环磷酰胺对布鲁氏菌侵染小鼠过程的影响
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摘要
目的为了探究布鲁氏菌在环磷酰胺(CP)诱导的免疫抑制小鼠的侵染过程和炎症反应。方法使用40 mg/kg剂量CP腹腔连续7 d注射昆明小鼠,之后侵染1×10~6CFU布鲁氏菌M5,检测小鼠体重食量,脏器质量,脏器相对质量。用CFU计数法计算组织细菌数,石蜡切片进行免疫组织化学观察,ELISA法检测血清IFN-γ、TNF-α和NO细胞因子的释放。结果我们发现与正常小鼠相比,CP显著降低了受试小鼠的体重和免疫器官的重量,并且显著增加了组织细菌数量、巨噬细胞的浸润和细胞因子的含量。结论环磷酰胺为布鲁氏菌M5在小鼠体内的增殖创造了更加适宜的环境,并且增加了布鲁氏菌的存活数量和小鼠的炎症反应。为布鲁氏菌引起宿主炎症反应以及免疫逃避研究提供新的思路,为进一步研究宿主细胞与病原菌的相互作用提供数据支持并奠定基础。
Objective The purpose of this research was to investigate the infection process and inflammatory response of brucella in immunosuppressive mice induced by cyclophosphamide( CP). Methods KM mice were i.p injected with 40 mg/kg dose CP for 7 days,and then infected with 1×10~6 CFU brucella M5. The body weight,food intake,and organ quality of the mice were measured,as well as the coefficient of viscera. Quantification of viable bacteria burden in viscera was determined by measuring CFU,paraffin section was used for immunohistochemical observation,and the IFN-γ、TNF-α and NO cytokines was detected by ELISA. We found that compared to the normal mice,CP significantly reduced the body weight and the weight of the immune organs of the tested mice,and significantly increased the number of bacterial burden in the tissues,infiltration of macrophages and the content of cytokines.Results Cyclophosphamide created a more suitable environment for brucella M5 proliferation in mice,increased the number of brucella survival and inflammation in mice.Conclusion It provides a new idea for brucella to induce host inflammatory response and immune escape,and provides data and foundation for further study on the interaction between host cells and pathogens.
Objective The purpose of this research was to investigate the infection process and inflammatory response of brucella in immunosuppressive mice induced by cyclophosphamide( CP). Methods KM mice were i.p injected with 40 mg/kg dose CP for 7 days,and then infected with 1×10~6 CFU brucella M5. The body weight,food intake,and organ quality of the mice were measured,as well as the coefficient of viscera. Quantification of viable bacteria burden in viscera was determined by measuring CFU,paraffin section was used for immunohistochemical observation,and the IFN-γ、TNF-α and NO cytokines was detected by ELISA. We found that compared to the normal mice,CP significantly reduced the body weight and the weight of the immune organs of the tested mice,and significantly increased the number of bacterial burden in the tissues,infiltration of macrophages and the content of cytokines.Results Cyclophosphamide created a more suitable environment for brucella M5 proliferation in mice,increased the number of brucella survival and inflammation in mice.Conclusion It provides a new idea for brucella to induce host inflammatory response and immune escape,and provides data and foundation for further study on the interaction between host cells and pathogens.
引文
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[16]Monika M S,Paulina K,Dominika B,et al.Cyclophosphamide modified murine peritoneal macrophages induce CD4+T contrasuppressor cells that protect contact sensitivity T effector cells from suppression[J].Pharmacological Reports,2018,796-803.
[17]Xavier M N,Winter M G,Spees A M,et al.CD4+T cellderived IL-10 promotes Brucella abortus persistence via modulation of macrophage function[J].PLoS pathogens,2013,9(6):1-17.
[18]Rocha V Z,Folco E J,Sukhova G,et al.Interferon-gamma,a th1 cytokine,regulates fat inflammation:a role for adaptive immunity in obesity[J].Circ Res,2008,103(5):467-476.
[19]Caron E,Gross A,Liautard J P,et al.Brucella species release a specific,protease-sensitive,inhibitor of TNF-alpha expression,active on human macrophage-like cells[J].Journal of immunology,1996,156(8):2885-2893.
[20]Christopher N,Julie S,Maura G,et al.Is nitric oxide an essential mediator in cervical inflammation and preterm birth?[J].The Journal of Maternal-Fetal&Neonatal Medicine,2018,31(13).
[21]Bernhard B,Andreas K,Katrin B S.Nitric oxide(no):an effector of apoptosis[J].Cell Death Differ,1999,6(10):969-975.
[2]Elfaki M G,Alaidan A A,Ahokail A A.Host response to Brucella infection:review and future perspective[J].Journal of infection in developing countries,2015,9(7):697.
[3]Xu X,Zhang X.Effects of cyclophosphamide on immune system and gut microbiota in mice[J].Microbiological Research,2015,171:97-106.
[4]Jayesh M,Piya P M,Manas K,et al.Immunomodulatory role of chitosan-based nanoparticles and oligosaccharides in cyclophosphamide-treated mice[J].Scandinavian Journal of Immunology,2019,89(4):438-445.
[5]Sung M W,Woo R Ch,Dooly J,et al.Immune enhancement effect of an herb complex extract through the activation of natural killer cells and the regulation of cytokine levels in a cyclophosphamide-induced immunosuppression rat model[J].Asian Pacific Journal of Tropical Medicine,2018,11(12):653-658.
[6]杨琴.益气补肾方对环磷酰胺致小鼠免疫抑制抵抗作用及药理学分析[J].四川中医,2018,36(8):47-49.Yang Q.Immunosuppressive effect and pharmacological analysis of Yiqi bushen recipe for Mice with cyclophosphamide induced immunosuppression[J].Journal of Sichuan of Traditional Chinese Medicine,2018,36(8):47-49.
[7]Manepalli S,Gandhi J A,Ekhar V V,et al.Characterization of a cyclophosphamide-induced murine model of immunosuppression to study acinetobacter baumannii pathogenesis[J].Journal of Medical Microbiology.2013,62(11):1747-1754.
[8]赵弋清,罗霞,陈东辉,等.不同剂量环磷酰胺诱导正常小鼠免疫抑制的对比研究[J].免疫学杂志,2005(1):(128-134).Zhao Y Q,Luo X,Cheng D H,et al.Comparison of immunosuppression induced by different doses of cyclophosphamide in normal mice[J].Immunological Journal,2005,(1):128-134.
[9]吴建良,汪培欢,王翔,等.红景天苷对环磷酰胺致骨髓损伤小鼠造血功能的影响[J].中药新药与临床药理,2013,24(4):371-374.Wu J L,Wang P H,Wang X,et al.Effects of salidroside on hematopoietic recovery in cyclophosphamide-induced marrow injury mice[J].Traditional Chinese Drug Research and Clinical Pharmacology,2013,24(4):371-374.
[10]宋雁,贾旭东,崔文明,等.不同途径和剂量环磷酰胺建立小鼠免疫抑制模型的对比研究[J].中国食品卫生杂志,2013,25(3):218-225.Song Y,Jia X D,Cui W M,et al.Comparison research of immunosuppression models induced by different ways and doses of cyclophosphamide in mice[J].Chinese Journal of Food Hygiene,2013,25(3):218-225.
[11]马增春,谭洪玲,肖成荣,等.环磷酰胺损伤小鼠骨髓造血的机制[J].毒理学杂志,2007,(4):332.Ma Z C,Tan H L,Xiao C R.et al.Mechanism of bone marrow hematopoiesis induced by cyclophosphamide in mice[J].Journal of Toxicology,2007,(4):332.
[12]何雅馨.环磷酰胺对小鼠脾脏的病理学影响[D].雅安:四川农业大学,2015.
[13]胡东.环磷酰胺对小鼠胸腺病理学影响的研究[D].雅安:四川农业大学,2015.
[14]Machelart A,K A,Demars A,et al.Chronic Brucella infection induces selective and persistent IFN-γ-dependent alterations of marginal zone macrophages in the spleen[J].Infection&Immunity,2017,85(11):115-117.
[15]李霞,李殿俊,富东旭.免疫抑制剂对小鼠胸腺及外周血、脾脏T淋巴细胞亚群的影响[J].哈尔滨医科大学学报:2005,39(4):325-326.Li X,Li J D,Fu D X.et al.Effect of immunosuppressant on T cell subpopulations in thymus,peripheral clood and spleen of mice[J].Journal of Harbin Medical University,2005,39(4):325-326.
[16]Monika M S,Paulina K,Dominika B,et al.Cyclophosphamide modified murine peritoneal macrophages induce CD4+T contrasuppressor cells that protect contact sensitivity T effector cells from suppression[J].Pharmacological Reports,2018,796-803.
[17]Xavier M N,Winter M G,Spees A M,et al.CD4+T cellderived IL-10 promotes Brucella abortus persistence via modulation of macrophage function[J].PLoS pathogens,2013,9(6):1-17.
[18]Rocha V Z,Folco E J,Sukhova G,et al.Interferon-gamma,a th1 cytokine,regulates fat inflammation:a role for adaptive immunity in obesity[J].Circ Res,2008,103(5):467-476.
[19]Caron E,Gross A,Liautard J P,et al.Brucella species release a specific,protease-sensitive,inhibitor of TNF-alpha expression,active on human macrophage-like cells[J].Journal of immunology,1996,156(8):2885-2893.
[20]Christopher N,Julie S,Maura G,et al.Is nitric oxide an essential mediator in cervical inflammation and preterm birth?[J].The Journal of Maternal-Fetal&Neonatal Medicine,2018,31(13).
[21]Bernhard B,Andreas K,Katrin B S.Nitric oxide(no):an effector of apoptosis[J].Cell Death Differ,1999,6(10):969-975.