ABSTRACT
Rift Valley fever virus (RVFV) is a zoonotic pathogen with pandemic potential. RVFV entry is mediated by the viral glycoprotein (Gn), but host entry factors remain poorly defined. Our genome-wide CRISPR screen identified low-density lipoprotein receptor-related protein 1 (mouse Lrp1/human LRP1), heat shock protein (Grp94), and receptor-associated protein (RAP) as critical host factors for RVFV infection. RVFV Gn directly binds to specific Lrp1 clusters and is glycosylation independent. Exogenous addition of murine RAP domain 3 (mRAPD3) and anti-Lrp1 antibodies neutralizes RVFV infection in taxonomically diverse cell lines. Mice treated with mRAPD3 and infected with pathogenic RVFV are protected from disease and death. A mutant mRAPD3 that binds Lrp1 weakly failed to protect from RVFV infection. Together, these data support Lrp1 as a host entry factor for RVFV infection and define a new target to limit RVFV infections.
Subject(s)
Host-Pathogen Interactions , Low Density Lipoprotein Receptor-Related Protein-1/metabolism , Rift Valley fever virus/physiology , Virus Internalization , Animals , Antibody Specificity/immunology , Base Sequence , Brain/pathology , Brain/virology , CRISPR-Cas Systems/genetics , Cell Membrane/metabolism , Cells, Cultured , Glycoproteins/metabolism , Glycosaminoglycans/metabolism , Glycosylation , Humans , LDL-Receptor Related Protein-Associated Protein/metabolism , Ligands , Low Density Lipoprotein Receptor-Related Protein-1/deficiency , Membrane Glycoproteins/metabolism , Mice , Protein Binding , Protein Denaturation , Rift Valley Fever/pathology , Rift Valley Fever/prevention & control , Rift Valley Fever/virology , Rift Valley fever virus/immunologyABSTRACT
Rift Valley fever virus (RVFV) infection causes abortions in ruminant livestock and is associated with an increased likelihood of miscarriages in women. Using sheep and human placenta explant cultures, we sought to identify tissues at the maternal-fetal interface targeted by RVFV. Sheep villi and fetal membranes were highly permissive to RVFV infection resulting in markedly higher virus titers than human cultures. Sheep cultures were most permissive to wild-type RVFV and ΔNSm infection, while live-attenuated RVFV vaccines (LAVs; MP-12, ΔNSs, and ΔNSs/ΔNSm) exhibited reduced replication. The human fetal membrane restricted wild-type and LAV replication, and when infection occurred, it was prominent on the maternal-facing side. Type I and type III interferons were induced in human villi exposed to LAVs lacking the NSs protein. This study supports the use of sheep and human placenta explants to understand vertical transmission of RVFV in mammals and whether LAVs are attenuated at the maternal-fetal interface.IMPORTANCEA direct comparison of replication of Rift Valley fever virus (RVFV) in sheep and human placental explants reveals comparative efficiencies and permissivity to infection and replication. Vaccine strains of RVFV demonstrated reduced infection and replication capacity in the mammalian placenta. This study represents the first direct cross-host comparison of the vertical transmission capacity of this high-priority emerging mosquito-transmitted virus.
Subject(s)
Infectious Disease Transmission, Vertical , Placenta , Rift Valley Fever , Rift Valley fever virus , Vaccines, Attenuated , Viral Vaccines , Virus Replication , Rift Valley fever virus/physiology , Rift Valley fever virus/immunology , Animals , Female , Pregnancy , Sheep , Placenta/virology , Humans , Rift Valley Fever/virology , Rift Valley Fever/transmission , Viral Vaccines/immunology , Sheep Diseases/virologyABSTRACT
Rift Valley fever virus (RVFV) is a mosquito-borne bunyavirus that causes severe and potentially fatal hemorrhagic fever in humans. Autophagy is a self-degradative process that can restrict viral replication at multiple infection steps. In this study, we evaluated the effects of RVFV-triggered autophagy on viral replication and immune responses. Our results showed that RVFV infection triggered autophagosome formation and induced complete autophagy. Impairing autophagy flux by depleting autophagy-related gene 5 (ATG5), ATG7, or sequestosome 1 (SQSTM1) or treatment with autophagy inhibitors markedly reduced viral RNA synthesis and progeny virus production. Mechanistically, our findings demonstrated that the RVFV nucleoprotein (NP) C-terminal domain interacts with the autophagy receptor SQSTM1 and promotes the SQSTM1-microtubule-associated protein 1 light chain 3 B (LC3B) interaction and autophagy. Deletion of the NP C-terminal domain impaired the interaction between NP and SQSTM1 and its ability to trigger autophagy. Notably, RVFV-triggered autophagy promoted viral infection in macrophages but not in other tested cell types, including Huh7 hepatocytes and human umbilical vein endothelial cells, suggesting cell type specificity of this mechanism. It was further revealed that RVFV NP-triggered autophagy dampens antiviral innate immune responses in infected macrophages to promote viral replication. These results provide novel insights into the mechanisms of RVFV-triggered autophagy and indicate the potential of targeting the autophagy pathway to develop antivirals against RVFV. IMPORTANCE We showed that RVFV infection induced the complete autophagy process. Depletion of the core autophagy genes ATG5, ATG7, or SQSTM1 or pharmacologic inhibition of autophagy in macrophages strongly suppressed RVFV replication. We further revealed that the RVFV NP C-terminal domain interacted with SQSTM1 and enhanced the SQSTM1/LC3B interaction to promote autophagy. RVFV NP-triggered autophagy strongly inhibited virus-induced expression of interferon-stimulated genes in infected macrophages but not in other tested cell types. Our study provides novel insights into the mechanisms of RVFV-triggered autophagy and highlights the potential of targeting autophagy flux to develop antivirals against this virus.
Subject(s)
Autophagy , Immunity, Innate , Nucleoproteins , Rift Valley fever virus , Immunity, Innate/immunology , Rift Valley fever virus/immunology , Nucleoproteins/immunology , Nucleoproteins/metabolism , Autophagy/immunology , Virus Replication , Cell Line , Rift Valley Fever/immunology , Humans , Animals , Macrophages/virologyABSTRACT
Rift Valley fever virus (RVFV), an emerging arboviral and zoonotic bunyavirus, causes severe disease in livestock and humans. Here, we report the isolation of a panel of monoclonal antibodies (mAbs) from the B cells of immune individuals following natural infection in Kenya or immunization with MP-12 vaccine. The B cell responses of individuals who were vaccinated or naturally infected recognized similar epitopes on both Gc and Gn proteins. The Gn-specific mAbs and two mAbs that do not recognize either monomeric Gc or Gn alone but recognized the hetero-oligomer glycoprotein complex (Gc+Gn) when Gc and Gn were coexpressed exhibited potent neutralizing activities in vitro, while Gc-specific mAbs exhibited relatively lower neutralizing capacity. The two Gc+Gn-specific mAbs and the Gn domain A-specific mAbs inhibited RVFV fusion to cells, suggesting that mAbs can inhibit the exposure of the fusion loop in Gc, a class II fusion protein, and thus prevent fusion by an indirect mechanism without direct fusion loop contact. Competition-binding analysis with coexpressed Gc/Gn and mutagenesis library screening indicated that these mAbs recognize four major antigenic sites, with two sites of vulnerability for neutralization on Gn. In experimental models of infection in mice, representative mAbs recognizing three of the antigenic sites reduced morbidity and mortality when used at a low dose in both prophylactic and therapeutic settings. This study identifies multiple candidate mAbs that may be suitable for use in humans against RVFV infection and highlights fusion inhibition against bunyaviruses as a potential contributor to potent antibody-mediated neutralization.
Subject(s)
Antibodies, Neutralizing/immunology , Antibodies, Viral/immunology , Rift Valley fever virus/immunology , Viral Fusion Proteins/immunology , Animals , Antibodies, Monoclonal/immunology , Cell Line, Tumor , Cells, Cultured , Chlorocebus aethiops , Epitopes/chemistry , Epitopes/immunology , HEK293 Cells , Humans , Mice , Mice, Inbred C57BL , Vero Cells , Viral Fusion Proteins/chemistryABSTRACT
Autophagy has been implicated as a component of host defense, but the significance of antimicrobial autophagy in vivo and the mechanism by which it is regulated during infection are poorly defined. Here we found that antiviral autophagy was conserved in flies and mammals during infection with Rift Valley fever virus (RVFV), a mosquito-borne virus that causes disease in humans and livestock. In Drosophila, Toll-7 limited RVFV replication and mortality through activation of autophagy. RVFV infection also elicited autophagy in mouse and human cells, and viral replication was increased in the absence of autophagy genes. The mammalian Toll-like receptor adaptor, MyD88, was required for anti-RVFV autophagy, revealing an evolutionarily conserved requirement for pattern-recognition receptors in antiviral autophagy. Pharmacologic activation of autophagy inhibited RVFV infection in mammalian cells, including primary hepatocytes and neurons. Thus, autophagy modulation might be an effective strategy for treating RVFV infection, which lacks approved vaccines and therapeutics.
Subject(s)
Autophagy/immunology , Rift Valley Fever/immunology , Rift Valley fever virus/immunology , Allyl Compounds/pharmacology , Animals , Antiviral Agents/pharmacology , Autophagy/drug effects , Cells, Cultured , Drosophila , Evolution, Molecular , Hepatocytes/drug effects , Hepatocytes/immunology , Hepatocytes/virology , Humans , Infection Control/methods , Mammals , Mice , Myeloid Differentiation Factor 88/metabolism , Neurons/drug effects , Neurons/immunology , Neurons/virology , Quinazolines/pharmacology , Rats , Rift Valley Fever/drug therapy , Toll-Like Receptor 7/metabolism , Virus ReplicationABSTRACT
Rift Valley fever virus (RVFV) is an arbovirus found throughout Africa. It causes disease that is typically mild and self-limiting; however, some infected individuals experience severe manifestations, including hepatitis, encephalitis, or even death. Reports of RVFV encephalitis are notable among immunosuppressed individuals, suggesting a role for adaptive immunity in preventing this severe complication. This phenomenon has been modeled in C57BL/6 mice depleted of CD4 T cells prior to infection with DelNSs RVFV (RVFV containing a deletion of nonstructural protein NSs), resulting in late-onset encephalitis accompanied by high levels of viral RNA in the brain in 30% of animals. In this study, we sought to define the specific type(s) of CD4 T cells that mediate protection from RVFV encephalitis. The viral epitopes targeted by CD4 and CD8 T cells were defined in C57BL/6 mice, and tetramers for both CD4 and CD8 T cells were generated. RVFV-specific CD8 T cells were expanded and of a cytotoxic and proliferating phenotype in the liver following infection. RVFV-specific CD4 T cells were identified in the liver and spleen following infection and phenotyped as largely Th1 or Tfh subtypes. Knockout mice lacking various aspects of pathways important in Th1 and Tfh development and function were used to demonstrate that T-bet, CD40, CD40L, and major histocompatibility complex class II (MHC-II) mediated protection from RVFV encephalitis, while gamma interferon (IFN-γ) and interleukin-12 (IL-12) were dispensable. Virus-specific antibody responses correlated with protection from encephalitis in all mouse strains, suggesting that Tfh/B cell interactions modulate clinical outcome in this model. IMPORTANCE The prevention of RVFV encephalitis requires intact adaptive immunity. In this study, we developed reagents to detect RVFV-specific T cells and provide evidence for Tfh cells and CD40/CD40L interactions as critical mediators of this protection.
Subject(s)
CD40 Antigens , CD40 Ligand , Encephalitis, Viral/prevention & control , Rift Valley Fever/immunology , Rift Valley fever virus/immunology , Rift Valley fever virus/physiology , T-Lymphocytes/immunology , Africa , Animals , Antibody Formation , B-Lymphocytes/immunology , Brain/virology , CD8-Positive T-Lymphocytes/immunology , Disease Models, Animal , Encephalitis, Viral/immunology , Encephalitis, Viral/virology , Epitopes , Female , Liver/immunology , Mice , Mice, Inbred C57BL , Mice, KnockoutABSTRACT
The epidemiology of Rift Valley fever virus (RVFV) and Crimean-Congo hemorrhagic fever virus (CCHFV) in Jordan is unknown. Our investigation showed 3% of 989 tested dairy cattle, sheep, and goats were RVFV seropositive and 14% were CCHFV seropositive. Ongoing surveillance is needed to assess risk to humans and protect public health.
Subject(s)
Hemorrhagic Fever Virus, Crimean-Congo , Hemorrhagic Fever, Crimean/epidemiology , Rift Valley Fever/epidemiology , Rift Valley fever virus , Animals , Antibodies, Viral , Cattle , Hemorrhagic Fever Virus, Crimean-Congo/immunology , Hemorrhagic Fever, Crimean/veterinary , Jordan/epidemiology , Rift Valley fever virus/immunology , Ruminants , Sheep , ZoonosesABSTRACT
Rift Valley fever virus (RVFV) causes severe disease in livestock concurrent with zoonotic transmission to humans. A subset of people infected with RVFV develop encephalitis, and significant gaps remain in our knowledge of how RVFV causes pathology in the brain. We previously found that, in Lewis rats, subcutaneous inoculation with RVFV resulted in subclinical disease while inhalation of RVFV in a small particle aerosol caused fatal encephalitis. Here, we compared the disease course of RVFV in Lewis rats after each different route of inoculation in order to understand more about pathogenic mechanisms of fatal RVFV encephalitis. In aerosol-infected rats with lethal encephalitis, neutrophils and macrophages were the major cell types infiltrating the CNS, and this was concomitant with microglia activation and extensive cytokine inflammation. Despite this, prevention of neutrophil infiltration into the brain did not ameliorate disease. Unexpectedly, in subcutaneously-inoculated rats with subclinical disease, detectable viral RNA was found in the brain along with T-cell infiltration. This study sheds new light on the pathogenic mechanisms of RVFV encephalitis.
Subject(s)
Brain/immunology , Encephalitis, Viral/immunology , Macrophages/immunology , Neutrophil Infiltration , Neutrophils/immunology , Rift Valley Fever/immunology , Rift Valley fever virus/immunology , Aerosols , Animals , Brain/pathology , Brain/virology , Cell Line , Cytokines/immunology , Encephalitis, Viral/pathology , Female , Humans , Macrophages/pathology , Neutrophils/pathology , Rats , Rats, Inbred Lew , Rift Valley Fever/pathologyABSTRACT
BACKGROUND: Prior to the first recorded outbreak of Rift Valley fever (RVF) in Uganda, in March 2016, earlier studies done until the 1970's indicated the presence of the RVF virus (RVFV) in the country, without any recorded outbreaks in either man or animals. While severe outbreaks of RVF occurred in the neighboring countries, none were reported in Uganda despite forecasts that placed some parts of Uganda at similar risk. The Ministry of Agriculture, Animal Industry and Fisheries (MAAIF) undertook studies to determine the RVF sero-prevalence in risk prone areas. Three datasets from cattle sheep and goats were obtained; one from retrospective samples collected in 2010-2011 from the northern region; the second from the western region in 2013 while the third was from a cross-sectional survey done in 2016 in the south-western region. Laboratory analysis involved the use of the Enzyme Linked Immunosorbent Assays (ELISA). Data were subjected to descriptive statistical analyses, including non-parametric chi-square tests for comparisons between districts and species in the regions. RESULTS: During the Yellow Fever outbreak investigation of 2010-2011 in the northern region, a total sero-prevalence of 6.7% was obtained for anti RVFV reacting antibodies (IgG and IgM) among the domestic ruminant population. The 2013 sero-survey in the western region showed a prevalence of 18.6% in cattle and 2.3% in small ruminants. The 2016 sero-survey in the districts of Kabale, Kanungu, Kasese, Kisoro and Rubirizi, in the south-western region, had the respective district RVF sero-prevalence of 16.0, 2.1, 0.8, 15.1and 2.7% among the domestic ruminants combined for this region; bovines exhibited the highest cumulative sero-prevalence of 15.2%, compared to 5.3 and 4.0% respectively for sheep and goats per species for the region. CONCLUSIONS: The absence of apparent outbreaks in Uganda, despite neighboring enzootic areas, having minimal restrictions to the exchange of livestock and their products across borders, suggest an unexpected RVF activity in the study areas that needs to be unraveled. Therefore, more in-depth studies are planned to mitigate the risk of an overt RVF outbreak in humans and animals as has occurred in neighboring countries.
Subject(s)
Animal Diseases/epidemiology , Rift Valley Fever/epidemiology , Rift Valley fever virus/immunology , Animal Diseases/virology , Animals , Cattle , Enzyme-Linked Immunosorbent Assay/veterinary , Goats , Immunoglobulin G/blood , Immunoglobulin M/blood , Prevalence , Rift Valley fever virus/isolation & purification , Seroepidemiologic Studies , Sheep , Uganda/epidemiologyABSTRACT
BACKGROUND: Rift Valley fever (RVF) is a vector-borne emerging zoonotic disease of animals and humans, characterized by socioeconomic losses to livestock farmers and global public health threat. The study determined RVFV seroprevalence in cattle, assessed pastoralists' knowledge about RVF, and factors that influence its occurrence in pastoral cattle herds of Nigeria. A cross-sectional study was conducted in pastoral herds of North-central Nigeria from 2017 to 2018. Data were collected using serology and questionnaire tools. Descriptive statistics were used to analyze the obtained data. Categorical variables were presented as proportions and their associations determined by Chi-square tests. Associations of risk factors were analyzed by univariable and multivariable logistic regressions analyses at 95% confidence level. RESULTS: The overall IgM seropositivity of RVFV in pastoral cattle herds was 5.6%. This was higher in nomadic herds (7.4%) than in agro-pastoral herds (3.8%). All animal demographic characteristics of age, sex and breeds were not significantly (p > 0.05) associated with RVFV occurrence in pastoral herds. All the 403 pastoralists selected participated in the study, with the majorities of them being male, married and have no formal education. Majority of the pastoralists had low knowledge levels about zoonotic RVFV infection. All identified socio-ecological factors significantly (p < 0.05) influenced RVFV occurrence in herds. Mosquitoes availability in cattle environment (OR = 7.81; 95% CI: 4.85, 12.37), presence of rivers and streams at grazing fields (OR = 10.80; 95% CI: 6.77, 17.34), high rainfall (OR = 4.30; 95% CI: 2.74, 6.59), irrigated rice fields (OR = 5.14; 95% CI: 3.21, 7.79), bushy vegetation (OR = 6.11; 95% CI: 3.96, 9.43), animal movement (OR = 2.2; 95% CI: 1.45, 3.25), and seasons (OR = 2.34; 95% CI: 1.55, 3.51) were more likely to influenced RVFV occurrence in cattle herds. CONCLUSIONS: Results of this study had illustrated recent circulation of RVFV in pastoral cattle herds in Nigeria and needs urgent interventions. The surveyed pastoralists had low knowledge level about RVF while the socio-ecological factors significantly influenced RVFV occurrence in herds. To address these gaps, pastoralists should be educated on clinical manifestations and modes of transmission of the disease in animals and humans, and mitigation measures. Adequate knowledge about RVF epidemiology will assure food security and public health.
Subject(s)
Animal Husbandry/methods , Cattle Diseases/epidemiology , Rift Valley Fever/epidemiology , Adult , Aged , Animals , Cattle , Cattle Diseases/transmission , Cross-Sectional Studies , Culicidae , Female , Humans , Immunoglobulin M/blood , Male , Middle Aged , Nigeria/epidemiology , Rift Valley Fever/transmission , Rift Valley fever virus/immunology , Risk Factors , Seroepidemiologic Studies , Socioeconomic Factors , ZoonosesABSTRACT
A clear distinction can be made regarding the susceptibility to and the severity of lesions in young lambs when compared to adult sheep. In particular, there are important differences in the lesions and tropism of Rift Valley fever virus (RVFV) in the liver, kidneys, and lymphoid tissues of young lambs. A total of 84 lambs (<6 weeks old), necropsied during the 2010 to 2011 Rift Valley fever (RVF) outbreak in South Africa, were examined by histopathology and immunohistochemistry (IHC). Of the 84 lambs, 71 were positive for RVFV. The most striking diagnostic feature in infected lambs was diffuse necrotizing hepatitis with multifocal liquefactive hepatic necrosis (primary foci) against a background of diffuse hepatocellular death. Lymphocytolysis was present in all lymphoid organs except for the thymus. Lesions in the kidney rarely progressed beyond hydropic change and occasional pyknosis or karyolysis in renal tubular epithelial cells. Viral antigen was diffusely present in the cytoplasm of hepatocytes, but this labeling was noticeably sparse in primary foci. Immunolabeling for RVFV in young lambs was also detected in macrophages, vascular smooth muscle cells, adrenocortical epithelial cells, renal tubular epithelial cells, renal perimacular cells, and cardiomyocytes. RVFV immunolabeling was also often present in capillaries and small blood vessels either as non-cell-associated viral antigen, as antigen in endothelial cells, or intravascular cellular debris. Specimens from the liver, spleen, kidney, and lungs were adequate to confirm a diagnosis of RVF. Characteristic lesions were present in these organs with the liver and spleen being the most consistently positive for RVFV by IHC.
Subject(s)
Antibodies, Viral/immunology , Antigens, Viral/immunology , Disease Outbreaks/veterinary , Rift Valley Fever/diagnosis , Rift Valley fever virus/physiology , Sheep Diseases/diagnosis , Tropism , Age Factors , Animals , Animals, Newborn , Immunohistochemistry/veterinary , Kidney/pathology , Kidney/virology , Liver/pathology , Liver/virology , Lung/pathology , Lung/virology , Organ Specificity , Rift Valley Fever/epidemiology , Rift Valley Fever/pathology , Rift Valley Fever/virology , Rift Valley fever virus/immunology , Sheep , Sheep Diseases/epidemiology , Sheep Diseases/pathology , Sheep Diseases/virology , South Africa/epidemiology , Spleen/pathology , Spleen/virologyABSTRACT
Rift Valley fever (RVF) is a vector-borne viral disease widespread in Africa. The primary cycle involves mosquitoes and wild and domestic ruminant hosts. Humans are usually contaminated after contact with infected ruminants. As many environmental, agricultural, epidemiological, and anthropogenic factors are implicated in RVF spread, the multidisciplinary One Health approach was needed to identify the drivers of RVF epidemics in Madagascar. We examined the environmental patterns associated with these epidemics, comparing human and ruminant serological data with environmental and cattle-trade data. In contrast to East Africa, environmental drivers did not trigger the epidemics: They only modulated local Rift Valley fever virus (RVFV) transmission in ruminants. Instead, RVFV was introduced through ruminant trade and subsequent movement of cattle between trade hubs caused its long-distance spread within the country. Contact with cattle brought in from infected districts was associated with higher infection risk in slaughterhouse workers. The finding that anthropogenic rather than environmental factors are the main drivers of RVF infection in humans can be used to design better prevention and early detection in the case of RVF resurgence in the region.
Subject(s)
Cattle Diseases/epidemiology , Rift Valley Fever/epidemiology , Abattoirs , Animals , Cattle , Cattle Diseases/blood , Cattle Diseases/immunology , Cattle Diseases/transmission , Commerce , Epidemics , Humans , Immunoglobulin G/blood , Immunoglobulin M/blood , Madagascar/epidemiology , Rift Valley Fever/blood , Rift Valley Fever/immunology , Rift Valley Fever/transmission , Rift Valley fever virus/immunology , Seroepidemiologic Studies , WeatherABSTRACT
Phylogenetic analysis of Rift Valley fever virus partial genomic sequences from a patient infected in South Africa in May 2018 suggests reemergence of an endemic lineage different from that of the epidemic in South Africa during 2010-2011. Surveillance during interepidemic periods should be intensified to better predict future epidemics.
Subject(s)
Communicable Diseases, Emerging/epidemiology , Communicable Diseases, Emerging/virology , Disease Outbreaks , Rift Valley Fever/epidemiology , Rift Valley Fever/virology , Rift Valley fever virus , Communicable Diseases, Emerging/history , History, 21st Century , Humans , Phylogeny , Population Surveillance , Rift Valley Fever/history , Rift Valley fever virus/classification , Rift Valley fever virus/genetics , Rift Valley fever virus/immunology , Seasons , South Africa/epidemiology , Viral Proteins/geneticsABSTRACT
The increasing risk of Rift Valley fever virus (RVFV) infection as a global veterinary and public health threat demands the development of safe and accurate diagnostic tests. The aim of this study was to assess the suitability of a baculovirus expression system to produce recombinant RVFV nucleoprotein (N) for use as serodiagnostic antigen in an indirect enzyme-linked immunosorbent assay (ELISA). The ability of the recombinant N antigen to detect RVFV antibody responses was evaluated in ELISA format using antisera from sheep and cattle experimentally infected with two genetically distinct wild-type RVFV strains and sera from indigenous sheep and goat populations exposed to natural RVFV field infection in The Gambia. The recombinant N exhibited specific reactivity with the N-specific monoclonal antibody and various hyperimmune serum samples from ruminants. The indirect ELISA detected N-specific antibody responses in animals with 100% sensitivity compared to the plaque reduction neutralization test (6 to 21 days postinfection) and with 97% and 100% specificity in sheep and cattle, respectively. There was a high level of correlation between the indirect N ELISA and the virus neutralization test for sheep sera (R2 = 0.75; 95% confidence interval [CI] = 0.73 to 0.92) and cattle sera (R2 = 0.80; 95% CI = 0.67 to 0.97); in addition, the N-specific ELISA detected RVFV seroprevalence levels of 26.1% and 54.3% in indigenous sheep and goats, respectively, in The Gambia. The high specificity and correlation with the virus neutralization test support the idea of the feasibility of using the recombinant baculovirus-expressed RVFV N-based indirect ELISA to assess RVFV seroprevalence in livestock in areas of endemicity and nonendemicity.
Subject(s)
Antigens, Viral/immunology , Enzyme-Linked Immunosorbent Assay , Nucleoproteins/immunology , Recombinant Proteins/immunology , Rift Valley Fever/diagnosis , Rift Valley Fever/immunology , Rift Valley fever virus/immunology , Animals , Antibodies, Viral/immunology , Baculoviridae/genetics , Genetic Vectors/genetics , Immunoglobulin G/immunology , Livestock , Neutralization Tests , Nucleoproteins/genetics , Recombinant Proteins/genetics , Sensitivity and Specificity , Sheep , Sheep Diseases/diagnosis , Sheep Diseases/immunologyABSTRACT
Rift Valley fever virus (RVFV) is an arbovirus that causes disease in livestock and humans in Africa and the Middle East. While human disease is typically mild and self-limiting, some individuals develop severe manifestations, such as hepatitis, hemorrhagic fever, or encephalitis. Encephalitis occurs 2 to 3 weeks after acute illness; therefore, we hypothesized that it was a result of an inadequate adaptive immunity. To test this hypothesis in vivo, we used an attenuated virus (DelNSsRVFV) that does not typically cause disease in mice. We first characterized the normal immune response to infection with DelNSsRVFV in immunocompetent mice and noted expansion of natural killer cells and monocytes, as well as activation of both CD8 and CD4 T cells. Depleting C57BL/6 mice of CD4 T cells prior to DelNSsRVFV infection resulted in encephalitis in 30% of the mice; in encephalitic mice, we noted infiltration of T cells and inflammatory monocytes into the brain. CD4 and CD8 codepletion in C57BL/6 mice, as well as CD4 depletion in CCR2 knockout mice, increased the frequency of encephalitis, demonstrating that these cell types normally contributed to the prevention of disease. Encephalitic mice had similar viral RNA loads in the brain regardless of which cell types were depleted, suggesting that CD4 T cells, CD8 T cells, and inflammatory monocytes did little to control viral replication in the brain. CD4-depleted mice exhibited diminished humoral and T cell memory responses, suggesting that these immune mechanisms contributed to peripheral control of virus, thus preventing infection of the brain.IMPORTANCE RVFV is found in Africa and the Middle East and is transmitted by mosquitos or through contact with infected animals. Infected individuals can develop mild disease or more severe forms, such as hepatitis or encephalitis. In order to understand why some individuals develop encephalitis, we first need to know which immune functions protect those who do not develop this form of disease. In this study, we used a mouse model of RVFV infection to demonstrate that CD4 T cells, CD8 T cells, and monocytes all contribute to prevention of encephalitis. Their likely mechanism of action is preventing RVFV from ever reaching the brain.
Subject(s)
CD4-Positive T-Lymphocytes/metabolism , CD8-Positive T-Lymphocytes/metabolism , Encephalitis, Viral/prevention & control , Monocytes/metabolism , Rift Valley Fever/prevention & control , Rift Valley fever virus/immunology , Animals , Brain/immunology , Brain/virology , Encephalitis, Viral/immunology , Immunity, Humoral , Immunity, Innate , Lymphocyte Activation , Mice , Mice, Inbred C57BL , Mice, Knockout , Receptors, CCR2/genetics , Rift Valley Fever/immunology , Rift Valley fever virus/pathogenicityABSTRACT
The risk of exposure of slaughterhouse workers to Rift Valley fever (RVF) virus-infected animals in Nigeria was assessed by determining the prevalence of anti-RVF IgM in cattle, goats and sheep slaughtered in a major abattoir in Ibadan, Nigeria. Blood samples were collected from 290 animals in Bodija Municipal abattoir, Ibadan, Nigeria in January and February 2017 and analyzed for the presence of RVF virus using IgM Enzyme-Linked Immunosorbent Assay (ELISA) and Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) for detection of the virus RNA. Descriptive statistics was used to analyze data. Overall, an IgM prevalence of 0.7% (2/290) was found among the blood samples of the animals, suggesting recent exposure to the virus. Antibody was detected in the sera from a cow and one goat. RVF virus RNA was not detected in the 2 IgM positive blood samples. There was no statistically significant relationship between RVF IgM infection and some variables of the animals, including age, sex and breed (p ≥ 0.05). Results of this study indicate active RVF virus transmission in domestic livestock in Nigeria. The study emphasizes the need to embark on monitoring of human and animal populations to prevent outbreak of the virus in the country.
Subject(s)
Rift Valley Fever/blood , Animals , Cattle , Enzyme-Linked Immunosorbent Assay , Female , Goats , Immunoglobulin M/blood , Immunoglobulin M/immunology , Male , Nigeria , Rift Valley Fever/immunology , Rift Valley fever virus/immunology , SheepABSTRACT
BACKGROUND & OBJECTIVES: Rift Valley fever virus (RVFV) is a vector-borne pathogen that causes serious outbreaks among livestock, and severe symptoms and mortality in humans. The virus is known to be widespread throughout African countries and Arabian peninsula. The aim of the present study was to investigate the seroprevalence of RVFV infection among human populations of Mersin province, Turkey. METHODS: A region-wide serological survey was conducted on humans residing in rural and urban areas of Mersin province located in the subtropical mediterranean region of Turkey from July 2011- January 2014. Plasma samples were tested for the presence of anti-RVFV antibodies using commercially available indirect immunofluorescence assay. RESULTS: The overall past infections were detected in 48 (4.9%) of the 977 human blood samples. The RVF virus- specific IgG positivity was detected in 33 (4.9%) of the 677 blood samples obtained from the urban area and in 15 (5%) of the 300 samples obtained from the rural area. There was no statistically significant difference in the distribution of RVFV IgG positivity rates between urban and rural areas (p = 0.933); though difference was significant between the rural areas (p = 0.029). INTERPRETATION & CONCLUSION: The study confirmed for the first time, the presence of the RVFV antibody in the urban and rural areas of mediterranean province of Mersin in Turkey, suggesting wide circulation of RVFV in the human population.
Subject(s)
Antibodies, Viral/blood , Rift Valley Fever/blood , Rift Valley fever virus/immunology , Adolescent , Adult , Aged , Aged, 80 and over , Disease Outbreaks , Female , Humans , Immunoglobulin G/blood , Male , Middle Aged , Rift Valley Fever/epidemiology , Rift Valley Fever/virology , Rift Valley fever virus/isolation & purification , Rural Population , Seroepidemiologic Studies , Turkey/epidemiology , Urban Population , Young AdultABSTRACT
Rift Valley fever virus is an arbovirus found in Africa and the Middle East. Most infected individuals experience a mild self-limiting illness; however, some develop severe disease including hepatitis, hemorrhagic fever, or encephalitis. The biological reasons for these marked differences in disease manifestation are unknown. In this study, we evaluate 32 biomarkers in serum of 26 patients from an outbreak that occurred in Saudi Arabia in 2000-2001. Eleven biomarkers correlated with viral RNA. Thirteen biomarkers were associated with a fatal outcome. No associations of biomarkers and hemorrhage or central nervous system disease were identified in this cohort.
Subject(s)
Biomarkers/blood , Rift Valley Fever/blood , Rift Valley Fever/epidemiology , Rift Valley fever virus , Adolescent , Adult , Aged , Aged, 80 and over , Cohort Studies , Cytokines/blood , Female , Humans , Male , Middle Aged , RNA, Viral/blood , Rift Valley Fever/immunology , Rift Valley Fever/virology , Rift Valley fever virus/genetics , Rift Valley fever virus/immunology , Young AdultABSTRACT
BACKGROUND: Rift Valley Fever Phlebovirus (RVFV) and Crimean-Congo Hemorrhagic Fever Orthonairovirus (CCHFV) specific antibodies had been documented among humans in urban settings of the southwestern and northern Cameroon in the late 1980s. Recently, evidence for enzootic circulation of RVFV was reported among livestock in both rural and urban settings in Cameroon. However, current estimates of human exposure to RVFV and CCHFV are still to be documented in Cameroon, especially in rural areas. The aim of this study was to assess the seroprevalence of RVFV and CCHFV in rural settings in the Southeastern rain forest of Cameroon. RESULTS: Using Enzyme-linked Immunosorbent Assays, the presence of RVFV and CCHFV Immunoglobulin G antibodies was investigated in plasma samples originating from 137 Pygmies from four villages of the East region of Cameroon. The studied population was found to be 12.4% (17/137) and 4.4% (6/137) seropositive for RVFV and CCHFV, respectively. The rates of RVFV IgG were comparable between the age groups and sex. Conversely, the rate of CCHFV IgG was significantly higher among the 41-60 years old participants (p = 0.02). CONCLUSIONS: This study provides a substantial evidence of the circulation of RVFV and CCHFV among rural inhabitants of the East region of Cameroon.
Subject(s)
Ape Diseases/epidemiology , Ape Diseases/virology , Hemorrhagic Fever Virus, Crimean-Congo/immunology , Hemorrhagic Fever, Crimean/veterinary , Pan paniscus , Rift Valley Fever/epidemiology , Rift Valley Fever/virology , Rift Valley fever virus/immunology , Adolescent , Adult , Aged , Aged, 80 and over , Animals , Antibodies, Viral/immunology , Cameroon/epidemiology , Female , Humans , Immunoglobulin G/immunology , Male , Middle Aged , Public Health Surveillance , Seroepidemiologic Studies , Young AdultABSTRACT
BACKGROUND: Rift Valley Fever (RVF) is a mosquito-borne viral zoonosis. To detect RVF virus (RVFV) infection, indirect immunoglobulin G (IgG) and immunoglobulin M (IgM) enzyme linked immunosorbent assays (ELISAs) which utilize recombinant RVFV nucleocapsid (RVFV-N) protein as assay antigen, have reportedly been used, however, there is still a need to develop more sensitive and specific methods of detection. METHODS: RVFV-N protein was expressed in Escherichia coli (E. coli) and purified by histidine-tag based affinity chromatography. This recombinant RVFV-N (rRVFV-N) protein was then used as antigen to develop an IgG sandwich ELISA and IgM capture ELISAs for human sera. Ninety six serum samples collected from healthy volunteers during the RVF surveillance programme in Kenya in 2013, and 93 serum samples collected from RVF-suspected patients during the 2006-2007 RVF outbreak in Kenya were used respectively, to evaluate the newly established rRVFV-N protein-based IgG sandwich ELISA and IgM capture ELISA systems in comparison with the inactivated virus-based ELISA systems. RESULTS: rRVFV-N protein-based-IgG sandwich ELISA and IgM capture ELISA for human sera were established. Both the new ELISA systems were in 100% concordance with the inactivated virus-based ELISA systems, with a sensitivity and specificity of 100%. CONCLUSIONS: Recombinant RVFV-N is a safe and affordable antigen for RVF diagnosis. Our rRVFV-N-based ELISA systems are safe and reliable tools for diagnosis of RVFV infection in humans and especially useful in large-scale epidemiological investigation and for application in developing countries.