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

Journal Article · Fri Jan 01 04:00:00 EST 1988 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.85.1.227· OSTI ID:6464555

Shimizu, Y; Geraghty, D E; Koller, B H; Orr, H T; DeMars, R

The HLA-A, -B, and -C class I human histocompatibility antigens and the genes that encode them have been isolated and characterized. The authors 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 ..cap 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../sub 2/-microglobulin and HLA class I ..cap 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 (/sup 35/S)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 ..cap alpha.. chain that coprecipitated with ..beta../sub 2/-microglobulin. These observations clearly show that these three cloned, nonallelic, non-HLA-A,B,C class I genes encode ..cap alpha.. chains that can be expressed in human cells.

• Research Organization: Univ. of Wisconsin, Madison (USA)
• OSTI ID: 6464555
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 85:1; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMAL CELLS
ANIMALS
ANTIBODIES
ANTIGENS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOXYLIC ACIDS
CELL FLOW SYSTEMS
CLONING
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DNA HYBRIDIZATION
DNA-CLONING
DRUGS
ELECTROMAGNETIC RADIATION
EVEN-ODD NUCLEI
GAMMA RADIATION
GENE RECOMBINATION
GENE REGULATION
GENES
HYBRIDIZATION
IMMUNOLOGY
IONIZING RADIATIONS
ISOTOPES
LEUKOCYTES
LIGHT NUCLEI
LIPOTROPIC FACTORS
LYMPHOCYTES
MAMMALS
MAN
MATERIALS
METHIONINE
MONOCLONAL ANTIBODIES
MUTAGENESIS
MUTANTS
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PRIMATES
RADIATIONS
RADIOINDUCTION
RADIOISOTOPES
SOMATIC CELLS
SULFUR 35
SULFUR ISOTOPES
VERTEBRATES