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PCV2对体外培养仔猪淋巴细胞TLRs的影响

DOI: 10.7685/j.issn.1000-2030.2013.05.016, PP. 93-98

Keywords: 仔猪淋巴细胞,猪圆环病毒2型,Toll样受体,荧光定量PCR,蛋白印迹法

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Abstract:

选取6头猪圆环病毒2型(PCV2)和猪繁殖与呼吸综合征病毒(PRRSV)抗原及抗体均为阴性的普通断奶仔猪,无菌取脾脏,制成单个细胞悬液进行体外培养,并分为PCV2组和对照组,PCV2组用PCV2与脾细胞共同培养,对照组加等量RPMI-1640培养液。分别于培养0、6、12、24和48h后收取悬浮的淋巴细胞,用间接免疫荧光法检测PCV2感染淋巴细胞的情况,相对定量RT-PCR法检测淋巴细胞中TLR2、TLR3、TLR4、TLR7、TLR8和TLR9mRNA的表达量,Westernblot检测TLR3和TLR4的蛋白表达情况。结果显示PCV2感染后,TLR2和TLR9mRNA的表达量在12、24和48h时均极显著高于对照组(P<0.01),TLR3和TLR4mRNA的表达量在24h时极显著高于对照组(P<0.01)。TLR3蛋白在6和12h显著高于对照组(P<0.05),TLR4蛋白在6、12、24和48h均显著高于对照组(P<0.05)。结果表明PCV2感染能显著上调体外培养的仔猪淋巴细胞TLR2、TLR3、TLR4和TLR9mRNA水平,并进一步显著上调仔猪淋巴细胞TLR3和TLR4蛋白的表达,从而导致机体炎性细胞因子的分泌紊乱。

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