Generation and characterization of chicken bone marrow-derived dendritic cells

Wu, Zhiguang, Rothwell, Lisa, Young, John R. , Kaufman, Jim, Butter, Colin and Kaiser, Pete (2010) Generation and characterization of chicken bone marrow-derived dendritic cells. Immunology, 129 (1). pp. 133-145. ISSN 0019-2805

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Dendritic cells (DCs) are bone marrow-derived professional antigen-presenting cells. The in vitro generation of DCs from either bone marrow or blood is routine in mammals. Their distinct morphology and phenotype and their unique ability to stimulate naïve T cells are used to define DCs. In this study, chicken bone marrow cells were cultured in the presence of recombinant chicken granulocyte-macrophage colony-stimulating factor (GM-CSF) and recombinant chicken interleukin-4 (IL-4) for 7 days. The cultured population showed the typical morphology of DCs, with the surface phenotype of major histocompatibility complex (MHC) class II+ (high), CD11c+ (high), CD40+ (moderate), CD1·1+ (moderate), CD86+ (low), CD83- and DEC-205-. Upon maturation with lipopolysaccharide (LPS) or CD40L, surface expression of CD40, CD1·1, CD86, CD83 and DEC-205 was greatly increased. Endocytosis and phagocytosis were assessed by fluorescein isothiocyanate (FITC)-dextran uptake and fluorescent bead uptake, respectively, and both decreased after stimulation. Non-stimulated chicken bone marrow-derived DCs (chBM-DCs) stimulated both allogeneic and syngeneic peripheral blood lymphocytes (PBLs) to proliferate in a mixed lymphocyte reaction (MLR). LPS-or CD40L-stimulated chBM-DCs were more effective T-cell stimulators in MLR than non-stimulated chBM-DCs. Cultured chBM-DCs could be matured to a T helper type 1 (Th1)-promoting phenotype by LPS or CD40L stimulation, as determined by mRNA expression levels of Th1 and Th2 cytokines. We have therefore cultured functional chBM-DCs in a non-mammalian species for the first time. © 2009 Blackwell Publishing Ltd.

Keywords:CD1 antigen, CD40 antigen, CD40 ligand, CD83 antigen, CD86 antigen, cytokine, fluorescein isothiocyanate dextran, glycoprotein p 15095, lipopolysaccharide, major histocompatibility antigen class 2, messenger RNA, recombinant granulocyte macrophage colony stimulating factor, recombinant interleukin 4, animal cell, animal tissue, antigen expression, article, bone marrow, bone marrow cell, cell culture, cell maturation, cell population, cell proliferation, cell stimulation, cell structure, chicken, controlled study, dendritic cell, endocytosis, mixed lymphocyte reaction, nonhuman, nucleotide sequence, peripheral lymphocyte, phagocytosis, phenotype, priority journal, protein expression, T lymphocyte activation, Th1 cell, Th2 cell, Animals, Antigens, CD, Cells, Cultured, Chickens, Cytokines, Dendritic Cells, Granulocyte-Macrophage Colony-Stimulating Factor, Histocompatibility Antigens Class II, Interleukin-4, Lymphocyte Activation, Lymphocyte Culture Test, Mixed, Th1 Cells, Th2 Cells
Subjects:C Biological Sciences > C700 Molecular Biology, Biophysics and Biochemistry
Divisions:College of Science > School of Life Sciences
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ID Code:19858
Deposited On:31 Dec 2015 21:27

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