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Publication type: Journal Article
Document type: Full Paper

Year: 1995

Author(s): Bensussan, A; Mansur, IG; Mallet, V; Rodriguez, AM; Girr, M; Weiss, EH; Brem, G; Boumsell, L; Gluckman, E; Dausset, J

Title: Detection of membrane-bound HLA-G translated products with a specific monoclonal antibody.

Source: Proc Natl Acad Sci U S A (92), 22 10292-10296.



Authors Vetmeduni Vienna:

Brem Gottfried

Vetmed Research Units
Institute of Animal Breeding and Genetics


Abstract:
A monomorphic anti-HLA-G monoclonal antibody (mAb) was obtained by immunization of HLA-B27/human beta(2)-microglobulin double-transgenic mice with transfected murine L cells expressing both HLA-G and human beta(2)-microglobulin, This mAb, designated BFL.1, specifically recognizes, by flow cytometry analysis, the immunizing HLA-G-expressing cells, whereas it does not bind to parental untransfected or to HLA-B7- and HLA-A3-transfected L cells, suggesting that it distinguishes between classical HLA-A and -B and nonclassical HLA-G class I molecules, This was further assessed by the absence of BFL.1 reactivity with a number of human cell lines known to express classical HLA class I proteins, In addition, we showed that the BFL.1 mAb also labels HLA-G-naturally-expressing JEG-3 and HLA-G-transfected JAR human choriocarcinoma cell lines as well as a subpopulation of first-trimester placental cytotrophoblast cells. Further biochemical studies mere performed by immunoprecipitation of biotinylated membrane lysates: BFL.1, like the monomorphic W6/32 mAb, immunoprecipitated a 39-kDa protein in HLA-G-expressing cell lines, a size corresponding to the predicted full-length HLA-G1 isoform, However, in contrast to W6/32, which immunoprecipitates both classical and nonclassical HLA class I heavy chains, BFL.1 mAb does not recognize the class Ia products, Such a mAb should he a useful tool for analysis of HLA-G protein expression in various normal and pathological human tissues and for determination of the function(s) of translated HLA-G products.

Keywords Pubmed: Animals - : Antibodies, Monoclonal - : Antibody Specificity - : Cell Line - : Cell Membrane - immunology : Female - immunology : Flow Cytometry - immunology : Fluorescent Antibody Technique, Indirect - immunology : HLA Antigens - analysis : Histocompatibility Antigens Class I - analysis : Humans - analysis : Mice - analysis : Mice, Inbred Strains - analysis : Mice, Transgenic - analysis : Placenta - analysis : Pregnancy - analysis : Pregnancy Trimester, First - analysis : Recombinant Proteins - analysis : Trophoblasts - immunology : Tumor Cells, Cultured - immunology

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