Baculovirus virions displaying infectious bursal disease virus VP2 protein protect chickens against infectious bursal disease virus infection

Infectious bursal disease (IBD) is an acute and contagious viral infection of young chickens caused by IBD virus (IBDV). The VP2 protein of IBDV is the only antigen for inducing neutralizing antibodies and protective immunity in the natural host. In the current study, we have succeeded in construction of one recombinant baculovirus BacSC-VP2 expressing His6-tagged VP2 with the baculovirus envelope protein gp64 transmembrane domain (TM) and cytoplasmic domain (CTD). The His6-tagged recombinant VP2 was expressed and anchored on the plasma membrane of Sf-9 cells, as examined by western blot and confocal microscopy. Immunogold electron microscopy demonstrated that the VP2 protein of IBDV was successfully displayed on the viral surface. Vaccination of chickens with the VP2-pseudotyped baculovirus vaccine (BacSC-VP2) elicited significantly higher levels of VP2-specific enzyme-linked immunosorbent assay antibodies and neutralizing antibodies than the control groups. IBDV-specific proliferation of lymphocytes was observed in chickens immunized with the recombinant BacSC-VP2. An in vivo challenge study of the recombinant baculovirus BacSC-VP2 showed effective protection against a very virulent (vv) IBDV infection in chickens. In addition, mortality and gross and histopathological findings in the bursa demonstrated the efficacy of the vaccine in reducing virulence of the disease. These results indicate that the recombinant baculovirus BacSC-VP2 can be a potential vaccine against IBDV infections.     

Experimental cryptosporidiosis and infectious bursal disease virus infection of specific-pathogen-free chickens

Specific-pathogen-free chickens orally inoculated at 4 days of age with a moderately pathogenic vaccine strain of infectious bursal disease virus (IBDV) and/or at 5 days of age with Cryptosporidium baileyi oocysts remained free of overt clinical signs throughout a 16-day period postinoculation (PI). The prepatency period for C. baileyi oocyst shedding was shorter in chickens receiving higher numbers of oocysts, but once shedding was detected, there were no obvious differences in shedding patterns among groups receiving 10(3) through 10(6) oocysts. On days 8 and 16 PI, cryptosporidia were located primarily in the bursae of Fabricius. IBDV exposure was associated with bursal follicle atrophy, whereas C. baileyi infection resulted in bursal epithelial hypertrophy and hyperplasia, mild follicle atrophy, and heterophil infiltration of the bursal mucosa. Examination of experimental groups of 30 birds each indicated that concurrent infection with both agents resulted in more severe bursal lesions, more infected birds, and greater numbers of cryptosporidia in infected tissues. At the termination of the trial, 16 days PI, Cryptosporidium infection was associated with a 6% decrease in mean body weight compared with controls.     

Serologic evidence of infectious bursal disease virus infection in Iowa turkeys

Two infectious bursal disease viruses (IBDVs)--a cell-culture-adapted chicken strain designated Edgar strain and a recent isolate from turkeys in Missouri--were used to assay sera from 10 Iowa turkey flocks collected between 1 and 16 weeks of age. The two viruses were serologically distinct in cross-neutralization tests. For all flocks, a similar serologic pattern was found consisting of (1) low maternal antibody titers to turkey IBDV and occasionally to Edgar strain IBDV between 1 and 3 weeks of age, (2) a period of very low or no detectable titers between 3 and 7 weeks of age, and (3) sharply rising high titers to turkey IBDV with low titers to Edgar IBDV beginning at 5 to 8 weeks of age. These findings indicate that infection with IBDV of the serotype represented in this study by the turkey isolate is common in Iowa turkey flocks, whereas infection with IBDV represented by Edgar strain is uncommon. Infection occurred between 3 and 7 weeks of age during the late brooding period or after birds had been moved to an intermediate growing facility. All flocks developed complicated respiratory disease with excessive mortality caused by Escherichia coli septicemia, typically between 3 and 6 weeks of age. Although there was a temporal relationship between IBDV infection and respiratory disease, the possible role of IBDV in the process is unknown.     

An age-related coagulation disorder associated with experimental infection with infectious bursal disease virus

Specific-pathogen-free White Leghorn chickens were inoculated with a field strain of infectious bursal disease virus. One group (A) was inoculated at 17 days after the chicks were hatched, and the other groups (C and E) were inoculated at posthatch day 42. Blood samples were obtained for determination of clotting times (whole blood recalcification, prothrombin, and activated partial thromboplastin times), virus-neutralizing antibody, and total hemolytic complement. There were significant increases in clotting times for groups C and E at 3 and 5 days after they were inoculated. There were no significant increases in clotting times at 3 days after inoculation in the group A chickens (inoculated at 17 days after hatching). There were no significant decreases in total complement activity in any of these chickens (groups A, C, and E). This study indicates that the mortality and clinical symptoms observed in chickens experimentally infected with infectious bursal disease virus may be associated with a clotting abnormality because it was noted only in chickens that developed severe clinical disease (inoculated at 42 days after hatching) and was not noted in chickens that remained clinically normal (inoculated at 17 days).     

Response of B congenic chickens to infection with infectious bursal disease virus.

Six congenic lines of chickens that differ from the parental inbred line RPRL-15I5 for genes in the major histocompatibility (B) complex were used to study the influence of the B haplotypes on the response of chickens to infection with virulent infectious bursal disease virus (IBDV) at 1 day or 4 weeks of age, and on the antibody response to vaccination with live or inactivated oil-emulsion (OE) IBDV vaccines at 7 weeks of age. IBDV-induced immunodepression and lesions in the bursa, spleen, and thymus in chickens infected with virus at 1 day of age were of the same degree of severity, regardless of line of chickens used. The response of blood cells to the mitogens phytohemagglutinin-M and concanavalin A was elevated in chickens infected with IBDV at 1 day of age. In an experiment conducted to study the effect of the B haplotype on IBDV infection in 4-week-old chickens, B congenic line C-12 (B12B12) showed the highest susceptibility to clinical IBD, with mortality of 79%. No detectable difference in the serological response to vaccination with live or OE IBDV vaccines was noted among chickens of various congenic lines. We conclude that the B haplotypes may influence IBDV-induced mortality, but not immunodepression or severity of lesions in lymphoid organs, or the antibody response to live or OE IBDV vaccines.     

传染性法氏囊病病毒感染细胞内源性microRNA表达差异分析

细胞内源性microRNA（miRNA）在宿主与病原相互作用过程中发挥着重要的调节功能。为了分析细胞内源性miRNA与传染性法氏囊病病毒（IBDV）的相互作用,用IBDV弱毒和强毒分别感染鸡胚成纤维细胞（CEF）和SPF鸡,24h后,取感染CEF和法氏囊组织,提取细胞总RNA,用Hy3/Hy5双色荧光标记,与miRNA芯片杂交,进行芯片内标准化、芯片间标准化、表达差异比较以及聚类分析。结果显示：在IBDV弱毒感染的CEF中,有17个细胞内源性miRNA表达上调,17个miRNA表达下调;在IBDV强毒感染的鸡法氏囊组织细胞中,有30个细胞内源性miRNA表达上调,18个miRNA表达下调。根据表达差异显著的miRNA序列设计引物,用荧光定量RT-PCR方法验证芯片检测结果,2种方法的检测结果一致。结果表明,IBDV感染可诱导细胞内源性miRNA表达变化,这些上调或下调的miRNA能调控细胞内多种代谢和信号传导途径发生异常,引起细胞及组织器官发生病理学变化。     

传染性法氏囊病病毒凋亡调节蛋白表达细胞株的建立

构建传染性法氏囊病病毒非结构蛋白基因（NS）的重组表达质粒pEGFP-C2-NS,然后在LipofectamineTM 2000介导下转染Vero细胞,活细胞状态下直接观察EGFP-NS蛋白在细胞中的表达与定位。结果显示,转染后4~5 h出现荧光蛋白表达,24 h达到高峰,呈小点样环核膜与细胞膜附近沉积,经G418筛选2~3周后稳定表达的荧光蛋白主要分布于细胞膜上。G418抗性细胞,经RT-PCR和Western-blot验证NS mRNA及其蛋白表达,提示已建立能稳定表达EGFP-NS蛋白的细胞株,所表达蛋白具有NS和EGFP双重活性,为进一步研究NS蛋白凋亡调节机制打下了基础。     

传染性法氏囊病病毒广西分离株血清亚型的确定

应用兔制备的抗传染性法氏囊病病毒(IBDV)高免血清,在鸡胚成纤维细胞(CEF)上对从广西发病鸡群分离的4个IBDV代表性流行毒株和2个常用疫苗株进行交叉中和试验。结果表明6个毒株被分为2个血清亚型;根据试验所得的R值,应用聚类分析法分析了各亚型毒株之间的亲缘关系,结果显示目前在广西流行的IBDV野毒株之间以及其与疫苗株间的抗原性存在一定的差异。研究结果对及时掌握广西IBDV流行毒株的抗原变异并为研制更有效的适合本地使用的IBD疫苗提供了科学依据。     

鸡传染性法氏囊病病毒的快速检测

随着 IBDV变异株的出现 ,发病鸡常不产生法氏囊的肉眼病变 ,由于传染性和非传染性因素亦可引起淋巴细胞减少和法氏囊坏死 ,所以 ,采取组织病理学方法诊断 IBDV感染并不完全可靠。由于主动性抗体应答需要一定的时间及大多数雏鸡都带有母源抗体 ,因此血清学早期诊断也不完全可靠。该试验采用本所研究的 IBD快速试纸与经典的琼扩试验 ,在对 IBDV的检测效果上进行了详细的比较 ,介绍如下。1　材料与方法1 .1　 IBD快速检测试纸　由河南省农业科学院生物技术研究所制备提供。1 .2　试验鸡　 1日龄试验鸡 30 0只由河南农业大学试验鸡场提…     

In vitro infection of fractionated chicken lymphocytes by infectious bursal disease virus

Lymphocytes from bursa of Fabricius and thymus of chickens were purified and separated into the three cell subsets--T, B, and null cells--by the techniques of nylon fiber columns and cytotoxicity tests. The in vitro susceptibility of the fractionated lymphocytes to a virulent strain of infectious bursal disease virus (IBDV) was studied by using immunofluorescence as the infection criterion. B cells were highly susceptible. By contrast, T cells and null cells were insusceptible to infection by IBDV. The relationship between the target cells for virus infection and those B cells that possessed surface immunoglobulin (SIg) was tested. B cells were further divided into SIg(M)- and SIg(G)-bearing cells by immunoadsorbent columns employing anti-immunoglobulin M(IgM) (mu-specific) or anti-IgG (gamma-specific) sera coated with Sephadex. The SIg(M)-bearing cells were highly susceptible. These results suggest strongly that SIg(M)-bearing B cells were the target cells for infection by IBDV.     

雏鸡传染性法氏囊病与大肠杆菌混合感染的诊断

传染性法氏囊病 ,近几年来在新疆地区时有流行 ,我们对 1 998年上半年对新疆乌鲁木齐、昌吉市某些专业养鸡户发病雏鸡群进行了诊断。1　临诊症状　发病突然 ,发病率高 ,死亡集中地发生于很短的几天之内 ,有典型死亡高峰曲线图 ,病雏啄肛 ,肛门周围羽毛粘着白色稀粪 ,病雏排出灰白色或水样稀粪 ,厌食 ,口渴 ,精神沉郁 ,羽毛竖立 ,眼闭或半闭 ,发抖 ,驱赶不动 ,极度虚弱 ,最后衰竭死亡。病死率 2 0 %左右 ,如发生多种混合感染 ,病死率可达 90 %。2　病理变化　共剖检 2 0 0余只 ,主要病理变化见法氏囊显著肿大 ,浆膜水肿 ,呈黄色胶冻样 ,切面…     

Serologic evidence of infectious bursal disease virus serotype II infection in Minnesota turkeys

Serum samples from seven randomly selected Minnesota turkey flocks were tested for antibodies to infectious bursal disease virus serotype I (Lukert strain, isolated from chickens, and North Carolina strain, isolated from turkeys) using a virus-neutralization (VN) test. All flocks were found to have low antibody titers to both Lukert and North Carolina strains. Five out of the seven flocks had high VN titers to the Missouri strain, a serotype II virus isolated from turkeys.     

Persistent infection with chicken anaemia virus and some effects of highly virulent infectious bursal disease virus infection on its persistency

Chicken anaemia virus (CAV) infectivity and the effect of highly virulent infectious bursal disease virus (hv IBDV) infection on CAV's infectivity were examined in chickens inoculated with CAV or inoculated dually with CAV and hv IBDV. Five chickens inoculated dually with hv IBDV at 35 days old and then with CAV at 40 days old exhibited no clinical signs of disease, but showed atrophic bursae of Fabricius when necropsied 4 weeks later. Upon examining the chickens at 7 days postinoculation (dpi) with CAV, it was found that hv IBDV infection had inhibited production of virus neutralising (VN) antibody to CAV, and that it was possible to recover CAV from plasma of these chickens. Although VN antibody to CAV appeared after 14 dpi, CAV was recovered from blood cells (BC s) at high titres ranging from 10(2.5)to 10(5.5)TCID(50)/0.1 ml, 7 to 28 dpi in IBDV -induced immunosuppressed chickens. In addition, CAV was sporadically recovered, using rectal swabs, from the dually inoculated chickens at low titers, ranging from 10(1.0)to 10(2. 0)TCID(50)/0.1 ml). In contrast, although CAV was recovered from BC s in most of the chickens inoculated with CAV alone, the titers were lower (10(1.0)to 10(2.5)TCID(50)/0.1 ml). No CAV was detected from the rectal swabs of these chickens. The results of virus recovery were confirmed by polymerase chain reaction. This study first examined the persistency of CAV in BC s and the effective enhancement of primary CAV infection as a result of immunosuppression caused by hv IBDV infection.     

The effect of infectious bursal disease virus infection on local and systemic antibody responses following infection of 3-week-old broiler chickens

Broiler chickens infected at 3 weeks of age with infectious bursal disease virus (IBDV) were given Brucella abortus (BA) or sheep red blood cell (SRBC) antigens before, during, and after the acute phase of the infection. Gland of Harder (GH) extracts and serum samples were used to assay local and systemic antibody titer to each antigen 7 days after antigen was administered. Antibody titers to both BA and SRBC antigens were lower (P less than 0.05) in GH extracts and serum of IBDV-infected broilers than uninfected controls. The responses to BA, a thymus-independent antigen, took longer to become depressed than the responses to SRBC, a thymus-dependent antigen. The depression of antibody titers following IBDV inoculation suggests compromise of both local and systemic immune function, a finding of importance to the broiler industry.     

Unexpected pattern of antibody response of hens following infectious bursal disease virus vaccine

The antibody titers to infectious bursal disease virus (IBDV) of a group of hens were determined every 2 weeks during the laying period using a kinetic-based enzyme-linked immunosorbent assay (ELISA). When the titers of the flock were regressed against time, the flock titer decayed with statistically significant linearity. However, when the antibody titers of individual hens were measured, their titers regressed on time in a significant quartic curvilinear fashion. Since these hens were not reimmunized, this suggests that a anamnestic response was stimulated from an unknown external source.     

Response of white leghorn chickens to infection with avian leukosis virus subgroup J and infectious bursal disease virus

The effects of viral-induced immunosuppression on the infectious status (viremia and antibody) and shedding of avian leukosis virus (ALV) were studied. Experimental white leghorn chickens were inoculated with ALV subgroup J (ALV-J) and infectious bursal disease virus (IBDV) at day of hatch with the ALV-J ADOL prototype strain Hcl, the Lukert strain of IBDV, or both. Appropriate groups were exposed a second time with the Lukert strain at 2 wk of age. Serum samples were collected at 2 and 4 wk of age for IBDV antibody detection. Samples for ALV-J viremia, antibody detection, and cloacal shedding were collected at 4, 10, 18, and 30 wk of age. The experiment was terminated at 30 wk of age, and birds were necropsied and examined grossly for tumor development. Neoplasias detected included hemangiomas, bile duct carcinoma, and anaplastic sarcoma of the nerve. Control birds and IBDV-infected birds were negative for ALV-J-induced viremia, antibodies, and cloacal shedding throughout experiment. By 10 wk, ALV-J-infected groups began to develop antibodies to ALV-J. However, at 18 wk the incidence of virus isolation increased in both groups, with a simultaneous decrease in antibody levels. At 30 wk, 97% of birds in the ALV-J group were virus positive and 41% were antibody positive. In the ALV-J/IDBV group, 96% of the birds were virus positive at 30 wk, and 27% had antibodies to ALV-J. In this study, infection with a mild classic strain of IBDV did not influence ALV-J infection or antibody production.