Zamani Fatemeh, Zare Shahneh Fateme, Aghebati-Maleki Leili, Baradaran Behzad
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. ; Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
Adv Pharm Bull. 2013;3(2):329-32. doi: 10.5681/apb.2013.053. Epub 2013 Aug 20.
CD14, one of the main differentiation markers on the surface of myeloid lineage cells, acts as a key role in activation of LPS-induced monocytes. LPS (lipopolysaccharide) binds to LPS-binding protein in plasma and are delivered to the cell surface receptor CD14. In this study, Various stimuli [Dimethyl Sulfoxide (DMSO), active 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and LPS], either alone or in combination, have been recognized that have an effect on the level of CD14 expression in the human HL-60 and U937 promonocytic cell lines and therefore induce their terminal differentiation into monocytes or mature macrophages.
U937 and HL-60 cells were cultured in RPMI 1640 supplemented with 10% FBS. For each cell line, 1Ã106 cells were seeded for 72 hours with DMSO, 14 days with LPS and 18 days with 1, 25-D3 in each well plate; then ELISA method was used to study their responses to the factors by means of anti-CD14.
ELISA assay demonstrated that U937 and HL-60 cells were induced by both [1,25(OH)2D3] and DMSO to obtain characteristics of adherent cells and express CD14 protein; moreover, LPS at a low dose increased CD14 expression on surface of this cells.
According to the our results, it is speculated that CD14 gene expression may be induced in human U937 and HL-60 cell lines by different factors including 1,25-D3, DMSO and LPS.
CD14是髓系细胞表面主要的分化标志物之一,在脂多糖(LPS)诱导的单核细胞激活中起关键作用。LPS与血浆中的LPS结合蛋白结合,并被递送至细胞表面受体CD14。在本研究中,已认识到各种刺激因素[二甲基亚砜(DMSO)、活性1,25 - 二羟基维生素D3 [1,25(OH)2D3]和LPS]单独或联合使用时,会对人HL - 60和U937原单核细胞系中CD14的表达水平产生影响,从而诱导它们终末分化为单核细胞或成熟巨噬细胞。
U937和HL - 60细胞在补充有10%胎牛血清的RPMI 1640培养基中培养。对于每个细胞系,在每个孔板中接种1×10⁶个细胞,分别用DMSO培养72小时,用LPS培养14天,用1,25 - D3培养18天;然后采用ELISA法,通过抗CD14来研究它们对这些因素的反应。
ELISA分析表明,[1,25(OH)2D3]和DMSO均可诱导U937和HL - 60细胞获得贴壁细胞特性并表达CD14蛋白;此外,低剂量的LPS可增加这些细胞表面CD14的表达。
根据我们的结果推测,包括1,25 - D3、DMSO和LPS在内的不同因素可能诱导人U937和HL - 60细胞系中CD14基因的表达。