RESUMEN
OBJECTIVE: The laboratory diagnostic detection of herpes simplex virus (HSV) from eye samples must be practical, timely, and definitive for appropriate therapy. Although polymerase chain reaction (PCR) and/or cell culture can be definitive, HSV results can be delayed. Enzyme Linked Virus Inducible System (ELVIS) is a test that can provide results within 24 to 48 hr. We evaluated "AmpliVue HSV 1+2 Assay" as a molecular colorimetric test that can detect HSV (1 or 2) DNA within 1 hr. METHODS: Cornea/conjunctival samples were tested retrospectively with AmpliVue against 53 true-positive and 20 true-negative specimens collected in chlamydial transport medium. All clinical specimens were tested by cell culture isolation, PCR, and ELVIS for routine patient care. RESULTS: The sensitivity of AmpliVue against ocular samples that were both culture-positive and PCR-positive was 84%. The specificity of AmpliVue was 100%. Only one clinical sample was HSV-2 positive, whereas all others tested positive for HSV-1. Based on PCR-positive and cell culture-negative samples, AmpliVue (11 of 17) tested more positive than ELVIS (0 of 17) (P=0.003, Fisher Exact). CONCLUSIONS: AmpliVue is moderately sensitive and highly specific as a practical and timely diagnostic test for detecting ocular HSV. Expertise is readily achieved and the test is straightforward with easy interpretation. Negative AmpliVue testing must be confirmed with PCR. AmpliVue has potential as an office-based diagnostic test.
Asunto(s)
Conjuntiva/virología , Córnea/virología , ADN Viral/análisis , Infecciones Virales del Ojo/diagnóstico , Queratitis Herpética/diagnóstico , Simplexvirus/genética , Proteínas Bacterianas , Células Cultivadas , Conjuntiva/patología , Córnea/patología , Infecciones Virales del Ojo/virología , Humanos , Queratitis Herpética/virología , Curva ROC , Proteínas Represoras , Estudios RetrospectivosRESUMEN
OBJECTIVE:: Bacterial keratitis occurs worldwide, and despite recent developments, it remains a potentially blinding condition. This study assesses the presence of herpes simplex virus (HSV-1 and -2) and varicella zoster virus (VZV) by quantitative real-time polymerase chain reaction (qPCR) in corneal scrapings from patients with bacterial keratitis. METHODS:: A total of 65 patients with clinical diagnoses of infectious corneal ulcers prospectively underwent clinical eye examinations. Corneal scrapings were investigated by Gram staining, Giemsa staining, culture, and qPCR (the study group). Risk factors and epidemiological data were recorded. The control group comprising 25 eyes with typical herpes dendritic keratitis was also analyzed by qPCR. RESULTS:: From the study group (n=65), nine patients (13.8%) had negative smears, cultures, and qPCR findings. Fifty-six (86.2%) patients had positive cultures: 51 for bacteria, 4 for fungi, and 1 for amoebae. Of the patients who had positive bacterial cultures, qPCR identified 10 patients who were also positive for virus: one for VZV and nine for HSV-1. Of the 25 patients in the control group, 21 tested positive for HSV-1 by qPCR analysis. CONCLUSIONS:: Herpes may be present in patients with bacterial corneal ulcers, and qPCR may be useful in its detection.
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Córnea/virología , Herpesvirus Humano 1/aislamiento & purificación , Herpesvirus Humano 2/aislamiento & purificación , Herpesvirus Humano 3/aislamiento & purificación , Queratitis Dendrítica/microbiología , Queratitis/microbiología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Sondas de ADN , Infecciones Bacterianas del Ojo/microbiología , Femenino , Humanos , Queratitis/diagnóstico , Queratitis/virología , Queratitis Dendrítica/diagnóstico , Queratitis Dendrítica/virología , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Adulto JovenRESUMEN
ABSTRACT Objective: Bacterial keratitis occurs worldwide, and despite recent developments, it remains a potentially blinding condition. This study assesses the presence of herpes simplex virus (HSV-1 and -2) and varicella zoster virus (VZV) by quantitative real-time polymerase chain reaction (qPCR) in corneal scrapings from patients with bacterial keratitis. Methods: A total of 65 patients with clinical diagnoses of infectious corneal ulcers prospectively underwent clinical eye examinations. Corneal scrapings were investigated by Gram staining, Giemsa staining, culture, and qPCR (the study group). Risk factors and epidemiological data were recorded. The control group comprising 25 eyes with typical herpes dendritic keratitis was also analyzed by qPCR. Results: From the study group (n=65), nine patients (13.8%) had negative smears, cultures, and qPCR findings. Fifty-six (86.2%) patients had positive cultures: 51 for bacteria, 4 for fungi, and 1 for amoebae. Of the patients who had positive bacterial cultures, qPCR identified 10 patients who were also positive for virus: one for VZV and nine for HSV-1. Of the 25 patients in the control group, 21 tested positive for HSV-1 by qPCR analysis. Conclusions: Herpes may be present in patients with bacterial corneal ulcers, and qPCR may be useful in its detection.
RESUMO Objetivo: Ceratites bacterianas ocorrem mundialmente e apesar dos novos desenvolvimentos permanece como uma condição que pode levar à cegueira. Avaliar a presença de herpes simples (-1 e -2) e vírus varicella zoster (VZV) por reação em cadeia quantitativa de polimerase em tempo real (qPCR) em raspados corneanos de pacientes com ceratite bacteriana. Métodos: Sessenta e cinco pacientes com ceratite infecciosa foram submetidos a raspados corneanos estudados para gram, Giemsa, cultura e qPCR (grupo de estudo). Foram avaliados fatores de risco e epidemiológicos. O grupo controle foi composto por 25 casos de úlcera dendrítica típica por herpes analisados por qPCR. Resultados: Do grupo de estudo (n=65), nove pacientes (13,8%) apresentaram cultura, qPCR e raspado negativos. Cinquenta e seis (86,2%) pacientes apresentaram cultura positiva, 51 para bacteria, 4 para fungo e 1 para ameba. A qPCR identificou 10 pacientes do grupo de cultura positiva para bactéria que também foram positivos para vírus, um VZV e 9 para HSV-1. Dos 25 pacientes que compunham o grupo controle, 21 apresentaram qPCR positivo para HSV-1. Conclusão: Herpes pode estar presente em pacientes com úlceras de córnea bacterianas e a qPCR pode ser útil na sua detecção.
Asunto(s)
Humanos , Masculino , Femenino , Niño , Adolescente , Adulto , Persona de Mediana Edad , Anciano , Anciano de 80 o más Años , Adulto Joven , Queratitis Dendrítica/microbiología , Herpesvirus Humano 2/aislamiento & purificación , Herpesvirus Humano 1/aislamiento & purificación , Herpesvirus Humano 3/aislamiento & purificación , Córnea/virología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Queratitis/microbiología , Sondas de ADN , Infecciones Bacterianas del Ojo/microbiología , Queratitis Dendrítica/diagnóstico , Queratitis Dendrítica/virología , Estudios Prospectivos , Queratitis/diagnóstico , Queratitis/virologíaRESUMEN
Objetive: The purpose of this study is to map the serological profile of candidates to corneal donation at Irmandade Santa Casa de Misericórdia de Porto Alegre, identifying the percentage of disposal by serology and the marker involved. Methods: There have been analised – retrospectively – the results of serology of all corneal donors, made between the period of 1st january 2006 and 31st december 2012. Data analised were related to age, gender and the results of serology pertinent to viral markers (HBsAg, anti-HBc, anti-HCV and anti-HIV), these, determined by immunosorbent tests (ELISA). Results: In the period of the study, there were 2476 corneal donors at the institution, with a major incidence on the male gender, on an average of 58.7 years old. 23% of retention because of serological unfitness was also identified, that is, 570 samples were non-negative to any of the used tests. The marker anti- HBc was the most prevalent on the studied population, followed by the Hepatitis C virus (HCV) and by the Human Immunodeficiency Virus (HIV). Conclusion: From the data found through this study, it is essential to have the participation of an efficient service on the serological evaluation of the candidates to corneal donation, once the security of the receptor must be taken into consideration in a population of donors with 23% of unfitness prevalence, in which the most prevalent marker is the one of Hepatits B. .
Objetivo: O intento deste desígnio é mapear o perfil sorológico dos candidatos a doação de córneas na Irmandade Santa Casa de Misericórdia de Porto Alegre, identificando o percentual de descarte por sorologia e o marcador envolvido. Métodos: Foram analisados – retrospectivamente – os resultados da sorologia de todos os doadores de córneas compreendidos entre 01 de janeiro de 2006 a 31 de dezembro de 2012. Foram avaliados os dados de idade, sexo e os resultados da sorologia pertinentes aos marcadores virais (HBsAg, anti-HBc, anti-HCV e anti-HIV) determinados por testes imunoenzimáticos (ELISA). Resultados: No período coberto pelo estudo, houve 2476 doadores de córneas na instituição, com maior ocorrência do sexo masculino e média de 58,7 anos de idade. Foram verificados 23,0% de retenção por inaptidão sorológica, ou seja, 570 amostras mostraram-se não-negativas para qualquer dos testes empregados. O marcador anti-HBc foi o mais prevalente na população aferida, seguido pelo vírus da hepatite C (HCV) e pelo vírus da imunodeficiência humana (HIV). Conclusão: Diante dos dados encontrados por este estudo, torna-se decisiva a participação de um serviço eficaz no tangente à avaliação sorológica dos candidatos à doação de córnea, uma vez que a segurança do receptor deve ser considerada numa população de doadores com prevalência de retenção por inaptidão sorológica de 23,0%, donde o marcador mais prevalente é o de hepatite B. .
Asunto(s)
Humanos , Masculino , Femenino , Persona de Mediana Edad , Donantes de Tejidos/estadística & datos numéricos , Biomarcadores , Trasplante de Córnea , Córnea/inmunología , Córnea/virología , Manejo de Especímenes/métodos , Obtención de Tejidos y Órganos/métodos , Obtención de Tejidos y Órganos/estadística & datos numéricos , Virosis/diagnóstico , Pruebas Serológicas/métodos , Anticuerpos Anti-VIH/sangre , Centrifugación , Técnicas para Inmunoenzimas/métodos , Anticuerpos contra la Hepatitis C/sangre , Recolección de Tejidos y Órganos/estadística & datos numéricos , Selección de Donante/normas , Selección de Donante/estadística & datos numéricos , Anticuerpos contra la Hepatitis B/sangre , Antígenos del Núcleo de la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/sangreRESUMEN
PURPOSE: To investigate discarded corneas due to positive serologic tests in donors from the Hospital São Paulo Eye Bank (BOHSP) during a two-year period. METHODS: Retrospective study of records from cornea donors between January 2006 and December 2007. Information such as serologic test results (Hepatitis B, C, and HIV), source of corneal tissue, donor's gender and age were tested for correlation. RESULTS: 902 corneas were processed by BOHSP; 12.9% (116) were discarded due to donor's positive test for hepatitis B, C, or HIV; 20.5% (185) were also discarded due to inconclusive result of serological tests; percentage of corneas discarded due to positive or inconclusive serological tests during this period was 33.4% (301). CONCLUSION: This study confirms the importance of serological tests in order to prevent disease transmission to corneal transplant recipients. However, new tests are necessary to decrease the number of inconclusive tests and decrease the number of discarded corneas.
Asunto(s)
Córnea/virología , Bancos de Ojos/estadística & datos numéricos , Infecciones por VIH/diagnóstico , Hepatitis B/diagnóstico , Hepatitis C/diagnóstico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Bancos de Ojos/normas , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Pruebas Serológicas , Donantes de Tejidos/estadística & datos numéricos , Recolección de Tejidos y Órganos/estadística & datos numéricos , Adulto JovenRESUMEN
Objetivos: Investigar descarte de córneas por sorologia positiva de doadores no Banco de Olhos do Hospital São Paulo (BOHSP) durante período de dois anos. Métodos: Revisão retrospectiva de prontuários de doadores de córneas do BOHSP entre janeiro de 2006 a dezembro de 2007.Correlações de dados como sorologias dos doadores (Hepatites B, C e HIV), procedência, sexo e faixa etária dos mesmos foram testados. Resultados:Das 902 córneas processadas no BOHSP, 12,9% (116) apresentaramresultado positivo para hepatite B, hepatite C ou HIV em seus doadores;20,5% (185) apresentaram testes sorológicos inconclusivos perfazendouma porcentagem de 33,4% (301) de córneas descartadas no período estudado devido a resultado positivo ou inconclusivo da sorologia dos doadores. Conclusão: O estudo confirma a validade e a importância da realização de testes sorológicos para prevenção de doenças que poderiam ser transmitidas aos eventuais receptores de córneas. Entretanto, testes mais eficazes são necessários para diminuir o número de casos de sorologias inconclusivas, visando diminuir o descarte de córneas que poderiam ser utilizadas.
Purpose: To investigate discarded corneas due to positive serologic tests in donors from the Hospital São Paulo EyeBank (BOHSP) during a two-year period. Methods: Retrospective study of records from cornea donors between January 2006 and December 2007. Information such as serologic test results (Hepatitis B, C, and HIV), source of corneal tissue, donors gender and age were tested for correlation. Results:902 corneas were processed by BOHSP; 12.9% (116) were discarded due to donors positive test for hepatitis B, C, orHIV; 20.5% (185) were also discarded due to inconclusive result of serological tests; percentage of corneas discardeddue to positive or inconclusive serological tests during this period was 33.4% (301). Conclusion: This study confirms the importance of serological tests in order to prevent disease transmission to corneal transplant recipients. However, new tests are necessary to decrease the number of inconclusive tests and decrease the number of discarded corneas.
Asunto(s)
Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Córnea/virología , Bancos de Ojos/estadística & datos numéricos , Infecciones por VIH/diagnóstico , Hepatitis B/diagnóstico , Hepatitis C/diagnóstico , Bancos de Ojos/normas , Estudios Retrospectivos , Pruebas Serológicas , Donantes de Tejidos/estadística & datos numéricos , Recolección de Tejidos y Órganos/estadística & datos numéricos , Adulto JovenRESUMEN
Meliacine (MA), an antiviral principle present in partially purified leaf extracts of Melia azedarach L., prevents the development of herpetic stromal keratitis (HSK) in mice by diminishing the viral load in the eye and the severity of lesions caused by a virus-induced immunopathological reaction. The tetranortriterpenoid 1-cinnamoyl-3,11-dihydroxymeliacarpin (CDM), obtained from MA purification, displays anti-herpetic activity and impedes nuclear factor kappaB (NF-kappaB) activation in HSV-1 infected conjunctival cells. To extend our understanding about CDM biological properties, we investigated its anti-HSV-1 activity as well as the effect on NF-kappaB activation and cytokine secretion induced by viral (HSV-1) and no-viral (LPS) stimuli, in corneal cells and macrophages. CDM exerted a potent anti-HSV-1 effect on corneal cells and inhibited NF-kappaB translocation to the nucleus, leading to a decrease in IL-6 production. Besides, CDM seemed to modulate IL-6 and TNF-alpha responses in macrophages, whether they were infected with HSV-1 or stimulated with LPS. However, CDM did not affect NF-kappaB activation in these cells, suggesting that an alternative NF-kappaB cell signaling pathway would be involved in the modulation of cytokine production. We conclude that, in addition to its antiviral effect, CDM would be acting as an immunomodulating compound which would be responsible for the improvement of murine HSK already reported.
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Antivirales/farmacología , Herpesvirus Humano 1/efectos de los fármacos , Queratitis Herpética/tratamiento farmacológico , Queratitis Herpética/inmunología , Limoninas/farmacología , Melia azedarach/química , Animales , Antivirales/química , Línea Celular , Células Cultivadas , Córnea/inmunología , Córnea/virología , Citocinas/inmunología , Humanos , Queratitis Herpética/virología , Limoninas/química , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , RatonesRESUMEN
PURPOSE: To document etiology and predictive value of clinical diagnosis in laboratory confirmed viral diseases. METHODS: Reports of culture-positive cases of samples collected from patients presenting from January 1987 - December 2001 were evaluated. RESULTS: One thousand nine hundred and sixty-four (1964) cultures were submitted during 1987-2001. Twenty-six percent were positive (514). Human herpesvirus 1 was the most frequent agent isolated from all positive culture (56%). Adenovirus was the most common virus isolated from conjunctiva (66%), human herpesvirus 1 from lid and cornea (76%, 88%) and cytomegalovirus from vitreous (27%). Some unusual pathogens were recovered from conjunctiva as cytomegalovirus and from cornea as adenovirus, enterovirus and cytomegalovirus. Recognition of common viral syndromes was human herpesvirus 1 (88%), epidemic keratoconjunctivitis (88%), acute hemorrhagic conjunctivitis (70%) and varicella zoster virus (100%). However, some misdiagnosed cases were observed. Thirteen percent of conjunctivitis thought to be caused by herpes were due to adenovirus, 3.2% to Enterovirus, 3.2% to varicella zoster virus and 3.2% to human cytomegalovirus. Also, 5% of cases with a clinical diagnosis of herpes keratitis were caused by adenovirus and 2.7% by enterovirus. Finally, 4.8% of cases thought to be adenovirus conjunctivitis were herpes conjunctivitis. CONCLUSIONS: Human herpesvirus 1 remains the most frequently isolated virus from ocular sites in general (56%). Nonherpetic corneal isolates were in decreasing order: adenovirus, enterovirus and cytomegalovirus. Clinical and laboratory correlation was less than 90%. The most misdiagnosed cases were herpes conjunctivitis and keratitis, some cases of adenovirus conjunctivitis some cases of acute hemorrhagic conjunctivitis. It is essential that a rapid and specific diagnosis is offered under atypical viral presentation for the institution of specific antiviral therapy and to avoid complications that can be a result of misdiagnosis and inappropriate treatment. Also it is important to do viral testing in order to confirm clinical diagnosis, report emerging infections, resistance and change in the epidemiology.
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Infecciones Virales del Ojo/diagnóstico , Queratoconjuntivitis/diagnóstico , Brasil , Conjuntivitis Hemorrágica Aguda/diagnóstico , Conjuntivitis Hemorrágica Aguda/virología , Córnea/virología , Virus ADN/aislamiento & purificación , Errores Diagnósticos/prevención & control , Enterovirus/aislamiento & purificación , Infecciones Virales del Ojo/virología , Reacciones Falso Positivas , Herpes Zóster/virología , Humanos , Queratoconjuntivitis/virología , Valor Predictivo de las Pruebas , Estaciones del Año , Factores de Tiempo , Cultivo de VirusRESUMEN
PURPOSE: To document etiology and predictive value of clinical diagnosis in laboratory confirmed viral diseases. METHODS: Reports of culture-positive cases of samples collected from patients presenting from January 1987 - December 2001 were evaluated. RESULTS: One thousand nine hundred and sixty-four (1964) cultures were submitted during 1987-2001. Twenty-six percent were positive (514). Human herpesvirus 1 was the most frequent agent isolated from all positive culture (56 percent). Adenovirus was the most common virus isolated from conjunctiva (66 percent), human herpesvirus 1 from lid and cornea (76 percent, 88 percent) and cytomegalovirus from vitreous (27 percent). Some unusual pathogens were recovered from conjunctiva as cytomegalovirus and from cornea as adenovirus, enterovirus and cytomegalovirus. Recognition of common viral syndromes was human herpesvirus 1 (88 percent), epidemic keratoconjunctivitis (88 percent), acute hemorrhagic conjunctivitis (70 percent) and varicella zoster virus (100 percent). However, some misdiagnosed cases were observed. Thirteen percent of conjunctivitis thought to be caused by herpes were due to adenovirus, 3.2 percent to Enterovirus, 3.2 percent to varicella zoster virus and 3.2 percent to human cytomegalovirus. Also, 5 percent of cases with a clinical diagnosis of herpes keratitis were caused by adenovirus and 2.7 percent by enterovirus. Finally, 4.8 percent of cases thought to be adenovirus conjunctivitis were herpes conjunctivitis. CONCLUSIONS: Human herpesvirus 1 remains the most frequently isolated virus from ocular sites in general (56 percent). Nonherpetic corneal isolates were in decreasing order: adenovirus, enterovirus and cytomegalovirus. Clinical and laboratory correlation was less than 90 percent. The most misdiagnosed cases were herpes conjunctivitis and keratitis, some cases of adenovirus conjunctivitis some cases of acute hemorrhagic conjunctivitis. It is essential that a rapid and...
OBJETIVOS: Documentar a etiologia e prever a importância do diagnóstico clínico em doenças virais oculares confirmadas em laboratório. MÉTODOS: Todos os relatórios de pacientes com cultura viral positiva durante o período de janeiro 1987 - dezembro 2001 foram analisados. RESULTADOS: Quinhentos e quatorze (514) casos foram encontrados. Em geral, herpesvírus 1 humano foi o agente mais freqüentemente isolado. Adenovírus foi o vírus mais comumente isolado da conjuntiva (66 por cento), herpesvírus 1 humano das pálpebras e córnea (76 por cento, 88 por cento respectivamente) e citomegalovírus do vítreo (27 por cento). Alguns agentes não usuais foram isolados da conjuntiva como citomegalovírus e da córnea como adenovírus, enterovírus e citomegalovírus. Reconhecimento das síndromes virais comuns foi: herpervírus 1 humano (88 por cento), ceratoconjuntivite epidêmica (88 por cento), conjuntivite aguda hemorrágica (70 por cento). Porém, alguns casos com diagnóstico incorreto foram observados. Treze por cento das conjuntivites com diagnóstico de herpes foram causadas por adenovírus, 3,2 por cento por enterovírus, 3,2 por cento por varicella zoster vírus e 3,2 por cento por citomegalovírus. Também, 5 por cento dos casos com diagnóstico clínico de ceratite herpética eram adenovírus e 2,7 por cento enterovírus. Além disso, 4,8 por cento dos casos em que se pensaram em conjuntivite por adenovírus, eram conjuntivite por herpes. Finalmente, 30 por cento dos casos em que se diagnosticaram conjuntive hemorrágica aguda, o agente etiológico era adenovírus. CONCLUSÃO: Em geral herpesvírus humano 1 continua a ser o vírus mais comum encontrado nas infecções oculares (56 por cento). Agentes não herpéticos isolados da córnea foram em ordem decrescente: adenovírus, enterovírus e citomegalovírus. A correlação entre o diagnóstico clínico e laboratorial foi menor do que 90 por cento. Um diagnóstico rápido e específico é essencial em casos de apresentações virais atípicas...
Asunto(s)
Humanos , Infecciones Virales del Ojo/diagnóstico , Infecciones Virales del Ojo/etiología , Queratoconjuntivitis/diagnóstico , Queratoconjuntivitis/etiología , Brasil/epidemiología , Técnicas de Laboratorio Clínico , Conjuntivitis Hemorrágica Aguda/diagnóstico , Conjuntivitis Hemorrágica Aguda/epidemiología , Conjuntivitis Hemorrágica Aguda/etiología , Córnea/virología , Virus ADN/aislamiento & purificación , Errores Diagnósticos/prevención & control , Enterovirus/aislamiento & purificación , Infecciones Virales del Ojo/epidemiología , Reacciones Falso Positivas , Herpes Zóster/virología , Queratoconjuntivitis/epidemiología , Valor Predictivo de las Pruebas , Estaciones del Año , Factores de Tiempo , Cultivo de VirusRESUMEN
AIMS: To determine whether herpes simplex keratitis (HSK) has declined as an indication for penetrating keratoplasty (PKP) at the University of California San Francisco (UCSF) over the past 30 years. METHODS: Records of the Hogan Eye Pathology Laboratory were reviewed to determine the incidence of PKP performed for HSK from 1972 through 2001. Archived corneal tissue with the diagnosis of HSK was evaluated for herpes simplex virus (HSV) DNA by polymerase chain reaction (PCR) based assays. RESULTS: The number of corneal buttons submitted with the clinical diagnosis of HSK decreased from 1972 to 2001, while the overall number of PKPs performed did not. The percentage of corneal buttons with a clinical diagnosis of HSK that contained detectable HSV DNA did not change over the course of the study period. CONCLUSION: HSK declined as an indication for PKP from 1972 to 2001 at UCSF. It is unlikely that this decline was the result of improved diagnostic accuracy since detection of HSV DNA in corneal buttons with a clinical diagnosis of HSK was similar at the beginning and end of the study period.
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Queratitis Herpética/epidemiología , Queratoplastia Penetrante/estadística & datos numéricos , Acanthamoeba/aislamiento & purificación , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Animales , California/epidemiología , Niño , Córnea/parasitología , Córnea/virología , ADN Protozoario/análisis , ADN Viral/análisis , Humanos , Queratitis Herpética/cirugía , Queratitis Herpética/virología , Persona de Mediana Edad , Simplexvirus/aislamiento & purificaciónRESUMEN
In this study, the immune-modulatory and vaccine effects of using an interleukin (IL)-18 expression plasmid as a genetic adjuvant to enhance DNA vaccine-induced immune responses were investigated in a mouse herpes simplex virus 1 (HSV-1) challenge model. BALB/c mice were immunized by three intramuscular inoculations of HSV-1 glycoprotein D (gD) DNA vaccine alone or in combination with a plasmid expressing mature IL-18 peptide. Both the serum IgG2a/IgG1 ratio and T helper 1-type (Th1) cytokines [IL-2 and interferon (IFN)-gamma] were increased significantly by the co-injection of the IL-18 plasmid compared with the injection of gD DNA alone. However, the production of IL-10 was inhibited by IL-18 plasmid co-injection. Furthermore, IL-18 plasmid co-injection efficiently enhanced antigen-specific lymphocyte proliferation and the delayed-type hypersensitivity response. When mice were challenged with HSV-1 at the cornea, co-injection of IL-18 plasmid with gD DNA vaccine showed significantly better protection, manifested as lower corneal lesion scores and faster recovery. These experiments indicate that co-injection of an IL-18 plasmid with gD DNA vaccine efficiently induces Th1-dominant immune responses and improves the protective effect against HSV-1 infection.
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Adyuvantes Inmunológicos , Herpes Simple/inmunología , Herpesvirus Humano 1/inmunología , Interleucina-18/inmunología , Plásmidos/genética , Vacunas de ADN/inmunología , Animales , Anticuerpos Antivirales/sangre , Córnea/inmunología , Córnea/patología , Córnea/virología , Citocinas/inmunología , Modelos Animales de Enfermedad , Femenino , Expresión Génica , Herpes Simple/prevención & control , Hipersensibilidad Tardía , Interleucina-18/genética , Ratones , Ratones Endogámicos BALB C , Bazo/citología , Bazo/inmunología , Células TH1/inmunología , Células Th2/inmunología , Vacunas de ADN/genéticaRESUMEN
Two different procedures for inoculation of HSV on corneas of BALB/c mice were evaluated. The first was by the use of HSV suspensions directly on the corneas and the other was after corneal scarification. Animals by this later method presented greater morbidity and mortality than those of first group, suggesting that inoculation of HSV without scarification of the cornea should be the method of choice for the study of HSV ophthalmic infection. This model showed also be an efficient experimental system to testing antiviral drugs.
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Modelos Animales de Enfermedad , Queratitis Herpética/virología , Simplexvirus/patogenicidad , Animales , Córnea/virología , Queratitis Herpética/patología , Masculino , Ratones , Ratones Endogámicos BALB CRESUMEN
PURPOSE: To study the effect of aminoguanidine (AMG), an inhibitor of nitric oxide production, on the ocular infection of Balb/c mice with herpes simplex virus (HSV) type 1 strain F and HSV-2 strain G. METHODS: Animals were treated with different amounts of AMG (0.5, 0.1, and 0.05 mg/mouse) by topical application in the eye from postinfection (PI) days -2 through +5, considering 0 the day of infection. At different PI days, development of herpetic keratitis was evaluated in treated and control mice. RESULTS: Treated animals showed a dose-dependent increase in ocular disease after viral infection, compared with control animals. Viral titers in ocular washings were higher in AMG-treated mice (PI day 2, HSV-1: AMG 0.5 mg, 1.3 x 10(3) plaque-forming units (PFU)/ml; control, 0. 22 x 10(2) PFU/ml, P < 0.025). At PI day 3, control corneas had only scattered inflammatory cells, whereas those from treated animals showed a conspicuous infiltrate consisting primarily of neutrophils. Viral titers were also higher in brains of treated mice. These animals died earlier and in a greater proportion than control animals (percentage of mortality, PI day 12, HSV-1: AMG 0.5 mg, 40% +/- 4%; control, 18% +/- 3%, P < 0.05). CONCLUSIONS: These data indicate an inhibitory effect of nitric oxide on HSV ocular infection.
Asunto(s)
Córnea/efectos de los fármacos , Inhibidores Enzimáticos/uso terapéutico , Guanidinas/uso terapéutico , Queratitis Herpética/tratamiento farmacológico , Óxido Nítrico Sintasa/antagonistas & inhibidores , Administración Tópica , Animales , Córnea/enzimología , Córnea/virología , ADN Viral/análisis , Progresión de la Enfermedad , Relación Dosis-Respuesta a Droga , Herpesvirus Humano 1/aislamiento & purificación , Herpesvirus Humano 1/fisiología , Herpesvirus Humano 2/aislamiento & purificación , Herpesvirus Humano 2/fisiología , Queratitis Herpética/patología , Queratitis Herpética/virología , Ratones , Ratones Endogámicos BALB C , Óxido Nítrico Sintasa/genética , Óxido Nítrico Sintasa/metabolismo , Soluciones Oftálmicas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Ganglio del Trigémino/virologíaRESUMEN
Herpetic stromal keratitis is caused by ocular infection with herpes simplex virus type 1 (HSV-1) and constitutes a leading cause of human blindness. The effect of meliacine, an antiviral compound isolated from leaves of Melia azedarach L. that inhibits HSV-1 replication in vitro, was examined on experimental corneal HSV-1 inoculation in Balb/c mice. Mice were inoculated with HSV-1 strain KOS at their corneas after abrasion. Meliacine was administered topically 3 times a day for 4 days beginning 1 day before inoculation. Infected animals treated or not with meliacine were observed carefully for the development of stromal keratitis and the clinical scoring was done 14 days post-infection. Histological examination of corneas and viral isolation from eyes from HSV-1 infected mice treated or not with meliacine were also carried out. It was found that the treatment of HSV-1-induced ocular disease in Balb/c mice with meliacine reduced significantly the development of clinical disease, as well as the histological damage in corneas. The viral titers detected in eyes of infected and treated mice were 2-orders-of-magnitude lower than those corresponding to HSV-1 infected control animals. Mock-infected and treated mice did not reveal any corneal alteration due to the administration of the compound. Meliacine was found to exert a strong antiviral action on HSV-1-induced ocular disease in mice with no evidence of toxic effects.