Processing of a viral glycoprotein in the endoplasmic reticulum for class II presentation

Shirley M. Bartido, Stephanie Diment, Carol S. Reiss

Research output: Contribution to journalArticle

Abstract

Endogenous processing of viral glycoproteins for presentation to CD4+ T cells is a poorly investigated aspect of antigen processing and presentation. This pathway may involve not only pathogens, but also self proteins, and may thus be involved in self-tolerance. We have characterized the processing of the endoplasmic reticulum-restricted glycoprotein (G) of vesicular stomatitis virus, termed poison tail (Gpt), biochemically and enzymatically, and by T cell recognition assays. Expressed with a vaccinia vector, Gpt remains endoglycosidase H-sensitive and does not mature to endoglycosidase D sensitivity. The protein is degraded in the ER with a T1/2 of 4 h. Gpt peptides are not secreted since Gpt-infected cells are unable to sensitize uninfected antigen-presenting cells in an innocent bystander assay. Using flow cytometry, Gpt is undetectable on the plasma membrane; in contrast, wild-type G is readily found on the surface or secreted into the milieu as soluble G following infection of A20 cells with a vaccinia recombinant expressing G. The degradation of Gpt is sensitive to the thiol reagent diamide and occurs optimally at physiological pH. A series of proteolytic inhibitors were tested: 3,4-dichloroisocoumarin and 1-chloro-3-tosylamido-7-amino-2-heptanone inhibited degradation, which suggests the involvement of a serine protease. The degradation does not require transport to the Golgi complex, and is not sensitive to a variety of lysosomotropic agents. We show that the degradation products include the immunogenic epitopes recognized by a panel of T cell clones and hybridomas.

Original languageEnglish (US)
Pages (from-to)2211-2219
Number of pages9
JournalEuropean Journal of Immunology
Volume25
Issue number8
DOIs
StatePublished - Aug 1995

Fingerprint

Endoplasmic Reticulum
Vaccinia
Glycoproteins
Antigen Presentation
T-Lymphocytes
Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase
Diamide
Self Tolerance
Sulfhydryl Reagents
Vesicular Stomatitis
Glycoside Hydrolases
Poisons
Hybridomas
Golgi Apparatus
Serine Proteases
Antigen-Presenting Cells
Epitopes
Flow Cytometry
Proteins
Clone Cells

Keywords

  • Class II
  • Endogenous processing
  • Glycoprotein
  • Vesicular stomatitis virus

ASJC Scopus subject areas

  • Immunology

Cite this

Processing of a viral glycoprotein in the endoplasmic reticulum for class II presentation. / Bartido, Shirley M.; Diment, Stephanie; Reiss, Carol S.

In: European Journal of Immunology, Vol. 25, No. 8, 08.1995, p. 2211-2219.

Research output: Contribution to journalArticle

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