Characterization of monoclonal antibodies recognizing HLA-G or HLA-E: new tools to analyze the expression of nonclassical HLA class I molecules.
Nonclassical major histocompatibility complex (MHC) class I human leukocyte antigen E (HLA-E) and HLA-G molecules differ from classical ones by specific patterns of transcription, protein expression, and immunotolerant functions. The HLA-G molecule can be expressed as four membrane-bound (HLA-G1 to -G4) and three soluble (HLA-G5 to -G7) proteins upon alternative splicing of its primary transcript. In this study, we describe a new set of monoclonal antibodies (mAbs) called MEM-G/01, -G/04, -G/09, -G/13, MEM-E/02, and -E/06 recognizing HLA-G or HLA-E. The pattern of reactivity of these mAbs were analyzed on transfected cells by flow cytometry, Western blotting, and immunochemistry. MEM-G/09 and -G/13 mAbs react exclusively with native HLA-G1 molecules, as the 87G mAb. MEM-G/01 recognizes (similar to the 4H84 mAb) the denatured HLA-G heavy chain of all isoforms, whereas MEM-G/04 recognizes selectively denatured HLA-G1, -G2, and -G5 isoforms. MEM-E/02 and -E/06 mAbs bind the denatured and cell surface HLA-E molecules, respectively. These mAbs were then used to analyze the expression of HLA-G and HLA-E on freshly isolated cytotrophoblast cells, on the JEG-3 placental tumor cell line, and on cryopreserved and paraffin-embedded serial sections of trophoblast tissue. These new mAbs represent valuable tools to study the expression of HLA-G and HLA-E molecules in cells and tissues under normal and pathologic conditions.
Menier
Catherine
C
Service de Recherches en Hémato-Immunologie, Hôpital Saint-Louis, IUH, Paris, France.
Saez
Berta
B
Horejsi
Vaclav
V
Martinozzi
Silvia
S
Krawice-Radanne
Irene
I
Bruel
Sylvie
S
Le Danff
Caroline
C
Reboul
Murielle
M
Hilgert
Ivan
I
Rabreau
Michèle
M
Larrad
Mur Luis
ML
Pla
Marika
M
Carosella
Edgardo D
ED
Rouas-Freiss
Nathalie
N
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D005786:Gene Expression Regulation; D005805:Genes, MHC Class I; D006680:HLA Antigens / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D007120:Immunochemistry / Q000379:methods; D010920:Placenta / Q000276:immunology; D011247:Pregnancy; D001613:beta 2-Microglobulin / Q000276:immunology; D000097625:HLA-E Antigens
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D006680:HLA Antigens; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I; D001613:beta 2-Microglobulin; D000097625:HLA-E Antigens