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Alcohol and Alcoholism (Oxford, Oxfordshire) (1)
Cellular immunology (1)
Chervenak, Robert (2)
Wang, Hao (2)
Zhou, Huijuan (2)
Brunson, Lee Ellen (1)
Chervenak, Deborah C. (1)
Mahler, Simon (1)
Moscatello, Kim M. (1)
Wolcott, Michael (1)
Wolcott, R. Michael (1)
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Alcohol Affects the Late Differentiation of Progenitor B Cells
Alcohol and Alcoholism (Oxford, Oxfordshire)
Aims: Previous studies show that alcohol exposure can affect the differentiation of progenitor B cells. Before final commitment to a B lineage, progenitor B cells usually undergo several important stages. However, it is still unclear whether alcohol alters B cell differentiation at which stages. The aim of this study was to determine which stage(s) of progenitor cell differentiation are affected by alcohol and to elucidate the mechanism(s) responsible for the effect of alcohol on B cell differentiation. Methods: Oligoclonal-neonatal-progenitor (ONP) cells from bone marrow cells of 2-week-old mice were cultured under different conditions in vitro with or without the exposure of 100 mM alcohol. Phenotype analysis was performed at different time points and expression levels of transcription factors (TFs) and cytokine receptors were measured quantitatively and kinetically. Results: After 3 days in vitro culture, ONP cells differentiated into two populations: B220−CD11b− and B220−CD11b+ cells. B220−CD11b− cells can further differentiate into B lineage cells only with the support of B220−CD11b+ cells. Cells exposed to 100 mM of alcohol during the first 3 days of culture showed no statistically significant difference in B cell formation after 12 days compared with the control group. However, cells exposed to alcohol from Day 4 till the end of culture yield very few B cells. Expression levels of TFs and cytokine receptors were down-regulated kinetically among ONP cells co-cultured with the addition of 100 mM alcohol. Conclusions: Alcohol affects the ONP cell differentiation into B lineage at a late stage. Alcohol also down-regulates the expression level of TFs and cytokine receptors resulting in the impairment of B cell differentiation.
Ethanol exhibits specificity in its effects on differentiation of hematopoietic progenitors1
Moscatello, Kim M.
Brunson, Lee Ellen
Chervenak, Deborah C.
Wolcott, R. Michael
Ethanol is a known teratogen but the mechanisms by which this simple compound affects fetal development remain unresolved. The goal of the current study was to determine the mechanism by which ethanol affects lymphoid differentiation using an in vitro model of ethanol exposure. Primitive hematopoietic oligoclonal neonatal progenitor cells (ONP), with the phenotype Lin−HSAloCD43loSca-1−c-Kit+ that are present in neonatal but not adult bone marrow were sorted from the bone marrow of 2-week-old C57BL/6J mice and cultured under conditions that favor either B cell or myeloid cell differentiation with or without addition of ethanol. The overall growth of the ONP cells was not significantly affected by inclusion of up to 100mM ethanol in the culture medium. However, the differentiation of the progenitor cells along the B-cell pathway was significantly impaired by ethanol in a dose dependent manner. Exposure of ONP cells to 100mM ethanol resulted in greater than 95% inhibition of B cell differentiation. Conversely, ethanol concentrations up to and including 100mM had no significant effect on differentiation along the myeloid pathway. The effect of ethanol on transcription factor expression was consistent with the effects on differentiation. ONP cells grown in 100mM ethanol failed to up-regulate Pax5 and EBF, transcriptional regulators that are necessary for B cell development. However, ethanol had no significant effect on the up-regulation of PU.1, a transcription factor that, when expressed in high concentration, favors myeloid cell development. Taken together, these results suggest that ethanol has specificity in its effects on differentiation of hematopoietic progenitors.
Rodent; B cells; Monocytes/macrophages; Cell differentiation; Hematopoiesis
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