The assembly of the classical, polymorphic major histocompatibility complex class I molecules in the endoplasmic reticulum requires the presence of peptide ligands and beta 2-microglobulin (beta 2m). Formation of this trimolecular complex is a prerequisite for efficient 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 difficult 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 beta 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.
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1 October 1992
Article|
October 01 1992
Impaired intracellular transport and cell surface expression of nonpolymorphic HLA-E: evidence for inefficient peptide binding.
M Ulbrecht,
M Ulbrecht
Institut für Immunologie, Ludwig-Maximilians-Universität München, Germany.
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J Kellermann,
J Kellermann
Institut für Immunologie, Ludwig-Maximilians-Universität München, Germany.
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J P Johnson,
J P Johnson
Institut für Immunologie, Ludwig-Maximilians-Universität München, Germany.
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E H Weiss
E H Weiss
Institut für Immunologie, Ludwig-Maximilians-Universität München, Germany.
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M Ulbrecht
Institut für Immunologie, Ludwig-Maximilians-Universität München, Germany.
J Kellermann
Institut für Immunologie, Ludwig-Maximilians-Universität München, Germany.
J P Johnson
Institut für Immunologie, Ludwig-Maximilians-Universität München, Germany.
E H Weiss
Institut für Immunologie, Ludwig-Maximilians-Universität München, Germany.
Online ISSN: 1540-9538
Print ISSN: 0022-1007
J Exp Med (1992) 176 (4): 1083–1090.
Citation
M Ulbrecht, J Kellermann, J P Johnson, E H Weiss; Impaired intracellular transport and cell surface expression of nonpolymorphic HLA-E: evidence for inefficient peptide binding.. J Exp Med 1 October 1992; 176 (4): 1083–1090. doi: https://doi.org/10.1084/jem.176.4.1083
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