Quiescent CD34+ early erythroid progenitors are resistant to several erythropoietic 'inhibitory' cytokines; role of FLIP.
In this study, quiescent bone marrow-derived CD34+ erythroid burst-forming units (BFU-E) were found to be resistant to the inhibitory effects of tumour necrosis factor (TNF)-alpha and -beta as well as interferon (IFN)-alpha, -beta and -gamma, in contrast to those stimulated by a combination of erytrhropoietin (Epo) plus kit ligand (KL). Unexpectedly, we found that TNF-alpha also inhibited the apoptosis of quiescent normal human CD34+ BFU-E cells. Accordingly, TNF-alpha added to CD34+ cells cultured for 2 d in serum-free medium protected clonogeneic BFU-E from undergoing serum deprivation-mediated apoptosis. Furthermore, the prosurvival effect of TNF-alpha in quiescent CD34+ cells was consistent with its ability to induce phosphorylation of mitogen-activated protein kinase (MAPK) p42/44. However, when added to CD34+ cells that were stimulated by Epo + KL, TNF-alpha induced apoptosis and inhibited proliferation of BFU-E. To explain this intriguing differential sensitivity between unstimulated CD34+ cells versus those stimulated by Epo + KL, we examined the expression of apoptosis-regulating genes (FLIP, BCL-2, BCL-XL, BAD and BAX) in these cells. Of all the genes tested, FLIP became rapidly downregulated in CD34+ cells 24 h after stimulation with Epo + KL, suggesting that it may protect quiescent CD34+ BFU-E progenitors residing in the bone marrow from the inhibitory effects of inflammatory cytokines. Thus, we hypothesize that cycling cells may become more sensitive to proapoptotic stimuli (e.g. chemotherapy, inhibitory cytokines) than quiescent ones because of the downregulation of protective FLIP.
D018952:Antigens, CD34 / Q000276:immunology*; D017209:Apoptosis / Q000235:genetics; D015153:Blotting, Western / Q000379:methods; D002478:Cells, Cultured; D016207:Cytokines / Q000276:immunology*; D004268:DNA-Binding Proteins; D015672:Erythroid Precursor Cells / Q000276:immunology*; D004921:Erythropoietin / Q000494:pharmacology; D005786:Gene Expression Regulation; D006801:Humans; D008213:Lymphocyte Activation; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019089:Stem Cell Factor / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics*; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology; D051720:Upstream Stimulatory Factors
D018952:Antigens, CD34; D016207:Cytokines; D004268:DNA-Binding Proteins; D019089:Stem Cell Factor; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; D051720:Upstream Stimulatory Factors; D004921:Erythropoietin; D019950:Mitogen-Activated Protein Kinase 1