结核分枝杆菌抗原对人单核细胞核苷酸结合寡聚化结构域样受体蛋白12表达的影响

doi:10.3969/j.issn.1000-6621.2015.04.002

中国防痨杂志 ›› 2015, Vol. 37 ›› Issue (4): 339-343.doi: 10.3969/j.issn.1000-6621.2015.04.002

结核分枝杆菌抗原对人单核细胞核苷酸结合寡聚化结构域样受体蛋白12表达的影响

刘艳华王若程小星

100091 北京，解放军第三〇九医院全军结核病研究所全军结核病防治重点实验室结核病诊疗新技术北京市重点实验室

收稿日期:2014-07-02 出版日期:2015-04-10 发布日期:2015-04-03
通信作者: 程小星 E-mail:xcheng2@139.com
基金资助:
国家自然科学基金青年基金项目（81101219）

M.tuberculosis antigens inhibit the NLRP12 expression in monocytes from patients with active pulmonary TB

LIU Yan-hua, WANG Ruo, CHENG Xiao-xing

Army Tuberculosis Prevention and Control Key Laboratory, Beijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute for Tuberculosis Research, The 309th Hospital of Chinese PLA, Beijing 100091, China

Received:2014-07-02 Online:2015-04-10 Published:2015-04-03
Contact: CHENG Xiao-xing E-mail:xcheng2@139.com

摘要/Abstract

摘要： 目的研究结核分枝杆菌（Mycobacterium tuberculosis, Mtb）抗原对人单核细胞中核苷酸结合寡聚化结构域样受体蛋白12（NLRP12）表达的影响，为深入研究NLRP12在结核病中的作用奠定实验基础。方法采集15例活动性肺结核患者和15名健康人的抗凝血，分离外周血中单核细胞，用Mtb抗原H37Rv全菌裂解液、早期分泌靶抗原6(EAST-6)和培养分泌蛋白10(CFP-10)的抗原多肽刺激，提取单核细胞的总RNA，采用荧光定量PCR方法检测NLRP12 mRNA的表达，采用配对t检验分析刺激前后单核细胞NLRP12的表达差异。以P<0.05为差异有统计学意义。结果全菌裂解液、EAST-6和CFP-10抗原多肽刺激后的活动性肺结核患者单核细胞与未刺激前的单核细胞相比，其NLRP12 mRNA的表达显著下调，由未刺激前的（1.8±1.26）×10^-2分别下降到（1.09±0.78）×10^-2（t=3.826，P<0.01）和（1.14±0.87）×10^-2（t=3.695，P<0.01）。全菌裂解液、EAST-6和CFP-10抗原多肽刺激后，健康人单核细胞中NLRP12 mRNA的表达与未刺激前相比也降低，由未刺激前的（1.65±0.99）×10^-2分别降至（1.2±0.87）×10^-2（t=1.909，P>0.05）和（1.38±1.02）×10^-2（t=1.151，P>0.05），差异无统计学意义。结论 Mtb抗原体外刺激能够抑制活动性肺结核患者单核细胞NLRP12的表达。

关键词: 结核, 肺, 结核分枝杆菌, 细胞内信号肽和蛋白质类：单核细胞, 重组融合蛋白质类

Abstract: Objective To study whether Mtb antigens have effect on the NLRP12 expression in peripheral blood monocytes, and provide experimental data to investigate the role of NLRP12 in tuberculosis. Methods Peripheral blood were collected from 15 healthy controls and 15 patients with active pulmonary tuberculosis. Monocytes were isolated from peripheral blood and then stimulated with Mtb antigens, including whole cell lysate of H37Rv strain and ESAT-6/CFP-10 peptide pool. Total RNA was extracted from unstimulated and stimulated cells, NLRP12 mRNA was detected by fluorescence quantitative PCR, and the difference of NLRP12 mRNA expression between stimulated and unstimulated cells was analyzed by paired t test.P value less than 0.05 was considered statistically significant. Results The reduction of NLRP12 mRNA in monocytes from active pulmonary TB was significant following stimulation by whole cell lysate and peptide pool, which reduced to (1.09±0.78)×10^-2 (t=3.826, P<0.01) and (1.14±0.87)×10^-2 (t=3.695, P<0.01) from (1.8±1.26)×10^-2, respectively. The expression of NLRP12 mRNA in monocytes from healthy subjects was no significant change following stimulation by whole cell lysate and peptide pool, although which reduced to (1.2±0.87)×10^-2 (t=1.909, P>0.05) and (1.38±1.02)×10^-2 (t=1.151, P>0.05) from (1.65±0.99)×10^-2, respectively. Conclusion Mtb antigens can inhibit the NLRP12 expression in monocytes from patients with active pulmonary Tin vitro.

Key words: Tuberculosis, pulmonary, Mycobacterium tuberculosis, Intracellular signaling peptides and proteins, Monocytes, Recombinant fusion proteins

刘艳华，王若，程小星. 结核分枝杆菌抗原对人单核细胞核苷酸结合寡聚化结构域样受体蛋白12表达的影响[J]. 中国防痨杂志, 2015, 37(4): 339-343. doi: 10.3969/j.issn.1000-6621.2015.04.002

LIU Yan-hua, WANG Ruo, CHENG Xiao-xing. M.tuberculosis antigens inhibit the NLRP12 expression in monocytes from patients with active pulmonary TB[J]. Chinese Journal of Antituberculosis, 2015, 37(4): 339-343. doi: 10.3969/j.issn.1000-6621.2015.04.002

参考文献

［1］Wang L, Manji GA, Grenier JM, et al. PYPAF7, a novel PYRIN-containing Apaf1-like protein that regulates activation of NF-kb and caspase-1-dependent cytokine processing. J Biol Chem, 2002, 277 (33): 29874-29880.
［2］Williams KI, Lich JD, Duncan JA, et al. The CATERPILLER protein monarch-1 is an antagonist of toll-like receptor-, tumor necrosis factor alpha-, and Mycobacterium tuberculosis-induced pro-inflammatory signals. J Biol Chem, 2005, 280 (48):39914-39924.
［3］Lich JD, Williams KL, Moore CB, et al. Monarch-1 suppresses non-canonical NF-kappaB activation and p52 dependent chemokine expression in monocytes. J Immunol, 2007, 178 (3):1256-1260.
［4］中华医学会结核病学分会. 肺结核诊断和治疗指南. 中华结核和呼吸杂志，2001, 24(2): 70-74.
［5］刘艳华, 翟斐, 王心静, 等. 活动性结核病人PBMCs中PLZF的表达降低与iNKT细胞的减少相关.中国免疫学杂志，2012, 28(9): 831-834.
［6］Shami PJ, Kanai N, Wang LY,et al.Identification and characterization of a novel gene that is upregulated in leukaemia cells by nitric oxide. Br J Haematol, 2001,112 (1): 138-147.
［7］Nowak K, Kuczek M, Ostropolska L, et al. The covalent structure of glyceraldehyde-phosphate dehydrogenase from human muscles. Isolation and amino acid sequences of peptides from tryptic digest. Hoppe Seylers Z Physiol Chem, 1975, 356 (7): 1181-1183.
［8］Kanneganti TD, Lamkanfi M, Núňez G. Intracellular NOD-like receptors in host defense and disease. Immunity, 2007, 27(4):549-559.
［9］Chamaillard M, Hashimoto M, Horie Y, et al. An essential role for NOD1 in host recognition of bacterial peptidoglycan containing diaminopimelic acid. Nat Immunol, 2003, 4(7):702-707.
［10］Inohara N, Ogura Y, Fontalba A, et al. Host recognition of bacterial muramyl dipeptide mediated through NOD2. Implications for Crohn’s disease. J Biol Chem, 2003, 278(8):5509-5512.
［11］Anand PK, Malireddi RK, Lukens JR, et al. NLRP6 negatively regulates innate immunity and host defence against bacterial pathogens. Nature, 2012, 488(7411):389-393.
［12］Allen IC, Wilson JE, Schneider M, et al. NLRP12 supresses colon inflammation and tumorigenesis through the negative regulation of noncanonical NF-kB signaling. Immunity, 2012, 36 (5):742-754.
［13］Williams KL, Taxman DJ, Linhoff MW, et al. Cutting edge: Monarch-1: A pyrin/nucleotide-binding domain/leucine-rich repeat protein that controls classical and nonclassical MHC class I genes. J Immunol, 2003, 170(11):5354-5358.
［14］Vladimer GI, Weng D, Paquette SW, et al. The NLRP12 inflammasome recognizes Yersinia pestis. Immunity, 2012, 37(1):96-107.
［15］Zaki MD, Man SM, Vogel P, et al. Salmonella exploits NLRP12-dependent innate immune signaling to suppress host defenses duing infection. Proc Natl Acad Sci U S A, 2014, 111(1):385-390.
［16］Allen IC, McElvania-Tekippe E, Wilson JE, et al. Characteri-zation of NLRP12 during the in vivo host immune response to Klebsiella pneumoniae and Mycobacterium tuberculosis. PLoS One, 2013, 8(4):e60842.
［17］Dwivedi V, Manickam C, Patterson R, et al. Intranasal deli-very of whole cell lysate of Mycobacterium tuberculosis induces protective immune responses to a modified live porcine reproductive and respiratory syndrome virus vaccine in pigs. Vaccine, 2011, 29(23): 4067-4076.
［18］Manickam C, Dwivedi V, Miller J, et al. Mycobacterium tuberculosis whole cell lysate enhances proliferation of CD8 positive lymphocytes and nitric oxide secretion in the lungs of live porcine respiratory and reproductive syndrome virus vaccinated pigs. Viral Immunol, 2013, 26(1):102-108.
［19］Pathak SK, Basu S, Basu KK, et al. Direct extracellular interaction between the early secreted antigen ESAT-6 of Mycobacterium tuberculosis and TLR2 inhibits TLR signaling in macrophages. Nat Immunol, 2007, 8(6):610-618.
［20］Chatterjee S, Dwivedi VP, Singh Y, et al. Early secreted antigen ESAT-6 of Mycobacterium tuberculosis promotes protective T helper 17 cell responses in a Toll-like responses in a Toll-like receptor-2-dependent manner. PLoS Pathog, 2011, 7(11):e1002378.
［21］Mir SA, Verma I, Sharma S. Immunotherapeutic potential of recombinant ESAT-6 protein in mouse model of experimental tuberculosis. Immunol Lett, 2014, 158(1/2):88-94.

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结核分枝杆菌抗原对人单核细胞核苷酸结合寡聚化结构域样受体蛋白12表达的影响

M.tuberculosis antigens inhibit the NLRP12 expression in monocytes from patients with active pulmonary TB

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