Role of natural killer cells, in comparison with T lymphocytes and monocytes, in the regulation of normal human megakaryocytopoiesis in vitro.

Natural killer (NK) cells are thought to play an important role in host defense against virus-infected and neoplastic cells. Recent reports suggest that these cells may also influence developmental events in the course of normal erythropoiesis and granulopoiesis. The role of NK cells in the regulation of normal human megakaryocytopoiesis has not been reported, but clinical observations suggest that NK cell effects on megakaryocyte progenitors might differ from those of other cell lineages. We therefore carried out in vitro studies designed to assess the influence of NK cells on the growth of autologous megakaryocyte colony-forming units (CFU-Meg). To provide a frame of reference for these experiments, the effect of T lymphocytes, and monocyte-macrophages (M luminal diameter) on autologous CFU-Meg cloning efficiency was also studied. Purified immune effector cells were co-cultured in plasma clots with both unseparated, and progenitor cell-enriched marrow mononuclear cells (MNC) at target to effector cell ratios varying from 100:1 to 1:1. Resulting megakaryocyte colonies were enumerated by indirect fluorescence microscopy by using a rabbit anti-human platelet glycoprotein antiserum as probe for cells of the megakaryocyte lineage. The addition of NK cells to both unseparated (n = 12), and progenitor-enriched (n = 3) MNC at target to effector cell ratios of 2:1 and 1:1 resulted in a significant (p less than 0.05) augmentation in CFU-Meg-derived colony formation. Augmentation of colony formation was blocked by incubating the NK cells in Leu-11b monoclonal antibody. Stimulation appeared to be carried out by the production of a soluble growth factor which was detectable in NK cell-conditioned medium. Exposure of NK cells for 18 hr to highly purified or recombinant gamma-interferon (500 U/10(6) cells), a putative NK cell stimulator, neither increased nor abrogated the stimulatory effect. The latter could be accomplished, however, by centrifuging (200 X G for 5 min), and then preincubating the target and effector cells together for 3 hr before plating. At no time was significant inhibition of CFU-Meg demonstrated. In contrast to these results, when tested at the same ratios, and under the same conditions, no consistent effect on CFU-Meg cloning efficiency could be demonstrated by the addition of whole T cells, T cell subsets, or M luminal diameter. These results suggest that NK cells could play a role in the basal regulation of megakaryocytopoiesis.(ABSTRACT TRUNCATED AT 400 WORDS)