重组结核分枝杆菌抗原ESAT-6、CFP-10以及ESAT-6/CFP-10融合蛋白对巨噬细胞免疫反应的抑制作用

Macrophage Immune Response Suppression by Recombinant Mycobacterium tuberculosis Antigens, the ESAT-6, CFP-10, and ESAT-6/CFP-10 Fusion Proteins.

作者信息

Seghatoleslam Atefeh, Hemmati Mina, Ebadat Saeedeh, Movahedi Bahram, Mostafavi-Pour Zohreh

机构信息

Recombinant Protein Laboratory, Biochemistry Department, Medical School, Shiraz University of Medical Sciences, Shiraz, Iran; Histomorphometry and Stereology Research Center, Medical School, Shiraz University of Medical Sciences, Shiraz, Iran.

Recombinant Protein Laboratory, Biochemistry Department, Medical School, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Biochemistry, Faculty of Medicine, Birjand University of Medical Sciences, Birjand, Iran.

出版信息

Iran J Med Sci. 2016 Jul;41(4):296-304.

BACKGROUND

Macrophage immune responses are affected by the secretory proteins of Mycobacterium tuberculosis (Mtb). This study aimed to examine the immune responses of macrophages to Mtb secretory antigens, namely ESAT-6, CFP-10, and ESAT-6/CFP-10.

METHODS

THP-1 cells (a human monocytic cell line) were cultured and differentiated to macrophages by phorbol 12-myristate 13-acetate. The cytotoxicity of the recombinant Mtb proteins was assessed using the MTT assay. Two important immune responses of macrophages, namely NO and ROS production, were measured in response to the ESAT-6, CFP-10, and ESAT-6/CFP-10 antigens. The data were analyzed using one-way ANOVA with SPSS, version 16, and considered significant at P<0.05.

RESULTS

The results showed that the ESAT-6, CFP-10, and ESAT-6/CFP-10 proteins markedly reduced macrophage immune response. The treatment of the THP-1-differentiated cells with ESAT-6, CFP-10, and ESAT-6/CFP-10 reduced NO and ROS production. The treated THP-1-differentiated cells exhibited less inducible NO synthase activity than did the untreated cells. No toxic effect on macrophage viability was observed for the applied proteins at the different concentrations.

CONCLUSION

It seems that the decline in macrophage immune response is due to the suppression of NO and ROS production pathways without any effect on cell viability.

背景

巨噬细胞免疫反应受结核分枝杆菌（Mtb）分泌蛋白的影响。本研究旨在检测巨噬细胞对Mtb分泌抗原（即ESAT-6、CFP-10和ESAT-6/CFP-10）的免疫反应。

方法

培养THP-1细胞（一种人单核细胞系），并用佛波酯12-肉豆蔻酸酯13-乙酸酯将其分化为巨噬细胞。使用MTT法评估重组Mtb蛋白的细胞毒性。检测巨噬细胞对ESAT-6、CFP-10和ESAT-6/CFP-10抗原产生的两种重要免疫反应，即一氧化氮（NO）和活性氧（ROS）的产生。使用SPSS 16版软件通过单因素方差分析对数据进行分析，P<0.05认为差异有统计学意义。

结果

结果显示，ESAT-6、CFP-10和ESAT-6/CFP-10蛋白显著降低巨噬细胞免疫反应。用ESAT-6、CFP-10和ESAT-6/CFP-10处理THP-1分化细胞可降低NO和ROS的产生。与未处理细胞相比，经处理的THP-1分化细胞诱导型一氧化氮合酶活性较低。在所应用的不同浓度蛋白中，未观察到对巨噬细胞活力的毒性作用。