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Identification of an Anti‐Human Osteogenic Sarcoma Monoclonal‐Antibody‐Defined Antigen on Mitogen‐Stimulated Peripheral Blood Mononuclear Cells
Author(s) -
PRICE M. R.,
CAMPBELL D. G.,
BALDWIN R. W.
Publication year - 1983
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1983.tb00873.x
Subject(s) - microbiology and biotechnology , phytohaemagglutinin , antigen , antibody , peripheral blood mononuclear cell , monoclonal antibody , biology , immunoprecipitation , chemistry , immunology , lymphocyte , biochemistry , in vitro
An anti‐human osteogenic sarcoma monoclonal antibody (mouse IgG2b) termed 79 IT/36 was found to exert complement‐dependent cytotoxicity against phytohaemagglutinin (PHA)‐stimulated peripheral blood mononuclear (PBMN) cells. This reaction was examined by flow cytofluorimetry using indirect membrane immunofluorescence to detect cell‐bound antibody and by measurement of the binding of fluroscreen‐isothiocyanate‐conjugated 791T/36 antibody to cells. The antibody reacted strongly wiih peripheral blood blast cells induced by PHA, and there was negligible reactivity wiih resting lymphocytes. Maximum binding was observed after 3 days' culture with PHA, coinciding with maximum DNA synthesis, and this represented of the order of 2 × 10 5 antibody molecules bound per cell. After cell surface radioiodination of PHA‐stimulated PBMN cells, detergent lysis and immunoprecipitation of antigen wiih 791T/36 antibody and Sepharose‐protein A, the apparent molecular weight of this antigen was determined to be 72,000 by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis. This is identical to that of the 791T/36‐defined antigen expressed on various osteogenic sarcoma cell lines [3, 17], and by this criterion the antigen is distinguishable from other cell surface markers of activated human lymphocytes.