Pesce Viglietti Ayelén Ivana, Arriola Benitez Paula Constanza, Gentilini María Virginia, Velásquez Lis Noelia, Fossati Carlos Alberto, Giambartolomei Guillermo Hernán, Delpino María Victoria
Instituto de Inmunología, Genética y Metabolismo, Hospital de Clínicas José de San Martín, Facultad de Medicina, CONICET-Universidad de Buenos Aires, Buenos Aires, Argentina.
Instituto de Medicina Experimental, CONICET-Academia Nacional de Medicina, Buenos Aires, Argentina.
Infect Immun. 2015 Oct 12;84(1):11-20. doi: 10.1128/IAI.01049-15. Print 2016 Jan.
Osteoarticular brucellosis is the most common localization of human active disease. Osteocytes are the most abundant cells of bone. They secrete factors that regulate the differentiation of both osteoblasts and osteoclasts during bone remodeling. The aim of this study is to determine if Brucella abortus infection modifies osteocyte function. Our results indicate that B. abortus infection induced matrix metalloproteinase 2 (MMP-2), receptor activator for NF-κB ligand (RANKL), proinflammatory cytokines, and keratinocyte chemoattractant (KC) secretion by osteocytes. In addition, supernatants from B. abortus-infected osteocytes induced bone marrow-derived monocytes (BMM) to undergo osteoclastogenesis. Using neutralizing antibodies against tumor necrosis factor alpha (TNF-α) or osteoprotegerin (OPG), RANKL's decoy receptor, we determined that TNF-α and RANKL are involved in osteoclastogenesis induced by supernatants from B. abortus-infected osteocytes. Connexin 43 (Cx43) and the integrins E11/gp38, integrin-α, integrin-β, and CD44 are involved in cell-cell interactions necessary for osteocyte survival. B. abortus infection inhibited the expression of Cx43 but did not modify the expression of integrins. Yet the expression of both Cx43 and integrins was inhibited by supernatants from B. abortus-infected macrophages. B. abortus infection was not capable of inducing osteocyte apoptosis. However, supernatants from B. abortus-infected macrophages induced osteocyte apoptosis in a dose-dependent manner. Taken together, our results indicate that B. abortus infection could alter osteocyte function, contributing to bone damage.
骨关节布鲁氏菌病是人类活动性疾病最常见的发病部位。骨细胞是骨骼中数量最多的细胞。它们分泌在骨重塑过程中调节成骨细胞和破骨细胞分化的因子。本研究的目的是确定流产布鲁氏菌感染是否会改变骨细胞功能。我们的结果表明,流产布鲁氏菌感染诱导骨细胞分泌基质金属蛋白酶2(MMP-2)、核因子κB受体活化因子配体(RANKL)、促炎细胞因子和角质形成细胞趋化因子(KC)。此外,流产布鲁氏菌感染的骨细胞培养上清液可诱导骨髓来源的单核细胞(BMM)发生破骨细胞生成。使用针对肿瘤坏死因子α(TNF-α)或骨保护素(OPG,RANKL的诱饵受体)的中和抗体,我们确定TNF-α和RANKL参与了流产布鲁氏菌感染的骨细胞培养上清液诱导的破骨细胞生成。连接蛋白43(Cx43)以及整合素E11/gp38、整合素-α、整合素-β和CD44参与骨细胞存活所必需的细胞间相互作用。流产布鲁氏菌感染抑制Cx43的表达,但不改变整合素的表达。然而,流产布鲁氏菌感染的巨噬细胞培养上清液抑制了Cx43和整合素的表达。流产布鲁氏菌感染不能诱导骨细胞凋亡。然而,流产布鲁氏菌感染的巨噬细胞培养上清液以剂量依赖的方式诱导骨细胞凋亡。综上所述,我们的结果表明,流产布鲁氏菌感染可能改变骨细胞功能,导致骨损伤。