多重耐药菌老年肺炎患者Th与Treg细胞及其细胞因子的表达
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摘要
目的检测老年多重耐药菌(MDRO)肺炎患者辅助性T细胞(Th)和调节性T细胞(Treg)及其细胞因子干扰素-γ(IFN-γ)、白细胞介素-4(IL-4)、白细胞介素-17(IL-17)、转化生长因子-β(TGF-β)的表达水平,探讨Th和Treg细胞介导的免疫反应在老年MDRO肺炎患者中的作用。方法选取2016年8月-2018年4月,在昆明医科大学附一院住院期间确诊为院内MDRO肺炎的老年患者30例,再根据是否需要呼吸或循环支持,将患者分为老年重症肺炎组14例、老年普通肺炎组16例,纳入同期健康体检老年人11名作为对照组。分离30例老年MDRO肺炎患者及11例健康老年人外周血的单个核细胞,采用流式细胞术检测Th1、Th2、Th17、Treg细胞各占CD4+T细胞的比例。酶联免疫吸附测定(ELISA)法检测血清中相关细胞因子IFN-γ、IL-4、IL-17、TGF-β的表达水平。结果重症肺炎组Th1以及Treg细胞占CD4+T细胞的比例分别为(1.743±0.606)%、(0.978±0.271)%,较健康对照组低(P<0.05),血清中的IFN-γ及TGF-β的表达量较健康对照组低(P<0.05),且重症肺炎组的TGF-β表达量较普通肺炎组低(P<0.05);重症肺炎组的Th17细胞占CD4+T细胞的比例及IL-17A的表达量与健康对照组相比增加(P<0.05)。结论老年MDRO肺炎患者的Th1、Treg细胞所介导的免疫反应受抑制,Th17/Treg细胞失平衡,并与病情严重程度有关系。这为研究抗感染免疫治疗新策略奠定了基础。
OBJETIVE To detect the expression levels of Th and Treg cells and cytokines Interferon-γ(IFN-γ),Interleukin-4(IL-4),Interleukin-17(IL-17)and transforming growth factor-β(TGF-β)in elderly patients with pneumonia caused by multidrug-resistant organisms(MDROs),and explore the role of Th and Treg cell-mediated immune responses in elderly patients with pneumonia caused by MDROs.METHODS A total of 30 elderly patients with pneumonia caused by MDROs who were treated in the First Affiliated Hospital of Kunming Medical University during Aug.2016 to Apr.2018 were enrolled in the study.According to whether respiratory or circulatory support was needed,the patients were divided into the elderly severe pneumonia group and the elderly general pneumonia group,and 11 healthy elderly people in the same period were included as the control group.Peripheral blood mononuclear cells from the 30 elderly patients with pneumonia caused by MDRO and 11 healthy elderly peoples were isolated.The proportions of Th1,Th2,Th17 and Treg cells in CD4+ T cells were determined by flow cytometry.The expression levels of related cytokines IFN-γ,IL-4,IL-17 and TGF-βin serum were detected by enzyme-linked immune sorbent assay(ELISA).RESULTS The proportions of Th1 and Treg cells in CD4+T cells in severe pneumonia patients were(1.743±0.606)%and(0.978±0.271)%,respectively,significantly lower than those of the healthy control group(P<0.05),the expression levels of IFN-γand TGF-βin serum were significantly lower than those of the healthy control group(P<0.05),and the expression of TGF-βin the severe pneumonia group was significantly lower than that in the general pneumonia group(P<0.05).The proportion of Th17 cells in CD4+T cells and the expression of IL-17 Ain the severe pneumonia group increased significantly as compared with the healthy controls(P<0.05).CONCLUSIONTh1 and Treg-mediate cellular immunity was inhibited in elderly patients with pneumonia caused by MDRO,and Th17/Treg was imbalanced and related to the severity of the disease.This study lays foundation for the development of new strategies for anti-infective immunotherapy.
OBJETIVE To detect the expression levels of Th and Treg cells and cytokines Interferon-γ(IFN-γ),Interleukin-4(IL-4),Interleukin-17(IL-17)and transforming growth factor-β(TGF-β)in elderly patients with pneumonia caused by multidrug-resistant organisms(MDROs),and explore the role of Th and Treg cell-mediated immune responses in elderly patients with pneumonia caused by MDROs.METHODS A total of 30 elderly patients with pneumonia caused by MDROs who were treated in the First Affiliated Hospital of Kunming Medical University during Aug.2016 to Apr.2018 were enrolled in the study.According to whether respiratory or circulatory support was needed,the patients were divided into the elderly severe pneumonia group and the elderly general pneumonia group,and 11 healthy elderly people in the same period were included as the control group.Peripheral blood mononuclear cells from the 30 elderly patients with pneumonia caused by MDRO and 11 healthy elderly peoples were isolated.The proportions of Th1,Th2,Th17 and Treg cells in CD4+ T cells were determined by flow cytometry.The expression levels of related cytokines IFN-γ,IL-4,IL-17 and TGF-βin serum were detected by enzyme-linked immune sorbent assay(ELISA).RESULTS The proportions of Th1 and Treg cells in CD4+T cells in severe pneumonia patients were(1.743±0.606)%and(0.978±0.271)%,respectively,significantly lower than those of the healthy control group(P<0.05),the expression levels of IFN-γand TGF-βin serum were significantly lower than those of the healthy control group(P<0.05),and the expression of TGF-βin the severe pneumonia group was significantly lower than that in the general pneumonia group(P<0.05).The proportion of Th17 cells in CD4+T cells and the expression of IL-17 Ain the severe pneumonia group increased significantly as compared with the healthy controls(P<0.05).CONCLUSIONTh1 and Treg-mediate cellular immunity was inhibited in elderly patients with pneumonia caused by MDRO,and Th17/Treg was imbalanced and related to the severity of the disease.This study lays foundation for the development of new strategies for anti-infective immunotherapy.
引文
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[17]中华医学会呼吸病学分会感染学组.中国成人医院获得性肺炎与呼吸机相关性肺炎诊断和治疗指南(2018年版)[J].中华结核和呼吸杂志,2018,41(42):255-280.
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[19]林媛媛,吴乐程,孙成勇,等.病毒性心肌炎患者外周血中Th17细胞和调节性T细胞的变化研究[J].中华医院感染学杂志,2017,27(22):5129-5132
[20]Luty J,Ruckemann-Dziurdzinska K,Witkowski JM,et al.Immunological aspects of autoimmune thyroid diseaseComplex interplay between cells and cytokines[J].Cytokine,2019,116:128-133.
[21]赵瑾,冯宪真,冯丽丽,等.Th17/Treg失衡在重症肺炎中的作用及其意义[J].医学分子生物学杂志,2016,13(1):17-20.
[22]黎焯基,成俊芬,陈大勇,等.IL-17在重症肺炎患者外周血的表达及意义[J].国际医药卫生导报,2014,20(23):3528-3530.
[23]Wang L,Guan N,Jin Y,et al.Subcutaneous vaccination with Porphyromonas gingivalis ameliorates periodontitis by modulating Th17/Treg imbalance in murine model[J].Int Immunopharmacol,2015,25(1):65-73.
[24]Adamzik M,Broll J,Steinmann J,et al.An increased alveolar CD4+CD25+Foxp3+T-regulatory cell ratio in acute respiratory distress syndrome is associated with increased 30-day mortality intensive[J].Intensive Care Med,2013,39(10):1743-1751
[2]王春喜,张纳新,陶丝煦,等.老年社区获得性肺炎病情评估系统和预后的相关性研究及药物治疗新进展[J].天津药学,2018,30(6):57-63.
[3]黄勋,邓子德,倪语星,等.多重耐药菌医院感染预防与控制中国专家共识[J].中国感染控制杂志,2015,14(1):1-9.
[4]Chang HH,Cohen T,Grad YH,et al.Origin and proliferation of multiple-drug resistance in bacterial pathogens[J].Microbiol Mol Biol Rev,2015,79(1):101-116.
[5]Kalil AC,Metersky ML,Klompas M,et al.Management of adults with hospital-acquired and ventilator-associated pneumonia:2016 Clinical Practice Guidelines by the Infectious Diseases Society of America and the American Thoracic Society[J].Clin Infect Dis,2016,63(5):e61-e111.
[6]Gasper DJ,Tejera MM,Suresh M.CD4T-cell memory generation and maintenance[J].Crit Rev Immunol,2014,34(2):121-146.
[7]Vella LA,Herati RS,Wherry EJ.CD4+T cell differentiation in chronic viral infections:the tfh perspective[J].Trends Mol Med.2017,23(12):1072-1087.
[8]刘娟,曹雪涛.2014年国内外免疫学研究重要进展[J].中国免疫学杂志,2015,31(1):1-11.
[9]王润超,万哲,李若瑜.21例侵袭性肺曲霉病患者Th以及Treg细胞的检测[J].中国真菌学杂志,2014,9(5):287-292.
[10]Keigo U,Makoto U,Kayo O,et al.Dendritic cell-based vaccine against fungal infection[J].Vaccine Design,2015,1403(8):537-549.
[11]Tangye SG,Pillay B,Randall KL,et al.DOCK8-Deficient CD4+T cells are biased to a Th2effector fate at the expense ofTh1and Th17cells[J].J Allergy Clin Immunol,2017,139(3):933-949.
[12]Rahman MA,Sobia P,Dwivedi VP,et al.Mycobacterium tuberculosis TlyA protein negatively regulates T helper(Th)1and Th17differentiation and promotes tuberculosis pathogenesis[J].J Biol Chem,2015,290(23):14407-14417.
[13]Fu YR,Gao KS,Ji R.Differential transcriptional response in macrophages infected with cell wall deficient versus normal Mycobacterium tuberculosis[J].Int J Biol Sci,2015,11(1):22-30.
[14]张影,陆坚,侯志平,等.肺结核患者疗程中外周血辅助性和调节性T细胞的动态研究[J].中国防痨杂志,2013,35(6):427-432.
[15]Kolls JK.CD4+T‐cell subsets and host defense in the lung[J].Immunol Rev,2013,252(1):156-163.
[16]American Thoracic Society;Infectious Diseases Society of A-merica.Guidelines for the management of adults with hospital-acquired,ventilator-associated,and healthcare-associated pneumonia[J].Am J Respir Crit Care Med,2005,171(4):388-416.
[17]中华医学会呼吸病学分会感染学组.中国成人医院获得性肺炎与呼吸机相关性肺炎诊断和治疗指南(2018年版)[J].中华结核和呼吸杂志,2018,41(42):255-280.
[18]董薇,白伟民.T淋巴细胞亚群与合并重症肺炎昏迷患者预后的关系[J].中国实用神经疾病杂志,2018,21(10):1106-1109.
[19]林媛媛,吴乐程,孙成勇,等.病毒性心肌炎患者外周血中Th17细胞和调节性T细胞的变化研究[J].中华医院感染学杂志,2017,27(22):5129-5132
[20]Luty J,Ruckemann-Dziurdzinska K,Witkowski JM,et al.Immunological aspects of autoimmune thyroid diseaseComplex interplay between cells and cytokines[J].Cytokine,2019,116:128-133.
[21]赵瑾,冯宪真,冯丽丽,等.Th17/Treg失衡在重症肺炎中的作用及其意义[J].医学分子生物学杂志,2016,13(1):17-20.
[22]黎焯基,成俊芬,陈大勇,等.IL-17在重症肺炎患者外周血的表达及意义[J].国际医药卫生导报,2014,20(23):3528-3530.
[23]Wang L,Guan N,Jin Y,et al.Subcutaneous vaccination with Porphyromonas gingivalis ameliorates periodontitis by modulating Th17/Treg imbalance in murine model[J].Int Immunopharmacol,2015,25(1):65-73.
[24]Adamzik M,Broll J,Steinmann J,et al.An increased alveolar CD4+CD25+Foxp3+T-regulatory cell ratio in acute respiratory distress syndrome is associated with increased 30-day mortality intensive[J].Intensive Care Med,2013,39(10):1743-1751