猪圆环病毒分子生物学研究进展

猪圆环病毒(porcine circovirus,PCV)是迄今发现的最小的动物病毒,对养猪业造成的损失日趋严重.主要对猪圆环病毒的基因组、致病基因、病毒受体等进行综述,旨在为猪圆环病毒的研究和猪圆环病毒病的预防提供参考.     

猪圆环病毒1-2型嵌合体、猪肺炎支原体二联灭活疫苗——瑞圆舒~?（Fostera~? PCV MH）的应用研究进展

正猪圆环病毒(Porcine circovirus, PCV)属于圆环病毒科(Circoviridae)、圆环病毒属(Circoviridae)。圆环病毒科病毒由单链环形DNA组成,目前已发现30多种圆环病毒,包含了目前已知的最小的动物DNA病毒,其中,猪圆环病毒包括猪圆环病毒1型(PCV1)、猪圆环病毒2型(PCV2)、猪圆环病毒3型(PCV3)和猪圆环病毒4型(PCV4)。通常认为PCV1对猪是无致病力的;PCV2是导致猪圆环病毒相关疾病(porcine circovirus associated diseases, PCVADs)的最主要病原,     

猪圆环病毒2型分子致病机理及相关疾病研究进展

猪圆环病毒(porcine circovirus,PCV)有两个基因型,分别为PCV1和PCV2。PCV2有致病性,它可以引发多种相关的疾病(PCVAD),如断奶后多系统衰竭综合征(PMWS)等。该病毒主要通过攻击猪的淋巴细胞,产生免疫抑制,诱发细胞凋亡,还可以和其他病毒协同感染,大大增加了动物的发病率及死亡率。文章对PCV2分子致病机理及引起的相关疾病和防治作一综述。     

猪圆环病毒及疫苗使用效果

猪圆环病毒型(porcine circovirus)是引起猪圆环病毒病(porcine circovirus desease,PCVD)的主要病原,其分为PCV1和PCV2。猪圆环病毒主要引发断奶仔猪多系统衰竭综合征(PMWS)、育肥猪皮炎与肾病综合征(PDNS)等无法治愈的疾病,该病的感染给全世界养猪业造成巨大损失。破坏猪体的免疫系统的猪圆环病毒2型,可以引起严重的免疫抑制。发病后常能引起病猪出现混合感染与继发感染,其中包括细菌和病毒、病毒与病毒之间相互感染,因此该病不容易进行诊断和治疗,此病的发生严重威胁猪只的健康。本文对病毒的致病机理、临床症状、病理变化、诊断和防制对策等方面的研究进展进行了简要综述。     

猪圆环病毒特点

<正>猪圆环病毒(porcine circovirus,PCV)有2种不同的血清型,即猪圆环病毒1型(PCV1)和猪圆环病毒2型(PCV2)。PCV1暂无致病性,PCV2被认为是引起猪圆环疾病(PCVDs)的主要病原。现就PCV的病原特点进行综述。1结构与特性     

我国猪圆环病毒2型感染的危害

<正>猪圆环病毒(porcine circo virus,PCV)属于圆环病毒科圆环病毒属的成员,根据致病性、抗原性及核苷酸序列的差异,PCV可分为两个血清型,即PCV1和PCV2。其中源自污染猪源细胞PK-15的称为PCV1,对猪无致病性,但广泛存在猪体内及猪源细胞系。PCV2具有致病性,是仔猪断奶后多系统衰竭综     

猪圆环病毒2型对猪的免疫抑制机理

<正>猪圆环病毒2型(PCV2)是圆环病毒科(Circoviridae)圆环病毒属的一员,分PCV1型和PCV2型,PCV1型不致病,PCV2型致病。猪圆环病毒2型(Porcine circovirus type2,PCV2)     

我国猪圆环病毒2型感染的危害

<正>猪圆环病毒(porcine circo virus,PCV)属于圆环病毒科圆环病毒属的成员,根据致病性、抗原性及核苷酸序列的差异,PCV可分为两个血清型,即PCV1和PCV2。其中源自污染猪源细胞PK-15的称为PCV1,对猪无致病性,但广泛存在猪体内及猪源细胞系。PCV2具有致病性,是仔猪断奶后多系统衰竭综     

猪圆环病毒3型在英国的感染情况调查

正猪圆环病毒(porcine?circovirus,PCV)系单股环状DNA病毒,是已知能自主复制的最小的动物病毒之一。根据其性质不同最初分为2种基因型:猪圆环病毒1型(PCV1)和猪圆环病毒2型(PCV2),其中PCV1不具有致病性,PCV2是在20世纪90年代后期被分离和鉴定,与猪的多系统疾病综合征相关且在世界范围内广泛传播,给养猪业带来巨大的经济损失。在使用PCV2     

驻马店市猪圆环病毒病流行情况调查与分析

<正>猪圆环病毒(porcine circovirus,PCV)现已知有三个血清型,即猪圆环病毒1型(PCV1)、猪圆环病毒2型(PCV2)、猪圆环病毒3型(PCV3)。其中PCV1为非致病性的病毒;PCV2对猪具有较强的易感性,感染猪可自鼻液、粪便等废物中排出病毒,经口腔、呼吸道途径感染不同年龄的猪,是一种对养猪业危害较大的免疫抑制性疫病;2016年10月,美国 Kansas州立     

Porcine circovirus type 2 (PCV2) distribution and replication in tissues and immune cells in early infected pigs

Replication of porcine circovirus type 2 (PCV2) in pigs, as measured by spliced capsid mRNA (Cap mRNA) and viral DNA, was investigated following experimental infection. Peripheral blood mononuclear cells (PBMCs), and tissue from bronchial lymph nodes (BLN), inguinal lymph nodes (ILN), tonsils, lungs, liver, kidneys, spleen and thymus from infected pigs on different days post-infection (DPI) were assessed. PCV2 replication differed dramatically between tissues from the same infected pig. The virus actively replicated in most tested tissues at 14DPI in association with increased PCV2 associated lesions and PCV2 antigen levels, although no clinical signs correlated with PCV2 associated disease were observed in infected pigs during the course of the study. The PCV2 Cap mRNA was detected only at 13DPI in PBMCs from infected pigs, suggesting replication of the virus in circulating blood is transient and not a major site for PCV2 replication in vivo. Evaluation of the Cap mRNA and viral DNA synthesis in T and B lymphocyte and monocyte populations from PBMCs and BLN at various intervals post-inoculation revealed replication of PCV2 in all cell subpopulations; however, viral replication in B lymphocytes was greater than observed in mononuclear cells isolated from BLN at 14DPI indicating that B lymphocytes may be an important cell population for PCV2 replication. These findings further our understanding of the cell types permissive for PCV2 replication and the pathogenesis of PCV2 infection in vivo.     

Distribution and characterization of IL-10-secreting cells in lymphoid tissues of PCV2-infected pigs

Distribution and characterization of interlukin-10 (IL-10)-secreting cells in lymphoid tissues of pigs naturally infected with porcine circovirus type 2 (PCV2) were evaluated in accordance with PCV2 antigen detection. After screening a total of 56 pigs showing the symptoms of postweaning multisystemic wasting syndrome (PMWS), 15 pigs were PCV2 positive and 5 pigs, which showed stronger positive signals over multiples tissues were further investigated. This study showed that in PCV2-infected lymphoid tissues, particularly mandibular lymph node, spleen and tonsil, IL-10 expression was mainly localized in T-cell rich areas but rarely in B cell rich areas. IL-10 was highly expressed in bystander cells but rarely in PCV2-infected cells. Elevated IL-10 expression was predominantly associated with T cells, but rarely with B cells or with macrophages. The results of this study provide evidence for the role of IL-10 in chronic PCV2 infection and its relation to PCV2 antigen in affected tissues. Constantly elevated levels of IL-10 lead to immunosuppression in persistent and chronic viral infections. The increased IL-10 expression observed in PCV2 infection in this study suggests that IL-10-mediated immunosuppression may play an important role in the pathogenesis and maintenance of naturally occurring PCV2 infection.     

Porcine circovirus type 2 (PCV2) viral components immunomodulate recall antigen responses

Porcine circovirus type 2 (PCV2) is a single-stranded circular DNA virus infecting domestic pigs worldwide. Interaction of this virus with the immune system apparently modulates the immune response of the host. In the present study, the implication of different components of PCV2 in the modulation of the immune response of the host were investigated by using PCV2 viral-like particles (VLPs) and 16 novel oligodeoxyribonucleotides containing CpG motifs (CpG-ODNs) based on the PCV2 genomic sequence. The role of these viral components was studied by evaluating the cytokine profiles (IFN-alpha, IFN-gamma, IL-10, IL-2 and IL-12) on porcine peripheral mononuclear cell (PBMC) and bone marrow-derived dendritic cell (BMDC) cultures. Also, the effect of PCV2 and its elements were examined in recall antigen (pseudorabies virus, PRV) responses. While PCV2 was a potent inducer of IL-10 by PBMCs, such effect was not observed using CpG-ODNs or VLPs. However, IFN-gamma and IL-2 production by recall antigen was repressed in presence of PCV2 and most of the studied CpG-ODNs. VLPs did not have such repressive effect. In BMDC cultures, PCV2 and most of CpG-ODNs were able to inhibit IFN-alpha secretion induced by PRV. Interestingly, CpG-ODNs with inhibitory effect were located within the PCV2 Rep gene. Additionally, PCV2 virus was a very strong IL-12 inducer in BMDC cultures. Whereas, IFN-alpha modulation on BMDC after PCV2 VLP treatment was neglectable, PCV2 VLPs were potent IL-12 inducers. Our data shows that PCV2 viral elements can distinctly regulate cytokine production depending on the cell population studied. Thus, the final immune response upon PCV2 infection seems to depend on the fine balance between the regulatory elements present in viral DNA and structural protein within the host immune system.     

猪圆环病毒2型感染PK-15细胞中外泌体的鉴定及其对细胞增殖和凋亡的影响

旨在分离感染PCV2的PK-15细胞分泌的外泌体,探讨其在PCV2感染淋巴细胞中的作用。提取PCV2感染PK-15细胞上清液中的外泌体,利用透射电镜观察外泌体形态、纳米颗粒跟踪分析测定外泌体粒径、Western blot检测外泌体标记蛋白分子（CD81、TSG101）;PKH67标记外泌体后检测细胞对外泌体的摄取;提取PCV2-外泌体基因组,检测PCV2 Rep和Cap基因;利用间接免疫荧光试验检测PCV2-外泌体感染PK-15细胞后PCV2病毒定位;通过绝对定量PCR检测PCV2-外泌体对淋巴细胞的感染率;利用CCK-8法检测淋巴细胞增殖;利用Annexin V-FITC/PI检测淋巴细胞凋亡率。透射电镜和外泌体粒径分析结果显示,PK-15细胞分泌的外泌体为双层膜的囊泡,直径30~200 nm;Western blot结果显示PK-15细胞分泌的外泌体存在特异性标记蛋白CD81和TSG101;外泌体摄取试验结果显示PCV2-外泌体能够被细胞摄取;PCR结果表明PCV2-外泌体基因组中含有PCV2 Rep和Cap基因;间接免疫荧光结果显示,与PCV2直接感染相比,PCV2-外泌体感染的淋巴细胞中,PCV2病毒拷贝显著升高,差异明显（P<0.01）,外泌体裂解后感染淋巴细胞能力与PCV2病毒无显著差异;PCV2-外泌体可显著抑制淋巴细胞增殖（P<0.01或P<0.05）;PCV2-外泌体显著提高淋巴细胞凋亡率。PCV2感染PK-15细胞的外泌体中携带病毒基因,感染淋巴细胞后使细胞增殖降低和细胞凋亡增加。     

Detection of neutralizing antibodies in postweaning multisystemic wasting syndrome (PMWS)-affected and non-PMWS-affected pigs

The notion that postweaning multisystemic wasting syndrome (PMWS)-affected pigs develop an impaired humoral response against porcine circovirus type 2 (PCV2) has been reported in several studies. However, little information is available regarding the presence of neutralizing antibodies (NA) in PCV2-infected pigs and their role in the pathogenesis of the disease. The aim of the present work was to further characterize the humoral response, and in particular the production of NA, in pigs with different PCV2-infection status. Seventy-two conventional pigs from different farms were classified into three groups based on PCV2 infection and clinico-pathological status, namely: PCV2-negative, non-PMWS PCV2-positive and PMWS-affected animals. In addition, 9-week old pigs from an experimental infection (6 controls and 14 PCV2-inoculated pigs) were also studied. NA and total PCV2 antibodies (TA) as well as viral load in serum were determined and correlated with the clinico-pathological status of pigs. Results indicated that PMWS-affected pigs had lower NA titres, if any, than healthy animals. NA titres were also inversely correlated with PCV2 load in serum. NA and TA titres were positively correlated; however, correlation differed among infection status, being lower in PCV2-positive pigs. Also, the diagnostic performance of each test was evaluated, indicating that the combination of viral neutralization and quantitative PCR in serum was useful to discard PMWS (specificity 92%). In experimentally infected animals, the evolution of NA paralleled the course TA, although a slight delay in NA production was seen in some animals. The increase of NA coincided with the drop in viral load. Results from this work further support that PMWS-affected pigs show an impaired humoral immune response and, particularly, an inefficient NA response against PCV2.     

Co-infection of porcine dendritic cells with porcine circovirus type 2a (PCV2a) and genotype II porcine reproductive and respiratory syndrome virus (PRRSV) induces CD4(+)CD25(+)FoxP3(+) T cells in vitro

Porcine circovirus associated disease (PCVAD) is currently one of the most economically important diseases in the global swine industry. Porcine circovirus type 2 (PCV2) is the primary causative agent, however co-infection with other swine pathogens such as porcine reproductive and respiratory syndrome virus (PRRSV) is often required to induce the full spectrum of clinical PCVAD. While the specific mechanisms of viral co-infection that lead to clinical disease are not fully understood, immune modulation by the co-infecting viruses likely plays a critical role. We evaluated the ability of dendritic cells (DC) infected with PRRSV, PCV2, or both to induce regulatory T cells (T(regs)) in vitro. DCs infected with PCV2 significantly increased CD4(+)CD25(+)FoxP3(+) T(regs) (p<0.05) and DCs co-infected with PRRSV and PCV2 induced significantly higher numbers of T(regs) than with PCV2 alone (p<0.05). Cytokine analysis indicated that the induction of T(regs) by co-infected DCs may be dependent on TGF-β and not IL-10. Our data support the immunomodulatory role of PCV2/PRRSV co-infection in the pathogenesis of PCVAD, specifically via T(reg)-mediated immunosuppression.     

Characterization of porcine circovirus type 2 (PCV2) infection in swine lymphocytes using mitogen-stimulated peripheral blood lymphocytes from healthy PCV2-carrier pigs

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.     

猪圆环病毒2型基因组DNA的结构与功能研究进展

猪圆环病毒2型(PCV2)是引起猪圆环病毒病相关疾病的主要致病原,可损伤宿主的免疫系统,加剧一些细菌病和病毒病感染的临床症状,在全球养猪业中造成巨大损失。PCV2的基因组DNA含有11个开放阅读框(ORFs),其中至少有7个ORFs编码的蛋白质分子质量大于5ku,但目前只有5种编码的蛋白(Rep、Rep'、Cap、ORF3和ORF4)发现于PCV2复制过程中。作者概述了当前PCV2基因组DNA的结构与功能,阐明PCV2的病原学,为防控PCV2感染提供科学依据。     

Advances on Structure and Function of Genomic DNA of Porcine Circovirus Type 2

Porcine circovirus type 2 (PCV2) is a disease caused by porcine circovirus, which can damage the host's immune system and exacerbate the clinical symptoms of many bacterial and viral infections.It has caused increasingly larger losses in the pig industry wordwide.The genomic DNA of PCV2 contains 11 open reading frames (ORFs) and at least seven potential ORFs-encoding proteins larger than 5 ku.However, only five virally encoded proteins, including Rep, Rep', Cap, ORF3 and ORF4, had been identified in PCV2 replication.In this review, we discussed the progress on structure and function of genomic DNA of PCV2 in order to provide insight into the scientific basis of the pathogenesis of PCV2 and prevent its infection.     

Porcine circovirus type 2 (PCV2) induces cell proliferation,fusion, and chemokine expression in swine monocytic cells in vitro

Granulomatous lymphadenitis is one of the pathognomonic lesions in post-weaning multisystemic wasting syndrome (PMWS)-affected pigs. This unique lesion has not been reported in direct association with viral infection in pigs. The objective of the present study was to evaluate whether porcine circovirus type 2 (PCV2) alone is able to induce functional modulation in porcine monocytic cells in vitro to elucidate its possible role in the development of granulomatous inflammation. It was found that the proliferation activity of blood monocytes (Mo) and monocyte-derived macrophages (MDM) was significantly enhanced by PCV2. During monocyte-macrophage differentiation, the PCV2 antigen-containing rate and formation of multinucleated giant cells (MGC) were significantly increased in MDM when compared to those in Mo. The MDM-derived MGC displayed a significantly higher PCV2 antigen-containing rate than did the mono-nucleated MDM. Supernatants from PCV2-inoculated MDM at 24 h post-inoculation induced an increased tendency of chemotactic activity for blood Mo. At the same inoculation time period, levels of mRNA expression of the monocytic chemokines, monocyte chemoattractant protein-1 and macrophage inflammatory protein-1, also significantly increased in PCV2-inoculated MDM. The results suggest that PCV2 alone may induce cell proliferation, fusion, and chemokine expression in swine monocytic cells. Thus, PCV2 itself may play a significant role in the induction of granulomatous inflammation in PMWS-affected pigs.