STAT6介导的巨噬细胞极化对布鲁氏菌胞内存活的影响

doi:10.11843/j.issn.0366-6964.2022.01.026

Abstract

Abstract: This study aimed to investigate the effect of STAT6 mediated macrophage polarization on intracellular survival of Brucella. Brucella smooth strain S2308(S2308) and rough vaccine strain RB51(RB51) were used to infect macrophages.M1-type macrophage marker factors p65, NOS2 and IL-1β were detected by qRT-PCR. mRNA expression levels of M2-type macrophage marker factors STAT6, ARG1 and IL-10 were detected by qRT-PCR. The expression of M1-type marker CD86 and M2-type marker CD206 was detected by flow cytometry. The inhibitory effect of p-STAT6 protein and inhibitor AS on the protein was detected by Western Blot.The expression levels of M1-type cytokines TNF-α and IL-12 and M2-type cytokines IL-4 and IL-10 were detected by ELISA. Finally, CFU count of intracellular colonies was carried out. The qRT-PCR results showed that the mRNA expression of M1 type factor was significantly induced at 8 h and 12 h after infection, the expression of M2 type factor was low at 72 h, and the expression of M2 type factor was high at 72 h. Flow cytometry showed that S2308 could significantly induce CD86 expression at 12 h after infection, and CD206 expression at 72 h after infection, but RB51 had no effect on both.Western Blot results showed that strain S2308 activated STAT6 signaling pathway 72 h after infection, while RB51 hardly activated STAT6 signaling pathway. The inhibitory effect of inhibitor AS was the best at 2 μmol·L^-1 concentration. ELISA results showed that AS inhibitors could significantly inhibit the release of IL-4 and IL-10, and promote the release of TNF-α and IL-12.CFU count results showed that intracellular bacteria in S2308 group were firstly decreased and then significantly increased, and S2308+IL-4 group was significantly higher than S2308 group, S2308+AS group was significantly lower than S2308 group, and there was no significant difference in RB51 group. Brucella S2308 can induce the transformation of M1 type macrophages to M2 type macrophages through STAT6 in the late stage of infection, and promote the release of Th2 type cytokines, which is conducive to the intracellular survival of Brucella. RB51 does not activate this pathway and does not affect intracellular survival.

Key words: Brucella, macrophage polarization, STAT6 signal path, intracellular survival

CLC Number:

S852.614

XI Jing, WANG Yueli, DENG Xiaoyu, YANG Qin, LI Peidong, ZHANG Jiangwei, SUN Tianhao, ZHU Liangquan, YI Jihai, CHEN Chuangfu. Effect of STAT6 Mediated Macrophage Polarization on Intracellular Survival of Brucella[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(1): 263-271.

References 30

[1]	TIAN M X, YIN Y, LIAN Z M, et al. A rough Brucella mutant induced macrophage death depends on secretion activity of T4SS, but not on cellular Txnip- and Caspase-2-mediated signaling pathway[J]. Vet Microbiol, 2020, 244:108648.
[2]	SMITH J A. Brucella lipopolysaccharide and pathogenicity:the core of the matter[J]. Virulence, 2018, 9(1):379-382.
[3]	易继海, 王月丽, 王震, 等. 布鲁氏菌胞内存活机制与巨噬细胞极化关系研究进展[J]. 中国畜牧兽医, 2018, 45(4):1082-1088.YI J H, WANG Y L, WANG Z, et al. Research progress on the relationship between intracellular survival mechanism of Brucella and macropohage polarization[J]. China Animal Husbandry & Veterinary Medicine, 2018, 45(4):1082-1088. (in Chinese)
[4]	DENG Y M, LIU X Y, DUAN K F, et al. Research progress on brucellosis[J]. Curr Med Chem, 2019, 26(30):5598-5608.
[5]	KAZEMI S, VAISI-RAYGANI A, KERAMAT F, et al. Evaluation of the relationship between IL-12, IL-13 and TNF-α gene polymorphisms with the susceptibility to brucellosis:a case control study[J]. BMC Infect Dis, 2019, 19(1):1036.
[6]	CHEN Q L, LAI S J, YIN W W, et al. Epidemic characteristics, high-risk townships and space-time clusters of human brucellosis in Shanxi Province of China, 2005-2014[J]. BMC Infect Dis, 2016, 16(1):760.
[7]	FUNES S C, RIOS M, ESCOBAR-VERA J, et al. Implications of macrophage polarization in autoimmunity[J]. Immunology, 2018, 154(2):186-195.
[8]	周琦, 孙慧娟, 于栋华, 等. 巨噬细胞M1/M2型极化在不同疾病中的作用机制[J]. 中国药理学通报, 2020, 36(11):1502-1506.ZHOU Q, SUN H J, YU D H, et al. Mechanisms of M1/M2 macrophage polarization in different diseases[J]. Chinese Pharmacological Bulletin, 2020, 36(11):1502-1506. (in Chinese)
[9]	BENOIT M, DESNUES B, MEGE J L. Macrophage polarization in bacterial infections[J]. J Immunol, 2008, 181(6):3733-3739.
[10]	EISELE N A, RUBY T, JACOBSON A, et al. Salmonella require the fatty acid regulator PPARδ for the establishment of a metabolic environment essential for long-term persistence[J]. Cell Host Microbe, 2013, 14(2):171-182.
[11]	NOVAK M L, KOH T J. Macrophage phenotypes during tissue repair[J]. J Leukoc Biol, 2013, 93(6):875-881.
[12]	XAVIER M N, WINTER M G, SPEES A M, et al. PPARγ-mediated increase in glucose availability sustains chronic Brucella abortus infection in alternatively activated macrophages[J]. Cell Host Microbe, 2013, 14(2):159-170.
[13]	TARIQ M, ZHANG J Q, LIANG G K, et al. Gefitinib inhibits M2-like polarization of tumor-associated macrophages in Lewis lung cancer by targeting the STAT6 signaling pathway[J]. Acta Pharmacol Sin, 2017, 38(11):1501-1511.
[14]	SHAPOURI-MOGHADDAM A, MOHAMMADIAN S, VAZINI H, et al. Macrophage plasticity, polarization, and function in health and disease[J]. J Cell Physiol, 2018, 223(9):6425-6440.
[15]	SOUZA-NETO J A, SIM S, DIMOPOULOS G. An evolutionary conserved function of the JAK-STAT pathway in anti-dengue defense[J]. Proc Natl Acad Sci U S A, 2009, 106(42):17841-17846.
[16]	BENSINGER S J, TONTONOZ P. Integration of metabolism and inflammation by lipid-activated nuclear receptors[J]. Nature, 2008, 454(7203):470-477.
[17]	LI J, RODRIGUEZ J P, NIU F F, et al. Structural basis for DNA recognition by STAT6[J]. Proc Natl Acad Sci U S A, 2016, 113(46):13015-13020.
[18]	董炳梅, 王金良, 唐娜, 等. 布鲁氏菌的致病机制与细胞免疫机制研究进展[J]. 中国人兽共患病学报, 2012, 28(6):635-639, 643.DONG B M, WANG J L, TANG N, et al. Research progress on pathogenic mechanism and cellular immunity of Brucella[J]. Chinese Journal of Zoonoses, 2012, 28(6):635-639, 643. (in Chinese)
[19]	BRAMBILLA P, BELLANI M, ISOLA M, et al. Increased M1/decreased M2 signature and signs of Th1/Th2 shift in chronic patients with bipolar disorder, but not in those with schizophrenia[J]. Transl Psychiatry, 2014, 4:e406.
[20]	SOBUE S, NOMURA T, ISHIKAWA T, et al. Th1/Th2 cytokine profiles and their relationship to clinical features in patients with chronic hepatitis C virus infection[J]. J Gastroenterol, 2001, 36(8):544-551.
[21]	BAI J Y, LI Y, XUE G H, et al. Requirement of Gαi1 and Gαi3 in interleukin-4-induced signaling, macrophage M2 polarization and allergic asthma response[J]. Theranostics, 2021, 11(10):4894-4909.
[22]	李智伟. 布鲁氏菌感染中巨噬细胞极化通过PD-1/PD-L2途径对T细胞亚群的调控作用研究[D]. 乌鲁木齐:新疆医科大学, 2020.LI Z W. Study on regulation of T-cell subpopulations by PD-1/PD-L2 pathway of macrophage polarization in brucella infection[D]. Urumqi:Xinjiang Medical University, 2020. (in Chinese)
[23]	沈家鲲, 唐倩, 崔洋洋, 等. 猪舍细颗粒物促进猪原代肺泡巨噬细胞向M1极化[J]. 畜牧兽医学报, 2021, 52(6): 1717-1726.SHEN J K, TANG Q, CUI Y Y, et al. Fine particulate matter from pig house promote M1 polarization in porcine primary alveolar macrophage[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(6):1717-1726. (in Chinese)
[24]	张晓东. 分枝杆菌感染免疫应答中巨噬细胞极化分型和多核巨细胞形成及功能的研究[D]. 北京:北京协和医学院, 2015.ZHANG X D. Formation mechanisms and functions of macrophage polarization and multinucleated giant cell in mycobacterial infection[D]. Beijing:Peking Union Medical College, 2015. (in Chinese)
[25]	王月丽, 易继海, 王震, 等. 布鲁菌对不同分化类型巨噬细胞JAK-STAT6信号通路的影响[J]. 中国预防兽医学报, 2017, 39(10):836-841.WANG Y L, YI J H, WANG Z, et al. Effects of Brucella on JAK-STAT6 signaling pathway in different differentiation types of macrophages[J]. Chinese Journal of Preventive Veterinary Medicine, 2017, 39(10):836-841. (in Chinese)
[26]	CLASS C K, NATOLI G. Molecular control of activation and priming in macrophages[J]. Nat Immunol, 2016, 17(1):26-33.
[27]	文志, 韩艳秋, 王俊瑞. 布鲁氏菌毒力因子研究进展[J]. 微生物学通报, 2021, 48(3):842-848.WEN Z, HAN Y Q, WANG J R. Virulence factors of Brucella:a review[J]. Microbiology China, 2021, 48(3):842-848. (in Chinese)
[28]	SHIREY K A, COLE L E, KEEGAN A D, et al. Francisella tularensis live vaccine strain induces macrophage alternative activation as a survival mechanism[J]. J Immunol, 2008, 181(6):4159-4167.
[29]	DABRAL N, BURCHAM G N, JAIN-GUPTA N, et al. Overexpression of wbkF gene in Brucella abortus RB51 WboA leads to increased O-polysaccharide expression and enhanced vaccine efficacy against B. abortus 2308, B. melitensis 16M, and B. suis 1330 in a murine brucellosis model[J]. PLOS One, 2019, 14(3):e0213587.
[30]	OUAHRANI-BETTACHE S, DE BAGÜÉS M P J, DE LA GARZA J, et al. Lethality of Brucella microti in a murine model of infection depends on the wbkE gene involved in O-polysaccharide synthesis[J]. Virulence, 2019, 10(1):868-878.

Effect of STAT6 Mediated Macrophage Polarization on Intracellular Survival of Brucella

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 30

Related Articles 11

Recommended Articles

Metrics

Comments

[1]	XU Zhenyu, DENG Xiaoyu, WANG Yueli, SUN Can, WU Aodi, CAO Jian, YI Jihai, WANG Yong, WANG Zhen, CHEN Chuangfu. Biological Characteristics of Brucella abortus A19ΔBtpA Deletion Strain and Its Immunogenicity Study [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2135-2145.
[2]	WU Shangjie, LUAN Yuanyuan, WANG Mingkun, ZHANG Hechun, YU Bo, MA Yuehui, JIANG Lin, HE Xiaohong. Advances of Disease-Resistant Breeding on Ovine Brucellosis [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 882-893.
[3]	ZHAI Yunyi, YUAN Ye, LI Junmei, TIAN Lulu, DIAO Ziyang, LI Bin, CHEN Jialu, ZHOU Dong, JIN Yaping, WANG Aihua. Preparation and Preliminary Application of Monoclonal Antibody to Brucella Outer Membrane Protein 16 [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2083-2091.
[4]	XIANG Caixia, WANG Xiangguo, LI Junmei, ZHI Feijie, FANG Jiaoyang, ZHENG Weifang, CHEN Jialu, JIN Yaping, WANG Aihua. The Influence of Brucella Type IV Secretes System Effector Protein VceC on Endoplasmic Reticulum Stress and Gonadal Hormone Secretory in Goat Trophoblast Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1210-1220.
[5]	LI Yang, ZHOU Dong, YIN Yanlong, ZHANG Guangdong, XIANG Caixia, ZHI Feijie, BAI Furong, LIN Pengfei, JIN Yaping, WANG Aihua. Effects of Brucella Outer Membrane Protein 16 on Apoptosis and Immune Activity of RAW264.7 Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2642-2651.
[6]	YANG Qin, DENG Xiaoyu, XIE Shanshan, YI Jihai, WANG Yong, ZHANG Qian, WANG Zhen, CHEN Chuangfu. Effects of Brucella bovis Type IV Secretion System on Endoplasmic Reticulum Stress and Apoptosis of Macrophages [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1192-1200.
[7]	WANG Fei, YANG Jie, Lü Qingjie, WANG Mixue, LIU Peng, ZHANG Ruoyu, SHI Congcong, WANG Xueying, LIN Lin, HUA Lin, SONG Wenbo, LIANG Wan, CHEN Huanchun, WU Bin, PENG Zhong. Isolation and Genomic Characterization of a Meningitis Causing Pasteurella multocida [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4346-4355.
[8]	MEI Li, WANG Yingchao, CHENG Rujia, YU Guoji, FAN Xuezheng, GAO Xiaolong, GAO Min, QIN Yuming, LI Xiaoying, LI Qiaoling, ZHU Liangquan, FENG Xiaoyu. A Droplet Digital PCR Method for Detection of Brucella [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(6): 1753-1759.
[9]	SHEN Jiakun, TANG Qian, CUI Yangyang, JIN Xiaoming, LI Yansen, LI Chunmei. Fine Particulate Matter from Pig House Promote M1 Polarization in Porcine Primary Alveolar Macrophage [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(6): 1717-1726.
[10]	JIANG Hui, FENG Yu, LI Xiaoying, FAN Xuezheng, PENG Xiaowei, DING Jiabo. Comparative Research of Four High Throughput Antibody Detection Methods for Brucella [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(11): 3208-3214.
[11]	WANG Shuli, ZHANG Huiru, BI Yanqi, WANG Dejuan, CHEN Lin, ZHANG Xiaoting, LI Zhiqiang. Analysis of Immune Responses Induced by Brucella Transcriptional Regulatory Factor HFQ [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(8): 1977-1984.