猪圆环病毒Ⅱ型和巨噬细胞对体外培养仔猪淋巴细胞白介素-6和白介素-10及其受体表达的影响  

Influence of Porcine Circovirus Type 2 and Macrophages on the Expression of IL-6,IL-10 and Their Receptors of Piglet Lymphocytes In Vitro

作　　者：陈耿[1] 华利忠[1] 张书霞[1] 

机构地区：[1]南京农业大学动物医学院,江苏南京210095

出　　处：《江苏农业学报》2010年第5期993-998,共6页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金资助项目(30471302;30871848)

摘　　要：选取5头猪圆环病毒Ⅱ型(PCV2)和猪繁殖与呼吸综合征病毒(PRRSV)血清抗体阴性的普通断奶仔猪,无菌取其脾脏制成单细胞悬液后,分设淋巴细胞(L组)、淋巴细胞+PCV2(L+V组)、淋巴细胞+巨噬细胞(L+M组)和淋巴细胞+巨噬细胞+PCV2(L+M+V组)4组进行体外培养。分别在培养后0 h、6 h、12 h和24 h收获淋巴细胞及其上清液。用ELISA检测上清液中猪淋巴细胞白介素-6(IL-6)和白介素-10(IL-10)的含量,用RT-PCR检测淋巴细胞中IL-6、IL-6R、IL-10和IL-10RmRNA的表达。结果显示:与L组相比,L+V组中IL-6的含量在攻毒后24 h显著升高(P<0.05);PCV2在攻毒后12 h对IL-6RmRNA的表达呈显著抑制(P<0.05);PCV2在攻毒后6 h显著促进IL-10的表达(P<0.05)。上述结果表明,PCV2能促进淋巴细胞中IL-6和IL-10的表达;而巨噬细胞能抑制PCV2对淋巴细胞中IL-6表达的上调作用,同时协同PCV2促进淋巴细胞中IL-10及其受体持续性高表达,导致持续的免疫抑制,在PCV2的致病过程中扮演重要角色。Five post-weaning piglets which were free of porcine circovirus type 2（PCV2） and porcine reproductive and respiratory syndrome virus（PRRSV）antibodies were chosen as the experimental animals.The single cell suspension isolated from spleens of these piglets with pinhead of injectors were distributed into four groups： lymphocytes（group L）,lymphocytes inoculated with PCV2（group L＋V）,macrophages and lymphocytes（group L＋M）,macrophages and lymphocytes inoculated with PCV2（group L＋M＋V）.Supernatants and lymphocytes were respectively harvested after the single cell suspension was treated in vitro with or without PCV2 for 0 h,6 h,12 h and 24 h.The contents of interleukin-6（IL-6） and interleukin-10（IL-10） in cell culture supernatants and the mRNA levels of IL-6,IL-6R,IL-10 and IL-10R of the splenic lymphocytes were detected by pig-specific ELISA kit and RT-PCR,respectively.The results showed that compared with group L,the contents of IL-6 in L＋V group were all increased after inoculated with PCV2,especially that at 24 h post-inoculation（HPI） being significantly increased（P〈0.05）;the mRNA level of IL-6 in L＋V group at 12 HPI was significantly decrease（P〈0.05）,the expression of IL-10 in L＋V group at 6 HPI was significantly enhanced（P〈0.05）.In conclusion,PCV2 could enhance the expressions of IL-6 and IL-10 in lymphocyte;macrophages could inhibit the up-regulation of PCV2 on the expression of IL-6 in lymphocytes,and macrophages with PCV2 could promote the high expressions of IL-10 and its receptor in lymphocyte,which would result in persistent immunosuppression and play an important role in the pathogenesis of PCV2.

关 键 词：猪圆环病毒Ⅱ型 巨噬细胞 淋巴细胞 白介素-6 白介素-10 

分 类 号：S852.33[农业科学—基础兽医学]