PCV2通过激活MyD88调控体外培养PAMs分泌IL-1β、IL-6和IL-10

doi:10.11843/j.issn.0366-6964.2014.04.020

Abstract

Abstract:

Six 30-day-old piglets (post-weaned) were selected as experiment animals，which were free of porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV) antibody and antigen as detected by ELISA and RT-PCR.The alveolar macrophages (AMs) isolated aseptically were divided into four groups，i.e.，control group，molecular myeloid differentiation 88 (MyD88) interference group (interference group)，PCV2 infection group (PCV2 group) and PCV2 infection-MyD88 interference group (PCV2-interference group)，and cultured in vitro.The MyD88 gene was knocked down by siRNA (small interfering RNA).After PCV2 infection，the cells and the cultural supernatants were collected at 0，6，12，24 and 48 h respectively.The gene-silencing efficiency of siRNA was detected by fluorescence quantitative PCR.The intracellular expression of MyD88 protein was detected by Western blot.The dynamic changes of IL-1β，IL-6 and IL-10 in the supernatants were assayed by ELISA.The results showed that siRNA could lead to 78% of the MyD88 gene silencing，and significantly affected the expression of MyD88 protein (P<0.01).The amount of MyD88 expressed in AMs in PCV2 group increased significantly after 6 h (P <0.05)，which was significantly higher compared to that in PCV2-interference group (P <0.01) at 12，24 and 48 h.The yield of IL-1β secreted in AMs at 6 h infected with PCV2，was notably higher compared with those in control group (P<0.01) and PCV2-interference group (P<0.05).The level of the secreted IL-6 in the PCV2 group was significantly higher than those in the control group (P<0.01) and PCV2-interference group (P<0.05) at 6 and 12 h.The yield of IL-10 secreted in AMs significantly increased (P<0.01) at 12，24 and 48 h infected with PCV2; the yield of IL-10 in PCV2-interference group significantly decreased (P<0.01) at 24 and 48 h.The results indicated that the infection of PCV2 contributed to different changes of the secretion of IL-1β，IL-6 and IL-10 in PAMs cultured in vitro，where MyD88 was involved as an important regulator.

CLC Number:

S852.33

HAN Jun-yuan，GUO Hua，ZHANG Ya-qun，DUAN Dian-ning，LI Xiao-lin，CHEN Meng-meng，ZHANG Shu-xia. PCV2 Regulates PAMs Secreting IL-1β，IL-6 and IL-10 through the Activation of MyD88 in vitro[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(4): 647-653.

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References

［1］HARDING J C.Postweaning multisystemic wasting syndrome：Epidemiology and clinical presentation ［J］.Swine Health Prod，1998，6(6):249-254.
［2］郎洪武，张广川，吴发权，等.断奶猪多系统衰弱综合征血清抗体检测［J］.中国兽医科技，2000，30(3)：3-5.
［3］CHAE C.A review of porcine circovirus 2-associated syndromes and diseases ［J］.Vet J, 2005，169(3):326-336.
［4］SEGALéS J，ROSELL C，DOMINGO M.Pathological findings associated with naturally acquired porcine circovirus type 2 associated disease ［J］.Vet Microbiol，2004，98(2):137-149.
［5］SCHMITZ N，KURRER M，BACHMANN M F，et al.Interleukin-1 is responsible for acute lung immunopathology but increases survival of respiratory influenza virus infection ［J］. J Virol，2005，79(10):6441-6448.
［6］LENZ A，FRANKLIN G A，CHEADLE W G，et al.Systemic inflammation after trauma ［J］.Injury，2007，38(12):1336-1345.
［7］FICKENSCHER H，HÖR S，KÜPERS H，et al.The interleukin-10 family of cytokines ［J］.Trends Immunol，2002，23(2):89-96.
［8］陈耿，张书霞，华利忠．PCV2对仔猪胸腺IL-2、IL-6和IL-10及其受体mRNA的影响及NO-NOS 系统的调控［J］.南京农业大学学报，2011，34(3):101-106.
［9］TAKEDA K，KAISHO T，AKIRA S.Toll-like receptor ［J］.Annu Rev Immunol，2003，21:335-376.
［10］KAISHO T，AKIRA S.Toll-like receptor function and signaling ［J］.J Allergy Clin Immunol，2006，117(5):979-987.
［11］LIU J，BAI J，LU Q，et al.Two-dimensional liqiud chromatography-tandem mass spectrometry coupled with isobric tages for relative and absolute quantification(iTRAQ) labeling approach revealed first proteome profiles of pulmonary alveolar macrophages infected with porcine circovirus type 2 ［J］.J Proteomics，2013，79:72-86.
［12］周谦军，范慧敏，刘中民.MyD88siRNA诱导猪调节性树突状细胞的实验研究［J］.中国细胞生物学学报，2010，32(3):415-421.
［13］CHANG H W，JENG C R，LIN T L，et al.Immunopathological effects of porcine circovirus type 2 (PCV2) on swine alveolar macrophages by in vitro inoculation ［J］.Vet Immunol Immunopathol，2006，110(3-4):207-219.
［14］CHAUNG H C，CHEN C W，HSIEH B L，et al.Toll-like receptor expressions in porcine alveolar macrophages and dendritic cells in responding to polyIC stimulation and porcine reproductive and respiratory syndrome virus (PRRSV) infection ［J］.Comp Immunol Microbiol Infect Dis，2010，33（3）:197-213.
［15］李晓琳，郭华，刘子臣，等.PCV2对体外培养仔猪淋巴细胞TLRs的影响［J］.南京农业大学学报，2013，36(5)：93-98.
［16］TAKEDA K，AKIRA S.TLR signaling pathways ［J］.Semin Immunol，2004，16（1）:3-9.
［17］CHRISTMAN J W，SADIKOT R T，BLACKWELL T S.The role of nuclear factor-κb in pulmonary diseases ［J］.Chest，2000，117(5):1482-1487.
［18］STEPTOE A，HAMER M，CHIDA Y.The effects of acute psychological stress on circulating inflammatory factors in humans：A review and meta analysis ［J］.Brain Behav Immun，2007，21(7):901-912.
［19］STEVENSON L S，MCCULLOUGH K，VINCENT I，et al.Cytokine and C-reactive protein profiles induced by porcine circovirus type 2 experimental infection in 3-week-old piglets ［J］.Viral Immunol，2006，19(2):189-195.
［20］SHANLEY T P，SCHMAL H，FRIEDL H P，et al.Regulatory effects of intrinsic IL-10 in IgG immune complex-induced lung injury ［J］.J Immunol，1995，154(7):3454-3460.
［21］胡健才，刘维永，梁继河，等.抗肿瘤坏死因子单克隆抗体对兔创伤性急性呼吸窘迫综合征的保护作用［J］.中华实验外科杂志，1998，15(6)：489-490.
［22］FICKENSCHER H，HÖR S，KÜPERS H，et al.The interleukin-10 family of cytokines ［J］.Trends Immunol，2002，23(2):89-96.
［23］DARWICH L，BALASCH M，PLANA-DURáN J，et al.Cytokine profiles of peripheral blood mononuclear cells from pigs with postweaning multisystemic wasting syndrome in response to mitogen，superantigen or recall viral antigens ［J］.J Gen Virol，2003，84(Pt 12):3453-3457.
［24］SIPOS W，DUVIGNEAU J C，WILLHEIM M，et al.Systemic cytokine profile in feeder pigs suffering from natural postweaning multisystemic wasting syndrome (PMWS) as determined by semiquantitative RT-PCR and flow cytometric intracellular cytokine detection ［J］.Vet Immunol Immunopathol，2004，99(1-2):63-71.
［25］DARWICH L，SEGALÉS J，RESENDES A，et al.Transient correlation between viremia levels and IL-10 expression in pigs subclinically infected with porcine circovirus type 2 (PCV2) ［J］.Res Vet Sci，2008，84(2):194-198.

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PCV2 Regulates PAMs Secreting IL-1β，IL-6 and IL-10 through the Activation of MyD88 in vitro

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