Lymphokine-activated killer cells in rats. III. A simple method for the purification of large granular lymphocytes and their rapid expansion and conversion into lymphokine-activated killer cells.
A simple method for the purification and rapid expansion of large granular lymphocytes into cells with efficient broad antitumor cytotoxicity after stimulation by human rIL-2 is described. Nylon-wool nonadherent splenic mononuclear leukocytes from Fischer 344 rats were cultured in medium containing 1,000 U/ml rIL-2. The initial response of a small subpopulation of cells (less than 2%) to rIL-2 was their adherence to the plastic surface. This response was noted as soon as 2 h after addition of rIL-2. 2-h rIL-2-activated plastic adherent lymphocytes were 90-98% LGL, expressed surface markers characteristic of rat NK cells (OX8 [CD8]+, asialo GM1, laminin+, OX19 [CD5]-, R1-3B3 [CD5]-, W3/25 [CD4]-, OX39 [CD25]-, Ia-, and Ig-), and expressed very high levels of cytotoxicity against YAC-1 target cells. In addition to the above markers, plastic-adherent LGLs obtained at 24, 48, or 72 h progressively expressed Ia surface antigens, but were not phagocytic and contained less than 1% monocytes/macrophages by morphology. When 24- or 48-h plastic-adherent LGL/NK cells were cultured over 3-4 d in rIL-2, the cells expanded between 30- and 100-fold, reaching densities between 2-3 X 10(6) cells/ml. These rapidly expanding LGL/NK cells also generated very high levels of LAK activity (including lysis of fresh NK-resistant solid tumor cells), expressed a phenotype characteristic of activated rat NK/LAK cells, and incorporated [3H]TdR into DNA. This technique not only provides a novel method for the purification of LGL/NK cells for in vitro studies but also provides a means for the rapid expansion of highly purified cells with high levels of broad antitumor (LAK) cytotoxicity.
Vujanovic
N L
NL
Pittsburgh Cancer Institute, Pennsylvania.
Herberman
R B
RB
Maghazachi
A A
AA
Hiserodt
J C
JC
D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D002469:Cell Separation; D002478:Cells, Cultured; D003601:Cytotoxicity Tests, Immunologic; D007376:Interleukin-2 / Q000494:pharmacology*; D007694:Killer Cells, Natural / Q000187:drug effects / Q000276:immunology*; D008297:Male; D010641:Phenotype; D051381:Rats; D011916:Rats, Inbred F344 / Q000276:immunology; D011994:Recombinant Proteins / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000954:Antigens, Surface; D007376:Interleukin-2; D011994:Recombinant Proteins