Co-expression of the human HLA-B27 class I antigen and the E3/19K protein of adenovirus-2 in insect cells using a baculovirus vector

Détails

ID Serval
serval:BIB_94B8E47BF50C
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Co-expression of the human HLA-B27 class I antigen and the E3/19K protein of adenovirus-2 in insect cells using a baculovirus vector
Périodique
International Immunology
Auteur(s)
Levy  F., Kvist  S.
ISSN
0953-8178 (Print)
Statut éditorial
Publié
Date de publication
1990
Volume
2
Numéro
10
Pages
995-1002
Notes
Journal Article
Résumé
We have expressed the human MHC class I HLA-B27 antigen, human beta 2-microglobulin, and the E3/19K protein of adenovirus-2 in Spodoptera frugiperda insect cells (Sf9) by using the Autographa californica nuclear polyhedrosis virus. All genes were inserted under the strong polyhedrin promoter in the vector pVL941. The proteins were expressed at high levels, ranging from 1 to 8 mg protein per 3 x 10(9) cells. Both a full-length and a truncated form of HLA-B27 were expressed. The latter was terminated at the border of the membrane-spanning segment at the extracellular side of the membrane. The HLA-B27 antigens and the E3/19K protein showed considerable heterogeneity with respect to glycosylation. Only a small fraction of HLA-B27 was assembled (less than 5%) with beta 2-microglobulin. Nevertheless, we were able to find the heavy chain at the cell surface, and by co-infection with the recombinant virus for beta 2-microglobulin we observed an increase in cell surface expression. The E3/19K protein of adenovirus-2 blocks cell surface expression of HLA class I antigens in human cells and has a similar effect in insect cells. In contrast to beta 2-microglobulin it assembles efficiently to the heavy HLA-B27 chain, both the full-length and truncated forms. The E3/19K protein does not stimulate assembly of HLA-B27-beta 2-microglobulin. Due to these problems of assembly and the heterogeneity of glycosylation we predict that it will be difficult to use HLA antigens produced in insect cells for X-ray crystallographic studies.
Mots-clé
Adenovirus E3 Proteins Adenoviruses, Human/genetics Animals Baculoviridae/genetics Cell Membrane/immunology/metabolism Cells, Cultured DNA/genetics Gene Expression Genetic Vectors HLA-B27 Antigen/biosynthesis/*genetics Humans Moths Viral Proteins/biosynthesis/*genetics beta 2-Microglobulin/biosynthesis/genetics
Pubmed
Web of science
Création de la notice
28/01/2008 12:17
Dernière modification de la notice
03/03/2018 19:38
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