目的 研究白念珠菌磷脂甘露聚糖( PLM)诱导人急性单核细胞白血病细胞系细胞(THP-1)产生的炎症反应是否依赖Toll样受体(TLR)2.方法 实时荧光定量逆转录PCR分析PLM体外刺激THP-1细胞TLR2、TLR4、前炎症因子[白介素(IL)-6]和趋化因子(IL-8)的mRNA表达水平.酶联免疫吸附法检测IL-6、IL-8分泌含量.免疫印迹法分析TLR2的蛋白表达.结果 PLM可升高THP-1细胞的IL-6和IL-8 mRNA表达和分泌水平(均P＝0.0000).PLM上调THP-1细胞的TLR2 mRNA和蛋白表达水平(P＝0.0000),但对TLR4 mRNA表达无影响.PLM经β-D-甘露糖苷水解酶处理后,不能诱导上述受体及因子的表达.TLR2中和抗体能抑制PLM诱导的IL-6和IL-8产生(P ＝0.0003,P＝0.0010).结论 白念珠菌胞壁PLM依赖TLP2介导激活人THP-1细胞产生炎症反应.%Objective To investigate whether Candida albicans-native phospholipomannan ( PLM ) induce an inflammation response through Toll-like receptor ( TLR) 2 in human acute monocytic leukemia cell line ( THP-1 ) cells. Methods Human THP-1 monocytes were challenged with PLM in vitro. The mRNA expressions of TLR2, TLR4, proinflammatory cytokine [ interleukin (IL)-6], and chemokine (IL-8) were assayed by real time reverse transcription polymerase chain reaction. The secretions of IL-6 and IL-8 were measured by enzyme-linked immunosorbent assay. The expression of TLR2 was analyzed with Western blot. Results PLM increased the mRNA expressions and secretions of proinflammatory cytokines ( IL-6) and che-mokines (IL-8) in THP-1 cells (all P-0.0000). PLM up-regulated the mRNA and protein levels of TLR2 (P = 0. 0000), whereas the mRNA level of TLR4 was not altered. PLM hydrolyzed with |J-D-mannosidemannohydrolase failed to induce gene and protein expressions of TLR2, IL-6, and IL-8. Anti-TLR2-neutralizing antibody blocked the PLM-induced secretions of IL-6 and IL-8 in THP-1 cells ( P = 0. 0003, P = 0.0010). Conclusion Candida alblcans-native PLM may contribute to the inflammatory responses during Candida infection in a TLR2-dependent manner.
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