Antibody-mediated enhancement of disease in feline infectious peritonitis: Comparisons with dengue hemorrhagic fever

Non-immune kittens passively immunized with feline serum containing high-titered antibodies reactive with feline infectious peritonitis virus (FIPV) developed a more rapid disease after FIPV challenge than did kittens pretreated with FIPV antibody-negative serum. Antibody-sensitized, FIPV challenged—kittens developed earlier clinical signs (including pyrexia, icterus, and thrombocytopenia) and died more rapidly than did non-sensitized, FIPV-challenged kittens. Mean survival time in sensitized kittens was significantly (P < 0.05) reduced compared to non-sensitized kittens (mean ± SEM, 10.0 ± 0.6 days vs. 28.8 ± 8.3 days, respectively). Lesions induced included fibrinous peritonitis, disseminated pyogranulomatous inflammation and necrotizing phlebitis and periphlebitis. FIPV antigen, immunoglobulin G, complement (C3) and fibrinogen were demonstrated in lesions by immunofluorescence microscopy.The pathogenesis of dengue hemorrhagic fever (DHF) in persons bears striking resemblance to that of FIP in experimental kittens. In both FIP and DHF, non-neutralizing antibody may promote acute disease by enhancement of virus infection in mononuclear phagocytes or by formation of immune complexes, activation of complement and secondary vascular disturbances.     

Neutralising antibodies to wildebeest-derived malignant catarrhal fever virus in African wildlife

A total of 2,722 sera collected between 1963 and 1983, from 43 different species of wildlife in 11 African countries was examined for neutralising antibodies against the wildebeest-derived strain of malignant catarrhal fever (MCF) virus. Antibodies were demonstrated in 10 species of Bovidae which included eight species from the sub-family Hippotraginae and one species each from Bovinae and Antilopinae. Neutralising antibodies were also recorded in hippopotamus. It is suggested that the high prevalence of antibodies recorded in sera from waterbuck and reedbuck indicate infection with MCF. However, titres in other species may be due to antigenically related viruses.     

Effect of malignant catarrhal fever virus infection on the immune response of rabbits to sheep red blood cells

Rabbits infected with African Malignant catarrhal fever virus mounted a depressed antibody response to sheep red blood cells compared to the antibody response of uninfected rabbits. Immunodepression was observed in rabbits immunised 0, 3, 5, 7 and 10 days after infection. Antibody response was not depressed in the rabbits immunised 1 day after infection. Despite the depressed antibody response to SRBC, the rabbits died with rising virus neutralising antibody to the virus. The possible causes of the immunodepression are discussed.     

Neotropical primary bat cell lines show restricted dengue virus replication

Dengue is the most widespread arboviral disease affecting humans. Bats are recognized carriers of emerging viral zoonoses and have been proposed as dengue reservoirs, since RNA/NS1 and/or antiviral antibodies have been detected. Yet, experimental inoculation of Artibeus bats failed to show virus replication. This conflicting results prevent drawing further conclusions of whether bats sustain dengue infection. To test bat cellular permissivity to dengue infection, we established primary bat embryonic cells from diverse organs and tissues of Artibeus jamaicensis, Molossus sinaloae, and Desmodus rotundus. We observed a limited serotype-, organ-, and bat species- specific dengue susceptibility. Only some Molossus-derived primary cells sustained poorly initial Dengue serotype-1 replication, though it was latter absent. To elucidate if Molossus bats may play a role in dengue replication, ecological or in vivo experiments must be performed. Taken together our results show that Dengue did not replicate efficiently in cell lines derived from Neotropical bat species.     

Malignant catarrhal fever virus specific secretory IgA in nasal secretions of wildebeest calves

Wildebeest IgA was isolated from nasal secretions and precolostrum. It was indentified by cross-reaction with anti-human and anti-bovine IgA sera.

Nasal secretions collected from wildebeest calves over 3 months old had malignant catarrhal fever virus neutralizing antibody activity. They also contained specific IgA to the virus as detected by indirect immunofluorescence. It is suggested that production of malignant catarrhal fever virus specific IgA in the nasal cavity, contributes to the elimination and cassation of the virus shed in the nasal secretions of wildebeest calves over 3 months. old.     

非洲猪瘟病毒免疫逃逸分子机制研究进展

非洲猪瘟(African swine fever，ASF)是由非洲猪瘟病毒(African swine fever virus，ASFV)感染猪引起的一种急性、烈性、高度接触性传染病，至今没有研发出安全有效的疫苗，一旦暴发会造成重大经济损失。ASFV在和宿主长期作用过程中，通过抑制干扰素和炎症反应，调节凋亡、自噬及细胞免疫等多种途径逃逸机体免疫反应促进自身复制，但具体的机制仍不完全清楚。ASFV复杂的免疫逃逸机制可能是阻碍有效疫苗研发的关键因素之一。借助生物信息学技术对ASFV的基因组和蛋白质组深入分析，筛选病毒的免疫调控关键基因和保护性抗原表位，将在ASFV免疫逃逸分子机制的研究与疫苗研发中发挥重要作用。本文主要对ASFV感染引起的免疫应答反应及可能的免疫逃逸机制研究进行概述，以期为ASF疫苗研制及综合防控提供思路。     

非洲猪瘟的风险分析和防控措施

非洲猪瘟(African swine fever, ASF)是一类动物传染病.由于该病死亡率较高,一旦发生将会给养殖业带来巨大的经济损失.该病因保护性免疫反应的复杂性,目前尚无有效的疫苗进行预防.为了防范非洲猪瘟病毒传入我国,根据该病毒的病原学和流行病学特性,尽可能的降低该病的感染和传播途径.并应用血清学和分子生物学无感染性的诊断方法快速、灵敏的检测该病毒,从而将该病传入我国的风险降至最低.     

Climate Factors as Important Determinants of Dengue Incidence in Curaçao

Macro‐ and microclimates may have variable impact on dengue incidence in different settings. We estimated the short‐term impact and delayed effects of climate variables on dengue morbidity in Curaçao. Monthly dengue incidence data from 1999 to 2009 were included to estimate the short‐term influences of climate variables by employing wavelet analysis, generalized additive models (GAM) and distributed lag nonlinear models (DLNM) on rainfall, temperature and relative humidity in relation to dengue incidence. Dengue incidence showed a significant irregular 4‐year multi‐annual cycle associated with climate variables. Based on GAM, temperature showed a U‐shape, while humidity and rainfall exhibited a dome‐shaped association, suggesting that deviation from mean temperature increases and deviation from mean humidity and rainfall decreases dengue incidence, respectively. Rainfall was associated with an immediate increase in dengue incidence of 4.1% (95% CI: 2.2–8.1%) after a 10‐mm increase, with a maximum increase of 6.5% (95% CI: 3.2–10.0%) after 1.5 month lag. A 1°C decrease of mean temperature was associated with a RR of 17.4% (95% CI: 11.2–27.0%); the effect was inversed for a 1°C increase of mean temperature (RR= 0.457, 95% CI: 0.278–0.752). Climate variables are important determinants of dengue incidence and provide insight into its short‐term effects. An increase in mean temperature was associated with lower dengue incidence, whereas lower temperatures were associated with higher dengue incidence.     

Establishment and characterization of dengue virus type 2 nonstructural protein 1 specific T cell lines

Immunity against dengue viruses (DENV) infection may include cellular immune responses which involve in the immunopathology of DENV infection hosts. This study was to establish short-term dengue virus type 2 (DENV2) nonstructural protein 1 (NS1) specific T cells from splenocytes from BALB/c mice immunized with DENV2 NS1 in vitro, which may be used to identify immunopathologic mechanism of dengue. Nine DENV2 NS1 specific T cell lines were successfully established by using limiting dilution methods and maintained for 20 weeks by re-stimulated with DENV2 NS1, recombinant mouse IL-2 and antigen presenting cell weekly. Phenotypically, these cells were mainly composed of CD3⁺CD4⁺ T cells. The culture supernatants of these cells contained large amounts of TNF-α and IFN-γ. Vascular tissue pathological change could be found in the mice adoptive transferred with DENV2 NS1 specific T cells. The results indicate that DENV2 NS1 specific T cells could be established and maintained with syngeneic T cell growth factors in vitro. Meanwhile, DENV2 NS1 specific T cells might contribute to the immunopathology of vascular leakage of dengue.     

Serological Diagnosis of Classical Swine Fever: A Comparison of a Modified Direct Complement Fixation Test with an Immunofluorescence Plaque Neutralization Test in the Diagnosis of Experimental Subclinical Infection

Antibody levels in post-infection sera from a pig inoculated with a low virulent strain of classical swine fever virus (Hannover 62) and in sera from two pigs inoculated with another low virulent strain (Spielbach 66) and from an in-contact pig were assayed by complement fixation and immunofluorescence using classical swine fever virus (ALD strain) and bovine virus diarrhoea virus (UG 59 strain) as antigens. The complement fixation test used was modified by addition of a preparation of porcine Glq to the complement and by mercaptoethanol treatment of the immune serum before use. The mercaptoethanol treatment of the immune serum resulted in complete elimination of a haemolytic prozone often seen with porcine immune sera.In the sera from the inoculated animals complement-fixing antibodies appeared earlier than neutralizing antibodies. A few weeks after inoculation there was a correlation between the presence of complement-fixing and neutralizing antibodies.During the entire observation period of 13 weeks it was not possible to demonstrate complement-fixing or neutralizing antibodies in serum from a pig exposed to infection by contact with the two pigs inoculated with the Spièlbach 66 strain of classical swine fever virus.     

The pathogenesis and immunology of bluetongue virus infection of ruminants

Bluetongue (BLU) virus is transmitted from infected to susceptible ruminants by hematophagous vector midges (Culicoides species). Cattle are important reservoir hosts of the virus because infection typically is asymptomatic and characterized by prolonged cell associated viremia, and because at least some species of insect vector preferentially feed on cattle. Interaction of BLU virus with the cell membrane of erythrocytes in infected cattle likely facilitates both prolonged viremia as well as infection of the insect vector. BLU disease is most common in sheep and some wildlife species. A variety of host, agent and environmental factors clearly can influence expression of disease in these species. The pathogenesis of BLU virus infection of cattle and sheep is remarkably similar, thus the basis for expression of disease in sheep but not cattle remains to be firmly established. Some difference in susceptibility of endothelial cells to infection in the two species is one potential explanation.

Ruminants develop a variety of antiviral responses after BLU virus infection. Antibodies to outer capsid protein VP2 are responsible for virus neutralization, and confer resistance to reinfection with the homologous serotype of BLU virus. Antibodies to epitopes on proteins which are common to all viruses of the BLU serogroup form the basis of current diagnostic serologic tests. Cell mediated responses have been incompletely characterized, in part because BLU virus replicates within dividing lymphocytes and virus-mediated cytolysis inhibits in vitro blastogenesis. Immunological competence of ruminants to BLU virus arises prior to midgestation, and suggestions that persistent immune tolerant BLU virus infection occurs after in utero exposure of cattle have not been substantiated and are not consistent with recent findings.     

非洲猪瘟病毒免疫及基因工程疫苗研究进展

非洲猪瘟（African swine fever,ASF）是由非洲猪瘟病毒（African swine fever virus,ASFV）引起家猪的一种急性、出血性、高度接触性蜱传病毒病,可致家猪网状内皮系统出血及高死亡率,是危害世界养猪业健康发展最严重的传染病之一。ASFV是一种大型的DNA病毒,结构复杂,其基因组编码大量蛋白。该论文介绍了ASFV的特性,免疫应答机制如免疫逃逸、体液免疫和细胞免疫,以及基因工程疫苗如减毒活疫苗、亚单位疫苗、病毒载体活疫苗及DNA疫苗研究的最新进展,并对中国ASF的疫苗研制、防控进行了展望。     

Recent progress in molecular biology of Crimean-Congo hemorrhagic fever

Crimean–Congo hemorrhagic fever (CCHF) is a severe hemorrhagic fever in humans with a case fatality rate of up to 50%. A causative agent of CCHF is CCHF virus, which is a tick-borne virus in the family Bunyaviridae, genus Nairovirus. The virus is transmitted to humans through infected tick bites, squashed ticks or from direct contact with viremic animals or humans. Outbreaks of CCHF have been documented in Africa, the Middle East, Eastern Europe and Western Asia where the vector and/or reservoir ticks of Hyalomma spp. are distributed. Recent advances in molecular and biochemical analyses of CCHF virus revealed that the virus encodes larger proteins compared to other genus of Bunyavirus and the processing of viral proteins are complicated. Recent studies also showed that the CCHF viruses are relatively divergent in its genome sequence and the viruses are grouped in seven different clades. In general, these phylogenetic analyses based on sequences of S-RNA and L-RNA segment of CCHF viruses indicate that the seven clades correlate with their geographical location. The phylogenetic topology based on M-RNA segment sequences of CCHF viruses is different from those based on S-RNA and L-RNA segments. These analyses indicate that M-RNA segment reassortment events occur more frequently than those in S- and L-RNA segments.     

Improved sero-monitoring assay for classical swine fever (CSF) using the recombinant E2 protein of a recent Korean isolate

Classical swine fever (CSF) is a highly contagious disease of pigs that causes fever, diarrhea and paralysis, often resulting in death. E2 is the major structural protein of the CSF virus (CSFV) and mediates the entrance of the virus, subsequently inducing a neutralizing immune response. In this study, the E2 gene of a recent Korean isolate of CSF, SW03, was cloned and the DNA sequence was compared to other strains via phylogenetic analysis. With the purified E2 protein, an enzyme-linked immunosorbent assay (ELISA) was developed for the serodiagnosis of CSFV infection. The sensitivity and specificity of the E2-ELISA were 96.1% and 94.8%, respectively. A total of 17 out of 485 field-collected pig sera tested demonstrated conflicting results between two ELISA methods, a commercial kit and the E2-ELISA. Of these sera, 60% were determined to be CSFV positive by a virus neutralization test (VNT), suggesting involvement of different immune responses in the cases of CSFV infection. As the E2-ELISA was developed using a recent Korean isolate, SW03, this assay is capable of rapidly identifying newly emerging CSFV strains.     

Surveillance of classical swine fever in wild boar in South Korea from 2010–2014

Classical swine fever (CSF) is a highly contagious systemic hemorrhagic viral disease of pigs. Wild boar plays a crucial role in the epidemiology of CSF. Between 2010 and 2014, samples were collected nationwide from 6,654 wild boars hunted in South Korea. Anti-CSF antibodies were identified in 0.59% (39 of 6,654) of the wild boar samples using a virus neutralization test and were primarily detected in wild boars living close to the demilitarized zone and the area of the Taebaek Mountains surroundings. The CSF virus (subgroup 2.1b) was isolated from two wild boars captured in a nearby border area. The criteria used to define high-risk areas for targeted CSF surveillance in South Korea should be further expanded to include other regions nationwide.     

No Evidence of Dengue Virus Infections in Several Species of Bats Captured in Central and Southern Mexico

Bats are reservoirs for viruses with zoonotic potential in the Americas, and scattered evidence exists suggesting that bats may act as reservoirs for dengue virus (DENV). To explore further the role of bats as part of DENV sylvatic cycles, 240 bats of 18 species were captured in 2 states of Mexico with contrasting ecological characteristics but concurrent DENV activity in humans. RT‐PCR analysis of RNA extracted from liver or spleen tissue from de bats failed to show evidence for the presence of DENV nucleic acids in these organs. In addition, plasma assayed by plaque reduction neutralization test showed no evidence of neutralizing anti‐DENV antibodies. These results suggest that American bats may not be reservoirs or amplification host for DENV infection.     

Bluetongue: Laboratory diagnosis

Definitive diagnosis of bluetongue virus (BTV) infection, often subclinical in domestic and wild ruminant relies heavily on laboratory techniques for BTV isolation and demonstration of BTV antigens, viral nucleic acids and antibodies. The virus can be isolated from blood components, mainly the erythrocyte fraction, collected from affected animals during the period of febrile response. Semen collected from male animals at the peak of viremia and tissues from affected animals and fetuses may also be used for BTV isolation. The primary procedure for BTV isolation is inoculation of embryonated chicken eggs with a subpassage onto cell cultures (e.g. BKH-21, Vero cell lines). In addition to the conventional techniques such as fluorescent antibody staining and virus neutralization procedures for sero-grouping and serotyping of BTV isolates, immunohistochemical, immunoenzymatic and immunoelectron microscopic techniques, using monoclonal antibodies (MAb), offer more rapid, specific and sensitive approaches for BTV identification and antigen detection. The progress of molecular biology, especially the development of genetic probes for hybridization analysis and polymerase chain reaction techniques for detection of BTV nucleic acids hold the promise of most efficient diagnostic assays. Among the various serogroup-specific assays for antibody detection, the agar gel immunodiffusion (AGID) and competitive (C) ELISA are the most widely used tests. Because of its limitations (i.e. anticomplementary serum and complexity of the procedure) the complement fixation (CF) test is virtually abandoned and is used in only a few laboratories. Although the AGID test is simple to perform and rapid, it is not highly sensitive or quantitative and has limitations in its specificity. Sera containing antibodies to other group of Orbiviruses (e.g. epizootic hemorrhagic disease) may result in non-specific reaction in the AGID test. Among several ELISAs that have recently been developed, the C.ELISA in which a group-specific MAb to BTV is used, has proved to be the most sensitive and specific assay for detection of antibodies to BTV. Following extensive national and international validation, the C.ELISA is gradually replacing the AGID as a universal test to certify ruminants for trade purposes and to diagnose BT infection in domestic and wild animals. The cell culture-based microtiter serum neutralization (MTSN) is the most commonly used assay for the detection of serotype-specific antibodies to the recognized BTVs in animal sera. The MTSN may be used to type virus isolates and also to monitor animal population for specific serotypes of BTV in epidemiological investigations.     

Factors critical for successful vaccination against classical swine fever in endemic areas

Classical swine fever (CSF) or hog cholera, caused by the classical swine fever virus (CSFV), is one of the most important viral diseases that cause serious economic loss to the swine industry worldwide. During the past 5 years, several techniques for measuring porcine cell-mediated immunity (CMI) were applied, in conjunction with other conventional techniques, to study factors that influence the induction of CSFV-specific immunity. Information, obtained from a series of experiments, demonstrated cell-mediated immune responses in providing protective immunity against CSF infection. Although it has been confirmed that commercially available modified live CSF vaccines are able to induce complete protection in vaccinated pigs, several factors including maternal immunity, the age of primary vaccination, vaccination protocol and complications caused by other pathogens, can greatly affect the effectiveness of CSF vaccines in the field.     

Wesselsbron virus infection in West African Dwarf Sheep: Effect of pre-infection serum flavivirus antibodies on severity of disease

West African Dwarf Sheep inoculated sequentially with two flaviviruses: yellow fever and Dakar bat or West Nile and yellow fever viruses, developed neutralizing (N) and haemagglutination inhibiting (HI) antibodies to several flaviviruses; dengue type 1, dengue type 2, yellow fever, West Nile and Wesselsbron viruses. Experimental infection of these animals with the indigenous strain of Wesselsbron virus resulted in the modification of the clinical disease. The disease was reduced in severity and there was complete absence of, or marked reduction in the level and duration of circulating virus and fever. Seronegative animals infected with the same strain of Wesselsbron virus developed the classical disease characterized by a high fever, anorexia, leucopenia and a high level of viraemia. The absence of clinical reports of Wesselsbron disease and lack of other isolations of the virus from Nigeria could possibly be explained by the presence of the flavivirus group antibodies in domestic animal sera.     

Classical swine fever virus infection modulates serum levels of INF-α, IL-8 and TNF-α in 6-month-old pigs

Several studies have highlighted the important role of cytokines in disease development of classical swine fever virus (CSFV) infection. In the present study, we examined the kinetics of 7 porcine cytokines in serum from pigs infected with 3 different CSFV strains. Based on the clinical picture in 6-month-old Danish pigs, the strains used for inoculation were classified as being of low (Bergen), low to moderate (Eystrup) and moderate to high (Lithuania) virulence. The cytokines interferon-alpha (INF-α), interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-α) showed increased levels after CSFV infection with more or less comparable course in the 3 groups. However, the cytokine level peaked with a 2–3 days delay in pigs infected with the low virulent strain compared to those infected with a moderately or highly virulent strain. These findings may indicate that INF-α, IL-8 and TNF-α are involved in the immune response during CSFV infection with strains of different virulence.