Mice were immunized with purified human monocytes or granulocytes obtained by leukapheresis and isolated on dextran gradients or by countercurrent centrifugation-elutriation. A monoclonal antibody, Mo95, was generated in response to monocytes and was found to react strongly with monocytes, large granular lymphocytes (LGL), granulocytes, eosinophils, and some myelomonocytic leukemia cells, but not with normal T or B lymphocytes, platelets, red cells, or leukemic cell lines. Mo95 is an IgG1 antibody, which precipitated a 95 kD molecular weight antigen. Addition of the Mo95 antibody to monocytes in the absence of complement did not inhibit lysozyme secretion nor did it affect superoxide production, C3b-rosetting, nitrotetrazolium blue reduction, phagocytosis, or chemotactic responses. A second antibody, PMN70, was found to react exclusively with granulocytes and not with monocytes, lymphocytes, LGL, platelets, red cells, or any of the myelomonocytic, T-cell-derived or B-cell-derived leukemic cell lines tested. The PMN70 antibody immunoprecipitated a 70 kD molecular weight antigen found only on mature granulocytes. Mo95 and PMN70 appear to be distinct from five other tested monoclonal antibodies reactive to monocytes and/or granulocytes on the basis of the fluorescent cell sorter and immunoprecipitation studies performed.

Animals↗

PubMed1983

Determination of the optimal human cell lines for development of human hybridomas.

Six human cell lines were compared with each other and with murine myeloma NS-1 as to their sensitivity to HAT medium and their ability to form hybrids with human lymphocytes, secret monoclonal immunoglobulin, clone, and maintain detectable levels of monoclonal immunoglobulin secretion for a period of time after fusion. Fusion efficiencies varied from 0 to 50%, and the incidence of immunoglobulin secretion ranged between 1 and 78% of the hybrids. Immunoglobulin secretion of cloned hybrids varied from 0.8 to 1.6 micrograms/ml/10(6) cells among the human-human hybrids and was 2.4 micrograms/ml/10(6) cells by the human-mouse hybrid. Among the lines tested, UC729-6 and HF2 appeared optimal for pursuing further studies with human-human hybridomas. In addition, although only a small percentage of hybrids were produced with HMy2, a very high percentage secreted immunoglobulin, so that this line also warrants further investigation to improve the efficiency of hybrid formation. Implications for specific monoclonal antibody production and for therapy of leukemias and lymphomas by anti-idiotype antibodies are discussed.

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PubMedDate not supplied

Nursing solidarnösc.

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