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Mechanism of cell mediated cyto toxicity at the single cell level 6. direct assessment of the cyto toxic potential of human peripheral blood nonlytic effector target cell conjugates



Mechanism of cell mediated cyto toxicity at the single cell level 6. direct assessment of the cyto toxic potential of human peripheral blood nonlytic effector target cell conjugates



Journal of Immunology 132(2): 594-598



Single cell cytotoxicity assays reveal that a large percentage of lymphocytes are unable to kill attached targets in a 4 to 18 h assay. Additional signals (in the form of lectin or anti-target antibody) delivered to target-bound lymphocytes enable these previously non-lytic lymphocytes to kill attached target cells. This finding was obtained by using a modification of the single cell assay, in which lectin or target cell antibody is incorporated into agarose with performed lymphocyte-target conjugates. Human peripheral blood lymphocytes (PBL) or Percoll density gradient-enriched large granular lymphocytes (LGL) were used as effector cells in natural killer (NK), antibody-dependent cellular cytotoxicity (ADCC) and lectin-dependent cellular cytotoxicity (LDCC) assay systems. The targets used were NK-sensitive K562 and Molt-4 [human leukemia cell lines] and NK-insensitive Raji [human Burkitt's lymphoma cells]. The following findings were made in the modified single cell assay. The frequency of cytotoxic NK or ADCC effector cells was not augmented, suggesting that the initial trigger was sufficient for lytic expression in these instances. The NK sensitive targets used do not bind nonspecifically to the LDCC effector cells. K562 coated with Con A [concanavalin A] serve as LDCC targets. The frequency of 2 target conjugate lysis by NK/K [killer] effectors was not augmented by Con A. Evidently, Con A does not potentiate the killing of multiple targets bound to a single cytotoxic lymphocyte. Although conjugates formed between LGL or PBL and NK insensitive Raji are non-lethal, significant lysis was observed when these conjugates were suspended in Con A or antibody agarose. Raji bind to cytotoxic NK,K and LDCC effector cells, but are lysed only when the appropriate trigger is provided. The cytotoxic potential of non-lytic conjugates appears to lie within the low density Percoll fraction, although the high density lymphocytes are able to nonlethally bind to targets. Target recognition and/or binding by the effector cells is a distinct event from the trigger of lytic process. The implications of these findings are discussed.

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Accession: 005866830

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PMID: 6606675



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