猪圆环病毒2型(porcine circovirus type 2,PCV2)主要损伤猪的免疫细胞,引起淋巴细胞缺失和免疫功能抑制.白细胞介素15(IL-15)具有广泛的免疫调节功能,对自然杀伤(NK)、CD8+T细胞和NKT细胞等多种免疫细胞的存活、增殖、分化和免疫功能发挥着重要的调节作用.为了明确PCV2对IL-15表达的影响,用PCV2感染4周龄仔猪(n=4),同时设立阴性对照组(n=4).感染后7 d,采集感染组和对照组的腹股沟淋巴结,应用猪细胞因子抗体芯片(QAP-CYT-1)定量检测组织中细胞因子的表达量,筛选差异细胞因子,对IL-15进行GO和KEGG富集分析.应用实时荧光定量PCR(qPCR)和ELISA方法在mRNA和蛋白水平上对IL-15的差异表达进行验证,并同时检测猪外周血单个核细胞(PBMC)和血清中的IL-15水平.与对照组相比,感染组的IL-15水平显著降低(P＜0.05);IL-15主要参与细胞因子与细胞因子受体相互作用、免疫应答、细胞活化,以及JAK-STAT和TNF信号通路等的调节过程;感染组的腹股沟淋巴结IL-15 mRNA和蛋白水平均显著低于对照组(P＜0.05),与QAP-CYT-1的检测结果一致.但同时检测的PBMC和血清中的IL-15 mRNA与蛋白水平没有显著差异.上述结果表明,PCV2对仔猪腹股沟淋巴结微环境内IL-15具有抑制作用.该研究可以为进一步揭示PCV2的免疫抑制机制提供重要信息.
Inhibition of PCV2 on IL-15 in inguinal lymph nodes of piglets
Porcine circovirus type 2(PCV2)mainly damages the immune cells of pigs,causing lym-phocyte depletion and immune suppression.Interleukin(IL)-15 regulates immune functions wide-ly,and plays an important regulatory role in the survival,proliferation,differentiation and immune function of a variety of immune cells such as natural killer(NK),CD8+T cells and NKT cells.In this study,in order to determine the effect of PCV2 on IL-15 expression,4-week-old piglets(n=4)were infected with PCV2 and the negative control group(n=4)was set up.On day 7 post-infec-tion,the inguinal lymph nodes of the infected and control groups were collected,and porcine cyto-kine antibody microarray(QAP-CYT-1)was employed to quantify the expression of cytokines in the tissues,screen for differential cytokines,and GO and KEGG enrichment analysis for IL-15 were conducted.Real-time quantitative fluorescent PCR(qPCR)and ELISA were used to verify the level of IL-15 mRNA and protein,and those in porcine peripheral blood mononuclear cells(PBMC)and serum were simultaneously detected.Compared with the negative control group,the expression lev-el of IL-15 was significantly reduced in the infected group(P＜0.05);IL-15 was mainly involved in cytokine-cytokine receptor interactions,immune responses,cellular activation,and the regulation of JAK-STAT and TNF signaling pathways.The levels of IL-15 mRNA and protein in inguinal lymph nodes in the infected group were significantly lower than those in the control group(P＜0.05),which was consistent with the detection results of QAP-CYT-1.However,there was no significant difference in IL-15 mRNA and protein levels in PBMC and serum.These results indicate that PCV2 can inhibit IL-15 in the inguinal lymph node microenvironment of piglets.This study can provide important information for further revealing the immunosuppressive mechanism of PCV2.
porcine circovirus type 2pigletsinguinal lymph nodesIL-15
万方数据
维普
