Use of a monoclonal antibody (W6/32) in structural studies of HLA-A,B,C, antigens
Parham, P.; Barnstable, C.J.; Bodmer, W.F.
Journal of Immunology 123(1): 342-349
1979
ISSN/ISBN: 0022-1767 PMID: 87477 Document Number: 139945
W6/32 cell lines and peripheral blood lymphocytes was measured with W6/32 antibody. B cell lines expressed, on average, 9 times as much cell surface HLA-A,B,C antigens as peripheral lymphocytes, although there was appreciable variation within each group. The B cell line Bri 8, for example, expressed 1.5 .times. 106 W6/32 antigenic sites, and by inference, HLA-A,B,C, molecules per cell. Equal amounts of .beta.2m and HLA-A,B,C chain were found on both cell types. The isolated HLA-A chain from intact 125I-HLA-A2 antigens weakly bound to W6/32 antibody in contrast to 125I-.beta.2-microglobulin (.beta.2m) isolated from the same preparation of HLA-A2 antigens that showed no demonstrable binding. When an excess of cold .beta.2m was added to the isolated 125I-HLA-A2 chain, the binding to W6/32 antibody was considerably enhanced. The W6/32 antigenic determinant apparently involves only amino acids of the HLA-A,B,C chain and is a product of their 3 dimensional configuration. Stable maintenance of this configuration appears to be dependent on the association of the HLA-A,B,C, chain with .beta.2m.