Usefulness of aqueous and vitreous humor analysis in infectious uveitis

OBJECTIVE: To evaluate the role of intraocular fluid analysis as a diagnostic aid for uveitis. METHODS: Twenty-eight samples (27 patients including 3 HIV-infected patients) with active (n=24) or non-active (n=4) uveitis were submitted to aqueous (AH; n=12) or vitreous humor (VH) analysis (n=16). All samples were analyzed by quantitative PCR for herpes simplex virus (HSV), varicella zoster virus (VZV), cytomegalovirus (CMV), Epstein-Barr virus (EBV) and Toxoplasma gondii. RESULTS: The positivity of the PCR in AH was 41.7% (5/12), with 50% (2/4) in immunocompetent and 67% (2/3) in HIV+ patients. The positivity of the PCR in VH was 31.2% (5/16), with 13% (1/8) in immunocompetent and 50% (4/8) in immunosuppressed HIV negative patients. The analysis was a determinant in the diagnostic definition in 58% of HA and 50% of VH. CONCLUSION: Even in posterior uveitis, initial AH analysis may be helpful. A careful formulation of possible clinical diagnosis seems to increase the chance of intraocular sample analysis being meaningful.

Evaluation of a Novel Array-Based Toxoplasma, Rubella, Cytomegalovirus, and Herpes Simplex Virus IgG Enzyme Linked Immunosorbent Assay and Its Comparison with Virion/Serion Enzyme Linked Immunosorbent Assays

BACKGROUND: The dramatic increase in use of the IgG test for toxoplasma, rubella, cytomegalovirus (CMV), and herpes simplex virus (HSV) [TORCH] has led to the requirement for a high-efficiency method that can be used in the clinical laboratory. This study aimed to compare the results of BGI-Array ELISA TORCH IgG (BGI-GBI, China) screening method to those of Virion/Serion TORCH IgG ELISA (Virion/Serion, Germany). METHODS: Serum specimens (n=400) submitted for routine IgG testing by Virion/Serion ELISA were also tested using the BGI-Array ELISA method. The agreements of these two kinds of method were analyzed by kappa-coefficients calculation. RESULTS: Following repeat testing, the BGI-Array ELISA TORCH IgG assays demonstrated agreements of 99.5% (398/400 specimens), 98% (392/400 specimens), 99% (396/400 specimens), and 99.5% (398/400 specimens), respectively. The BGI-Array ELISA IgG assays provided results comparable to Virion/Serion ELISA results, with kappa-coefficients showing near-perfect agreement for the HSV (kappa=0.87), rubella (kappa=0.92) and CMV (kappa=0.93) and substantial agreement for the toxoplasma (kappa=0.80) IgG assays. The use of the BGI-Array ELISA TORCH IgG assays could reduce the turnaround time (1.5 hr vs. 5 hr by Virion/Serion ELISA for 100 specimens) and were easy to use. CONCLUSIONS: BGI-Array ELISA TORCH IgG shows a good agreement with Virion/Serion ELISA methods and is suitable for clinical application.

Enhanced anti-tumor effect of a gene gun-delivered DNA vaccine encoding the human papillomavirus type 16 oncoproteins genetically fused to the herpes simplex virus glycoprotein D

Anti-cancer DNA vaccines have attracted growing interest as a simple and non-invasive method for both the treatment and prevention of tumors induced by human papillomaviruses. Nonetheless, the low immunogenicity of parenterally administered vaccines, particularly regarding the activation of cytotoxic CD8+ T cell responses, suggests that further improvements in both vaccine composition and administration routes are still required. In the present study, we report the immune responses and anti-tumor effects of a DNA vaccine (pgD-E7E6E5) expressing three proteins (E7, E6, and E5) of the human papillomavirus type 16 genetically fused to the glycoprotein D of the human herpes simplex virus type 1, which was administered to mice by the intradermal (id) route using a gene gun. A single id dose of pgD-E7E6E5 (2 µg/dose) induced a strong activation of E7-specific interferon-γ (INF-γ)-producing CD8+ T cells and full prophylactic anti-tumor effects in the vaccinated mice. Three vaccine doses inhibited tumor growth in 70 percent of the mice with established tumors. In addition, a single vaccine dose consisting of the co-administration of pgD-E7E6E5 and the vector encoding interleukin-12 or granulocyte-macrophage colony-stimulating factor further enhanced the therapeutic anti-tumor effects and conferred protection to 60 and 50 percent of the vaccinated mice, respectively. In conclusion, id administration of pgD-E7E6E5 significantly enhanced the immunogenicity and anti-tumor effects of the DNA vaccine, representing a promising administration route for future clinical trials.

Diagnóstico microbiológico de infecciones de transmisión Sexual: Parte II. ITS virales / Microbiologic diagnosis of sexually transmitted diseases: Part II. STD of viral origin

Sexually transmitted diseases are highly prevalent and a public health concern worldwide. Herpes simplex virus (HSV) and human papilloma virus (HPV) are described. The development of rapid, sensible and specific diagnostic assays has been difficult because of their pathogenic features. In the last years, molecular biology based techniques allowed a better and wider range of diagnosis, as in the HPV-cancer association. In this article, standardized diagnostic methodologies for HSV and HPV are reviewed.

Las Infecciones virales de transmisión sexual son altamente prevalentes y constituyen un problema de salud pública en el mundo. Entre los agentes que se contagian por esta vía, se describe acerca de virus herpes simplex (HSV) y virus papiloma humano (HPV). Las características patogénicas de estas infecciones han dificultado la implementación de técnicas diagnósticas rápidas, sensibles y específicas para el diagnóstico clínico habitual. En los últimos años las metodologías diagnósticas sustentadas en la biología molecular han permitido mej orar y ampliar el rango de diagnóstico posible para estos agentes infecciosos y relacionarlos con otras patologías, como es el caso de HPV y cáncer. En el presente artículo se revisan metodologías diagnósticas implementadas para el diagnóstico microbiológico de HSV y de HPV.

Modulation of protective immunity against herpes simplex virus via mucosal genetic co-transfer of DNA vaccine with beta2-adrenergic agonist

Cholera toxin, which has been frequently used as mucosal adjuvant, leads to an irreversible activation of adenylyl cyclase, thereby accumulating cAMP in target cells. Here, it was assumed that beta2-adrenergic agonist salbutamol may have modulatory functions of immunity induced by DNA vaccine, since beta2-adrenergic agonists induce a temporary cAMP accumulation. To test this assumption, the present study evaluated the modulatory functions of salbutamol co-administered with DNA vaccine expressing gB of herpes simplex virus (HSV) via intranasal (i.n.) route. We found that the i.n. co-administration of salbutamol enhanced gB-specific IgG and IgA responses in both systemic and mucosal tissues, but optimal dosages of co-administered salbutamol were required to induce maximal immune responses. Moreover, the mucosal co-delivery of salbutamol with HSV DNA vaccine induced Th2-biased immunity against HSV antigen, as evidenced by IgG isotypes and Th1/Th2-type cytokine production. The enhanced immune responses caused by co-administration of salbutamol provided effective and rapid responses to HSV mucosal challenge, thereby conferring prolonged survival and reduced inflammation against viral infection. Therefore, these results suggest that salbutamol may be an attractive adjuvant for mucosal genetic transfer of DNA vaccine.

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.

Nuevos métodos diagnósticos para virus herpes simplex (HSV): importancia de la serología / New diagnostic methods for herpes simplex virus (HSV): the importance of serology

En los últimos años el gran avance de la biología molecular aplicada al diagnóstico de agentes infecciosos ha dado como resultado nuevas metodologías, más sensibles, específicas y rápidas, al alcance del médico clínico. Para el diagnóstico de los virus herpes simplex (HSV), los métodos de detección genómica y las nuevas técnicas específicas de serología han cubierto nuevas posibilidades de confirmación etiológica frente a la sospecha clínica, todo lo cual resulta finalmente en un mejor manejo terapéutico y epidemiológico.

In recent years, the great advances in the field of molecular biology, applied to diagnosis of infectious agents have resulted in new, more sensible, specific and faster methods that are more accessible to the physician. In the case of herpes simplex virus (HSV), genomic methods and specific serology have opened new possibilities for etiologic confirmation, resulting in better therapeutic and epidemiological management.

Inmunovaloración de anticuerpos IgM en muestras sospechosas de sarampión clínicamente

Se realiza el estudio de 60 sueros pareados sospechosos de Sarampión clínicamente llegados al Laboratorio Diagnóstico del Instituto de Medicina Tropical "Pedro Kourí" entre enero y mayo de 1996, procedentes de la vigilancia seroepidemiológica de la vacuna triple viral (sarampion, rubéola y parotiditis), a los cuales se les realizó la detección de anticuerpos hemaglutinantes a sarampión y rubéola, así como de IgM, con el estuche diagnóstico Clark Laboratories INC Measles IgM ELISA. Los casos positivos se confirmaron por las técnicas de inmunofluorescencia indirecta-IgM y neutralización. Se obtuvieron por inhibición de la hemaglutinación, 3 casos positivos a sarampión y rubéola, los cuales negativos a IgM de sarampión y a los que se les determinaron anticuerpos a Epstein Barr, Citomegalovirus y al virus herpes simple mediante inmunifluorescencia indirecta (IFI) la capacidad de estos virus de inducir respuesta policlonal. Por medio del ELISA IgM Clark se detectaron 6 casos positivos los cuales resultaron negativos por IFI.

60 matched sera clinically syspicious of measles that were received at the Diagnostic Laboratory of the "Pedro Kourí" Tropical Medicine Institute between January and May, 1996, coming from the seroepidemiological surveillance of the MPR vaccine were studied. The detection of measles and rubella hemagglutinant antibodies, as well as of IgM, was carried out with the Clark Laboratories INC. Measles IgM ELISA diagnostic kit. The positive cases were confirmed by the IgM-indirect immunofluorescence and neutralization. 3 positive cases to measles and rubella, which were negative to measles IgM, were obtained by hemagglutination inhibition. Antibodies against Epstein Barr, cytomegalovirus and herpes simplex virus were also determined by indirect immunofluorescence (IIF) due to the capacity of these viruses to induces polyclonal responses. 6 positive cases, which were negative by IIF, were detected by means of the above mentioned diagnostic kit.

Detection of herpes simplex virus and herpes simplex virus-antibodies in encephalitis and similar infectious diseases

The purpose of this study was to use the polymerase chain reaction [PCR] technique to detect herpes simplex virus genome in cerebrospinal fluid [CSF], together with enzyme immunoassay and antigen immunoblotting [AIB] to detect antibodies against herpes simplex virus, in 41 patients with encephalitis and similar infectious diseases of the central nervous system. None of the patients had the diagnosis of herpes simplex encephalitis. Cerebrospinal fluid and matched serum specimens were obtained within the first week of clinical symptoms. Oligoclonal IgG bands were found by isoelectric focusing in the cerebrospinal fluid of 20/41 patients of whom 9 had local synthesis and 11 had a systemic immune response. Antibody specific to herpes simplex virus was found by enzyme-linked immunosorbent assay [ELISA] in 10/33 patients tested. Herpes simplex virus-specific antibodies were found in the cerebrospinal fluid by antigen immunoblotting in 5/31 patients investigated, but these were of low affinity. There was no correlation between findings on ELISA and either the clinical diagnosis, the presence of oligoclonal IgG by isoelectric focusing, or the detection of specific antibodies by antigen immunoblotting. Careful interpretation of specific antibodies is necessary for they may result from non-specific activation by an unrelated agent. Polymerase chain reaction for herpes simplex virus deoxyribonucleic acid [DNA] may be used to help in the diagnosis of early cases of HSE and should also be considered in cases presenting with atypical clinical features and inconclusive results from laboratory tests

Medición de anticuerpos contra HSV-1 / Measurement of antibodies against HSV-1

El herpes simple tipo I (HSV I) es una de las patologías de mayor prevalencia en la población humana siendo los odontólogos un grupo de los más afectados. El presente estudio se realizó en 56 estudiantes de décimo semestre de la Pontificia Universidad Javeriana durante el primer período de 1997. Se hizo a través de un inmunotest y relacionaba la presencia de anticuerpos contra el HSV I y las manifestaciones clínicas, observando los anticuepos IgG específicos para el virus. En los hombres se encontró una alta incidencia de la enfermedad con títulos variables de IgG en suero sin que existiera una respuesta protectora en el caso de recurrencia. Asimismo en el estudio se evidencian individuos que presentan la enfermedad sin que existan títulos séricos de la inmunoglobulina estudiada.

Herpes simplex virus shedding in bone marrow transplant recipients during low-dose oral acyclovir prohylaxis

A 400mg dose twice-a-day oral acyclovir prophylaxis regimen was evaluated in 50 allogenic transplant recipients. Twenty (40 percent) patients experienced 24 episodes of herpes simplex virus (HSV) shedding; 17 (70.8 percent) occurring during prophylaxis. Thirteen of such episodes were asymptomatic and, in three, it was difficult to differentiate severe mucositis from viral lesions. In the remaining one, HSV pneumonia was suspected after a bronchoalveolar lavage (BAL) procedure performed in an attempt to early detection of cytomegalovirus (CMV). All cases responded to acyclovir therapy or dose adjustment suggesting that acyclovir resistance did not account for the occurrence of infection in our patients. These data demonstrated that oral acyclovir prophylaxis, 400mg dose twice-a-day, was inadequate to suppress viral shedding. The bronchoalveolar lavage procedure in a patient with HSV shedding could precipitate HSV spread to the lungs and the occurrence of pneumonia.

Herpes simplex hepatitis.

We report a 76-year-old man who presented with hepatitis. IgM antibodies to herpes simplex virus were positive and scraping from skin lesions showed presence of herpetic inclusion bodies. The patient died 4 days after the onset of illness.

Estudio serológico evolutivo a herpes simple y citomegalovirus en pacientes cubanos infectados con el virus de la inmunodeficiencia humana (VIH) / Serological study of herpes simplex and cytomegalovirus in VIH infected Cuban patients

Se estudió la respuesta serológica evolutiva frente a los virus herpes simple y Citomegalovirus en pacientes infectados con el VIH, agrupados en diferentes estadios de la enfermedad. Se observaron fluctaciones en los TPG de anticuerpos contra HSV en el grupo de pacientes asintomáticos con respecto a los pacientes SIDA y a los fallecidos. Existe una marcada caída en el TPG de anticuerpos contra HSV y CMV aproximadamente 1 año antes del fallecimiento de los pacientes

Herpes simples genital, uma doença sexualmente transmissível / Genital herpes simplex, a sexually transmitted disease

Na avaliaçäo da transmissibilidade sexual dos herpes simples genital, foi estudada a prevalência de anticorpos específicos contra o herpes simples vírus do tipo 2 (HSV-2), em populaçöes de baixo e de alto risco para doenças sexualmente transmissíveis (DST). A populaçäo de baixo risco para a aquisiçäo das DST foi constituída por 155 doadores voluntários de sangue no HUCFF/UFRJ, entre fevereiro e junho de 1994. Todos foram submetidos a um questionário acerca dos fatores de risco para a aquisiçäo das DST e que traçou o perfil sócio-epidemiológico desta populaçäo. Os pacientes de alto risco para a aquisiçäo das DST incluíram 85 portadores do vírus da imunodeficiência humana (HIV), tendo como fatores de risco o homossexualismo, a promiscuidade heterossexual ou contato sexual com o parceiro soropositivo para o HIV. Foram avaliadas, sorologicamente, 20 prostitutas, entre maio e julho de 1994. O teste usado para a análise de todas as amostras de soro, provenientes dos pacientes, foi o ELISA convencional específico para a detecçäo de IgG anti-HSV-2. Para os doadores de sangue, a prevalência do HSV-2 foi de 53,8 por cento, superior à observada em países ocidentais desenvolvidos. Entre os pacientes HIV positivos, ela atingiu 73 por cento (p<0,01). Para todo o grupo de alto risco para a aquisiçäo das doenças sexualmente transmissíveis atingiu 72 por cento de prevalência (p<0,05, razäo de chance = 6,27). As variáveis que mais se associaram à soroprevalência do HSV-2 foram a multiplicidade de parceiros sexuais, relaçöes sexuais com portador de herpes genital, história prévia de abortos e de relaçöes homossexuais. Os nossos achados permitem sugerir a realizaçäo de estudos adicionais de soroprevalência em nosso meio, especialmente entre os pacientes HIV positivos. Os pacientes soropositivos para o HSV-2 devem ser orientados quanto aos mecanismos de transmissäo do vírus

Necrotizing myelopathy--a report of two cases with review of literature.

Necrotizing myelopathy is an uncommon neurological disorder. Till 1991, only 31 cases have been described in the literature. In this report, clinical and neuropathological features in two patients with necrotizing myelopathy are described. The precise aetiological agent in first patient was undetermined, However in the second patient there was serological evidence, suggestive of Herpes simplex virus infection.

Seroprevalence of toxoplasma gondii, rubella virus, cytomegalovirus herpes simplex virus (TORCH) and syphilis in Jamaican pregnant women

Seroprevalence of toxoplasma gondii, rubella virus, cytomegalovirus, herpes simplex virus infections (TORCH) and syphilis were determined in order to assess the immune/susceptibility status in Jamaican pregnant women in 1986. The positive rates were 57%(T. gondii), 69%(rubella), 97%(CMV), 91%(HSV), and 4.9%(syphilis), respectively. The rate of reactivity for rubella was over 50%in all parishes, the highest being 85%in St. Thomas. The seroprevalence of T. gondii was lowest in Trelawny (37.5%). There were no significant differences in seropositivity of CMV and HSV infections between women from various parishes. The importance of seroprevalence of the TORCH group of agents and syphilis on perinatal morbidity and mortality in Jamaican women is discussed, and appropriate recommendations for prevention and control of congenital infections in Jamaica are suggested.