Lymphokine-activated killer cells in rats: Generation of natural killer cells and lymphokine-activated killer cells from bone marrow progenitor cells

Abstract

The coculture of rat bone marrow cells with recombinant interleukin-2 induced the generation of cells mediating natural killer (NK) activity and subsequent lymphokine-activated killer (LAK) activity depending upon the dose of IL-2 and time of culture. NK activity was detected as early as 4 to 5 days after the addition of IL-2 and could be evoked with as little as 5 to 50 U/ ml. The induced NK cells had large granular lymphocyte (LGL) morphology and expressed 0X8 and asialo GM 1 surface markers but did not express OX19 or W3/25 markers. LAK activity was detected only after 5 days of culture, and required above 100 U/ml IL-2. Cells mediating LAK activity also expressed OX8 and asialo GM 1 but not OX19. The generation of detectable NK and subsequent LAK activity was due to induction of early progenitor cells and not contaminating mature LGL/NK cells within the bone marrow population since of removal of such mature NK cells with l-leucine methyl ester (L-LME) did not affect the subsequent generation of either activity. Moreover, the removal of actively dividing cells as well as mature NK cells from the bone marrow by treatment with 5-fluorouracil (5-FU) in vivo enriched the remaining bone marrow population for both NK and LAK progenitor cells. The phenotype of the L-LME- and 5-FU-resistant NK and LAK progenitor cells within populations of bone marrow was determined by antibody plus complement depletion analysis. Although treatment of normal bone marrow with anti-asialo GM 1 + C reduced the induction of NK and LAK activity in 5-day cultures, treatment of 5-FU marrow with anti-asialo GM 1 + C did not affect either activity. Treatment with a pan-T cell antibody + C did not affect the development of NK or LAK activity under any conditions. Thus, the 5-FU-resistant NK/LAK progenitors were asialo GM 1 negative but became asialo GM 1 + after induction by IL-2. Finally, evidence that bone marrow-derived LAK cells were generated directly from the IL-2-induced NK cells was obtained by treating the IL-2-induced LGL/NK cells with L-LME. This eliminated the induced NK activity as well as the capacity of the remaining bone marrow cells to generate LAK activity. Thus, in rat bone marrow, NK cells appear to be derived from asialo GM 1 −, OX19 −, nongranular, 5-FU- and L-LME-resistant progenitor cells which can be induced to generate LGL with NK activity with low levels of IL-2. The IL-2-induced NK cells became asialo GM 1 +, OX8 +, OX19 − and have a granular morphology. Moreover these IL-2-induced NK cells can be further activated to mediate LAK activity by higher of IL-2