过敏毒素C5a和C3a在微小隐孢子虫感染中的表达分析
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摘要
为明确过敏毒素C5a和C3a在小鼠感染和清除隐孢子虫(Cryptosporidium)感染过程中的作用,本研究以微小隐孢子虫(C.parvum)为研究对象,以BALB/c小鼠为感染模型,采用实时荧光定量PCR分析了C.parvum感染小鼠的小肠相关淋巴组织和脾脏中过敏毒素C3a、C5a及其相应受体(C3aR和C5aR)的mRNA水平。结果显示,在脾脏和小肠中,不仅过敏毒素C5a和C3a在感染后出现了显著上调表达,其受体C5aR和C3aR的表达水平在感染后亦显著上调表达(P<0.05),并呈现动态变化。这些研究结果提示C5a/C5aR和C3a/C3aR信号参与了宿主抗微小隐孢子虫感染的过程。
In order to evaluate the role of anaphylatoxins C5 aand C3aduring Cryptosporidiuminfection,the present study examined their mRNA levels and receptors(C5aR and C3aR)in small intestines and spleens in C.parvum-infecting BABL/c mice by real-time quantitative PCR(qPCR).Both the expression levels of anaphylatoxins(C5aand C3a)and their receptors(C5aR and C3aR)were significantly upregulated(P<0.05)in spleens and small intestines,and dynamic changes of mRNA levels were also observed.These results suggested that signals of C5a/C5 aR and C3a/C3 aR would be involved in the anti-infection of hosts against C.parvum.
In order to evaluate the role of anaphylatoxins C5 aand C3aduring Cryptosporidiuminfection,the present study examined their mRNA levels and receptors(C5aR and C3aR)in small intestines and spleens in C.parvum-infecting BABL/c mice by real-time quantitative PCR(qPCR).Both the expression levels of anaphylatoxins(C5aand C3a)and their receptors(C5aR and C3aR)were significantly upregulated(P<0.05)in spleens and small intestines,and dynamic changes of mRNA levels were also observed.These results suggested that signals of C5a/C5 aR and C3a/C3 aR would be involved in the anti-infection of hosts against C.parvum.
引文
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[6]ZHOUW.The new face of anaphylatoxins in immune regulation[J].Immunobiology,2012,217(2):225-234.
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[8]KIM H,ERDMAN L K,LU Z,et al.Functional roles for C5aand C5aR but not C5L2in the pathogenesis of human and experimental cerebral malaria[J].Infect Immun,2014,82(1):371-379.
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[14]KEARNA A M,TURNER A J L,ELTRINGHAM GJ A,et al.Rapid detection and quantification of CMVDNA in urine using lightcycler based real-time PCR[J].J Clin Virol,2002,24(1-2):131-134.
[15]GERARD C,BAO L,OROZCO O,et al.Structure diversity in the extracellular faces of peptidergic G-protein-coupled receptors.Molecular cloning of the mouse C5aanaphylatoxin receptor[J].J Immunol,1992,149(8):2600-2606.
[16]RAMIREZ N E,WARD L A,SREEVATSAN S.Areview of the biology and epidemiology of cryptosporidiosis in humans and animals[J].Microbes Infect,2004,6(8):773-785.
[17]ZHAO G H,FANG Y Q,RYANU,et al.Dynamics of Th17associating cytokines in Cryptosporidium parvum-infected mice[J].Parasitol Res,2015,[Epub ahead of print].
[18]KIM A H,DIMITRIOU I D,HOLLAND M C,et al.Complement C5areceptor is essential for the optimal generation of antiviral CD8+T cell responses[J].JImmunol,2004,173(4):2524-2529.
[19]KHL J,BAELDER R,LEWKOWICH I P,et al.Aregulatory role for the C5aanaphylatoxin in type 2immunity in asthma[J].J Clin Invest,2006,116(3):783-796.
[20]FREGONESE L,SWAN F J,VAN SCHADEWIJKA,et al.Expression of the anaphylatoxin receptors C3aR and C5aR is increased in fatal asthma[J].J Allergy Clin Immunol,2005,115(6):1148-1154.
[2]FAYER R.Taxonomy and species delimitation in Cryptosporidium[J].Exp Parasitol,2010,124(1):90-97.
[3]LAPETA J.Cryptosporidiosis and Cryptosporidium species in animals and humans:a thirty colour rainbow[J].Int J Parasitol,2013,43(12-13):957-970.
[4]SAMIE A,MAKUWA S,MTSHALI S,et al.Parasitic infection among HIV/AIDS patients at Bela-Bela clinic,Limpopo province,South Africa with special reference to Cryptosporidium[J].Southeast Asian JTrop Med Public Health,2014,45(4):783-795.
[5]PETRY F,JAKOBI V,TESSEMA T S.Host immune response to Cryptosporidium parvum infection[J].Exp Parasitol,2010,126(3):304-309.
[6]ZHOUW.The new face of anaphylatoxins in immune regulation[J].Immunobiology,2012,217(2):225-234.
[7]SCHMUDDE I,LAUMONNIER Y,KHL J.Anaphylatoxins coordinate innate and adaptive immune responses in allergic asthma[J].Semin Immunol,2013,25(1):2-11.
[8]KIM H,ERDMAN L K,LU Z,et al.Functional roles for C5aand C5aR but not C5L2in the pathogenesis of human and experimental cerebral malaria[J].Infect Immun,2014,82(1):371-379.
[9]STRAINIC M G,LIU J,HUANG D,et al.Locally produced complement fragments C5aand C3aprovide both costimulatory and survival signals to naive CD4+T cells[J].Immunity,2008,28(3):425-435.
[10]KELLY P,JACK D L,NAEEM A,et al.Mannosebinding lectin is a component of innate mucosal defense against Cryptosporidium parvumin AIDS[J].Gastroenterology,2000,119(5):1236-1242
[11]方艳琴,杜帅之,郭雅旭,等.不同Ⅱd亚型微小隐孢子虫对BALB/c小鼠的感染性[J].西北农业学报,2014,12(23):38-42.
[12]LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T))Method[J].Methods,2001,25(4):402-408.
[13]NELLEMANN C,VINGGAARD A M,DAL-GAARDM,et al.Quantification of antiandrogen effect determined by light cycler technology[J].Toxicology,2001,163(1):329-381.
[14]KEARNA A M,TURNER A J L,ELTRINGHAM GJ A,et al.Rapid detection and quantification of CMVDNA in urine using lightcycler based real-time PCR[J].J Clin Virol,2002,24(1-2):131-134.
[15]GERARD C,BAO L,OROZCO O,et al.Structure diversity in the extracellular faces of peptidergic G-protein-coupled receptors.Molecular cloning of the mouse C5aanaphylatoxin receptor[J].J Immunol,1992,149(8):2600-2606.
[16]RAMIREZ N E,WARD L A,SREEVATSAN S.Areview of the biology and epidemiology of cryptosporidiosis in humans and animals[J].Microbes Infect,2004,6(8):773-785.
[17]ZHAO G H,FANG Y Q,RYANU,et al.Dynamics of Th17associating cytokines in Cryptosporidium parvum-infected mice[J].Parasitol Res,2015,[Epub ahead of print].
[18]KIM A H,DIMITRIOU I D,HOLLAND M C,et al.Complement C5areceptor is essential for the optimal generation of antiviral CD8+T cell responses[J].JImmunol,2004,173(4):2524-2529.
[19]KHL J,BAELDER R,LEWKOWICH I P,et al.Aregulatory role for the C5aanaphylatoxin in type 2immunity in asthma[J].J Clin Invest,2006,116(3):783-796.
[20]FREGONESE L,SWAN F J,VAN SCHADEWIJKA,et al.Expression of the anaphylatoxin receptors C3aR and C5aR is increased in fatal asthma[J].J Allergy Clin Immunol,2005,115(6):1148-1154.