Human anti-HLA-DQw2 monoclonal antibody secreted by an Epstein-Barr-virus--transformed lymphoblastoid cell line: assessment of the monoclonality, allospecificity, and target.
IgM molecules were purified by the use of anti-IgM antibody-coupled Sepharose from the culture supernatant of an Epstein-Barr-virus-transformed lymphoblastoid cell line, MP1, that secretes alloantibodies possessing HLA-DQw2 specificity as defined by the cytotoxicity assay. The obtained IgM preparation was labeled with radioactive iodine-125I and fractionated by gel filtration. It contained pentameric IgM and smaller oligomeric IgMs. When tested by the direct cellular binding assay against a panel of HLA-typed cell lines, they all showed the DR3 and DR7 association pattern characteristic of DQw2. A weak but significant binding was detected for DR1, DR6, and DR9. On isoelectrofocusing, MP1 pentameric IgM gave a restricted banding pattern comparable to monoclonal IgM obtained from a patient with Waldenström's syndrome. Moreover, the pattern was identical to that of IgM purified from the culture supernatant of a defined hybrid clone, 162, that was generated by fusing MP1 cells with heteromyeloma D33 cells. The target class II molecules showed the dimeric structure that conforms to DQw2 molecules.
Pistillo
M P
MP
Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy.
Tanigaki
N
N
Chua
R
R
Mazzoleni
O
O
Ferrara
G B
GB
D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000917:Antibody Formation; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D003601:Cytotoxicity Tests, Immunologic; D004591:Electrophoresis, Polyacrylamide Gel; D006683:HLA-DQ Antigens / Q000276:immunology*; D004854:Herpesvirus 4, Human; D007075:Immunoglobulin M / Q000276:immunology
D000911:Antibodies, Monoclonal; D006683:HLA-DQ Antigens; D007075:Immunoglobulin M