Louis Encephalitis Virus Infection Research Articles

Haemovigilance survey and screening strategy for arthropod-borneviruses in blood donors from Argentina.

Arthropod-borne viruses (arboviruses) count among emerging infections, which represent a major challenge for transfusion safety worldwide. To assess the risk of arboviruses-transmission by transfusion (ATT), we performed a survey to evaluate the potential threat for transfusion safety. Samples were retrospectively and randomly collected from donors who donated during the peak of dengue incidence in Cordoba (years: 2016and 2019-2022). A cost-efficient strategy for molecular screening was implemented with a nucleic acid test (NAT) configured with Flavivirus and Alphavirus-universal degenerated primers targeting conserved gene regions. Besides, we evaluated the neutralizing antibody (NAb) prevalence by plaque reduction neutralization test (PRNT). A total of 1438 samples were collected. Among the NAT-screened samples, one resulted positive for Flavivirus detection. Subsequent sequencing of the PCR product revealed Saint Louis Encephalitis Virus (SLEV) infection (GeneBank accession number OR236721). NAb prevalence was 2.95% for anti-Dengue, 9.94% anti-SLEV, 1.09% anti-West Nile Virus, and 0% anti-Chikungunya. One of the NAb-positive samples also resulted positive for IgM against SLEV but negative by ARN detection. This is the first haemovigilance study developed in Argentina that evaluates the potential risk of ATT and the first research to determine the prevalence of NAb against Flavivirus through PNRT to avoid possible cross-reactions between Ab against Flavivirus. Herein, the finding of one SLEV-viremic donor and the detection of anti-SLEV IgM in a different donor demonstrated a potential threat for transfusion safety and emphasized the need for increased vigilance and proactive measures to ensure the safety of blood supplies.

Type I interferons are essential while type II interferon is dispensable for protection against St. Louis encephalitis virus infection in the mouse brain

ABSTRACT St. Louis encephalitis virus (SLEV) is a neglected mosquito-borne flavivirus that causes severe neurological disease in humans. SLEV replication in the central nervous system (CNS) induces the local production of interferons (IFNs), which are attributed to host protection. The antiviral response to SLEV infection in the CNS is not completely understood, which led us to characterize the roles of IFNs using mouse models of St. Louis encephalitis. We infected mice deficient in type I IFN receptor (ABR−/-) or deficient in Type II IFN (IFNγ−/-) and assessed the contribution of each pathway to disease development. We found that type I and II IFNs play different roles in SLEV infection. Deficiency in type I IFN signaling was associated to an early and increased mortality, uncontrolled SLEV replication and impaired ISG expression, leading to increased proinflammatory cytokine production and brain pathology. Conversely, IFNγ−/- mice were moderately resistant to SLEV infection. IFNγ deficiency caused no changes to viral load or SLEV-induced encephalitis and did not change the expression of ISGs in the brain. We found that type I IFN is essential for the control of SLEV replication whereas type II IFN was not associated with protection in this model.

Reemergence of St. Louis Encephalitis Virus, California, 2015.

St. Louis encephalitis virus infection was detected in summer 2015 in southern California after an 11-year absence, concomitant with an Arizona outbreak. Sequence comparisons showed close identity of California and Arizona isolates with 2005 Argentine isolates, suggesting introduction from South America and underscoring the value of continued arbovirus surveillance.

Molecular evidence of St. Louis encephalitis virus infection in patients in Buenos Aires, Argentina

We report two cases of St. Louis encephalitis where the virus was detected in patients' sera directly by molecular techniques allowing subsequent typing. Phylogenetic analysis of both samples showed that NS5 sequences clustered with viruses previously classified as genotype III.

Immunoglobulin G subclasses in antibody responses to St. Louis encephalitis virus infections

The aim of this study was to recognize the specific antiviral response patterns of IgG1, IgG2, IgG3 and IgG4 subclasses, elicited during St. Louis encephalitis virus (SLEV) infection in humans. Eighty-five samples of human sera from 44 patients with SLEV infection were obtained between days 1 and 365 or later, after onset of the disease. These samples were processed by immunofluorescence assay for detection of IgG1-, IgG2-, IgG3- and IgG4-specific antibodies. We demonstrate the presence of all isotypes of IgG for more than a year in patients infected with SLEV. However; isotype IgG1 was present at the highest titers, with a peak between days 8 and 30 after onset of the disease.

Etiology of Acute Undifferentiated Febrile Illness in the Amazon Basin of Ecuador

We conducted a longitudinal observational study of 533 patients presenting to two hospitals in the Ecuadorean Amazon basin with acute undifferentiated febrile illness (AUFI) from 2001 through 2004. Viral isolation, reverse transcription-polymerase chain reaction (RT-PCR), IgM seroconversion, and malaria smears identified pathogens responsible for fever in 122 (40.1%) of 304 patients who provided both acute and convalescent blood samples. Leptospirosis was found in 40 (13.2%), malaria in 38 (12.5%), rickettsioses in 18 (5.9%), dengue fever in 16 (5.3%), Q fever in 15 (4.9%), brucellosis in 4 (1.3%), Ilhéus infection in 3 (1.0%), and Venezuelan equine encephalitis (VEE), Oropouche, and St. Louis encephalitis virus infections in less than 1% of these patients. Viral isolation and RT-PCR on another 229 participants who provided only acute samples identified 3 cases of dengue fever, 2 of VEE, and 1 of Ilhéus. None of these pathogens, except for malaria, had previously been detected in the study area.

The First Documented Case of Microbacterium sp. Pneumonia

© 2008 Elsevier 0196-4399/00 (see frontmatter) 45 (CMV IgG) avidity and relationship of avidity to CMV IgM levels. Clin. Diagn. Lab. Immunol. 9:824-827. 23. Prince, H.E. et al. 2005. Utilization of follow-up specimens from viremic blood donors to assess the value of West Nile virus immunoglobulin G avidity as an indicator of recent infection. Clin. Diagn. Lab. Immunol. 12:1123-1126. 24. Fox, J.L. et al. 2006. Immunoglobulin G avidity in differentiation between early and late antibody responses to West Nile virus. Clin. Vaccine Immunol. 13:33-36. 25. Wong S.J. et al. 2003. Immunoassay targeting nonstructural protein 5 to differentiate West Nile virus infection from dengue and St. Louis encephalitis virus infections and from flavivirus vaccination. J. Clin. Microbiol. 41:4217-4223.

Serologic Diagnosis of West Nile and St. Louis Encephalitis Virus Infections in Domestic Chickens

Adult domestic chickens were infected with West Nile virus (WNV) or St. Louis encephalitis virus (SLEV) and challenged with homologous or heterologous virus at 21 or 56 days postinfection (dpi). Sera were collected at selected time points after infection and assayed by enzyme immunoassay (EIA), plaque reduction neutralization test (PRNT), and a Western blot (WB) alternative to PRNT. EIA results were sensitive and accurate (few false positives) but not specific, requiring a confirmatory test to determine virus infection history. PRNT results generally were specific until challenge, after which test results were frequently equivocal and inadequate to determine first or second infecting virus. WB results confirmed the serologic cross-reactivity between WNV and SLEV envelope protein. Non-structural protein 1 and pre-membrane protein reactivities were highly specific for WNV during SLEV infection, but less specific for SLEV during WNV infection. WB and PRNT specificities were similar for both viruses from 6 to 14 dpi, and sensitivities to WNV were virtually identical.

Differentiation of West Nile and St. Louis Encephalitis Virus Infections by Use of Noninfectious Virus-Like Particles with Reduced Cross-Reactivity

Differential diagnosis of St. Louis encephalitis virus (SLEV) and West Nile virus (WNV) infections can be complicated due to the high degree of cross-reactivity observed in most serodiagnostic assays. In an effort to provide a more specific diagnostic test, we developed virus-like particle (VLP) antigens with reduced cross-reactivity for both SLEV and WNV by identifying and mutating envelope protein amino acids within the cross-reactive epitopes of VLP expression plasmids. To determine the serodiagnostic discriminatory ability of the antigens with reduced cross-reactivity, a panel of 134 human serum samples collected predominately from North American patients with SLEV or WNV infections was used to evaluate the performance of these novel antigens in imunoglobulin M antibody-capture enzyme-linked immunosorbent assays. Positive/negative ratios and the resulting diagnostic classifications were compared between the mutant and the wild-type (WT) VLPs. The mutant VLP antigens were more specific, with higher positive predictive values and higher likelihood ratios than the WT VLP antigens. Both the SLEV and WNV mutant VLPs greatly reduced the observed cross-reactivity, significantly increasing the specificity and sensitivity of the assay. The use of these novel VLP antigens with reduced cross-reactivity in these serodiagnostic assays and others should lead to more accurate diagnoses of current infections, thereby reducing the need for time-consuming and cumbersome confirmatory plaque-reduction neutralization tests to differentiate between SLEV and WNV infections in North America.

Post-infectious encephalomyelitis associated with St. Louis encephalitis virus infection

Neurologic illness associated with acute St. Louis encephalitis, West Nile, and Japanese encephalitis virus infection includes acute aseptic meningitis, encephalomyelitis, and a poliomyelitis-like syndrome. Few post-infectious immune-mediated neurologic events associated with flaviviral infection have been reported. The authors report on a woman with apparent post-infectious encephalomyelitis associated with recent St. Louis encephalitis virus infection, suggesting that neurologic illness from flaviviruses may also be seen in the post-infectious period following mild clinical illness.

Occlusive vasculitis in a patient with concomitant west nile virus infection

Purpose To describe a patient with occlusive, retinal vasculitis and concomitant, confirmed, acute West Nile virus (WNV) infection. Design Observational case report. Methods Main outcome measures included comprehensive ophthalmic examination with fluorescein angiography, color photography, and serologic testing for WNV and St. Louis encephalitis (SLE) virus including plaque reduction neutralization testing (PRNT). Results A 46-year-old woman developed a sudden decrease in vision in her left eye 2 weeks after confirmed WNV infection and demonstrated multiple, small, patchy areas of retinal edema with scattered microaneurysms. Fluorescein angiography showed multiple branch artery occlusions with extensive nonperfusion. Serologic titers for WNV were positive for acute infection. Plaque reduction neutralization testing confirmed WNV infection and excluded St. Louis encephalitis virus infection. Other etiologies of occlusive vasculitis were not present. Conclusion Occlusive, retinal vasculitis may occur in the setting of acute WNV infection.

Immunoassay targeting nonstructural protein 5 to differentiate West Nile virus infection from dengue and St. Louis encephalitis virus infections and from flavivirus vaccination.

West Nile virus (WNV) is an emerging flavivirus that has caused frequent epidemics since 1996. Besides natural transmission by mosquitoes, WNV can also be transmitted through blood transfusion and organ transplantation, thus heightening the urgency of development of a specific and rapid serologic assay of WNV infection. The current immunoassays lack specificity because they are based on detection of antibodies against WNV structural proteins and immune responses to structural proteins among flaviviruses cross-react to each other. Here, we describe microsphere immunoassays that detect antibodies to nonstructural proteins 3 and 5 (NS3 and NS5). In contrast to immunoassays based on viral envelope and NS3 proteins, the NS5-based assay (i) reliably discriminates between WNV infections and dengue virus or St. Louis encephalitis virus infections, (ii) differentiates between flavivirus vaccination and natural WNV infection, and (iii) indicates recent infections. These unique features of the NS5-based immunoassay will be very useful for both clinical and veterinary diagnosis of WNV infection.

Interferon-α protects mice against lethal infection with St Louis encephalitis virus delivered by the aerosol and subcutaneous routes

In common with other flaviviruses, there is no specific therapy for St Louis encephalitis (SLE) virus infections. A number of cases have occurred where infection may have been acquired by the aerosol route in laboratory accidents. The recombinant human interferon hybrids IFN-α A/D (Roche Laboratories) and IFN-α B/D (Ciba-Geigy) have activity in murine models. Given for several days around the time of exposure to the virus or shortly after, these compounds reduce the mortality from SLE virus administered to mice subcutaneously by up to 70%. In an aerosol model of SLE disease, the mortality was reduced to 30–50% compared to 100% in controls, depending on the challenge level of virus. These results suggest that interferon-α could be used to reduce the mortality from SLE infection after known exposure to the virus.

Effects of House Sparrow Age and Arbovirus Infection on Attraction of Mosquitoes

To test the hypothesis that arbovirus infection or advancing age increases the attractiveness of avian hosts to vector mosquitoes, we used an olfactometer that measures the response of mosquitoes to the passive, vertical diffusion of host-related cues. St. Louis encephalitis or western equine encephalomyelitis virus infection in house sparrows (Passer domesticus (L.], regardless of age, had no detectable effect on Culex quinquefasciatus Say or Cx. tarsalis Coquillett attraction, respectively, at close range. Sparrow age, however, was associated positively with increased Cx. quinquefasciatus attraction; i.e., nestlings were significantly less attractive than adults, and there was a trend for increased mosquito attraction as nestling age increased. When birds were examined in groups, attraction was not equal to the sum of the individual birds in the group. Nestlings, either alone or in a group, were less attractive than a single adult. One adult plus three older nestlings were more attractive than one adult with three young nestlings, but no more attractive than one adult alone. We speculate on the epidemiological significance of these laboratory results to arbovirus epizootic transmission.

Epidemiologic aspects of a St. Louis encephalitis outbreak in Mesa County, Colorado.

An epidemic of St. Louis encephalitis in 1985 in Mesa County, Colorado, led to 17 cases, including one fatality. Risk was associated with advanced age and residence in Grand Junction, the county's principal city. A trend was observed toward higher risk in females. However, increased risk in females was not associated with higher infection rates (increased exposure). Capture enzyme immunoassays detected specific immunoglobulin M (IgM) and immunoglobulin A after infection. A serosurvey of Grand Junction residents disclosed an infection rate of 4.0%, indicating that 1,123 epidemic St. Louis encephalitis infections may have occurred in the city. Evidence of previous St. Louis encephalitis virus infection was found in 11.2% of survey respondents who had neutralizing antibody to the virus without specific IgM. The prevalence of St. Louis encephalitis virus antibody was similar to rates observed in serosurveys undertaken 30 years earlier, indicating that the level of endemic St. Louis encephalitis transmission in the city had not changed appreciably in that interval.

Experimental St. Louis encephalitis virus infection of sloths and cormorants.

Experimental infection of 11 Bradypus variegatus and Choloepus hoffmanni sloths with St. Louis encephalitis (SLE) virus produced detectable viremias of seven to 27 (median 13) days duration and maximum titers of 2.7 to 6.5 (median 5.1) log10 median suckling mouse intracranial lethal doses (SMicLD50) per ml. Experimental SLE viremia onset was delayed and maximum titer depressed in two sloths concurrently infected with naturally acquired viruses. SLE viremias in four experimentally inoculated cormorants Phalacrocorax olivaceus were shorter, and of equal or lower titer, than in sloths. Colonized Culex pipiens quinquefasciatus mosquitoes were infected by feeding on sloths circulating at least 4.8 log10 SMicLD50 of SLE virus per ml, and subsequently transmitted the infection to mice and chicks. An uninoculated baby Bradypus became infected by contact transmission from its mother. The antibody response of sloths to SLE virus was slow, being undetectable until several weeks post-inoculation. However, both sloth species developed high and long-lasting neutralizing and hemagglutination-inhibition antibody titers. The complement-fixation antibody response in Bradypus was lower and slower to develop than in Choloepus. Sloths with naturally acquired SLE virus antibody did not become detectably viremic after experimental inoculation. Neither sloths nor cormorants become overly ill from SLE virus infection.

Visceral target organs in systemic St. Louis encephalitis virus infection of hamsters

Ultrastructural aspects of St. Louis encephalitis virus infection of the major extraneutral organs and tissues of suckling hamsters were examined. In the pancreas, both the exocrine and endocrine portions were equally affected by the virus. A feature apparently unique to flaviviruses was the accumulation of virus particles in all types of secretory granules in this organ. Virus particles were seen within myocardial fibers and within the smooth muscle cells and endothelial cells of small blood vessels of the heart. In the intestines, the lamina propria was the most severely infected, with virus particles accumulated in all cell types.

Evaluation of the solid-phase radioimmunoassay for diagnosis of St. Louis encephalitis infection in humans.

Since the use of highly purified viral proteins in the solid-phase radioimmunoassay (SPRIA) procedure is not practical for routine diagnostic serology, the procedure was adapted for use with the crude antigens now commonly used to diagnose St. Louis encephalitis (SLE) virus infections. SLE and various other alpha- and flavivirus antigens were tested by both the SPRIA procedure and conventional serological methods. Standards were established for use of the crude antigens in the SPRIA procedure. Reproducibility of the procedure was similar to that of conventional tests. The SPRIA procedure specifically differentiated SLE from clinically similar alphaviruses, but did not differentiate SLE from related flaviviruses very well. SPRIA diagnosis of SLE infections by the immunoglobulin G response in paired sera was good. Generally, the crude antigen SPRIA procedure was as sensitive as the conventional serological tests, but not as specific.

Influence of sex and age on natural resistance to St. Louis encephalitis virus infection in mice.

A difference was observed in susceptibility of adult male and female mice to St. Louis encephalitis (SLE) virus as measured by the death rate after intravenous challenge. Female mice that had susceptibility similar to that of males at 2 months of age had increased resistance to SLE virus at 3 and 4 months of age. The increased resistance occurred after sexual maturity, indicating that the resistance factor possibly was related to an aging process in the female. The susceptibility of male mice remained unchanged over the 2- to 4-month period. Neither pregnancy nor castration had any effect on resistance of adult mice to St. Louis encephalitis virus.

St. Louis encephalitis virus: An ultrastructural study of infection in a mosquito vector

The course of St. Louis encephalitis virus infection of Culex pipiens pipiens Linn, mosquitoes was followed sequentially by electron microscopy. At the site of initial viral invasion of mosquito parenchyma in the midgut, epithelial infection involved a rather constant proportion of cells that yielded only moderate numbers of virus particles. Virus was observed in the midgut at locations where spread via the hemolymph could occur. Tissues in intimate contact with hemolymph (abdominal muscles, malpighian tubules, ovarian sheath) became infected, but only modest numbers of virus particles were ever produced. In sharp contrast, an ever increasing number of of virus particles were formed in the epithelial cells of the salivary glands. Virus was primarily yielded into the cisternae of endoplasmic reticulum and then shed from the apical end of cells into the lumen of the glands. Very few particles were associated with lateral or basal margins of salivary gland epithelium, indicating a directional “preference” of virus for shedding through apical plasma membrane. So much virus was shed into the limited space of the glandular lumen and its diverticula that dispersed particles formed into crystalline arrays from day 25 onward; one of the larger of these crystals was estimated to contain more than 50,000 virus particles. Changes in the infected cells of all the mosquito organs examined were interpreted as physiologic variances relative to feeding time and not as specifically due to viral cytopathology.