West Nile Virus Specific Antibodies Research Articles

West Nile Virus in Slovenia.

West Nile virus (WNV) is a flavivirus transmitted by mosquitoes. Birds are the reservoir for the virus; humans, horses and other mammals are dead-end hosts. Infections caused by WNV in humans can vary from asymptomatic infections to West Nile fever (WNF) or West Nile neuroinvasive disease (WNND). In 1995, a serosurvey was performed in Slovenia on forest workers, and WNV specific IgG antibodies were confirmed in 6.8% of the screened samples, indicating that WNV is circulating in Slovenia. No human disease cases were detected in Slovenia until 2013, when the first case of WNV infection was confirmed in a retrospective study in a 79-year old man with meningitis. In 2018, three patients with WNND were confirmed by laboratory tests, with detection of IgM antibodies in the cerebrospinal fluid of the patients. In one of the patients, WNV RNA was detected in the urine sample. In 2017, 2018 and 2019, a mosquito study was performed in Slovenia. Mosquitoes were sampled on 14 control locations and 35 additional locations in 2019. No WNV was detected in mosquitoes in 2017 and 2019, but we confirmed the virus in a pool of Culex sp. mosquitoes in 2018. The virus was successfully isolated, and complete genome sequence was acquired. The whole genome of the WNV was also sequenced from the patient’s urine sample. The whole genome sequences of the WNV virus detected in Slovenian patient and mosquito indicate the virus most likely spread from the north, because of the geographic proximity and because the sequences cluster with the Austrian and Hungarian sequences. A sentinel study was performed on dog sera samples, and we were able to confirm IgG antibodies in 1.8% and 4.3% of the samples in 2017 and 2018, respectively. Though Slovenia is not a highly endemic country for WNV, we have established that the virus circulates in Slovenia.

West Nile Virus Infection with Neurological Disorders: A Case Report and a Brief Review of the Situation in Bulgaria

SUMMARYA case of a 66-year-old man with West Nile neuroinvassive disease manifested with fever, weakness, fatigue, consciousness disorders and underlying diabetes mellitus type 2 and cardiovascular diseases is presented. Laboratory data showed elevated erythrocyte sedimentation rate and fibrinogen. Serological tests revealed West Nile virus specific antibodies of class IgM and IgG in serum. West Nile virus RNA was detected in urine sample. Supportive therapy was applied.

Serological evidence of widespread West Nile virus and Japanese encephalitis virus infection in native domestic ducks (Anas platyrhynchos var domesticus) in Kuttanad region, Kerala, India

Birds can act as reservoirs of West Nile virus (WNV) with a key role in its epidemiology. WNV lineage 1 associated fatal cases of human encephalitis in 2011 and acute flaccid paralysis in 2013 were reported in Alappuzha district, Kerala, India. But no information is available on WNV circulation in domestic ducks, which are abundant, cohabit with humans and occupy wetlands and water bodies in the region. To determine the extent of WNV infection, we investigated 209 sera, 250 oral and 350 cloacal swab samples from local Chara and Chemballi domestic ducks (Anas platyrhynchos var domesticus) in the districts of Alappuzha, Kottayam, Kollam and Pathanamthitta collected during January and March 2015. The serum samples were tested for WNV antibodies first by a competition ELISA and then by a micro virus neutralization test (micro-VNT), while oral and cloacal swabs were subjected to WNV real-time RT-PCR. Ninety five ducks showed evidence of flavivirus antibodies by ELISA. End point neutralizing antibody titre against WNV and Japanese encephalitis virus (JEV) revealed WNV specific antibodies in 24 (11.5%) ducks in 3 districts, JEV specific antibodies in 21 (10%) ducks in 2 districts and flavivirus specific antibodies in 19 (9%) ducks. However, no WNV genomic RNA could be detected. The results of this study demonstrate evidence of widespread WNV and JEV infection in domestic ducks in Kuttanad region, Kerala with a higher seroprevalence to WNV than JEV. Additionally, it highlights the utility of domestic ducks as a surveillance tool to detect WNV/JEV circulation in a region.

Suspect West Nile virus encephalomyelitis in an imported horse in the UK

SummaryThis report describes a suspect case of West Nile virus (WNV) encephalomyelitis, reported for the first time in a horse in the UK. The affected gelding had been imported from the Republic of Cyprus and travelled through several WNV endemic areas in Europe before arriving at the premises in Lincolnshire. Clinical signs included muscle fasciculations, weakness of the hindlimbs and transient lip twitching that quickly progressed to depression and recumbency. West Nile virus specific antibodies were detected by serological tests in the absence of a previous history of vaccination. The horse improved clinically 10 days after the onset of disease and fully recovered in 12 weeks. Follow‐ups at 12 and 20 months post event did not reveal any sequela and the horse was performing adequately at novice level. This article refers to the same horse mentioned by Fooks et al. (2014) and it is an extension of the previous published work.

West Nile Virus Seroprevalence in Horses from Romania: First Step in the Infection Risk Assessment

West Nile virus (WNV) infections are known to be endemic in some areas of Romania. After the last epidemics registered in 2010 in the country, it is considered that WNV has changed its patterns of evolution. The surveillance of West Nile virus includes surveillance of host species and mosquitoes vectors in order to assess the risk of WNV infection in human population. In this paper we describe the preliminary investigations made for the establishment of mosquitoes collection sites consisting in a seroepidemiological study for the detection of WNV specific antibodies made in three counties from the eastern Romania. Horse blood samples were collected from Buzau, Braila and Tulcea Counties. From these, 60 samples were chosen randomly (at least 5 from each sampling area) and tested using a competitive ELISA. The results showed that in Tulcea and Braila Counties WNV is enzootic. In Tulcea, high values of seroconversion were registered not only in Danube Delta (eastern part of the County), but also in the central and southern area of the county. As for Buzau County, the tested sera were negative for specific anti WNV antibodies. In Braila and Tulcea Counties, the high rates of seroprevalence in certain areas permitted us to establish collecting sites for mosquitoes. In Buzau County West Nile virus specific antibodies were not detected until now. The collection of mosquitoes and further detection of WNV will gave us the possibility to characterize WNV circulating strains in Romania and to determine the risk areas for human infection.

Suspect West Nile encephalitis in a horse imported into the UK from Europe

The authors have detected West Nile virus-specific antibodies in a horse in the UK with signs of neurological disease. The horse had recently been imported from Europe and does not...

Infection with non-lethal West Nile virus Eg101 strain induces immunity that protects mice against the lethal West Nile virus NY99 strain.

Herein we demonstrate that infection of mice with West Nile virus (WNV) Eg101 provides protective immunity against lethal challenge with WNV NY99. Our data demonstrated that WNV Eg101 is largely non-virulent in adult mice when compared to WNV NY99. By day 6 after infection, WNV-specific IgM and IgG antibodies, and neutralizing antibodies were detected in the serum of all WNV Eg101 infected mice. Plaque reduction neutralization test data demonstrated that serum from WNV Eg101 infected mice neutralized WNV Eg101 and WNV NY99 strains with similar efficiency. Three weeks after infection, WNV Eg101 immunized mice were challenged subcutaneously or intracranially with lethal dose of WNV NY99 and observed for additional three weeks. All the challenged mice were protected against disease and no morbidity and mortality was observed in any mice. In conclusion, our data for the first time demonstrate that infection of mice with WNV Eg101 induced high titers of WNV specific IgM and IgG antibodies, and cross-reactive neutralizing antibodies, and the resulting immunity protected all immunized animals from both subcutaneous and intracranial challenge with WNV NY99. These observations suggest that WNV Eg101 may be a suitable strain for the development of a vaccine in humans against virulent strains of WNV.

Indigenous West Nile Virus Infections in Horses in Albania

Serum samples collected from 167 equines of 12 districts in Albania were tested for West Nile virus-specific antibodies by enzyme-linked immunosorbent assay and virus neutralization assay, using WNV lineage 1 and 2. In addition, 95 bird serum samples from Albania and 29 horse samples from Kosovo were tested in ELISA. An overall seroprevalence rate of 22% was found in horses from Albania, whereas no specific antibodies were found in the equine samples from Kosovo and the bird samples. This is the first report indicating WNV infections in animals in Albania, and the first reported seroprevalence study conducted for Kosovo. These results provide evidence for widespread infections of WNV in Albania.

Use of Competition ELISA for Monitoring of West Nile Virus Infections in Horses in Germany

West Nile virus (WNV) is a mosquito-borne viral pathogen of global importance and is considered to be the most widespread flavivirus in the World. Horses, as dead-end hosts, can be infected by bridge mosquito vectors and undergo either subclinical infections or develop severe neurological diseases. The aim of this study was to detect WNV specific antibodies in horses in Germany as an indicator for an endemic circulation of WNV. Sera from more than 5,000 horses (primarily fallen stock animals) were collected in eight different federal states of Germany from 2010 to 2012. Sera were screened by a competitive ELISA and positive reactions were verified by an indirect IgM ELISA and/or by virus neutralization tests (VNT) for WNV and Tick-borne encephalitis virus (TBEV) in order to exclude cross-reacting antibody reactions. In essence WNV specific antibodies could not be detected in any of the horse sera. Not surprisingly, a small number of sera contained antibodies against TBEV. It is noteworthy that equine sera were often collected from horse carcasses and therefore were of poor quality. Nonetheless, these sera were still suitable for WNV ELISA testing, i.e., they did not produce a high background reaction which is a frequently observed phenomenon. According to these data there is no evidence for indigenous WNV infections in horses in Germany at present.

Towards the Conservation of Endangered Avian Species: A Recombinant West Nile Virus Vaccine Results in Increased Humoral and Cellular Immune Responses in Japanese Quail (Coturnix japonica)

West Nile Virus (WNV) arrived in North America in 1999 and is now endemic. Many families of birds, especially corvids, are highly susceptible to WNV and infection often results in fatality. Avian species susceptible to WNV infection also include endangered species, such as the Greater Sage-Grouse (Centrocercus uropbasianuts) and the Eastern Loggerhead Shrike (Lanius ludovicianus migrans). The virus has been shown to contribute towards the likelihood of their extinction. Although a clear and present threat, there exists no avian WNV vaccine available to combat this lethal menace. As a first step in establishing an avian model for testing candidate WNV vaccines, avian antibody based reagents were assessed for cross-reactivity with Japanese quail (Coturnix japonica) T cell markers CD4 and CD8; the most reactive were found to be the anti-duck CD8 antibody, clone Du-CD8-1, and the anti-chicken/turkey CD4 antibody, clone CT4. These reagents were then used to assess vaccine performance as well as to establish T cell populations in quail, with a novel population of CD4/CD8 double positive T cells being identified in Japanese quail. Concurrently, non-replicating recombinant adenoviruses, expressing either the WNV envelope or NS3 ‘genes’ were constructed and assessed for effectiveness as avian vaccines. Japanese Quail were selected for testing the vaccines, as they provide an avian model that parallels the population diversity of bird species in the wild. Both the level of WNV specific antibodies and the number of T cells in vaccinated birds were increased compared to unvaccinated controls. The results indicate the vaccines to be effective in increasing both humoral and cellular immune responses. These recombinant vaccines therefore may find utility as tools to protect and maintain domestic and wild avian populations. Their implementation may also arrest the progression towards extinction of endangered avian species and reduce the viral reservoir that potentiates infection in humans.

Seroprevalence of West Nile Virus–Specific Antibodies in a Cohort of Blood Donors in Northeastern Italy

IgG and IgM levels against West Nile virus (WNV) were measured in 20,033 serum samples that were obtained between October 2008 to September 2009 from 9913 blood donors in the district of Ferrara, northeastern Italy. As confirmatory test, a microneutralization assay was used to detect the presence of neutralizing antibodies against WNV. Sixty-eight subjects (0.69%) were positive for anti-WNV by immunofluorescence assay. Large differences in the prevalence of antibodies to WNV were noted between towns in the area evaluated.

Persistence of West Nile Virus-Specific Antibodies in Viremic Blood Donors

We evaluated West Nile virus (WNV) antibody persistence by using follow-up plasma samples from 35 blood donors who made viremic donations in 2005. At 26 to 34 days of follow-up, all of the donors (n = 33) were positive for WNV immunoglobulin M (IgM), IgA, and IgG. At 1-year of follow-up, 17% of the donors (n = 23) were positive for WNV IgM, 57% were positive for WNV IgA, and 100% were positive for WNV IgG.

Transmission of West Nile Virus Through Human Breast Milk Seems to Be Rare

In September 2002, possible transmission of West Nile virus via human milk was reported for the first time. Since 2003, the Centers for Disease Control and Prevention collected reports of maternal or infant West Nile virus illness during the breastfeeding period. All of the reported instances were reviewed. In addition, milk samples from women infected during pregnancy were tested for West Nile virus RNA and West Nile virus-specific antibodies. Six infants were reported to have breastfed from mothers with West Nile virus fever. Five of the 6 infants had no illness or detectable antibodies to West Nile virus in serum after onset of maternal illness. One infant who was not tested and developed a rash was otherwise well 1 week after onset of maternal illness. In addition, 2 infants were reported to have developed West Nile virus illness while breastfeeding; preceding maternal illness was not documented. Two breastfed infants whose mothers acquired West Nile virus fever in the last week of pregnancy developed West Nile virus-specific antibodies; both infant infections could have been congenitally acquired. Of 45 milk samples from women infected with West Nile virus during pregnancy, 2 had West Nile virus RNA, and 14 had immunoglobin M antibodies to West Nile virus. Of 10 reported instances since 2003 of maternal or infant West Nile virus illness while breastfeeding, transmission of West Nile virus through human milk could neither be ruled out nor confirmed for 5 cases; in 5 others, serologic tests indicated no vertical transmission. Transmission of West Nile virus through breastfeeding seems to be rare, but more information is needed.

Crystal Structure of West Nile Virus Envelope Glycoprotein Reveals Viral Surface Epitopes

West Nile virus, a member of the Flavivirus genus, causes fever that can progress to life-threatening encephalitis. The major envelope glycoprotein, E, of these viruses mediates viral attachment and entry by membrane fusion. We have determined the crystal structure of a soluble fragment of West Nile virus E. The structure adopts the same overall fold as that of the E proteins from dengue and tick-borne encephalitis viruses. The conformation of domain II is different from that in other prefusion E structures, however, and resembles the conformation of domain II in postfusion E structures. The epitopes of neutralizing West Nile virus-specific antibodies map to a region of domain III that is exposed on the viral surface and has been implicated in receptor binding. In contrast, we show that certain recombinant therapeutic antibodies, which cross-neutralize West Nile and dengue viruses, bind a peptide from domain I that is exposed only during the membrane fusion transition. By revealing the details of the molecular landscape of the West Nile virus surface, our structure will assist the design of antiviral vaccines and therapeutics.

Geographic Factors Contributing to a High Seroprevalence of West Nile Virus-Specific Antibodies in Humans following an Epidemic

Sera of 624 blood donors were evaluated to determine seroprevalence of West Nile virus (WNV) antibodies following the 2003 WNV epidemic in Nebraska. Geographic factors contributing to differences in WNV seropositivity were evaluated. The overall prevalence of WNV in Nebraska was higher than reported previously in other U.S. locations (9.5% WNV immunoglobulin G seroprevalence rate), with the highest prevalence identified in the western part of the state (19.7%), followed by the central (13.8%) and the eastern (4.2%) parts. Regions of the state with the highest WNV-positive mosquito rates correlated with the highest human WNV seroprevalence rates. The results showed that both the western and central parts of the state, where mosquito positivity rates were highest, had significantly higher seroprevalence rates than the eastern region. Additional studies are needed to determine whether the high prevalence rates in Nebraska will be reflected in other states and what impact environmental and geographical factors may have on future outbreaks of WNV infection.

Pathogenicity of West Nile Virus for Turkeys

In the fall of 1999, West Nile virus (WNV) was isolated during an outbreak of neurologic disease in humans, horses, and wild and zoological birds in New York, Connecticut, and New Jersey. Turkeys could potentially be a large reservoir for WNV because of the high-density turkey farming and the presence of large wild turkey populations in the eastern seaboard of the United States. Little is known about the pathogenicity of WNV in domestic or wild turkeys. Specific-pathogen-free 3-wk-old turkeys were inoculated subcutaneously with 10(3.3) mean tissue culture infective doses of a WNV strain isolated fromthe index case in a New York crow. No clinical signs were observed in the turkeys over the 21 days of the experiment. One turkey died abruptly at 8 days postinoculation (DPI). Many turkeys developed viremia between 2 and 10 DPI, but the average level of virus was very low, less than needed to efficiently infect mosquitos. Low levels of WNV were detected in feces on 4 and 7 DPI, but no virus was isolated from oropharyngeal swabs. WNV wasnot transmitted from WNV-inoculated to contact-exposed turkeys. All WNV-inoculated poults seroconverted on 7 DPI. In the turkey that died, WNV was not isolated from intestine, myocardium, brain, kidney, or cloacal and oropharyngeal swabs, but sparse viral antigen was demonstrated by immunohistochemistry in the heart and spleen. Turkeys in contact with WNV-inoculated turkeys and sham-inoculated controls lacked WNV specific antibodies,and WNV was not isolated from plasma and cloacal and oropharyngeal swabs. These data suggest that WNV lacks the potential to be a major new disease of turkeys and that turkeys will not be a significant amplifying host for infecting mosquitos.