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PMID: 1655683 Published · ppublish English Journal Article

Efficient cDNA expression vectors for stable and transient expression of HLA-DR in transfected fibroblast and lymphoid cells.

Human immunology ·Vol. 31 ·No. 4 ·1991-08-00 ·Pages 229-35

Long EO, Rosen-Bronson S, Karp DR, Malnati M, Sekaly RP, Jaraquemada D

Abstract

cDNA expression vectors with several useful features were constructed. First, the long terminal repeat of Rous sarcoma virus was used as a promoter to obtain high levels of expression in various cells of human and mouse origin. Second, cis-linked expression units that confer resistance either to mycophenolic acid or the neomycin analog G418 were inserted to facitate the isolation of transfected cells expressing the cDNA of interest. Third, by replicating in simian COS cells, these vectors can be used for efficient transient expression. cDNA fragments encoding the DR alpha or DR beta chains of human class II major histocompatibility complex antigens were inserted into these vectors and high levels of cell surface HLA-DR antigen were obtained after cotransfection into mouse and human fibroblasts. These vectors were also successfully used to correct the inability of a class II-negative B cell line, derived from a patient with a congenital immunodeficiency, to present peptide antigen to DR-restricted T cells.

MeSH Terms
Animals Avian Sarcoma Viruses/genetics Cell Line, Transformed DNA/genetics Fibroblasts Flow Cytometry Fluorescent Antibody Technique Gene Expression Genetic Vectors HLA-DR Antigens/genetics,immunology Humans Immunologic Deficiency Syndromes/genetics Lymphocytes Mice Plasmids/genetics Repetitive Sequences, Nucleic Acid Transfection Tumor Cells, Cultured
Chemicals
HLA-DR Antigens DNA
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Long E O
Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
Rosen-Bronson S
Karp D R
Malnati M
Sekaly R P
Jaraquemada D
Article Info
Journal
Human immunology
Abbr.
Hum Immunol
ISSN
0198-8859
Published
1991-08-00
Pages
229-35
Language
English
Region
United States
NLM ID
8010936
Subset
IM
Databases
GENBANK
M83236, M83237, S73250, S73294, S73300, S73304, S73309, S73312, S73315, S73319
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