Purification and target cell range of in vivo elicited blast natural killer cells.
Blast natural killer (NK) cells were elicited in the spleens of mice by treatments with the interferon inducers lymphocytic choriomeningitis virus (LCMV) or polyinosinic-polycytidylic acid (poly I:C). The blast-NK cells, separated on the basis of size by centrifugal elutriation, were compared with blast cytotoxic T lymphocytes (CTL) generated during infection with LCMV. In vivo treatments with antibody to asialo GM1 (AGM1) blocked the appearance of blast-NK cells but not blast-CTL. Antibody and complement depletion experiments indicated that the blast-NK cells were AGM1+, NK 1.2+/-, Lyt-5+/-, Thy+/-, Qa-5/NK 1.1+, Lyt-2-, B23.1-, and J11d-. Blast-NK cells could be unequivocally distinguished from blast-CTL, because the blast-CTL were completely sensitive to treatments with anti-Lyt-2 and complement, whereas the blast-NK cells were completely resistant. The blast-NK cells were purified from populations of large-size cells by antibody and complement treatments that depleted the co-eluting monocyte/macrophages and polymorphonuclear leukocytes. The population resulting after separation from dead cells over Percoll gradients represented approximately 1% of the total spleen cells, contained greater than 60% large granular lymphocytes and mediated greater than 15% killing of YAC-1 target cells in a 4-hr 51Cr release assay at an effector to target cell ratio of 1:1. The purified blast-NK cells lysed a broad range of target cells at relatively low effector to target cell ratios. The order of sensitivity of the target cells was YAC-1 much greater than K562 approximately equal to L-929 much greater than P815, consistent with that reported for NK cell-mediated lysis. The ability of the blast-NK cells to mediate lysis of NK cells also was examined. The purified NK cells mediated significant levels of lysis against the NK-like cloned line, NK1B6B10, in a 51Cr release assay. Furthermore, the purified blast-NK cells mediated lysis of bound blast-NK cells in a single-cell agarose assay. These results indicate that highly purified blast-NK cells are exceptionally efficient at mediating lysis and suggest that NK cells may act to negatively regulate the proliferation of NK cells by lysing other NK cells.
D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D002469:Cell Separation / Q000379:methods; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods; D003602:Cytotoxicity, Immunologic; D005677:G(M1) Ganglioside; D006028:Glycosphingolipids / Q000276:immunology; D007106:Immune Sera / Q000008:administration & dosage; D007694:Killer Cells, Natural / Q000145:classification / Q000276:immunology*; D008213:Lymphocyte Activation; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D010641:Phenotype
D000954:Antigens, Surface; D006028:Glycosphingolipids; D007106:Immune Sera; D005677:G(M1) Ganglioside; C018835:asialo GM1 ganglioside