the endoplasmic reticulum requires the presence of peptide ligands and ~2-microglobulin (~2m). Formation of this trimolecular complex is a prerequisite for e~cient transport to the cell surface, where presented peptides are scanned by T lymphocytes. The function of the other class I molecules is in dispute. The human, nonclassical class I gene, HLA-E, was found to be ubiquitously transcribed, whereas cell surface expression was dif~cult to detect upon transfection. Pulse chase experiments revealed that the HLA-E heavy chain in transfectants, obtained with the murine myeloma cell line P3X63-Ag8.653 (X63), displays a significant reduction in oligosaccharide maturation and intracellular transport compared with HLA-B27 in corresponding transfectants. The accordingly low HLA-E cell surface expression could be significantly enhanced by either reducing the culture temperature or by supplementing the medium with human ~2m, suggesting inefficient binding of endogenous peptides to HLA-E. To analyze whether HLA-E binds peptides and to identify the corresponding ligands, fractions of acid-extracted material from HLA-E/X63 transfectants were separated by reverse phase HPLC and were tested for their ability to enhance HLA-E cell surface expression. Two fractions specifically increased the HLA class I expression on the HLA-E transfectant clone.

• Dokumententyp: Zeitschriftenartikel
• Keywords: Genetics
• Fakultät: Biologie
• Themengebiete: 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften; Biologie
• Sprache: Englisch