Supplementary characteristics of anti-MHC class II monoclonal antibodies elicited by an ALL cell line: immunofluorescence cytofluorometry, C-dependent cytotoxicity, two-dimensional analysis of antigen.
Monoclonal antibodies directed to MHC class II antigen(s), elicited by a non-T, non-B ALL cell line, were characterized by immunofluorescence flow cytofluorometry and ELISA immunofiltration measurements of their immunoreactivity with selected neoplastic hemopoietic cell lines, determination of their complement-dependent cytotoxic activity against isolated peripheral blood B and T lymphocytes and by two-dimensional electrophoretic analysis (isoelectric focusing, SDS-PAGE) of radiolabeled, immunoprecipitated by these antibodies cell surface antigens. Patterns of these immunological reactivities, as well as two-dimensional radioimmunoprecipitation patterns (acidic heavy chain p35 and basic light chain p30) of antigens recognized by these antibodies confirm their anti-MHC class II specificity. One of these antibodies (braFB6; IgG2b) displayed identical pattern of expression on cell lines and cell types as the typical anti-MHC class II antibodies, but immunoprecipitated only a single chain p30 radioiodinated cell surface protein (with two-dimensional pattern close to the beta-chain of MHC class II DR antigen). These properties indicate the ability of braFB6 monoclonal antibody to recognize a nonpolymorphic determinant of DP-MHC class II antigen.
Chorváth
B
B
Cancer Research Institute, Slovak Academy of Sciences, Bratislava, Czechoslovakia.
Duraj
J
J
Sedlák
J
J
Plesková
I
I
Munozová
H
H
Buc
M
M
D000911:Antibodies, Monoclonal / Q000276:immunology*; D002460:Cell Line; D003165:Complement System Proteins / Q000276:immunology; D003602:Cytotoxicity, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006681:HLA-D Antigens / Q000276:immunology*; D006801:Humans; D007945:Leukemia, Lymphoid / Q000276:immunology*
D000911:Antibodies, Monoclonal; D006681:HLA-D Antigens; D003165:Complement System Proteins