Structural characterization of two CD1A allelic variants.
CD1 molecules are specialized in presenting lipidic antigens to T lymphocytes. They are structurally and evolutionary related to MHC molecules and show very limited polymorphism. We have previously described and partially characterized a new human CD1A allele differing from the wild type CD1A by a substitution of Cysteine by Tryptophan at position 52 in the alpha1 domain of the CD1A molecule. The frequency of this allele varies from 10% in individuals of Caucasian origin to 56% in Chinese people. The aim of the present work was to structurally characterize this CD1A allele. To do this we have cloned and sequenced the full-length cDNA encoding the new CD1A allele. The cDNA sequence of this allele encodes a protein differing the wild type in two amino acids at positions 14 (Threonine versus Isoleucine) and 52 (Cysteine versus Tryptophan). The cDNAs encoding both wild type and mutant CD1A were cloned in the expression vector pSRalphaNeo and transfected into C1R and L721.221 cells. Cell surface expression of the protein products in transfected cell lines were analyzed by flow cytometry and immunoprecipitation using CD1a-specific monoclonal antibodies. Our results indicate that both allelic products are efficiently expressed on the cell surface.
Oteo
M
M
Inmunología, Facultad de Medicina, Universidad Complutense, 28040, Madrid, Spain.
Arribas
P
P
Setién
F
F
Parra
J F
JF
Mirones
I
I
Gómez del Moral
M
M
Martínez-Naves
E
E
D000483:Alleles; D000911:Antibodies, Monoclonal / Q000032:analysis; D018949:Antigens, CD1 / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics* / Q000276:immunology; D002461:Cell Line, Transformed; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000302:isolation & purification; D005434:Flow Cytometry; D014644:Genetic Variation / Q000276:immunology*; D005822:Genetic Vectors / Q000096:biosynthesis / Q000276:immunology; D006801:Humans; D011233:Precipitin Tests; D014162:Transfection; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D018949:Antigens, CD1; C419393:CD1a antigen; D018076:DNA, Complementary