Seroprevalencia contra agentes ToRCH en mujeres indígenas en edad fértil, estado Zulia, Venezuela / Prevalence of infectious agents in indigenous women of childbearing age in Venezuela

Introducción. El termino ToRCH comprende a los patógenos Toxoplasma gondii, virus de la rubéola, citomegalovirus y virus herpes simple 1 y 2. En mujeres embarazadas expuestas pueden ser causa de abortos y malformaciones congénitas en el neonato. Objetivo. Determinar la seroprevalencia de infección por los agentes causantes del síndrome ToRCH en mujeres en edad fértil de algunas comunidades indígenas yukpa de Venezuela. Materiales y métodos. En el año 2007 fueron seleccionadas 109 muestras de 151 mujeres, en edades comprendidas entre 14 y 40 años. La detección de anticuerpos se hizo por el método de inmunoensayo enzimático indirecto o ELISA de Smartest Diagnostics™. Resultados. El 85,5 % presentó anticuerpos contra T. gondii, el 95,4 % para rubéola, el 75,2 % para citomegalovirus y el 97,2 % para el virus herpes simple 1 y 2. Se observa que el 21,1 % y el 30,2 % presentaron relación entre la variable aborto y las infecciones por citomegalovirus y virus herpes simple 1 y 2, respectivamente. Conclusiones. Existe alta seroprevalencia de infecciones por los agentes causantes del síndrome ToRCH en mujeres en edad fértil de la etnia indígena yukpa. Las condiciones sanitarias precarias y el consumo de agua contaminada con ooquistes, favorecen la adquisición de la infección por T. gondii. El hacinamiento, el inicio a temprana de edad de la actividad sexual y el número de parejas, pueden incidir en la presencia de citomegalovirus y virus herpes simple 1 y 2.

Introduction. The ToRCH syndrome includes the following infectious pathogens: Toxoplasma gondii, rubella, cytomegalovirus and herpes simplex virus 1 and 2. In susceptible pregnant women, these pathogens can cause abortions and congenital malformation in the newborn babies. Objective. The seroprevalence of infection by ToRCH agents was determined in women of childbearing age in several Venezuelan Yukpa indigenous communities. Material and methods. In 2007, 109 samples were selected from 151 women with an age range of 14 to 40 years old. The determination of antibodies against ToRCH agents was carried out through the indirect enzyme immunoassay technique by ELISA´s technique of Smartest Diagnostics. Results. Of the 109 samples, 85.5% presented antibodies against T. gondii, 95.4% for rubella, 75.2% for cytomegalovirus and 97.2% for and herpes simplex virus 1 and 2. A relationship between abortion and infection by cytomegalovirus and herpes simplex virus 1and 2 was noted in 21.1% and 30.2% of women presented, respectively. Conclusions. The findings show a high prevalence of ToRCH agents in women in childbearing age in Yukpa indigenous communities in Venezuela. Poor sanitary conditions and consumption of water contaminated with oocysts may be an important way of transmission of T. gondii. Overcrowding in the communities, sexual activity at an early age and number of partners and may be related to the presence of cytomegalovirus and herpes simplex virus HSV-1 and 2.

Ativação da Resposta Imune Inata Mediada por Receptores do Tipo Toll em Infecção por Vírus Herpes Simplex tipo 1(HSV-1) em Modelo Murino

Este trabalho foi realizado no laboratório de Imunopatologia do Centro de Pesquisa RenéRachou (CPqRR) e no Laboratório de Vírus (Labvirus) da Universidade Federal de MinasGerais (UFMG). Os herpesvirus possuem genoma DNA e são envelopados.Aproximadamente 70% dos adultos já tiveram contato com Herpes Simplex tipo 1 (HSV-1).O HSV-1 causa úlceras ou até mesmo lesões mais graves como encefalite, e após a infecção,o vírus comumente fica latente e pode ser ou não reativado. Células do sistema immuneinato, através dos “Receptores do Tipo Toll” (TLRs), reconhecem “padrões molecularesassociados a patógenos” (PAMPs) orquestrando a resposta imune contra o vírus

Quandoativados, TLRs iniciam uma cascata de sinalização que culmina na ativação de genesrelacionados à defesa imune inata (óxido nítrico-NO, citocinas e quimiocinas). Outros estudosdo grupo mostraram que animais nocautes (KO) para TLR9 ou TLR2/9 apresentam altamortalidade na infecção por HSV-1, ao contrário dos C57BL/6 selvagens (WT) e TLR2KO,que são capazes de controlar o vírus. Objetivamos pesquisar a interrelação na expressão dosTLRs 1, 2, 3, 6, 7 e 9 em animais TLR2 KO, TLR9 KO e TLR2/9 KO infectados por HSV-1e principais citocinas, células e mecanismos imune celulares contra infecção por HSV-1. Nopresente trabalho camundongos WT, TLR2KO, TLR9KO e TLR2/9KO foram infectados viaintranasal com 106 u.f.p. de HSV-1, e seus controles receberam PBS. A eutanásia dos animaisfoi realizada no 5º dia após infecção (dpi), que previamente já foi demonstrado ser o pico dainfecção neste modelo, e então gânglios trigêmeos (TG) e cérebros foram coletados. Aexpressão de TLRs 1, 2, 3, 6, 7 e 9 foi avaliada por PCR em Tempo Real. Camundongos WTinfectados apresentaram diferença significativa na expressão destes TLRs comparados comseus respectivos controles no TG, mas não no cérebro. Nos KOs, houve aumento da expressãodos TLRs tanto nos TGs (mas com níveis mais moderados comparado aos WT) quanto noscérebros. Também foi avaliada a expressão de gp91phox, p22phox e iNOS. A expressão de iNOSem C57BL/6 WT infectados foi maior que nos outros grupos, mas o mesmo não ocorreu paragp91phox, p22phox. Adicionalmente, foi mensurada a produção de NO por macrófagosintraperitoneais, através da reação de Griess, que demonstrou maior produção por macrófagosWT, comparados aos grupos TLR KOs

Ensaios de histopatologia e imunofluorescênciademonstraram maior celularidade nos TGs infectados, além de confirmar a presença de CD8,macrófagos, e aumento da produção de iNOS nos C57BL/6 WT infectados, o que não ocorreucom os camundongos TLRs KO. Em ensaio de sobrevivência com C57BL/6 WT, CCL3 KO,CD8 KO, RAG KO e iNOS KO a importância das células T e do iNOS foi confirmada, umavez que CD8 KO, RAG KO e iNOS KO tiveram 0% de sobrevida. Camundongos C57BL/6WT, RAG KO, CCL3 KO e iNOS KO tiveram seus TGs analisados por PCR em tempo realpara verificar a expressão do gene viral VP-16 e MCP-1, iNOS, IP-10, Rantes e TNF- no 5ºdpi. Estes dados indicaram a relevância de iNOS e das célúlas T na resposta inata contra oHSV-1, uma vez que os RAG KO foram irresponsivos à expressão das citocinas, enquanto osiNOS KO apresentaram uma resposta exacerbada das mesmas. Ensaio de citometria indicouque IFN produzido pelas células T CD8, nos TGs, e produzido pelas células T CD4 eNatural Killer (NK), nos linfonodos regionais, são um possível mecanismo de controle contraa infecção de HSV-1 dos animais WT infectados. Hipotetizamos que a expressão coordenadade TLRs reflete na orquestração eficiente do sistema imune inato, através da produçãoequilibrada de quimiocinas e citocinas e da atividade imune celular adequada, com produçãode NO por macrófagos e ação de células T CD8, T CD4 e NK controlando a infecção porHSV-1

Ativação da resposta imune inata mediada por receptores do tipo toll em infecção por vírus herpes simplex tipo 1(HSV-1) em modelo murino / Activation of the innate immune response mediated by toll receptors type in infection herpes simplex virus type 1 (HSV-1) in murine model

Este trabalho foi realizado no laboratório de Imunopatologia do Centro de Pesquisa RenéRachou (CPqRR) e no Laboratório de Vírus (Labvirus) da Universidade Federal de MinasGerais (UFMG). Os herpesvirus possuem genoma DNA e são envelopados.Aproximadamente 70% dos adultos já tiveram contato com Herpes Simplex tipo 1 (HSV-1).O HSV-1 causa úlceras ou até mesmo lesões mais graves como encefalite, e após a infecção,o vírus comumente fica latente e pode ser ou não reativado. Células do sistema immuneinato, através dos “Receptores do Tipo Toll” (TLRs), reconhecem “padrões molecularesassociados a patógenos” (PAMPs) orquestrando a resposta imune contra o vírus. Quando ativados, TLRs iniciam uma cascata de sinalização que culmina na ativação de genesrelacionados à defesa imune inata (óxido nítrico-NO, citocinas e quimiocinas). Outros estudosdo grupo mostraram que animais nocautes (KO) para TLR9 ou TLR2/9 apresentam altamortalidade na infecção por HSV-1, ao contrário dos C57BL/6 selvagens (WT) e TLR2KO,que são capazes de controlar o vírus. Objetivamos pesquisar a interrelação na expressão dosTLRs 1, 2, 3, 6, 7 e 9 em animais TLR2 KO, TLR9 KO e TLR2/9 KO infectados por HSV-1e principais citocinas, células e mecanismos imune celulares contra infecção por HSV-1. Nopresente trabalho camundongos WT, TLR2KO, TLR9KO e TLR2/9KO foram infectados viaintranasal com 106 u.f.p. de HSV-1, e seus controles receberam PBS. A eutanásia dos animaisfoi realizada no 5º dia após infecção (dpi), que previamente já foi demonstrado ser o pico dainfecção neste modelo, e então gânglios trigêmeos (TG) e cérebros foram coletados. Aexpressão de TLRs 1, 2, 3, 6, 7 e 9 foi avaliada por PCR em Tempo Real. Camundongos WTinfectados apresentaram diferença significativa na expressão destes TLRs comparados comseus respectivos controles no TG, mas não no cérebro. Nos KOs, houve aumento da expressãodos TLRs tanto nos TGs (mas com níveis mais moderados comparado aos WT) quanto noscérebros. Também foi avaliada a expressão de gp91phox, p22phox e iNOS. A expressão de iNOSem C57BL/6 WT infectados foi maior que nos outros grupos, mas o mesmo não ocorreu paragp91phox, p22phox. Adicionalmente, foi mensurada a produção de NO por macrófagosintraperitoneais, através da reação de Griess, que demonstrou maior produção por macrófagosWT, comparados aos grupos TLR KOs. Ensaios de histopatologia e imunofluorescênciademonstraram maior celularidade nos TGs infectados, além de confirmar a presença de CD8,macrófagos, e aumento da produção de iNOS nos C57BL/6 WT infectados, o que não ocorreucom os camundongos TLRs KO. Em ensaio de sobrevivência com C57BL/6 WT, CCL3 KO,CD8 KO, RAG KO e iNOS KO a importância das células T e do iNOS foi confirmada, umavez que CD8 KO, RAG KO e iNOS KO tiveram 0% de sobrevida. Camundongos C57BL/6WT, RAG KO, CCL3 KO e iNOS KO tiveram seus TGs analisados por PCR em tempo realpara verificar a expressão do gene viral VP-16 e MCP-1, iNOS, IP-10, Rantes e TNF- no 5ºdpi. Estes dados indicaram a relevância de iNOS e das célúlas T na resposta inata contra oHSV-1, uma vez que os RAG KO foram irresponsivos à expressão das citocinas, enquanto osiNOS KO apresentaram uma resposta exacerbada das mesmas. Ensaio de citometria indicouque IFN produzido pelas células T CD8, nos TGs, e produzido pelas células T CD4 eNatural Killer (NK), nos linfonodos regionais, são um possível mecanismo de controle contraa infecção de HSV-1 dos animais WT infectados. Hipotetizamos que a expressão coordenadade TLRs reflete na orquestração eficiente do sistema imune inato, através da produçãoequilibrada de quimiocinas e citocinas e da atividade imune celular adequada, com produçãode NO por macrófagos e ação de células T CD8, T CD4 e NK controlando a infecção porHSV-1.

Role of IL-4 in an experimental model of encephalitis induced by intracranial inoculation of herpes simplex virus-1 (HSV-1) / Papel da IL-4 em modelo experimental de encefalite induzida pela inoculação intracraniana do herpes simplex vírus-1 (HSV-1)

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.

Soroprevalência de anticorpos contra vírus herpes simples 1-2 no Brasil / Seroprevalencia de anticuerpos contra virus herpes simples 1-2 en Brasil / Seroprevalence of herpes simplex 1-2 antibodies in Brazil

OBJETIVO: Estimar a soroprevalência de anticorpos por vírus herpes simples (HSV-1 e HSV-2) e analisar fatores associados no Brasil. MÉTODOS: Estudo transversal realizado entre 1996 e 1997 em 1.090 indivíduos com idade entre um e 40 anos da população geral, em quatro diferentes regiões geográficas no Brasil. Foram analisadas amostras sangüíneas para detecção de anticorpos para HSV-1 e HSV-2 com teste tipo-específico Elisa. Foram descritas freqüências e proporções, comparadas entre grupos utilizando o teste de Fisher bilateral exato. Foi realizada análise de regressão logística para avaliar influência das variáveis sociodemográficas e histórico de DST, sobre a soroprevalência de HSV-1 e/ou HSV-2. RESULTADOS: As soroprevalências de anticorpos para HSV-1 e HSV-2, ajustadas por idade, foram 67,2 por cento e 11,3 por cento respectivamente, sem diferença quanto ao sexo e maiores na Região Norte...

OBJECTIVE: To estimate the seroprevalence of HSV-1 and HSV-2 antibodies in Brazil and to analyze factors associated. METHODS: Cross-sectional study including subjects aged 1-40 years from the general population in four different geographical areas in Brazil between 1996 and 1997. All subjects were stratified by age and gender and 1,090 of them were included in the final analysis. Blood samples were tested for HSV-1 and HSV-2 antibodies by type-specific (gG1 and gG2) ELISA. Frequencies and proportions were described and compared among groups using two-sided Fisher's exact test. A logistic regression analysis was performed to assess the influence of the variables age, gender, geographical area, socioeconomic condition, past history of STD, seropositivity for anti-HSV-1 or anti-HSV-2 and interactions of any of these factors on the seroprevalence of HSV-1 and/or HSV-2. RESULTS: The age-adjusted seroprevalences of HSV-1 and HSV-2 antibodies were 67.2 percent and 11.3 percent, respectively, without sex differences and being higher in the North region...

OBJETIVO: Estimar la seroprevalencia de anticuerpos por virus herpes simples (HSV-1 y HSV-2) en diferentes áreas geográficas en Brasil y analizar factores asociados. MÉTODOS: Estudio transversal realizado entre 1996 y 1997 con individuos de la población en general en cuatro diferentes áreas geográficas en Brasil y estratificados por edad (de uno a 40 años) y sexo, de los cuales 1.090 fueron incluidos en el análisis final. Fueron analizadas muestras de sangre para detección de anticuerpos para HSV-1 y HSV-2 con prueba tipo-específica ELISA gG1-gG2. Fueron descritas frecuencias y proporciones y comparadas entre grupos utilizando la prueba de Fisher bilateral exacta. Fue realizado análisis de regresión logística para evaluar influencia de las variables edad, sexo, geografía, grupo económico, histórico de DST, seropositividad para anti-HSV-1 o anti-HSV-2 e interacciones de cualquiera de esos factores sobre la seroprevalencia de HSV-1 y/o HSV-2. RESULTADOS: La tasa de seroprevalencia de anticuerpos para HSV-1 ajustada por edad fue de 67,2 por ciento, sin diferencia con relación al sexo, siendo mayor en la Región Norte...

Elevada seroprevalencia de citomegalovirus, virus herpes simplex tipo 1 y virus Epstein Barr en adultos con virus de la inmunodeficiencia humana / High seroprevalence of cytomegalovirus, herpes simplex type 1 virus and Epstein Barr virus infection among human immunodefciency virus-infected adults

Background: Cytomegalovirus (CMV), herpes simplex type 1 (HSV-1) and Epstein Barr virus (EBV) are latent persistent infections. Their reactivation may cause illnesses and death in human immunodefciency virus-infected (HIV) people. World wide seroprevalence of these viruses is over 50 percent. In Chile, information is not available. Aim: To determine the seroprevalence of CMV, HSV-1 and EBV in Chilean HIV-infected adults. Patients and Methods: A total of 400 HIV- infected adults aged 17 to 67 years (340 males) were studied during 2005 and 2006. CMV, HSV-1 and EBV serum antibodies were measured by enzyme-linked immunoabsorbent assay. Results: The mean lapse from the diagnosis of HIV and serum testing was 67 months and 69.5 percent patients received antiretroviral therapy. Sixty seven percent of the sample were men who had sex with men (MSM). The seroprevalence for CMV, HSV-1 and EBV were 98.5, 92.2 and 99.7 percent, respectively. No patient had negative antibodies for all three viruses. Male patients that were negative for HSV-1 had a lower frequency of MSM than the rest of males (26 percent vs 62 percent, p < 0.01). Conclusions: There is a high prevalence of positive antibodies against CMV, HSV-1 and EBV in Chilean adults infected with HIV. Specifc diagnostic tests and antiviral therapy should be available for these patients.

Co-infection of herpes simplex virus (HSV) with human immunodeficiency virus (HIV) in women with reproductive tract infections (RTI).

In India, HSV seroprevalence and its coinfection with HIV among female patients with reproductive tract infections (RTI) are sparse. We aim to ascertain the seroprevalence of HSV and its coinfection with HIV and common sexually transmitted infections attending Obstetrics and Gynaecology outpatient department, RIMS. The study included 92 female patients with RTI. Diagnostic serology was done for HSV-1 and HSV-2 using group specific IgM indirect immunoassay using ELISA, HIV by 3 ELISA/Rapid/Simple (E/R/S) test of different biological antigen. Diagnosis of RTI was made on clinical grounds with appropriate laboratory investigations--microscopy, Gram stain smear etc. Bacterial vaginosis was diagnosed using Nugent's criteria, Syphilis by rapid plasma reagin (RPR) card test and Chlamydia trachomatis by IgG ELISA. Out of 92 sera tested for HSV, 18 (19.6%) were IgM HSV positive and 9 (9.8%) were HIV positive. Co-infection rate of HSV in HIV positive was 16.7%. None of the patients had clinical herpes genitalis, all were subclinical cases. 55.5% of HSV positives belongs to age group 21 to 30 years. Of the HSV-1 and HSV-2 IgM positives 3 (15%) had HIV, 4 (22.2%) bacterial vaginosis, 2 (11.1%) were RPR positive, 4 (22.2%) Chlamydia trachomatis, 3 (15%) were pregnant. 16 (88.8%) were unemployed, 14 (77.7%) had education level below 10 standard. Our study suggest that every case of RTI, be it an ulcerative or nonulcerative must be thoroughly evaluated by laboratory testing for primary subclinical genital HSV coinfection as this has profound implications on their judicious management and aversion of complications. Early diagnosis and treatment of HSV infection together with prophylaxis for recurrent HSV disease will prevent progression and spread of HIV disease.

An immunofluorescence test for diagnosis of ophthalmic herpes in a mouse corneal model

Herpes simplex virus type 1 (HSV-1) ophthalmic disease is the most common cause of corneal blindness in humans world-wide. Current culture techniques for HSV take several days and commercially available HSV laboratory based diagnostic techniques vary in sensitivity. Our study was conducted to evaluate the use of a quicker and simpler method to herpes ophthalmic diagnosis. Corneal smears were made by firm imprints of infected mouse eyes to glass slides, after smears were fixated with cold acetone, and an indirect immunofluorescence (IIF) method was performed using monoclonal antibodies in a murine model of ophthalmic herpes. Eye swabs from infected mice were inoculated in Vero cells for virus isolation. Cytology and histology of the eye were also performed, using hematoxylin-eosin routine. Mouse eyes were examined by slit-lamp biomicroscopy for evidence of herpetic disease at various times postinoculation. We made a comparative evaluation of sensitivity, specificity and speed of methods for laboratory detection of HSV. Our results indicate that this IIF method is quick, sensitive, specific and can be useful in the diagnosis of ophthalmic herpes as demonstrated in an animal model.

A doença oftálmica do vírus herpes simplex do tipo 1 (HSV-1) é a causa mais comum de cegueira córnea em humanos mundialmente. Técnicas de cultura atuais para HSV levam vários dias e laboratórios de HSV comercialmente disponíveis estabelecem que as técnicas diagnósticas variam em sensibilidade. Nosso estudo foi conduzido para avaliar a aplicação prática de um método mais rápido e simples para diagnosticar o herpes oftálmico. Decalques córneos foram feitos por impressões firmes de olhos de camundongos a lâminas de vidro, depois os decalques foram fixados com acetona fria, e um método de imunofluorescência indireta (IIF) foi executado empregando anticorpos monoclonais no modelo murino de herpes oftálmico. Swabs de córnea foram inoculados em células Vero para o isolamento de vírus a partir de camundongos infectados. A citologia e a histologia do olho foi feita pela rotina de hematoxilina e eosina. Os olhos de camundongos foram examinados através de oftalmomicroscopia para evidência de doença herpética em vários tempos pós-inoculação. A avaliação comparativa da sensibilidade, especificidade e velocidade de métodos para detecção laboratorial de HSV foi feita. Nossos resultados indicam que este método de IIF é rápido, sensível, específico e pode ser útil no diagnóstico de herpes oftálmico como demonstrado no modelo animal.

Seroprevalence of HSV1 and HSV2 infections in family planning clinic attenders.

Few studies from India documented seroprevalence of HSV 1 and 2 infection in different population groups. We determined HSV 1 and 2 seroprevalence in a cohort of adults 16-40 year of age, attending the family planning clinic. For the overall study population, 63% were seropositives, 33.3% for HSV 1 alone. 16.6% for HSV 2 and 13.3% had mixed infection. By the statistical analysis, the mean age difference between the two sexes for either infection was not significant. HSV-2 seroprevalence was associated with an increasing age. Men were more likely than women to be seropositive for HSV2. More studies from India are required to coroborate our findings.

Increase of genital HSV-1 and mixed HSV-1 and HSV-2 infection in Bangkok, Thailand.

From January 1998 to December 2004, 207 out of 1125 samples were HSV isolation positive and typed. Two groups of patients, Thai and foreigner, as well as site of infection, non-genital and genital area, were identified. The prevalence of non-genital HSV-1 infection from 27 samples of Thai patients was 81.84%. Out of 180 genital samples, 39.02% HSV-1 and 43.09% HSV-2 from 123 Thai patients and 36.84% HSV-1 and 49.12% HSV-2 of 57 foreigner patients were determined. Moreover mixed infection of HSV-1 and HSV-2 was found in both Thai and foreigner groups, 17.89% and 14.04%, respectively. The prevalence of genital HSV-1 infection in Thai patients chronologically increases from 1.6% to 56.91% from of 1985 to 2004. Increase of HSV-1 genital infection and mixed HSV-1 and HSV-2 infection in Thai people might probably be due to changing of sexual behavior in the AIDS era.

Induction of apoptosis by herpes simplex virus in Jurkat cells is partly through caspase-3, -8 and -9 activation.

Herpes simplex virus (HSV), a large DNA containing virus, is endemic in all human populations investigated. After infection of mucocutaneuos surfaces, HSV establishes a latent infection in nerve cells. Various immune evasion mechanisms have been shown to be utilized by HSV including apoptosis induction in Tlymphocytes. However, the mechanisms of T cell infection and apoptosis by HSV are still unknown. The present study investigated the molecular mechanisms of apoptosis induction in T cells by HSV The Jurkat T cell line was used as a representative for T cells. Apoptosis detection by Annexin Vassay demonstrated that both HSV-1 and HSV-2 induced apoptosis in Jurkat cells and caspase-3, -8, and -9 inhibitors blocked apoptosis induced by HSV-1 and HSV-2. The data suggested that HSV-1 and HSV-2 induced apoptosis in T lymphocytes by caspase-dependent pathway. However, apoptosis may occur through other mechanism(s) since caspase inhibitors used in the present study could not completely inhibit apoptosis induced by HSV infection. In addition, the data demonstrated that the number of apoptotic cells induced by HSV-2 was significantly higher than byHSV-1 at 12 hour post-infection (h p.i.) (p = 0.003). Further studies in peripheral blood T cells and the proteins of viruses involved in apoptosis induction should be further performed in order to elucidate the molecular mechanisms of apoptosis induced by these viruses.

Role of serology in the diagnosis of herpes simplex encephalitis.

Twenty one cerebrospinal fluid (CSF)/brain tissue (16 CSF and 5 brain tissue) from patients clinically suspected of herpes simplex encephalitis (HSE) were collected during one year period and was subjected for the detection of HSV type I and type II antigen by direct Immunofluorescence (DIF). Paired serum and CSF samples obtained from 12 patients were tested for herpes simplex virus antibodies by indirect immunofluorescence test. Of the 21 cases, two were positive for HSV-1 antigen in CSF and one in brain tissue by DIF. Virus specific IgG antibody in paired CSF and serum samples was positive in one case only. In none, virus could be grown in verocell line. In 2 Patients antemortem diagnosis was possible and parenteral acyclovir could be administered in one case that recovered following treatment. Thus, besides antibody detection, direct immunofluorescence is an useful and rapid method for the early diagnosis of HSE.

In vitro induced antibody production for the diagnosis of herpes simplex virus infection in immunosuppressed patients

We developed and evaluated a specific test for herpes simplex virus (HSV) infection based on the secretion of HSV-specific antibodies by lymphocytes stimulated in vitro with HSV-1 antigens. The in vitro induced antibody production (IVIAP) test was used for the diagnosis of HSV infection in 43 seropositive selected subjects: 9 healthy subjects (controls), 30 symptomatic patients (26 of them immunocompromised and 4 immunocompetent) and 4 patients with varicella zoster infection. Anti-HSV antibodies were detected by an immune assay using an anti-human IgG peroxidase conjugate. The test showed a sensitivity of 93 per cent (15/16) and specificity of 92 per cent (1/13) which were confirmed by positive culture or clinical and laboratory follow-up. One AIDS patient had a false-negative result and one false-positive result (1/9) was obtained among the healthy subjects. All patients infected with varicella zoster virus were negative to the IVIAP test. The test is rapid, inexpensive, easy to interpret and can be used for the diagnosis of HSV infections, especially in immunocompromised patients.

Clinical features and virologic studies in viral retinitis.

Viral retinitis is an important infectious disease of the retina which can occur in both healthy and immunocompromized or immunodeficient individuals. The clinical picture and the role of laboratory studies in diagnosis of viral retinitis are still not well-defined. We correlated the clinical picture and virologic study in the serum and vitreous specimens by Enzyme-linked Immunosorbent Assay (ELISA), rapid immunofluorescence technique and culture in five clinically suspected patients of viral retinitis. None of the patients had any evidence of systemic viral infections. In four patients, the virus was detected by immunofluorescence, ELISA or culture from the vitreous sample. Paired serum samples showed elevation of antiviral IgG titre in two cases and high antiviral IgM titre in all cases. Our study evaluated the role of virological investigations of vitreous aspirate and rising antibody titre in the paired serum samples in the diagnosis of active viral retinitis.

Monoclonal anti-idiotypes to herpes simplex virus type 1 capable of antigen specific priming & stimulatory activity.

Anti-idiotypic antibodies (Ab-2) to HSV-1 (herpes simplex virus 1) neutralizing monoclonal antibody were raised by hybridoma. These Ab-2 were found to represent an epitope of glycoprotein B (gB-1) of this virus. To further characterise this antibody for its ability to mimic the antigenic epitope, in vitro lymphoproliferation assays were done. In this assay (i) antigen specific lymphocyte priming activity of the three monoclonal Ab-2 and (ii) the in vitro stimulating ability of these Ab-2 for gb-1 primed mouse lymphocytes were tested. We could identify two monoclonal Ab-2 which were able to prime the mouse lymphocytes in vivo. These antibodies were able to recognise the in vitro stimulation signal of the antigen gB-1 and consequently could proliferate. The stimulation index was comparable to that with the antigen. These two Ab-2 were also recognized by the antigen primed mouse lymphocytes in a specific manner.

Experimental ulcerative herpetic keratitis: Humoral immune response and its role in corneal protection

Twenty five rabbits were inoculated with HSV type [1] to estimate the humoral immune response and corneal protection. The serum neutralizing antibody could be detected 15 days after inoculation. Ig[G], Ig[A] and Ig[M] specific to the virus were detected by immunofluorescence. Only corneae of 2 out of 14 immunized rabbits developed corneal ulcers, when challenged with the virus, as compared to 5 non-immunized that developed ulcer. The 2 immunized rabbits that developed corneal ulcers showed complete healing in 3 days to treatment with trifluorotrhymidine as compared to 6 days for the non-immunized control group [5 rabbits]