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PMID: 3257565 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Transfer and expression of three cloned human non-HLA-A,B,C class I major histocompatibility complex genes in mutant lymphoblastoid cells.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 85 ·No. 1 ·1988-01-00 ·Pages 227-31

Shimizu Y, Geraghty DE, Koller BH, Orr HT, DeMars R

Abstract

The HLA-A, -B, and -C class I human histocompatibility antigens and the genes that encode them have been isolated and characterized. Apparently complete class I non-HLA-A, B, C genes have been identified on HindIII-generated 5.4-kilobase (kb), 6.0-kb, and 6.2-kb DNA fragments derived from lymphoblastoid cell line (LCL) 721. We studied the expressibility of these genes by subcloning them into the nonintegrating pHeBo vector and transferring the chimeric plasmids into mutant LCL 721.221. This mutant was derived from LCL 721 by means of immunoselections following gamma-ray mutagenesis that eliminated expressions of the HLA-A, -B, and -C alpha chains. The HLA-A, B, C-null phenotype of mutant 721.221 made it possible to monitor the expression of class I genes transferred into it by assaying cell surface binding of monoclonal antibodies BBM.1 and W6/32, which recognize beta 2-microglobulin and HLA class I alpha-chain epitopes, respectively. Increased binding of BBM.1 and W6/32 was clearly observed in transferents containing the class I gene of the 6.0-kb DNA fragment but not in transferents containing the class I genes of the 5.4- and 6.2-kb DNA fragments. However, one-dimensional gel electrophoresis of BBM.1 and W6/32 immunoprecipitates made with [35S]methionine-labeled cell lysates showed that transfer of each non-HLA-A, B, C class I gene into 721.221 resulted in the appearance of an alpha chain that coprecipitated with beta 2-microglobulin. The three previously unreported alpha chains differed from each other in size and were smaller than HLA-A, -B, and -C alpha chains. These observations clearly show that these three cloned, nonallelic, non-HLA-A, B, C class I genes encode alpha chains that can be expressed in human cells.

MeSH Terms
B-Lymphocytes/immunology Cell Line Cloning, Molecular Genes, MHC Class I HLA Antigens/genetics Humans Mutation Transcription, Genetic
Chemicals
HLA Antigens
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Shimizu Y
Laboratory of Genetics, University of Wisconsin, Madison 53706.
Geraghty D E
Koller B H
Orr H T
DeMars R
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.85.1.227
Published
1988-01-00
Pages
227-31
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC279517
Subset
IM
Grants
NIGMS NIH HHS · 5T-32-GM07133 · United States
NIAID NIH HHS · AI-15486 · United States
NIAID NIH HHS · AI-18124 · United States
External Links
PubMed source DOI: 10.1073/pnas.85.1.227 PMC Full Text View journal details
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