GENERATION OF MONOCLONAL-ANTIBODIES TO A HUMAN NATURAL-KILLER CLONE - CHARACTERIZATION OF 2 NATURAL KILLER-ASSOCIATED ANTIGENS, NKH1A AND NKH2, EXPRESSED ON SUBSETS OF LARGE GRANULAR LYMPHOCYTES
GENERATION OF MONOCLONAL-ANTIBODIES TO A HUMAN NATURAL-KILLER CLONE - CHARACTERIZATION OF 2 NATURAL KILLER-ASSOCIATED ANTIGENS, NKH1A AND NKH2, EXPRESSED ON SUBSETS OF LARGE GRANULAR LYMPHOCYTES
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DOI：
10.1172/jci111794
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发表时间：
1985-01-01
影响因子：
15.9
通讯作者：
RITZ, J
中科院分区：
文献类型：
--
作者：
HERCEND, T;GRIFFIN, JD;RITZ, J
The initial characterization of 2 monoclonal antibodies directed at antigens selectively expressed on large granular lymphocytes (LGL) is reported in the present paper. These 2 reagents, anti-natural killer (NK) H1A and anti-NKH2, were obtained following immunization of mouse spleen cells with a cloned human NK cell lined termed JTE. In fresh human peripheral blood, both anti-NKH1A and anti-NKH2 selectively reacted with cells that appeared morphologically as large granular lymphocytes. Complement lysis studies and 2 color fluorescence analysis demonstrated that some LGL express both antigens and other cells express only NKH1A or NKH2. Functional analysis of these subsets indicated that the population of NKH1A+ cells contains the entire pool of NK active lymphocytes, whereas expression of NKH2 antigen appeared to delineate a unique subpopulation pf LGL which, in a resting state, display a low degree of spontaneous cytotoxicity. Expression of NKH1A and NKH2 was also investigated using a series of 9 well characterized human NK clones. All NK clones were found to be NKH1A+ and 4 out of 9 also expressed NKH2. These results strongly supported the view that NKH1A is a pan-NK associated antigen, and indicated that at least a fraction of cloned NKH2+ LGL are strongly cytotoxic. Anti-NKH1A was shown to have the same specificity as the previously described N901 antibody and was found here to precipitate a 200,000-220,000-MW molecule in SDS[sodium dodecyl sulfate]-polyacrylamide gel electrophoresis (PAGE) analysis. Anti-NKH2 was specific for a structure that migrates at 60,000 MW in SDS-PAGE analysis under reducing conditions. Two color immunofluorescence analysis of NKH1A, NKH2 and other NK-associated antigens (Leu7 and B73.1) demonstrated variable degrees of coexpression of these antigens, which confirmed that NKH1A and NKH2 define distinct cell surface structures. Anti-NKH1A and anti-NKH2 appear to be useful reagents for characterizing LGL present in human peripheral blood and for identifying functionally relevant subsets within this heterogeneous population of cytotoxic lymphocytes.
