RESUMO
Stray dogs and cats pose significant challenges for public health and animal welfare due to their potential involvement in zoonotic disease transmission, accidents, and aggressions. Large urban centers exacerbated challenges due to the presence of these animals in public areas with high human density. Ethical Population Management Programs (EPMP), rooted in the One Health approach, are crucial for addressing this issue comprehensively. This study aimed to demonstrate the approach on cats and dogs EPMP and evaluate the perceptions of academic community regarding EPMP implementation on a campus situated in urban territory. The study was conducted at the Pampulha campus of UFMG in Belo Horizonte, Brazil. In response to issues of animal abandonment and conflicts, the Permanent Commission for Animal Policies (CPPA-UFMG) was established in 2019 to manage the campus's dog, cat, and wildlife populations. The commission implemented the Trap-Neuter-Return (TNR) method, along with health assessments and vaccinations for animals. Interviews were conducted with campus staff to gauge their perception of animal management strategies. Retrospective and prospective analyses of the commission's actions were carried out to assess implementation processes and challenges. The animal population survey conducted on campus between July 2018 and September 2021 revealed a total of 266 animals recorded. Among these animals, 195 were cats (73.3%) and 71 were dogs (26.7%), with the majority being adults. Subsequent surveys in 2019 and 2021 showed a slight increase in the animal population, with measures such as sterilization contributing to population control. Perception analysis among campus users indicated strategies such as TNR were widely endorsed for population control. The employees perception questionnaire was applied to 115 individuals, representing 42 units/departments and five gates. Associations were found between these beliefs and support for institutional actions. The majority favored sterilization (92.17%) and agreed that TNR is an appropriate approach to population control. Overall, the study reflects a community concerned about animal welfare and supportive of measures to address population management and cruelty prevention. The continuous efforts of the university's CPPA have led to stability in the resident animal population, indicating success in achieving population control objectives.
RESUMO
Herpes simplex virus type 1 (HSV-1) is the main etiological agent of acute and sporadic encephalitis. Proteins of the suppressor of cytokine signaling (SOCS) family have shown to regulate the inflammation during HSV-1 infection in the brain. However, the effects of SOCS2 and SOCS3 in viral encephalitis remain unclear. The aim of the current study is to investigate the potential association between SOCS2, SOCS3, cytokines, and hippocampal damage, especially neuronal apoptosis, during acute intracranial HSV-1 infection in mice. Male C57BL/6 mice were infected by intracranial route with 102 plaque-forming units (PFU) inoculum of purified HSV-1. At three days post-infection (3 d.p.i.), mice were euthanized and their hippocampi were collected for histopathological analysis, immunohistochemical reaction against active caspase-3 and quantification of SOCS2, SOCS3 and cytokines (tumoral necrosis factor (TNF), interleukin (IL) 1ß, IL-6, IL-10; interferon (IFN) -α, IFN-ß, IFN-γ) mRNA expression. Infected mice exhibited neuronal loss and hemorrhagic focus in Cornu Ammonis (CA) region. The apoptotic index was higher in infected mice compared to controls. HSV-1 infection was associated with increased hippocampal expression of TNF, IL1-ß, IL-6 and IFNα/IFNß and decreased expression of IL-10, IFN-γ, SOCS2 and SOCS3. Our results suggest that down regulation of SOCS2 and SOCS3 contributes to a pro-inflammatory environment associated with hippocampal damage and neuronal apoptosis during acute HSV-1 infection in mice.
Assuntos
Encefalite por Herpes Simples/metabolismo , Hipocampo/virologia , Inflamação/metabolismo , Proteína 3 Supressora da Sinalização de Citocinas/metabolismo , Proteínas Supressoras da Sinalização de Citocina/metabolismo , Animais , Apoptose/fisiologia , Chlorocebus aethiops , Citocinas/metabolismo , Hipocampo/metabolismo , Masculino , Camundongos , Neurônios/metabolismo , Neurônios/virologia , Células VeroRESUMO
ABSTRACT: In order to detect and identify Campylobacter spp. in broiler chicken carcasses, and to compare detection methods, 43 chilled and 43 frozen carcasses were collected and analyzed. Three methodologies were evaluated: an automated Enzyme Linked Fluorescent Assay (ELFA) VIDAS®30, Polymerase Chain Reaction (PCR) and real-time PCR. Only four chilled carcasses (4.6%) were considered positive for Campylobacter spp. by VIDAS®30 and no sample was positive when the conventional PCR technique was used. However, real-time PCR showed a higher incidence of contamination by Campylobacter spp. in broiler carcasses, with 45 (52.3%) positive samples. C. jejuni was the species most frequently reported in the samples (88.8%). No differences in the frequencies of Campylobacter spp. were observed between the chilled and frozen broiler carcasses. In conclusion, real-time PCR was the most sensitive method for the detection of Campylobacter spp. in chilled or frozen broiler carcasses, which were mainly contaminated by C. jejuni.
RESUMO: Com o objetivo de detectar e identificar Campylobacter spp. em carcaças de frango de corte utilizando três metodologias distintas - ensaio imunoenzimático VIDAS®30, Reação em Cadeia da Polimerase (PCR) e PCR em tempo real - foram coletadas e analisadas 43 carcaças de frango resfriadas e 43 congeladas. Quatro carcaças refrigeradas (4,6%) foram consideradas positivas para Campylobacter spp. pelo VIDAS®30 e nenhuma amostra positiva foi identificada quando utilizada a técnica de PCR. Porém, ao analisar as carcaças pela metodologia da PCR em tempo real, foi observada uma maior incidência de Campylobacter spp., com 45 amostras (52,3%) positivas, sendo que Campylobacter jejuni foi a espécie mais frequentemente encontrada nas amostras (88,8%). Não foi observada diferença na frequência do micro-organismo entre carcaças de frangos resfriadas e congeladas. Concluiu-se que a técnica de PCR em tempo real apresentou maior sensibilidade na detecção de Campylobacter spp. em carcaças de frangos de corte e que foi encontrada elevada presença de carcaças contaminadas, especialmente por C. jejuni.
RESUMO
BACKGROUND: Herpes simplex virus type 1 (HSV-1) cause not only mild symptoms but also blindness and encephalitis. It was previously shown that the immune response against HSV-1 occurs mainly in the trigeminal ganglia (TG) and that Toll-like receptors 2 and 9 (TLR2/9) are important in mediating this response. It was also demonstrated that iNOS (nitric oxide synthase) and interleukin 1 beta (IL-1ß) play an essential role in the defense against HSV-1 infection. Importantly, the present work aimed to identify the primary cells responsible for iNOS and IL-1ß production and search for other important molecules and cells that might or might not depend on TLR2/9 receptors to mediate the immune response against HSV-1. METHODS: C57BL/6 (wild type, WT) and TLR2/9-/- mice were infected by the intranasal route with HSV-1 (1 × 106 p.f.u.). Cells were obtained from the TG and spleen tissues and the profile of immune cells was determined by flow cytometry in infected and mock infected WT and knockout mice. The percentage of cells producing iNOS, IL-1ß, granzyme B and perforin was also determined by flow cytometry. Chemokine monocyte chemoattractant protein-1 (MCP1) was measured by Cytometric Bead Array (CBA) in the TG, spleen and lung. Expression of type I interferons (IFNs), interleukins (IL) 5 and 10, IL-1ß and granzyme B were quantified by real time PCR. RESULTS: The results indicate that dendritic cells (DCs) and monocytes/macrophages (Mo/MÏ) were the main sources of IL-1ß and iNOS, respectively, which, together with type I IFNs, were essential for the immune response against HSV-1. Additionally, we showed that granzyme B produced by CD8+ T and NK lymphocytes and MCP-1 were also important for this immune response. Moreover, our data indicate that the robust production of MCP-1 and granzyme B is either TLR-independent or down regulated by TLRs and occurs in the TG of TLR2/9-/- infected mice. CONCLUSION: Taken together, our data provide strong evidence that the responses mediated by DCs, Mo/MÏ, NK and CD8+ T lymphocytes through IL-1ß, iNOS and granzyme B production, respectively, together with the production of type I IFN early in the infection, are crucial to host defense against HSV-1.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Células Dendríticas/imunologia , Herpesvirus Humano 1/imunologia , Células Matadoras Naturais/imunologia , Macrófagos/imunologia , Gânglio Trigeminal/imunologia , Gânglio Trigeminal/virologia , Animais , Citometria de Fluxo , Granzimas/metabolismo , Humanos , Interferon Tipo I/metabolismo , Interleucina-1beta/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Óxido Nítrico Sintase Tipo II/metabolismo , Receptor 2 Toll-Like/deficiência , Receptor 2 Toll-Like/metabolismo , Receptor Toll-Like 9/deficiência , Receptor Toll-Like 9/metabolismoRESUMO
The most severe manifestation of Herpes Simplex Type 1 virus (HSV-1) infection is encephalitis characterized by arousal impairment and seizures that can evolve to coma and death. Previous studies reported the involvement of suppressor of cytokine signaling (SOCS) proteins, specifically SOCS1 and SOCS3, in HSV-1 infection, suggesting that other members of this family could be involved in the immune response against HSV-1. No previous study has reported the role of SOCS2 in HSV-1 infection. In the current study, C57BL/6 wild-type mice (WT) and mice deficient in SOCS2 gene (SOCS2-/-) were subjected to intracranial inoculation with 102 plaque forming units (PFU) of HSV-1. Survival curve, neuroinflammatory parameters and neuropathology were evaluated. Infected SOCS2-/- mice had increased survival in comparison with infected WT animals. This better outcome was associated with reduced leukocyte infiltration, concentration of cytokines, and structural changes in the brain. SOCS2 seems to play a detrimental role in HSV-1 encephalitis. Moreover, the control of neuroinflammatory response in HSV-1 infection was of paramount importance to clinical outcome.
Assuntos
Encefalite por Herpes Simples/imunologia , Herpes Simples/imunologia , Herpesvirus Humano 1 , Proteínas Supressoras da Sinalização de Citocina/deficiência , Animais , Encéfalo/imunologia , Encéfalo/patologia , Quimiocinas/metabolismo , Chlorocebus aethiops , Citocinas/metabolismo , Modelos Animais de Doenças , Encefalite por Herpes Simples/patologia , Herpes Simples/patologia , Leucócitos/imunologia , Leucócitos/patologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteínas Supressoras da Sinalização de Citocina/genética , Análise de Sobrevida , Células Vero , Carga ViralRESUMO
Herpes simplex virus type 1 (HSV-1) is a human pathogen that may cause severe encephalitis. The exacerbated immune response against the virus contributes to the disease severity and death. Platelet activating factor (PAF) is a mediator capable of inducing increase in vascular permeability, production of cytokines on endothelial cells and leukocytes. We aimed to investigate the activation of PAF receptor (PAFR) and its contribution to the severity of the inflammatory response in the brain following HSV-1 infection. C57BL/6 wild-type (WT) and PAFR deficient (PAFR-/-) mice were inoculated intracranially with 104 plaque-forming units (PFU) of HSV-1. Visualization of leukocyte recruitment was performed using intravital microscopy. Cells infiltration in the brain tissue were analyzed by flow cytometry. Brain was removed for chemokine assessment by ELISA and for histopathological analysis. The pharmacological inhibition by the PAFR antagonist UK-74,505 was also analyzed. In PAFR-/- mice, there was delayed lethality but no difference in viral load. Histopathological analysis of infected PAFR-/- mice showed that brain lesions were less severe when compared to their WT counterparts. Moreover, PAFR-/- mice showed less TCD4+, TCD8+ and macrophages in brain tissue. This reduction of the presence of leukocytes in parenchyma may be mechanistically explained by a decrease in leukocytes rolling and adhesion. PAFR-/- mice also presented a reduction of the chemokine CXCL9 in the brain. In addition, by antagonizing PAFR, survival of C57BL/6 infected mice increased. Altogether, our data suggest that PAFR plays a role in the pathogenesis of experimental HSV-1 meningoencephalitis, and its blockade prevents severe disease manifestation.
Assuntos
Herpes Simples/metabolismo , Herpesvirus Humano 1 , Meningoencefalite/metabolismo , Glicoproteínas da Membrana de Plaquetas/antagonistas & inibidores , Glicoproteínas da Membrana de Plaquetas/deficiência , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Receptores Acoplados a Proteínas G/deficiência , Índice de Gravidade de Doença , Animais , Encéfalo/patologia , Encéfalo/virologia , Di-Hidropiridinas/farmacologia , Di-Hidropiridinas/uso terapêutico , Herpes Simples/patologia , Herpes Simples/prevenção & controle , Imidazóis/farmacologia , Imidazóis/uso terapêutico , Masculino , Meningoencefalite/patologia , Meningoencefalite/prevenção & controle , Camundongos , Camundongos Endogâmicos C57BL , Camundongos KnockoutRESUMO
This study presents the first description of Bovine herpesvirus 6 (BoHV-6) that was isolated from buffaloes of Amazon region in Brazil. Phylogenetic analysis showed that the BoHV-6 Brazilian strains clustered with the sequence of BoHV-6 from elsewhere available at the GenBank. It was observed in some buffaloes with lymphoproliferative disease in one herd, thus the animals were also tested for Bovine leukemia virus (BLV), which has been associated to lymphoma in bovines. All animals were negative to BLV. These results indicate that BoHV-6 is present in buffaloes in Brazil, but the importance and impact of this infection and its association with any illness is still undefined.
Assuntos
Infecções por Herpesviridae/veterinária , Varicellovirus/isolamento & purificação , Animais , Brasil/epidemiologia , Búfalos , DNA Viral/genética , Infecções por Herpesviridae/epidemiologia , Infecções por Herpesviridae/virologia , Filogenia , Reação em Cadeia da Polimerase/veterinária , Varicellovirus/genéticaRESUMO
The aims of this study were to address the protective immune response induced by S19 vaccination (n=10) and RB51 revaccination, in pregnant (n=9) and non-pregnant (n=10) S19 calfhood-vaccinated cattle as follows: evaluate the in vitro CD4(+) and CD8(+) T-lymphocytes specific proliferation, and in vitro expression of IFN-γ by CD4(+) and CD8(+) T-cells and IL-4 by CD4(+), CD8(+) and CD21(+) lymphocytes subset. Upon in vitro stimulation with γ-irradiated Brucella abortus 2308, blood mononuclear cells from S19 vaccinated and RB51 revaccinated cows exhibited significantly higher proliferation of CD4(+) and CD8(+) T-lymphocytes and CD4(+)IFN-γ(+) T-cells compared to non-vaccinated animals. RB51 revaccination, regardless of the pregnancy status, did not enhance the proliferation of CD4(+) or CD8(+) T-cells nor IFN-γ or IL-4 production. Data from the present study suggest that cattle's cellular immune response induced after brucellosis vaccination and revaccination is due to CD4(+) and CD8(+) T-lymphocytes, being CD4(+) T-cells the main source of IFN-γ.
Assuntos
Vacina contra Brucelose/imunologia , Brucelose Bovina/prevenção & controle , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Citocinas/imunologia , Animais , Brucella abortus , Bovinos , Feminino , Imunização Secundária , Interferon gama/imunologia , Interleucina-4/imunologia , Gravidez , Vacinação/veterináriaRESUMO
BACKGROUND: Herpes simplex 1 (HSV-1) causes various human clinical manifestations, ranging from simple cold sores to encephalitis. Innate immune cells recognize pathogens through Toll-like receptors (TLRs), thus initiating the immune response. Previously, we demonstrated that the immune response against HSV-1 is dependent on TLR2 and TLR9 expression and on IFN gamma production in the trigeminal ganglia (TG) of infected mice. In this work, we further investigated the cells, molecules, and mechanisms of HSV-1 infection control, especially those that are TLR-dependent. METHODS: C57BL/6 wild-type (WT), TLR2-/-, TLR9-/-, and TLR2/9-/- mice were intranasally infected with HSV-1. On the viral peak day, the TG and brains were collected from mice and TLR expression was measured in the TG and brain and inducible nitric oxide synthase (iNOS) expression was measured in the TG by real-time PCR. Immunofluorescence assays were performed in mice TG to detect iNOS production by F4/80+ cells. Intraperitoneal macrophages nitric oxide (NO) production was evaluated by the Griess assay. WT, CD8-/-, RAG-/-, and iNOS-/- mice were intranasally infected in a survival assay, and their cytokine expression was measured in the TG by real-time PCR. RESULTS: Infected WT mice exhibited significantly increased TLR expression, compared with their respective controls, in the TG but not in the brain. TLR-deficient mice had moderately increased TLR expression in the TG and brain in compare with the non-infected animals. iNOS expression in the WT infected mice TG was higher than in the other groups with increased production by macrophages in the WT infected mice, which did not occur in the TLR2/9-/- mice. Additionally, the intraperitoneal macrophages of the WT mice had a higher production of NO compared with those of the TLR-deficient mice. The CD8-/-, RAG-/-, and iNOS-/- mice had 100% mortality after the HSV-1 infection compared with 10% of the WT mice. Cytokines were overexpressed in the iNOS-/- infected mice, while the RAG-/- mice were nearly unresponsive to the virus. CONCLUSION: TLRs efficiently orchestrate the innate immune cells, eliciting macrophage response (with NO production by the macrophages), thereby controlling the HSV-1 infection through the immune response in the TG of mice.
Assuntos
Herpes Simples/patologia , Herpesvirus Humano 1/fisiologia , Óxido Nítrico Sintase Tipo II/metabolismo , Receptor 2 Toll-Like/metabolismo , Receptor Toll-Like 9/metabolismo , Gânglio Trigeminal/metabolismo , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Encéfalo/virologia , Citocinas/metabolismo , Modelos Animais de Doenças , Antígenos do Núcleo do Vírus da Hepatite B/metabolismo , Humanos , Linfócitos/efeitos dos fármacos , Linfócitos/virologia , Macrófagos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , RNA Mensageiro/metabolismo , Receptores Imunológicos/metabolismo , Tioglicolatos/uso terapêutico , Receptor 2 Toll-Like/genética , Receptor Toll-Like 9/genética , Proteínas do Core Viral/metabolismoRESUMO
BACKGROUND: The neuroinflammatory response aimed at clearance of herpes simplex virus-1 (HSV-1) plays a key role in the pathogenesis of neuroaxonal damage in herpetic encephalitis. Leukocytes activated in an adaptive immune response access brain tissue by passing through the blood-brain barrier. The chemokine CCL5/RANTES is involved in recruitment of these cells to the brain acting via the receptors CCR1, CCR3 and mainly CCR5. Here, we evaluated the role of CCR5 on traffic of leukocytes in the brain microvasculature, cellular and cytokines profile in a severe form of herpetic encephalitis. RESULTS: Wild type and mice lacking CCR5 (CCR5-/-) were inoculated intracerebrally with 104 PFU of neurotropic HSV-1. We evaluated the traffic of leukocytes in the brain microvasculature using intravital microscopy and the profile of cytokines by Enzyme-Linked Immunosorbent Assay at 1 day post infection. Flow cytometry and histopathological analyses were also carried out in brain tissue. Absence of CCR5 leads to lower viral load and an increased leukocyte adhesion in brain microvasculature, predominantly of neutrophils (CD11+ Ly6G+ cells). Moreover, there was a significant increase in the levels of MIP-1/CCL2, RANTES/CCL5, KC/CXCL1 and MIG/CXCL9 in the brain of infected CCR5-/- mice. CONCLUSIONS: These results suggest that the absence of CCR5 may boost the immune response with a high neutrophil recruitment which most likely helps in viral clearance. Nonetheless, the elevated immune response may be detrimental to the host.
Assuntos
Encefalite por Herpes Simples/imunologia , Encefalite por Herpes Simples/patologia , Regulação da Expressão Gênica/genética , Infiltração de Neutrófilos/fisiologia , Receptores CCR5/deficiência , Animais , Antígenos Ly/metabolismo , Encéfalo/imunologia , Encéfalo/patologia , Encéfalo/virologia , Adesão Celular/genética , Adesão Celular/imunologia , Citocinas/metabolismo , Modelos Animais de Doenças , Encefalite por Herpes Simples/genética , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Leucócitos/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores CCR5/genéticaRESUMO
Herpes simplex virus-1 (HSV-1) is a pathogen that may cause severe encephalitis in humans. In this study, we aimed to investigate the role of interleukin-4 (IL-4) in a model of HSV-1 brain infection. IL-4 knockout (IL-4-/-) and wild type (WT) C57BL/6 mice were inoculated with 10(4) plaque-forming units of HSV-1 by the intracranial route. Histopathologic analysis revealed a distinct profile of infiltrating cells at 3 days post-infection (dpi). Infected WT mice presented mononuclear inflammatory cells while IL-4-/- mice developed meningoencephalitis with predominance of neutrophils. IL-4-/- mice had diminished leukocyte adhesion at 3 dpi when compared to infected WT animals in intravital microscopy study. Conversely no differences were found in cerebral levels of CXCL1, CXCL9, CCL3, CCL5 and TNF-α between WT and IL-4-/- infected mice. IL-4 may play a role in the recruitment of cells into central nervous system in this acute model of severe encephalitis caused by HSV-1.
Assuntos
Quimiocinas/imunologia , Encefalite por Herpes Simples/imunologia , Herpesvirus Humano 1/imunologia , Interleucina-4/imunologia , Fator de Necrose Tumoral alfa/imunologia , Doença Aguda , Animais , Movimento Celular/imunologia , Modelos Animais de Doenças , Encefalite por Herpes Simples/patologia , Interleucina-4/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
Herpes simplex virus-1 (HSV-1) is a pathogen that may cause severe encephalitis in humans. In this study, we aimed to investigate the role of interleukin-4 (IL-4) in a model of HSV-1 brain infection. IL-4 knockout (IL-4-/-) and wild type (WT) C57BL/6 mice were inoculated with 10(4) plaque-forming units of HSV-1 by the intracranial route. Histopathologic analysis revealed a distinct profile of infiltrating cells at 3 days post-infection (dpi). Infected WT mice presented mononuclear inflammatory cells while IL-4-/- mice developed meningoencephalitis with predominance of neutrophils. IL-4-/- mice had diminished leukocyte adhesion at 3 dpi when compared to infected WT animals in intravital microscopy study. Conversely no differences were found in cerebral levels of CXCL1, CXCL9, CCL3, CCL5 and TNF-α between WT and IL-4-/- infected mice. IL-4 may play a role in the recruitment of cells into central nervous system in this acute model of severe encephalitis caused by HSV-1.
O vírus herpes simplex-1 (HSV-1) é um patógeno que pode causar encefalite grave em humanos. Neste estudo, buscamos investigar o papel da interleucina-4 (IL-4) no modelo de infecção intracerebral por HSV-1. Camundongos C57BL/6 selvagens (WT) e deficientes no gene IL-4 (IL-4-/-) foram inoculados com 10(4) unidades formadoras de placas de HSV-1 por via intracraniana. A análise histopatológica revelou um padrão distinto de infiltrado leucocitário. Camundongos WT infectados apresentaram infiltrado de células mononucleares, enquanto camundongos IL-4-/- desenvolveram meningoencefalite com predomínio de neutrófilos 3 dias pós-infecção (dpi). Animais IL-4-/- tiveram menor adesão de leucócitos 3 dpi quando comparados aos animais WT infectados à microscopia intravital. Em contrapartida, não foram encontradas diferenças nos níveis cerebrais de CXCL1, CXCL9, CCL3, CCL5 e TNF-α entre camundongos WT e IL-4-/- infectados. Esses resultados sugerem que IL-4 pode desempenhar um papel no recrutamento de células no sistema nervoso central neste modelo agudo de encefalite grave causada pelo HSV-1.
Assuntos
Animais , Masculino , Camundongos , Quimiocinas/imunologia , Encefalite por Herpes Simples/imunologia , Herpesvirus Humano 1/imunologia , /imunologia , Fator de Necrose Tumoral alfa/imunologia , Doença Aguda , Movimento Celular/imunologia , Modelos Animais de Doenças , Encefalite por Herpes Simples/patologia , /fisiologiaRESUMO
Herpes simplex virus 1 (HSV-1) is a neurotropic DNA virus that is responsible for several clinical manifestations in humans, including encephalitis. HSV-1 triggers toll-like receptors (TLRs), which elicit cytokine production. Viral multiplication and cytokine expression in C57BL/6 wild-type (WT) mice infected with HSV-1 were evaluated. Virus was found in the trigeminal ganglia (TG), but not in the brains of animals without signs of encephalitis, between 2 and 6 days postinfection (d.p.i.). Cytokine expression in the TG peaked at 5 d.p.i. TLR9-/- and TLR2/9-/- mice were more susceptible to the virus, with 60% and 100% mortality, respectively, as opposed to 10% in the WT and TLR2-/- mice. Increased levels of both CXCL10/IP-10 and CCL2/MCP-1, as well as reduced levels of interferon-γ and interleukin 1-ß transcripts, measured in both the TG and brains at 5 d.p.i., and the presence of virus in the brain were correlated with total mortality in TLR2/9-/- mice. Cytokine alterations in TLR2/9-/- mice coincided with histopathological changes in their brains, which did not occur in WT and TLR2-/- mice and occurred only slightly in TLR9-/- mouse brain. Increased cellularity, macrophages, CD8 T cells producing interferon-γ, and expression levels of TLR2 and TLR9 were detected in the TG of WT-infected mice. We hypothesize that HSV-1 infection is controlled by TLR-dependent immune responses in the TG, which prevent HSV-1 encephalitis.
Assuntos
Herpes Simples/imunologia , Herpes Simples/virologia , Herpesvirus Humano 1/imunologia , Receptor 2 Toll-Like/imunologia , Receptor Toll-Like 9/imunologia , Gânglio Trigeminal/imunologia , Gânglio Trigeminal/virologia , Animais , Encéfalo/imunologia , Encéfalo/patologia , Encéfalo/virologia , Linhagem Celular , Quimiocinas/sangue , Quimiocinas/imunologia , Citocinas/sangue , Citocinas/imunologia , Herpesvirus Humano 1/patogenicidade , Humanos , Subunidade p40 da Interleucina-12/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptor 2 Toll-Like/genética , Receptor Toll-Like 9/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Herpes simplex virus-1 (HSV-1) is a pathogen for humans that may cause severe encephalitis. Tumor necrosis factor alpha (TNF-alpha) plays a role in several viral diseases of the central nervous system (CNS). The classic proinflammatory activities of TNF-alpha are mediated mainly through activation of the receptor 1 for TNF-alpha (TNFR1). However, when HSV-1 is inoculated in the periphery, TNF-alpha seems to protect C57Bl/6 mice against encephalitis by a mechanism independent of TNFR1. This study aims to investigate the role of TNFR1 in HSV-1 encephalitis induced by the inoculation of the virus into the brain. Wild-type C57BL/6 (WT) and TNFR1(-/-) were inoculated with 10(2) plaque-forming units of HSV-1 by the intracranial route. Infection with HSV-1 was lethal in TNFR1(-/-) mice in early times after infection. TNFR1(-/-) mice had reduced expression of the chemokines CCL3 and CCL5, and decreased leukocyte adhesion in the brain vasculature compared to WT mice 4 days post-infection (dpi). At this time point TNFR1(-/-) infected mice also had higher HSV-1 viral replication and more injuries in the brain, especially in the hippocampus. In conclusion, TNFR1 seems to play a relevant role in the control of viral replication in the CNS when HSV-1 is inoculated by intracranial route.
Assuntos
Encefalite por Herpes Simples/metabolismo , Herpesvirus Humano 1 , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Animais , Encéfalo/irrigação sanguínea , Encéfalo/metabolismo , Encéfalo/patologia , Quimiocina CCL3/metabolismo , Quimiocina CCL5/metabolismo , Encefalite por Herpes Simples/imunologia , Encefalite por Herpes Simples/patologia , Endotélio Vascular/metabolismo , Endotélio Vascular/patologia , Leucócitos/metabolismo , Leucócitos/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores Tipo I de Fatores de Necrose Tumoral/genética , Índice de Gravidade de Doença , Análise de SobrevidaRESUMO
The Herpes simplex virus-1 (HSV-1) is responsible for several clinical manifestations in humans, including encephalitis. To induce encephalitis, C57BL/6 mice were inoculated with 10(4) plaque-forming cells of HSV-1 by the intracranial route. Met-RANTES (regulated upon activation, normal T cell expressed and presumably secreted) (10 microg/mouse), a CC chemokine family receptor (CCR)1 and CCR5 antagonist, was given subcutaneously the day before, immediately after, and at days 1, 2, and 3 after infection. Treatment with Met-RANTES had no effect on the viral titers. In contrast, intravital microscopy revealed that treatment with Met-RANTES decreased the number of leukocytes adherent to the pial microvasculature at days 1 and 3 after infection. The levels of the chemokines CCL3, CCL5, CXCL1, and CXCL9 increased after infection and were enhanced further by the treatment with Met-RANTES. Treatment with a polyclonal anti-CCL5 antibody 2 h before the intravital microscopy decreased leukocyte adhesion in the microcirculation of infected mice. In conclusion, CCL5, a chemokine that binds to CCR1 and CCR5, is essential for leukocyte adhesion during HSV-1 encephalitis. However, blocking of CCR1 and CCR5 did not affect HSV-1 replication, suggesting that other immune mechanisms are involved in the process of infection control.