Zhao Jingge, Shiratori Beata, Chagan-Yasutan Haorile, Matsumoto Makoto, Niki Toshiro, Tanaka Michinori, Takahashi Yayoi, Usami Osumu, Ashino Yugo, Hattori Toshio
Division of Disaster-Related Infectious Diseases, International Research Institute of Disaster Science, Tohoku University, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575, Japan.
KKR Tohoku Kosai Hospital, Kokubun-cho 2-3-11, Aoba-ku, Sendai-shi, Miyagi 980-0803, Japan.
Int J Mol Sci. 2017 Jun 28;18(7):1382. doi: 10.3390/ijms18071382.
In this study, we investigated the role of a matricellular protein galectin-9 (Gal-9) in pleural effusion related to tuberculosis (TB). Plasma and pleural fluid of a patient with extrapulmonary TB were analyzed for cytokine content by ELISA and Luminex. Peripheral blood mononuclear cells (PBMCs) and pleural fluid cells (PFCs) were examined for interferon-γ (IFN-γ) secretion by the enzyme-linked immunospot (ELISPOT) assay or IFN-γ ELISA, for apoptosis and necrosis by Cell Death Detection ELISA, and also underwent cell sorting. The results indicate that compared to plasma, pleural fluid had increased levels of IFN-γ (1.6 vs. 55.5 pg/mL), IL-10, IL-12p40, vascular endothelial growth factor (VEGF), and Gal-9 (3.0 vs. 936.0 pg/mL), respectively. PFCs culture supernatant exhibited higher concentration of Gal-9 compared to PBMCs in culture, consistent with enriched Gal-9 staining in the granuloma that is in closer vicinity to PFCs compared to PBMCs. PFCS displayed higher IFN-γ secretion after stimulation with TB antigens ESAT-6/CFP-10. Furthermore, in PFCs, Gal-9 alone could stimulate IFN-γ synthesis in culture or ELISPOT, which was inhibited by a Gal-9 antagonist lactose, and which may promote apoptosis and necrosis. These findings suggest that Gal-9 could modulate immune responses and participate in immunopathology of pleural effusion during TB.
在本研究中，我们调查了基质细胞蛋白半乳糖凝集素-9（Gal-9）在结核（TB）相关胸腔积液中的作用。通过酶联免疫吸附测定（ELISA）和Luminex对一名肺外结核患者的血浆和胸腔积液进行细胞因子含量分析。通过酶联免疫斑点（ELISPOT）测定或IFN-γ ELISA检测外周血单核细胞（PBMC）和胸腔积液细胞（PFC）的干扰素-γ（IFN-γ）分泌，通过细胞死亡检测ELISA检测细胞凋亡和坏死情况，并且进行细胞分选。结果表明，与血浆相比，胸腔积液中IFN-γ（1.6对55.5 pg/mL）、IL-10、IL-12p40、血管内皮生长因子（VEGF）和Gal-9（3.0对936.0 pg/mL）的水平分别升高。与培养中的PBMC相比，PFC培养上清液中Gal-9的浓度更高，这与肉芽肿中靠近PFC处的Gal-9染色比靠近PBMC处更丰富一致。在用结核抗原ESAT-6/CFP-10刺激后，PFC显示出更高的IFN-γ分泌。此外，在PFC中，单独的Gal-9可在培养或ELISPOT中刺激IFN-γ合成，并被Gal-9拮抗剂乳糖抑制，且这可能促进细胞凋亡和坏死。这些发现表明，Gal-9可调节免疫反应并参与结核期间胸腔积液的免疫病理学过程。
