CD163片段在PRRSV感染过程中的作用

使用Scanprosite软件对猪肺泡巨噬细胞的CD163蛋白结构和功能域进行预测,并对其进行分段克隆,构建原核表达载体pET-28a(Y-P1,Y-P2,Y-P3),片段位置Y-P1(160~798bp),Y-P2(790~20 146bp),Y-P3(2 143~3 084bp)。利用镍柱纯化重组蛋白,免疫小鼠获得特异性抗体。通过病毒阻断试验,验证Y-P1,Y-P2,Y-P3的PRRSV阻断情况;构建真核表达载体pCD163,P1(1~798bp),P2(790~2 046bp),P3(2 023~3 345bp)分别转染PRRSV非易感细胞系BHK-21,验证其PRRSV感染情况。结果显示,经PCR、双酶切及测序鉴定构建的原核和真核表达载体是正确的,SDS-PAGE检验Y-P1,Y-P2,Y-P3蛋白大小分别为27 000,46 000,39 000。Y-P2蛋白,抗Y-P2的小鼠血清能够阻断PRRSV感染Marc-145细胞;转染pCD163的细胞系能够感染PRRSV,而P1,P2,P3片段不感染PRRSV。CD163其790~2 046bp可能是PRRSV感染细胞与CD163受体作用位点。     

猪对PRRSV易感性差异的研究进展

猪繁殖与呼吸综合征是当前危害养猪业最严重的传染病之一,其病原为猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndromevirus,PRRSV)。由于PRRSV的不断变异,其感染力和致病力增强,现有疫苗的保护效果不佳,且弱毒活疫苗有变异成新强毒的风险。本文从宿主遗传变异的角度讨论猪对PRRSV的易感性,包括猪免疫应答的变异与易感性、猪对PRRSV易感性的品种(系)间差异以及PRRSV抗性育种亟待解决的问题,试图为PRRS的防制提供新的思路。     

Associations of natural variation in the CD163 and other candidate genes on host response of nursery pigs to porcine reproductive and respiratory syndrome virus infection

Pigs with complete resistance to porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV) have been produced by genetically knocking out the CD163 gene that encodes a receptor of the PRRSV for entry into macrophages. The objectives of this study were to evaluate associations of naturally occurring single nucleotide polymorphisms (SNPs) in the CD163 gene and in three other candidate genes (CD169, RGS16, and TRAF1) with host response to PRRSV-only infection and to PRRS vaccination and PRRSV/porcine circovirus 2b (PCV2b) coinfection. SNPs in the CD163 gene were not included on SNP genotyping panels that were used for previous genome-wide association analyses of these data. An additional objective was to identify the potential genetic interaction of variants at these four candidate genes with a mutation in the GBP5 gene that was previously identified to be associated with host response to PRRSV infection. Finally, the association of SNPs with expression level of the nearby gene was tested. Several SNPs in the CD163, CD169, and RGS16 genes were significantly associated with host response under PRRSV-only and/or PRRSV/PCV2b coinfection. The effects of all SNPs that were significant in the PRRSV-only infection trials depend on genetic background. The effects of some SNPs in the CD163, CD169, and RGS16 genes depend on genotype at the putative causative mutation in the GBP5 gene, which indicates a potential biological interaction of these genes with GBP5. In addition, genome-wide association results for the PRRSV-only infection trials revealed that SNPs located in the CDK5RAP2 or MEGF9 genes, near the TRAF1 gene, had suggestive effects on PRRS viral load, which indicates that these SNPs might contribute to PRRSV neuropathogenesis. In conclusion, natural genetic variants in the CD163, CD169, and RGS16 genes are associated with resistance to PRRSV and/or PCV2b infection and appear to interact with the resistance quantitative trait locus in the GBP5 gene. The identified SNPs can be used to select for increased natural resistance to PRRSV and/or PRRSV-PCV2b coinfection.     

In utero infection with porcine reproductive and respiratory syndrome virus modulates leukocyte subpopulations in peripheral blood and bronchoalveolar fluid of surviving piglets

It is well known that piglets congenitally infected with porcine reproductive and respiratory syndrome virus (PRRSV) can be viremic at birth, and that preweaning mortality due to secondary infections often increases during acute outbreaks of PRRS. Therefore, an immunosuppressive effect of in utero infection has been suggested. The aim of the present study was to characterise the changes of leukocyte populations in piglets surviving in utero infection with PRRSV. A total of 27 liveborn uninfected control piglets and 22 piglets infected transplacentally with a Danish strain of PRRSV were included. At 2 and 4 weeks of age, 21 of 22 (96%) and 7 of 14 (50%) examined infected piglets were still viremic, whereas PRRSV could not be detected in the six infected piglets examined at 6 weeks of age. Flow cytometry analysis was used to determine the phenotypic composition of leukocytes in peripheral blood and bronchoalveolar lavage fluid (BALF) of 2-, 4- and 6-week-old infected piglets and age-matched uninfected controls. The key observation in the present study is that high levels of CD8(+) cells constitute a dominant feature in peripheral blood and BALF of piglets surviving in utero infection with PRRSV. In BALF, the average high level of CD8(+) cells in 2-week-old infected piglets (33.4 +/- 12.6%) was followed by a decline to 7.3 +/- 3.0 and 11.1 +/- 3.0% at 4 and 6 weeks of age. BALF of control piglets contained 1.6 +/- 0.9, 2.3 +/- 1.8 and 1.9 +/- 0.5% CD8(+) cells, only. In peripheral blood, however, the average number of CD8(+) cells remained at high levels in the infected piglets throughout the post-natal experimental period (2.8 +/- 1.9, 2.9 +/- 1.8 and 3.2 +/- 1.7 x 10(6) CD8(+) cells/ml at 2, 4 and 6 weeks, respectively). In the controls, the average levels of CD8(+) cells were 0.9+/-0.2, 1.9 +/- 1.7 and 1.6 +/- 0.5 x 10(6)/ml, respectively. Furthermore, the numbers of CD2(+) , CD4(+)CD8(+) and SLA-classII(+) cells, respectively, in peripheral blood, together with the levels of CD2(+) and CD3(+) cells in BALF were increased in the infected piglets infected in utero compared to the uninfected controls.The kinetic analyses carried out in the present study reflect that in utero infection with PRRSV modulates immune cell populations in peripheral blood and BALF of surviving piglets. The observed changes are characterised by high levels of CD8(+) cells supporting an important role of these cells in PRRSV infection. The present results, however, do not support the existence of post-natal immunosuppression following in utero infection with PRRSV.     

Acute phase response in porcine reproductive and respiratory syndrome virus infection

This study was focused on the changes observed in the serum concentration of haptoglobin (Hp), C-reactive protein (CRP), serum amyloid A (SAA) and Pig-major acute protein (Pig-MAP), during experimental porcine reproductive and respiratory syndrome virus (PRRSV) infection and in their relationship with the expression of interleukin 1β (IL-1β), interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α). Hp and Pig-MAP serum levels were increased at 10 dpi, but CRP and SAA showed a delayed and highly variable increase. All three proinflammatory cytokines were poorly expressed, and only a mild increase in IL-1β was observed at 7 dpi. The increased expression of Hp coincided with the light enhancement observed in both IL-6 and TNF-α, and might be related with an increased expression of IL-10. The low expression of TNF-α might point to a possible mechanism of viral evasion of host-immune response. This issue and the delayed expression of CRP and SAA should be taken into account in future studies about modulation of the immune response by PRRSV infection.     

Pathogenesis of in utero infection in porcine fetuses with porcine reproductive and respiratory syndrome virus.

Porcine fetuses were exposed in utero to porcine reproductive and respiratory syndrome virus (PRRSV) at stages of gestation ranging from 34 to 85 days and examined 17 to 31 days later to determine the effect of gestational age on fetal susceptibility. For each of the 8 litters tested during the study, all of the fetuses of 1 horn of the uterus were exposed to virus by intraamniotic injection; those of the other horn were exposed similarly to a sham inoculum that consisted of sterile cell culture medium. Viral infectivity titers associated with fetal tissues collected at necropsy indicated that, regardless of gestational age, the virus had replicated in fetuses exposed intraamniotically. In addition, virus had also spread and replicated in sham-inoculated littermates in 3 litters. On the basis of these findings it appears that there may be little or no temporal difference in fetal susceptibility to infection with PRRSV. If so, the lack of early fetal death as a commonly recognized feature of naturally occurring cases of PRRS may be due to a greater resistance of early gestational fetuses to the lethal effects of PRRSV, as suggested by this study, and/or a greater likelihood of transplacental infection during late gestation.     

Pathogenesis of porcine reproductive and respiratory syndrome virus-induced increase in susceptibility to Streptococcus suis infection

Eighty 3-week-old crossbred pigs were randomly assigned to six groups (13-14 pigs/group). Group 1 pigs served as uninoculated controls, group 2 pigs were inoculated intranasally (i.n.) with Streptococcus suis serotype 2, group 3 pigs were inoculated i.n. with a modified live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine, group 4 pigs were inoculated i.n. with the same vaccine and with S. suis, group 5 pigs were inoculated i.n. with VR-2385 (a high-virulence strain of PRRSV), and group 6 pigs were inoculated i.n. with VR-2385 and S. suis. Pigs exposed to both PRRSV and S. suis were inoculated with PRRSV 7 days prior to S. suis inoculation. The pigs were 26 days old when inoculated with S. suis. Respiratory disease was significantly more severe in groups 5 and 6. Mortality rate was the highest in group 6 (87.5%). This rate was significantly higher than that observed in all other groups except group 4 (37.5%). The mortality rate in group 2, inoculated with S. suis alone, was 14.3%. No pigs from groups 1, 3, or 5 died prior to the scheduled necropsies at 10 and 28 days postinoculation with PRRSV (DPI). To study the effect of PRRSV and/or S. suis on pulmonary clearance by pulmonary intravascular macrophages, six pigs from each group were intravenously infused with 3% copper phthalocyanine tetrasulfonic acid in saline prior to necropsy at 10 DPI. Mean copper levels in the lungs of pigs in groups 2, 5, and 6 were significantly lower than those in control pigs. The mean percentage of lung tissue grossly affected by pneumonia at 10 DPI was 0%, 1%, 0%, 3%, 64%, and 62% for groups 1-6, respectively. Both gross and microscopic interstitial pneumonia lesions were significantly more severe in the VR2385-inoculated groups (5 and 6). PRRSV was isolated from bronchoalveolar lavage fluid collected at necropsy from 100% of the pigs in groups 5 and 6, 71.4% of pigs in group 4, 38.5% of pigs in group 3, and none of the pigs in groups 1 or 2. Streptococcus suis serotype 2 was cultured from the internal tissues of 7.7%, 28.6%, and 78.6% of the pigs in groups 2, 4, and 6, respectively. Streptococcus suis serotype 2 was isolated from whole blood at necropsy from 7.7%, 35.7%, and 78.6% of pigs in groups 2, 4, and 6, respectively. Significantly more pigs in group 6 had S. suis isolated from whole blood and internal tissues. In summary, both high-virulence PRRSV and S. suis decreased copper clearance, and the incidence of isolation of S. suis and PRRSV was higher in dually inoculated pigs. PRRSV-induced suppression of pulmonary intravascular macrophage function may in part explain PRRSV-associated increased susceptibility to S. suis infection.     

湖南高致病性猪蓝耳病隐性感染情况调查

用ELISA和RT-PCR的方法对采集自湖南省内20个规模场1007份血清和3个市级定点屠宰场50份猪肺门淋巴结进行蓝耳病血清抗体检测和高致病性猪蓝耳病病毒的检测。结果1007份血清中,蓝耳病抗体阳性率为72.9%(734/1007),高致病性猪蓝耳病病毒携毒率为3.2%(32/1007),50份肺门淋巴结病毒阳性率为16%(8/50),其中,蓝耳病免疫与非免疫猪血清其抗体阳性率相差不显著,种猪的抗体阳性率明显高于商品猪,而其病毒携毒率为0%。部分规模猪场和眼观健康的育肥猪存在高致病性猪蓝耳病病毒的隐性感染。     

Changes in peripheral blood leukocyte subpopulations in piglets co-infected experimentally with porcine reproductive and respiratory syndrome virus and porcine circovirus type 2

Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are pathogens, which can significantly affect the swine industry worldwide. Field surveys suggest that simultaneous PRRSV and PCV2 infection is common in pigs. The objective of this study was to measure the changes in peripheral blood leukocyte subpopulations in piglets co-infected experimentally with PRRSV and PCV2, in order to analyze the synergistic influence of co-infection on the immune system. Changes in peripheral blood leukocyte subpopulations were systematically measured by flow cytometry (FCM). The levels of antibodies to PRRSV and PCV2 were detected by indirect Enzyme-Linked ImmunoSorbent Assay (ELISA) and the indirect fluorescent antibody test (IFA), respectively. Serum viral loads were measured using real-time PCR. The results showed that piglets co-infected with PRRSV and PCV2 exhibited slower generation and lower levels of antibodies to PRRSV and PCV2, and increased amounts and a prolonged presence of both PRRSV and PCV2 in serum, in comparison to the piglets infected with either virus alone. The major finding in our study was that the total and differential leukocyte counts, including white blood cells (WBCs), monocytes, granulocytes and lymphocytes (T, B and NK cells, as well as T-cell subpopulations), dramatically decreased early during co-infection with PRRSV and PCV2 for about two weeks, in contrast with animals singly infected with either PRRSV or PCV2. These results suggest that PRRSV and PCV2 co-infection results in a synergistic decrease in immune cells in the peripheral blood of piglets. These data contribute to the understanding of the immunosuppressive effects resulting from PRRSV and PCV2 co-infection in pigs.     

广西猪繁殖与呼吸综合征病毒感染状况调查

采用RT-PCR技术,对2004年1月至2005年4月期间,广西13个市104个疑似猪繁殖与呼吸综合征病毒(PRRSV)感染猪场,无菌采取231头病、死猪的组织病料(肺脏、淋巴结、脾脏)进行了病毒检测。同时,对鉴定为PRRSV阳性的组织病料和猪场进行了猪圆环病毒2型(PCV2)、猪瘟病毒(CSFV)和猪伪狂犬病毒(PRV)的检测,以确定猪群中PRRSV与PCV2、CSFV和PRV混合感染情况。结果从12个市的115份组织病料中检出PRRSV,病料的平均阳性率为49.78%(115/231),猪场的平均阳性率为61.54%(64/104),不同地市有一定的差异。PRRSV与PCV2、CSFV和/或PRV二重或多重混和感染的组织病料总数为53份,猪场总数为39个,混合感染的组织病料和猪场的总阳性率分别为22.94%(53/231)和37.50%(39/104)。混合感染的组织病料占PRRSV阳性组织病料的46.09%(53/115),混合感染的猪场占PRRSV阳性猪场的60.94%(39/64)。其中以PCV2和PRRSV混合感染的组织病料和猪场数最多。由此可见,PRRSV感染在广西猪场已普遍存在,与其他病毒混合感染现象逐渐趋向复杂化。     

猪繁殖与呼吸综合征病毒感染的诊断及其分子特点

采用流行病学调查、临床症状观察、病理学检查、病毒分离、分离毒株测序以及遗传演化分析的方法,对北京及其周边地区4个猪场剖检的4头猪进行分析。结果:流行病学和临床症状表现为急性高热性传染病;病理学观察为非化脓性脑炎和间质性肺炎等多器官严重的病理变化;RT-PCR和病毒分离确定该疫情的主要病原是猪繁殖与呼吸综合征病毒(PRRSV)。与典型PRRSV感染不同的是:成年猪感染率达50%以上,病死率达80%以上;PRRSV分离株全基因序列分析表明属于PRRSV北美洲型,特别是PRRSV NSP2不连续缺失30个氨基酸,说明此次流行的PRRSV毒株可能为高致病性毒株。     

Experimental dual infection of specific pathogen-free pigs with porcine reproductive and respiratory syndrome virus and pseudorabies virus

Twenty 6-week-old specific pathogen-free pigs were divided into four groups. On day 0 of the experiment, PRRSV-PRV (n = 6) and PRRSV (n = 4) groups were intranasally inoculated with porcine reproductive and respiratory syndrome virus (PRRSV) (10(5.6) TCID50). On day 7, the PRRSV-PRV and PRV (n = 6) groups were intranasally inoculated with pseudorabies virus (PRV) (10(3.6) TCID50). Control pigs (n = 4) were kept as uninoculated negative controls. Half of the pigs in each group were euthanized and necropsied on day 14 or 21. Clinical signs such as depression and anorexia were observed in the PRRSV-PRV and PRV groups after inoculation with PRV. Although febrile response was observed after virus inoculations, the duration of that response was prolonged in the PRRSV-PRV group compared with the other groups. The lungs in the PRRSV-PRV group failed to collapse and were mottled or diffusely tan and red, whereas the lungs of the pigs in the other groups were grossly normal. Histopathologically, interstitial pneumonia was present in all PRRSV-inoculated pigs, but the pneumonic lesions were more severe in the PRRSV-PRV group. Mean PRRSV titres of tonsil and lung in the PRRSV-PRV group were significantly (P < 0.05) higher than that in the PRRSV group on day 21. These results indicate that dual infection with PRRSV and PRV increased clinical signs and pneumonic lesions in pigs infected with both viruses, as compared to pigs infected with PRRSV or PRV only, at least in the present experimental conditions.     

猪繁殖与呼吸综合征病毒接触感染仔猪模型的建立

为了建立猪繁殖与呼吸综合征病毒(PRRSV)接触感染模型,将易感仔猪与PRRSV人工感染仔猪在隔离器中同居饲养,每日观察记录体温变化和临床症状,定期采集血清和粪便样品,用荧光定量RT-PCR检测病毒血症和粪便排毒水平;对病死猪和试验结束迫杀猪进行剖检和肺脏组织病理学检查,并检测各器官病毒载量和组织分布。结果显示:接触感染仔猪出现高热和PRRS症状时间分别较人工接种猪推迟2~3d;第3天起血清检测PRRSV阳性,第10天病毒血症上升至较高水平(108.6拷贝/mL);第5天起粪便检测PRRSV阳性,第7天粪便排毒量上升至较高水平(103.9拷贝/0.1g);3头接触感染仔猪分别于同居后第12天和第13天死亡,较人工接种猪推迟1~2d,病死猪的主要器官病毒载量以及剖检和肺脏显微病理变化与人工接种病死猪相似。这些结果表明,本研究建立的接触感染仔猪模型可用于PRRS疫苗免疫效果评价。     

Differential immunity in pigs with high and low responses to porcine reproductive and respiratory syndrome virus infection

One hundred Hampshire x Duroc cross-bred pigs (HD) and 100 NE Index line (I) pigs were infected with porcine reproductive and respiratory syndrome (PRRS) virus and evaluated for resistance/susceptibility. Controls (100/line) were uninfected littermates to the infected pigs. Viremia, change in weight (WTdelta), and rectal temperature at 0, 4, 7, and 14 d postinfection were recorded. Lung, bronchial lymph node (BLN), and blood tissue were collected at necropsy (14 d postinfection). The first principal component from principal component analyses of all variables was used to rank the pigs for phenotypic response to PRRS virus. Low responders (low PRRS burden) had high WTdelta, low viremia, and few lung lesions; high responders (high PRRS burden) had low WTdelta, high viremia, and many lesions. The RNA was extracted from lung and BLN tissue of the 7 highest and 7 lowest responders per line and from each of their littermates. Expression of 11 innate and T helper 1 immune markers was evaluated with cDNA in a 2 x 2 x 2 factorial design. Significant upregulation in lung, lymph, or both of infected pigs relative to controls occurred for all but one gene. Expression differences were greater in HD than I pigs. Significant downregulation for certain immune genes in low pigs, relative to littermate controls, was detected in lung and BLN, particularly in line I. Serum levels of the immune cytokines affirmed the gene expression differences. High preinfection serum levels of IL 8 were significantly associated with PRRS virus-resistant, low pigs. After infection, low expression of interferon gamma in cDNA and in serum was also correlated with PRRS virus resistance. Important genetic associations were revealed for fine mapping of candidate genes for PRRS virus resistance and determining the causative alleles.     

Interaction of the European genotype porcine reproductive and respiratory syndrome virus (PRRSV) with sialoadhesin (CD169/Siglec-1) inhibits alveolar macrophage phagocytosis

ABSTRACT: Porcine reproductive and respiratory syndrome virus (PRRSV) is an arterivirus that shows a restricted in vivo tropism for subsets of porcine macrophages, with alveolar macrophages being major target cells. The virus is associated with respiratory problems in pigs of all ages and is commonly isolated on farms with porcine respiratory disease complex (PRDC). Due to virus-induced macrophage death early in infection, PRRSV hampers the innate defence against pathogens in the lungs. In addition, the virus might also directly affect the antimicrobial functions of macrophages. This study examined whether interaction of European genotype PRRSV with primary alveolar macrophages (PAM) affects their phagocytic capacity. Inoculation of macrophages with both subtype I PRRSV (LV) and subtype III PRRSV (Lena) showed that the virus inhibits PAM phagocytosis. Similar results were obtained using inactivated PRRSV (LV), showing that initial interaction of the virion with the cell is sufficient to reduce phagocytosis, and that no productive infection is required. When macrophages were incubated with sialoadhesin- (Sn) or CD163-specific antibodies, two entry mediators of the virus, only Sn-specific antibodies downregulated the phagocytic capacity of PAM, indicating that interaction with Sn, but not CD163, mediates the inhibitory effect of PRRSV on phagocytosis. In conclusion, this study shows that European genotype PRRSV inhibits PAM phagocytosis in vitro, through the interaction with its internalization receptor Sn. If similar events occur in vivo, this interaction may be important in the development of PRDC, as often seen in the field.     

Pathogenesis and prevention of placental and transplacental porcine reproductive and respiratory syndrome virus infection

Porcine reproductive and respiratory syndrome virus (PRRSV)-induced reproductive problems are characterized by embryonic death, late-term abortions, early farrowing and increase in number of dead and mummified fetuses, and weak-born piglets. The virus recovery from fetal tissues illustrates transplacental infection, but despite many studies on the subject, the means by which PRRSV spreads from mother to fetus and the exact pathophysiological basis of the virus-induced reproductive failure remain unexplained. Recent findings from our group indicate that the endometrium and placenta are involved in the PRRSV passage from mother to fetus and that virus replication in the endometrial/placental tissues can be the actual reason for fetal death. The main purpose of this review is to clarify the role that PRRSV replication and PRRSV-induced changes in the endometrium/placenta play in the pathogenesis of PRRSV-induced reproductive failure in pregnant sows. In addition, strategies to control placental and transplacental PRRSV infection are discussed.     

猪繁殖与呼吸综合征病毒的研究进展

猪繁殖与呼吸综合征 (Porcine reproductiveand respiratory syndrome,PRRS)是 1 987年发现的一种新的猪病 ,它主要是引起母猪的繁殖障碍 ,如流产、早产、死胎、木乃伊胎和呼吸道症状的一种由猪繁殖与呼吸综合征病毒 (PRRSV)引起的接触性传染病。此病最初在美国首先被发现 ,随后相继在世界各国报道 ,我国台湾省也有疫情的报道。 1 995年我国首次从进口猪中检出 PRRS阳性猪 ,并从中分离到 PRRSV。 1 996年郭宝清等首次从国内 PRRS血清阳性猪群中分离到 PRRSV,从而证实了我国也有此病的流行[1] 。由于 PRRS主要是引起母猪的繁殖…     

Persistence of porcine reproductive and respiratory syndrome virus infection in a swine operation.

A herd of Quebec seedstock pigs experienced in early 1992 a typical outbreak of porcine reproductive and respiratory syndrome (PRRS) associated with lesions of interstitial, proliferative and necrotizing pneumonia in weaned piglets. The nature of the infection was confirmed by serology using indirect immunofluorescence (IIF) and virus isolation in primary cultures of porcine alveolar macrophages (PAM). Farm production recovered after eight weeks of losses. In order to evaluate the persistence of infection in the herd, five SPF-piglets were introduced in two different sections of the PRRS-affected barn four months after the disappearance of clinical symptoms, and two others were placed in a neighboring building with apparently healthy farrow-to-finnish pigs. Clinical signs, body temperature, humoral immune response, virological and histopathological findings were recorded over a 42-day period. Clinical signs were evident in all of the sentinels and prolonged fever (> or = 40 degrees C) was recorded one day post-exposure (PE). Antibody titers to PRRS virus could be detected by IIF on PAM seven days PE, and reached 1:1024 by day 21 PE. Three of the sentinels developed significant virus neutralizing antibody titers (> 1:8 to < or = 1:128) by day 35 PE. In all cases, the virus could be isolated from the serum between day 7 and 42 PE. Thus, the virus and specific antibodies coexisted for several weeks. Lesions of interstitial pneumonia was demonstrated in few animals. In experimental inoculation studies, the viral strain isolated from the sentinel pigs produced severe reproductive disorders in two sows inoculated at 95 days of gestation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Serodiagnosis of porcine reproductive and respiratory syndrome virus infection with the use of glycoprotein 5 antigens

Glycoprotein 5 (GP5) of porcine reproductive and respiratory syndrome virus (PRRSV) has been studied extensively as a target for vaccine development. This study evaluated the serodiagnostic application of PRRSV GP5 by enzyme-linked immunosorbent assay (ELISA). Two immunodominant peptides (VR #1 and VR #2) and two neutralizing ectodomain-containing peptides (Ecto #1 and Ecto #2), as well as recombinant GP5 (rGP5) as a control, were prepared. Serum from unvaccinated pigs was screened for the antibodies that bind to these peptide and protein antigens. The results were compared with those from a commercially available diagnostic ELISA kit (HerdChek), which uses the nucleocapsid (N) protein as an antigen. Only VR #1+#2 showed a result statistically similar to that of N protein. Ecto #1 and Ecto #2 had a lower sensitivity than VR #1+#2 and rGP5. The peptides and rGP5 showed significant associations with the N protein (P < 0.05 or 0.01), which suggests that GP5 may also be a candidate serodiagnostic antigen. Since antibodies against GP5 persist much longer than those against the N protein, GP5 itself and some of its fragments are thought to be good targets for serodiagnosis. In addition, the presence of antibodies against the PRRSV structural antigens showed significant antigen-dependent differences.