Tumor necrosis factor-alpha expressed constitutively in erythroid cells or induced by erythropoietin has negative and stimulatory roles in normal erythropoiesis and erythroleukemia

Abstract

Binding of erythropoietin (EPO) to its receptor (EPOR) on erythroid cells induces the activation of numerous signal transduction pathways, including the mitogen-activated protein kinase Jun-N-terminal kinase (JNK). In an effort to understand the regulation of EPO-induced proliferation and JNK activation, we have examined the role of potential autocrine factors in the proliferation of the murine erythroleukemia cell line HCD57. We report here that treatment of these cells with EPO induced the expression and secretion of tumor necrosis factor alpha (TNF-α). EPO-dependent proliferation was reduced by the addition of neutralizing antibodies to TNF-α, and exogenously added TNF-α induced proliferation of HCD57 cells. EPO also could induce TNF-α expression in BAF3 and DA3 myeloid cells ectopically expressing EPOR. Addition of TNF-α activated JNK in HCD57 cells, and the activity of JNK was partially inhibited by addition of a TNF-α neutralizing antibody. Primary human and murine erythroid progenitors expressed TNF-α in either an EPO-dependent or constitutive manner. However, TNF-α had an inhibitory effect on both immature primary human and murine cells, suggestive that the proliferative effects of TNF-α may be limited to erythroleukemic cells. This study suggests a novel role for autocrine TNF-α expression in the proliferation of erythroleukemia cells that is distinct from the effect of TNF-α in normal erythropoiesis.