Anti-HLA-B7,B27,Bw42,Bw54,Bw55,Bw56, Bw67,Bw73 monoclonal antibodies: specificity, idiotypes, and application for a double determinant immunoassay.
The monoclonal antibodies (MoAbs) KS3 and KS4 are secreted by hybridomas constructed with splenocytes from a BALB/c mouse sequentially immunized with the cultured lymphoid cells JKu and LG-2 which share only the HLA-B27 specificity. Serologic and immunochemical assays have shown that the two MoAbs recognize the same (or spatially close) determinant expressed by HLA-B7,B27,Bw42,Bw54,Bw55,Bw56,Bw67, and Bw73 alloantigens. This determinant is spatially close but distinct from those defined by the anti HLA-B27 monoclonal antibodies described in the literature. The syngeneic antiidiotypic MoAb T12-105 and T12-211 elicited with MoAb KS4 were shown to recognize idiotopes within the antigen combining site of MoAb KS3 and KS4. Neither idiotope was detected on the anti HLA class I and anti HLA class II monoclonal antibodies tested. The MoAb KS4 in combination with the anti human beta 2-microglobulin MoAb NAMB-1 was utilized to develop a double determinant immunoassay (DDIA). The latter represents a sensitive method to detect and quantitate HLA-B27 antigens in spent culture medium of lymphoid cell lines and in serum. Typing for HLA-B27 antigens with the DDIA of sera from HLA typed donors yielded results highly correlated with those of the conventional lymphocytotoxicity assay.
Sakaguchi
K
K
Department of Microbiology and Immunology, New York Medical College, Valhalla 10595.
Ono
R
R
Tsujisaki
M
M
Richiardi
P
P
Carbonara
A
A
Park
M S
MS
Tonai
R
R
Terasaki
P I
PI
Ferrone
S
S
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay; D006680:HLA Antigens / Q000276:immunology*; D015235:HLA-B Antigens; D015796:HLA-B27 Antigen; D015793:HLA-B7 Antigen; D006801:Humans; D007130:Immunoglobulin Idiotypes / Q000276:immunology; D066298:In Vitro Techniques; D013167:Spondylitis, Ankylosing / Q000276:immunology
D000911:Antibodies, Monoclonal; D006680:HLA Antigens; D015235:HLA-B Antigens; D015796:HLA-B27 Antigen; C489636:HLA-B42 antigen; C113783:HLA-B54 antigen; C082944:HLA-B55 antigen; C056079:HLA-B56 antigen; C088036:HLA-B67 antigen; D015793:HLA-B7 Antigen; C088197:HLA-B73 antigen; D007130:Immunoglobulin Idiotypes