C5a/C5aR信号在微小隐孢子虫感染中的免疫调节作用研究

doi:10.11843/j.issn.0366-6964.2022.08.020

Abstract

Abstract: The aim of this study was to explore the regulation of C5a/C5aR signal in the immune response of host CD4⁺ T cell during Cryptosporidium parvum infection. We have utilized C. parvum as the research object, and selected BALB/c suckling mice and C5aR inhibited BALB/c suckling mice as infection models. The expression level of C5aR in the ileum tissues of BALB/c suckling mice was detected by using real-time PCR and immunohistochemistry. The expression patterns of C. parvum HSP70 gene and main effector cytokines (IFN-γ, IL-4, IL-17 and TGF-β) of CD4⁺ T cell subsets (Th1, Th2, Th17 cells and Treg cells) were analyzed by real-time PCR. The injury of ileal mucosa in suckling mice was observed by histopathology analysis. Compared with mice in the control group, C. parvum infection could significantly increase the expression levels of C5aR mRNA and protein (P<0.05), as well as IFN-γ in the ileum tissues of suckling mice (P<0.05). Compared with mice infected with C. parvum, the expression levels of main effector cytokines of Th1, Th2 and Treg cells, namely IFN-γ, IL-4 and TGF-β, significantly reduced in C5aR inhibited BALB/C suckling mice infected with C. parvum (P<0.05), and the expression level of IL-17, the main effector cytokine of Th17 cells, significantly increased in suckling mice treated with C5aR inhibitor (P<0.05). Histopathology analysis of ileal mucosa in suckling mice indicated inhibition of C5aR could significantly improve the changes of villus diameter and mucosal thickness of ileum caused by C. parvum infection (P<0.05), but not for the villus length and the ratio of villus length to crypt depth. Analysis of the mRNA level for Cryptosporidium HSP70 gene showed that inhibition of C5aR could significantly affect the proliferation of C. parvum in ileum (P<0.05). C5a/C5aR signal may participate in the process of interaction between host and Cryptosporidium by dynamically regulating the expression of major effector cytokines for CD4⁺ T cell subsets, providing reference basis for further understanding the interaction mechanism between Cryptosporidium spp. and host.

Key words: Cryptosporidium parvum, BALB/c sucking mouse, C5a/C5aR signal, CD4⁺ T cell subsets, regulation

CLC Number:

WU Xuemei, YANG Xin, YUAN Yajie, YIN Yanling, LAI Peng, SONG Junke, SHI Huaiping, ZHAO Guanghui. Immunomodulatory Effect of C5a/C5aR Signal during Cryptosporidium parvum Infection[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2621-2632.

References 41

[1]	YANG X, GUO Y Q, XIAO L H, et al. Molecular epidemiology of human cryptosporidiosis in low- and middle-income countries[J]. Clin Microbiol Rev, 2021, 34(2):e00087-19.
[2]	JEŽKOVÁ J, LIMPOUCHOVÁ Z, PREDIGER J, et al. Cryptosporidium myocastoris n. sp. (Apicomplexa:Cryptosporidiidae), the species adapted to the nutria (Myocastor coypus)[J]. Microorganisms, 2021, 9(4):813.
[3]	BOUZID M, HUNTER P R, CHALMERS R M, et al. Cryptosporidium pathogenicity and virulence[J]. Clin Microbiol Rev, 2013, 26(1):115-134.
[4]	CHECKLEY W, WHITE A C, JAGANATH D, et al. A review of the global burden, novel diagnostics, therapeutics, and vaccine targets for Cryptosporidium[J]. Lancet Infect Dis, 2015, 15(1):85-94.
[5]	PETRY F, JAKOBI V, TESSEMA T S. Host immune response to Cryptosporidium parvum infection[J]. Exp Parasitol, 2010, 126(3):304-309.
[6]	LAURENT F, LACROIX-LAMANDÉ S. Innate immune responses play a key role in controlling infection of the intestinal epithelium by Cryptosporidium[J]. Int J Parasitol, 2017, 47(12):711-721.
[7]	BARAKAT F M, MCDONALD V, DI SANTO J P, et al. Roles for NK cells and an NK cell-independent source of intestinal gamma interferon for innate immunity to Cryptosporidium parvum infection[J]. Infect Immun, 2009, 77(11):5044-5049.
[8]	IVANOVA D L, DENTON S L, FETTEL K D, et al. Innate lymphoid cells in protection, pathology, and adaptive immunity during Apicomplexan infection[J]. Front Immunol, 2019, 10:196.
[9]	PETRY F, JAKOBI V, WAGNER S, et al. Binding and activation of human and mouse complement by Cryptosporidium parvum (Apicomplexa) and susceptibility of C1q- and MBL-deficient mice to infection[J]. Mol Immunol, 2008, 45(12):3392-3400.
[10]	MCNAIR N N, MEAD J R. CD4+ effector and memory cell populations protect against Cryptosporidium parvum infection[J]. Microbes Infect, 2013, 15(8-9):599-606.
[11]	BORAD A, WARD H. Human immune responses in cryptosporidiosis[J]. Future Microbiol, 2010, 5(3):507-519.
[12]	RIGGS M W. Recent advances in cryptosporidiosis:the immune response[J]. Microbes Infect, 2002, 4(10):1067-1080.
[13]	TESSEMA T S, SCHWAMB B, LOCHNER M, et al. Dynamics of gut mucosal and systemic Th1/Th2 cytokine responses in interferon-gamma and interleukin-12p40 knock out mice during primary and challenge Cryptosporidium parvum infection[J]. Immunobiology, 2009, 214(6):454-466.
[14]	MCDONALD V, KORBEL D S, BARAKAT F M, et al. Innate immune responses against Cryptosporidium parvum infection[J]. Parasite Immunol, 2013, 35(2):55-64.
[15]	ZHAO G H, FANG Y Q, RYAN U, et. al. Dynamics of Th17 associating cytokines in Cryptosporidium parvum-infected mice[J]. Parasitol Res, 2016, 115(2):879-887.
[16]	PLANCHON S, FIOCCHI C, TAKAFUJI V, et al. Transforming growth factor-β1 preserves epithelial barrier function:identification of receptors, biochemical intermediates, and cytokine antagonists[J]. J Cell Physiol, 1999, 181(1):55-66.
[17]	RICKLIN D, HAJISHENGALLIS G, YANG K J, et al. Complement:a key system for immune surveillance and homeostasis[J]. Nat Immunol, 2010, 11(9):785-797.
[18]	HAAS K M, HASEGAWA M, STEEBER D A, et al. Complement receptors CD21/35 link innate and protective immunity during Streptococcus pneumoniae infection by regulating IgG3 antibody responses[J]. Immunity, 2002, 17(6):713-723.
[19]	DEMPSEY P W, ALLISON M E D, AKKARAJU S, et al. C3d of complement as a molecular adjuvant:bridging innate and acquired immunity[J]. Science, 1996, 271(5247):348-350.
[20]	KEMPER C, ATKINSON J P. T-cell regulation:with complements from innate immunity[J]. Nat Rev Immunol, 2007, 7(1):9-18.
[21]	DING P P, LI L, LI L Y, et al. C5aR1 is a master regulator in colorectal tumorigenesis via immune modulation[J]. Theranostics, 2020, 10(19):8619-8632.
[22]	STRAINIC M G, LIU J B, HUANG D P, et al. Locally produced complement fragments C5a and C3a provide both costimulatory and survival signals to naive CD4+ T cells[J]. Immunity, 2008, 28(3):425-435.
[23]	STRAINIC M G, SHEVACH E M, AN F Q, et al. Absence of signaling into CD4+ cells via C3aR and C5aR enables autoinductive TGF-β1 signaling and induction of Foxp3+ regulatory T cells[J]. Nat Immunol, 2013, 14(2):162-171.
[24]	AJONA D, ORTIZ-ESPINOSA S, PIO R. Complement anaphylatoxins C3a and C5a:emerging roles in cancer progression and treatment[J]. Semin Cell Dev Biol, 2019, 85:153-163.
[25]	MOULTON R A, MASHRUWALA M A, SMITH A K, et al. Complement C5a anaphylatoxin is an innate determinant of dendritic cell-induced Th1 immunity to Mycobacterium bovis BCG infection in mice[J]. J Leukoc Biol, 2007, 82(4):956-967.
[26]	SCHMUDDE I, STRÖVER H A, VOLLBRANDT T, et al. C5a receptor signalling in dendritic cells controls the development of maladaptive Th2 and Th17 immunity in experimental allergic asthma[J]. Mucosal Immunol, 2013, 6(4):807-825.
[27]	SUN S H, ZHAO G Y, LIU C F, et al. Treatment with anti-C5a antibody improves the outcome of H7 N9 virus infection in African green monkeys[J]. Clin Infect Dis, 2015, 60(4):586-595.
[28]	KIM S H, YANG I Y, JANG S H, et al. C5a receptor-targeting ligand-mediated delivery of dengue virus antigen to M cells evokes antigen-specific systemic and mucosal immune responses in oral immunization[J]. Microbes Infect, 2013, 15(13):895-902.
[29]	CALAME D G, MUELLER-ORTIZ S L, MORALES J E, et al. The C5a anaphylatoxin receptor (C5aR1) protects against Listeria monocytogenes infection by inhibiting type 1 IFN expression[J]. J Immunol, 2014, 193(10):5099-5107.
[30]	LIU T P, XU G L, GUO B, et al. An essential role for C5aR signaling in the optimal induction of a malaria-specific CD4+ T cell response by a whole-killed blood-stage vaccine[J]. J Immunol, 2013, 191(1):178-186.
[31]	SCHMITZ V, ALMEIDA L N, SVENSJÖ E, et al. C5a and bradykinin receptor cross-talk regulates innate and adaptive immunity in Trypanosoma cruzi infection[J]. J Immunol, 2014, 193(7):3613-3623.
[32]	GARCÉS-SANCHEZ G, WILDERER P A, HORN H, et al. Assessment of the viability of Cryptosporidium parvum oocysts with the induction ratio of hsp70 mRNA production in manure[J]. J Microbiol Methods, 2013, 94(3):280-289.
[33]	FONSECA M I, AGER R R, CHU S H, et al. Treatment with a C5aR antagonist decreases pathology and enhances behavioral performance in murine models of Alzheimer's disease[J]. J Immunol, 2009, 183(2):1375-1383.
[34]	李松瑞, 牛子文, 张素梅, 等. 微小隐孢子虫感染动物模型及其应用研究进展[J]. 中国病原生物学杂志, 2018, 13(7):785-788.LI S R, NIU Z W, ZHANG S M, et al. Advances in the study of and uses of animal models of Cryptosporidium parvum infection[J]. Journal of Pathogen Biology, 2018, 13(7):785-788. (in Chinese)
[35]	BENAMROUZ S, GUYOT K, GAZZOLA S, et al. Cryptosporidium parvum infection in SCID mice infected with only one oocyst:qPCR assessment of parasite replication in tissues and development of digestive cancer[J]. PLoS One, 2012, 7(12):e51232.
[36]	陈甫, 黄克和. 建立微小隐孢子虫感染小鼠模型方法的研究[J]. 中国人兽共患病学报, 2007, 23(2):195-198.CHEN F, HUANG K H. Study on the optimal conditions to establish the mouse model infected with Cryptosporidium parvum oocysts[J]. Chinese Journal of Zoonoses, 2007, 23(2):195-198. (in Chinese)
[37]	卢思奇, 罗晓冰, 陈小宁, 等. 实验感染昆明鼠粪便微小隐孢子虫18SrRNA基因检测[J]. 中国寄生虫病防治杂志, 2003, 16(4):196-199.LU S Q, LUO X B, CHEN X N, et al. Detection of Cryptosporidium parvum in stool specimens of experimentally infected km mice by 18SrRNA gene amplification[J]. Chinese Journal of Parasitic Disease Control, 2003, 16(4):196-199. (in Chinese)
[38]	CLARKE E V, TENNER A J. Complement modulation of T cell immune responses during homeostasis and disease[J]. J Leukoc Biol, 2014, 96(5):745-756.
[39]	郭雅旭, 吴飞, 方艳琴, 等. 过敏毒素C5a和C3a在微小隐孢子虫感染中的表达分析[J]. 中国兽医学报, 2017, 37(5):871-874, 908.GUO Y X, WU F, FANG Y Q, et al. Expression analysis of anaphylatoxins C5a and C3a in Cryptosporidium parvum-infecting mice[J]. Chinese Journal of Veterinary Science, 2017, 37(5):871-874, 908. (in Chinese)
[40]	KIM H, ERDMAN L K, LU Z Y, et al. Functional roles for C5a and C5aR but not C5L2 in the pathogenesis of human and experimental cerebral malaria[J]. Infect Immun, 2014, 82(1):371-379.
[41]	BRIUKHOVETSKA D, OHM B, MEY F T, et al. C5aR1, activation drives early IFN-γ production to control exprimental Toxoplasma gondii infection[J]. Front Immunol, 2020, 11:1397

Immunomodulatory Effect of C5a/C5aR Signal during Cryptosporidium parvum Infection

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