de Pinho Rosa Teixeira, da Silva Wellington Seguins, de Castro Côrtes Luzia Monteiro, da Silva Vasconcelos Sousa Periela, de Araujo Soares Renata Oliveira, Alves Carlos Roberto
Laboratório de Imunologia Clínica, Av. Brasil 4365 - Manguinhos, Rio de Janeiro, CEP 21040-360, Brasil.
Laboratório de Imunologia Clínica, Av. Brasil 4365 - Manguinhos, Rio de Janeiro, CEP 21040-360, Brasil; Fundação Ataulpho de Paiva, Av. Pedro II 260 - São Cristóvão, Rio de Janeiro, CEP 2094 1000, Brasil.
Exp Parasitol. 2014 Dec;147:72-80. doi: 10.1016/j.exppara.2014.09.003. Epub 2014 Oct 18.
Matrix metalloproteinases (MMPs) constitute a large family of Zn(2+) and Ca(2+) dependent endopeptidases implicated in tissue remodeling and chronic inflammation. MMPs also play key roles in the activation of growth factors, chemokines and cytokines produced by many cell types, including lymphocytes, granulocytes, and, in particular, activated macrophages. Their synthesis and secretion appear to be important in a number of physiological processes, including the inflammatory process. Here, we investigated the interaction between human and mouse macrophages with T. cruzi Colombian and Y strains to characterize MMP-9 and cytokine production in this system. Supernatants and total extract of T. cruzi infected human and mouse macrophages were obtained and used to assess MMP-9 profile and inflammatory cytokines. The presence of metalloproteinase activity was determined by zymography, enzyme-linked immunosorbent assay and immunoblotting assays. The effect of cytokines on MMP-9 production in human macrophages was verified by previous incubation of cytokines on these cells in culture, and analyzed by zymography. We detected an increase in MMP-9 production in the culture supernatants of T. cruzi infected human and mouse macrophages. The addition of IL-1β or TNF-α to human macrophage cultures increased MMP-9 production. In contrast, MMP-9 production was down-modulated when human macrophage cultures were treated with IFN-γ or IL-4 before infection. Human macrophages infected with T. cruzi Y or Colombian strains produced increased levels of MMP-9, which was related to the production of cytokines such as IL-1β, TNF-α and IL-6.
基质金属蛋白酶（MMPs）构成了一个庞大的锌离子（Zn(2+)）和钙离子（Ca(2+)）依赖性内肽酶家族，与组织重塑和慢性炎症有关。MMPs在多种细胞类型（包括淋巴细胞、粒细胞，特别是活化的巨噬细胞）产生的生长因子、趋化因子和细胞因子的激活中也起着关键作用。它们的合成和分泌在包括炎症过程在内的许多生理过程中似乎都很重要。在此，我们研究了人类和小鼠巨噬细胞与克氏锥虫哥伦比亚株和Y株之间的相互作用，以表征该系统中MMP-9和细胞因子的产生。获取克氏锥虫感染的人类和小鼠巨噬细胞的上清液和总提取物，用于评估MMP-9谱和炎性细胞因子。通过酶谱法、酶联免疫吸附测定和免疫印迹测定来确定金属蛋白酶活性的存在。通过预先在培养的这些细胞中孵育细胞因子来验证细胞因子对人类巨噬细胞中MMP-9产生的影响，并通过酶谱法进行分析。我们检测到克氏锥虫感染的人类和小鼠巨噬细胞培养上清液中MMP-9的产生增加。向人类巨噬细胞培养物中添加IL-1β或TNF-α会增加MMP-9的产生。相反，在感染前用IFN-γ或IL-4处理人类巨噬细胞培养物时，MMP-9的产生会被下调。感染克氏锥虫Y株或哥伦比亚株的人类巨噬细胞产生的MMP-9水平升高，这与IL-1β、TNF-α和IL-6等细胞因子的产生有关。
