The epitope recognized by pan-HLA class I-reactive monoclonal antibody W6/32 and its relationship to unusual stability of the HLA-B27/beta2-microglobulin complex.
A broadly used pan-HLA class I-reactive monoclonal antibody W6/32 is believed to recognize a conformational epitope dependent on association between heavy chains and beta2-microglobulin (beta2m). However, in the present study we report that W6/32 does recognize at least some free HLA class I heavy chains under the partially denaturating conditions of nonreducing Western blotting, namely nearly all HLA-B allelic products. Furthermore, we confirm and largely extend our previous observation that complexes of beta2m with heavy chains of a few HLA class I allelic forms (most notably HLA-B27) exhibit unusual resistance to dissociation by SDS, which is reminiscent of MHC class II molecules. In addition, our data indicate the existence of covalent (disulfide-linked) heterodimers of certain HLA class I heavy chains (namely Cw1 and Cw4) and beta2m.
Tran
T M
TM
Institute of Clinical and Experimental Medicine, HLA Centre, Vídenská 1958, 14000 Prague 4, Czech Republic.
Ivanyi
P
P
Hilgert
I
I
Brdicka
T
T
Pla
M
M
Breur
B
B
Flieger
M
M
Ivasková
E
E
Horejsí
V
V
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002460:Cell Line; D002478:Cells, Cultured; D000939:Epitopes / Q000276:immunology*; D015235:HLA-B Antigens / Q000276:immunology; D015796:HLA-B27 Antigen / Q000276:immunology / Q000378:metabolism*; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D006801:Humans; D046911:Macromolecular Substances; D051379:Mice; D011489:Protein Denaturation; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D001613:beta 2-Microglobulin / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015235:HLA-B Antigens; D015796:HLA-B27 Antigen; D015395:Histocompatibility Antigens Class I; D046911:Macromolecular Substances; D001613:beta 2-Microglobulin; D012967:Sodium Dodecyl Sulfate