Lin Chun-Ming, Jeng Chian-Ren, Chang Hui-Wen, Guo Ing-Cherng, Huang Yu-Liang, Tsai Yi-Chieh, Chia Mi-Yuan, Pang Victor Fei
Division of Pathobiology, Graduate Institute of Veterinary Medicine, National Taiwan University, Taipei 106, Taiwan, ROC.
Vet Immunol Immunopathol. 2008 Aug 15;124(3-4):355-66. doi: 10.1016/j.vetimm.2008.04.019. Epub 2008 May 4.
Information regarding the susceptibility of swine lymphocytes to PCV2 is rather limited. To further explore and characterize the PCV2 infection in swine lymphocytes, an in vitro model using concanavalin A (Con A)-stimulated peripheral blood lymphocytes (PBLs) obtained from clinically healthy PCV2-carrier pigs was introduced. It was found that the PCV2 antigen-containing rate was below 2% in PBLs from healthy PCV2-free pigs following treated simultaneously with Con A and PCV2. However, significantly higher PCV2 antigen- and nucleic acid-containing rates could be seen in Con A-stimulated PBLs from clinically healthy PCV2-carrier pigs. Prior to Con A treatment, both of the PCV2 antigen- and nucleic acid-containing rates in PBLs from healthy PCV2-carrier pigs were less than 1%; however, they reached 22.1+/-5.7% by flow cytometry and 27.1+/-6.5% by in situ hybridization, respectively, at 4-day post-incubation with Con A. Phenotyping of PCV2 antigen-containing cells revealed that PCV2-positive cells could be detected in both T and B lymphocyte populations within which IgM-positive B lymphocytes appeared to have a relatively higher positive rate. The Con A-stimulated PBLs also displayed a significantly higher viral load by the measurement of either PCV2 DNA copy number or viral titer when compared with the non-treated PBLs from healthy PCV2-carrier pigs. The results indicate that PBLs, especially IgM-bearing B lymphocytes, are indeed susceptible to PCV2 infection and PCV2 is capable of replicating in dividing lymphocytes. This activation-induced replication may explain in part the pathogenesis of lymphoid depletion in PMWS-affected pigs.
关于猪淋巴细胞对猪圆环病毒2型(PCV2)易感性的信息相当有限。为了进一步探究和表征PCV2在猪淋巴细胞中的感染情况,引入了一种体外模型,该模型使用从临床健康的PCV2携带猪获取的经刀豆蛋白A(Con A)刺激的外周血淋巴细胞(PBLs)。结果发现,在同时用Con A和PCV2处理的健康无PCV2猪的PBLs中,含PCV2抗原的比率低于2%。然而,在临床健康的PCV2携带猪经Con A刺激的PBLs中,可以观察到含PCV2抗原和核酸的比率显著更高。在Con A处理之前,健康PCV2携带猪的PBLs中含PCV2抗原和核酸的比率均小于1%;然而,在与Con A孵育4天后,通过流式细胞术检测其分别达到22.1±5.7%,通过原位杂交检测达到27.1±6.5%。对含PCV2抗原细胞的表型分析显示,在T和B淋巴细胞群体中均可检测到PCV2阳性细胞,其中IgM阳性B淋巴细胞似乎具有相对较高的阳性率。与来自健康PCV2携带猪的未处理PBLs相比,通过测量PCV2 DNA拷贝数或病毒滴度发现,经Con A刺激的PBLs也显示出显著更高的病毒载量。结果表明,PBLs,尤其是携带IgM的B淋巴细胞,确实对PCV2感染易感,并且PCV2能够在分裂的淋巴细胞中复制。这种激活诱导的复制可能部分解释了受断奶仔猪多系统衰竭综合征(PMWS)影响的猪中淋巴细胞耗竭的发病机制。