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猪圆环病毒2型在猪免疫系统细胞中的感染与复制的体外研究

In vitro studies on the infection and replication of porcine circovirus type 2 in cells of the porcine immune system.

作者信息

Gilpin D F, McCullough K, Meehan B M, McNeilly F, McNair I, Stevenson L S, Foster J C, Ellis J A, Krakowka S, Adair B M, Allan G M

机构信息

Department of Veterinary Science, Queens University Belfast, Stoney Road, Stormont, Belfast, BT4 3SD, UK.

出版信息

Vet Immunol Immunopathol. 2003 Aug 15;94(3-4):149-61. doi: 10.1016/s0165-2427(03)00087-4.

Abstract

Porcine circovirus type 2 (PCV2) nucleic acid and/or antigens are consistently observed in cells of monocytic morphology in lesions of pigs affected by post-weaning multisystemic wasting syndrome (PMWS). In this study, PCV2 antigen was detected in the cytoplasm of monocytes, pulmonary macrophages (PMs) and monocyte-derived macrophages exposed to the virus in vitro, by immunofluorescence analysis (IFA) and the phenotype of these cells confirmed by detection of monocytic cell surface markers using flow cytometry. Viral antigen was not observed in lymphocytic cells. Replication of the virus in PMs was investigated further by comparison to that observed in the continuous pig kidney cell line (PK15A) using quantitative virus titration, quantitative PCR and by the detection of double stranded DNA intermediates of viral replication by Southern blotting analyses. Although increases in viral DNA and levels of infectious virus progeny and the presence of replicative intermediates, indicative of viral replication, were observed in PK15A cells, no such changes were observed in PMs in spite of the fact that infectious virus, viral antigen and viral DNA persisted in the cells for at least the duration of the experiment. These results suggest that in vivo, monocytic cells may not represent the primary target for PCV2 replication.

摘要

在受断奶后多系统消耗综合征(PMWS)影响的猪的病变中,始终能在单核细胞形态的细胞中观察到2型猪圆环病毒(PCV2)核酸和/或抗原。在本研究中,通过免疫荧光分析(IFA)在单核细胞、肺巨噬细胞(PM)以及体外暴露于该病毒的单核细胞衍生巨噬细胞的细胞质中检测到PCV2抗原,并使用流式细胞术检测单核细胞表面标志物来确认这些细胞的表型。在淋巴细胞中未观察到病毒抗原。通过定量病毒滴定、定量PCR以及使用Southern印迹分析检测病毒复制的双链DNA中间体,进一步研究了该病毒在PM中的复制情况,并与在连续猪肾细胞系(PK15A)中观察到的情况进行比较。尽管在PK15A细胞中观察到病毒DNA增加、感染性病毒后代水平增加以及存在指示病毒复制的复制中间体,但在PM中未观察到此类变化,尽管感染性病毒、病毒抗原和病毒DNA在细胞中至少在实验期间持续存在。这些结果表明,在体内,单核细胞可能不是PCV2复制的主要靶细胞。

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