Inhibitory effects of tumor necrosis factor on hematopoiesis seen in vitro are translated to increased numbers of both committed and multipotent progenitors in TNF-deficient mice

Abstract

Objectives: The effects of TNF deficiency on myelopoiesis were evaluated in long-term (LTBMC) and short-term bone marrow cultures (STBMC) and compared to hematopoietic activity in vivo in TNF-deficient mice.

Methods: LTBMC and STBMC were established from bone marrow of TNF-deficient mice in the presence or absence of soluble TNF. Total cell production was measured over time, as well as the number of colony-forming units in culture (CFU-C). Morphology of nonadherent (NA) cells in LTBMC was assessed after 10 weeks. Bone marrow cells (BMC) and peripheral blood (PB) cells were used to determine lineage distribution within the hematopoietic system. BMC were sorted to obtain Lin(-)c-kit(+)Sca-1- and Lin(-)c-kit(+)Sca-1+ cells, which were plated in semisolid media to determine CFU-C numbers or injected into irradiated recipients to determine colony formation in the spleen (CFU-S).

Results: TNF-deficient LTBMC and STBMC show increased proliferative capacity, which can be inhibited by exogenous TNF to wild-type levels. Morphological analysis of NA cells from TNF-deficient LTBMC revealed increased numbers of cells at early stages of granulocytic differentiation (myeloblasts/promyelocytes) paralleled by a sharp decrease in the number of terminally differentiated polymorphonuclear neutrophils. Slightly elevated numbers of leukocytes, mainly neutrophils, were detected in PB of TNF-deficient mice. In bone marrow of TNF-deficient mice a significant increase in the number of both CFU-GM within Lin(-)c-kit(+)Sca-1- population and CFU-S within Lin(-)c-kit(+)Sca-1+ population was observed.

Conclusions: TNF has inhibitory effects on granulocyte-macrophage progenitors in vitro and on committed and primitive hematopoietic progenitors in vivo. However, in adult organism TNF deficiency is mostly compensated and controlled.