Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Propolis is a natural compound and has been highlighted for its antioxidant, antiinflammatory and antiviral properties. The purpose of this study was to investigate if brown hydroalcoholic propolis extract (HPE) protects against vaginal lesions caused by herpes simplex virus type 2 (HSV-2) in female BALB/c mice. The treatment was divided in 5 days of pre-treatment with HPE [50 mg/kg, once a day, intragastric (i.g.)], HSV-2 infection [10 ml of a solution 1x102 plaque-forming unit (PFU/ml 1 HSV-2), intravaginal inoculation at day 6] and post-treatment with HPE (50 mg/kg) for 5 days more. At day 11, in vivo (score of lesions) and ex vivo analysis [haematological and histological evaluation; superoxide dismutase (SOD), catalase (CAT) and myeloperoxidase (MPO) activities; reactive species (RS), tyrosine nitration, non-protein thiols (NPSH) and ascorbic acid (AA) levels] were carried out. HPE treatment reduced extravaginal lesions and the histological damage caused by HSV-2 infection in vaginal tissues of animals. HPE was able to decrease RS, tyrosine nitration, AA levels and MPO activity. Also, it protected against the inhibition of CAT activity in vaginal tissues of mice. HPE promoted protective effect on HSV-2 infected animals by acting on inflammatory and oxidative processes, and this effect probably is due its antioxidant and anti-inflammatory properties. / O própolis é um composto natural e se destaca por suas propriedades antioxidante, antiinflamatória e antiviral. O objetivo do presente estudo foi investigar se o extrato hidroalcoólico de própolis marrom (EHP) tem efeito protetor frente às lesões vaginais causadas pelo vírus do herpes simplex tipo 2 (HSV-2) em camundongos fêmeas BALB / c. O tratamento foi dividido em 5 dias de pré-tratamento com EHP [50 mg/kg, uma vez por dia, via intragástrica (i.g.)], infecção por HSV-2 [10 ml de uma solução de 1x102 de unidades formadoras de placas (PFU/ml-1 HSV-2), inoculação intravaginal no dia 6] e pós-tratamento com EHP (50 mg/kg) durante mais 5 dias. No dia 11, análises in vivo (escore de lesões) e ex vivo [avaliação hematológica e histológica; atividade das enzimas superóxido dismutase (SOD), catalase (CAT) e mieloperoxidase (MPO); espécies reativas (ER), níveis de nitração da tirosina, tióis não proteicos (NPSH) e ácido ascórbico (AA)] foram realizadas. O tratamento com o EHP reduziu as lesões extravaginais e os danos histológicos causados pelo HSV-2 no tecido vaginal dos animais infectados. O EHP foi capaz de diminuir os níveis de ER, de nitração da tirosina, de AA e a atividade da MPO. Além disso, protegeu contra a inibição de atividade da CAT nos tecidos vaginais. O EHP promoveu efeito protetor nos animais infectados com HSV-2 agindo sobre os processos inflamatórios e oxidativos, este efeito provavelmente ocorre devido as suas propriedades antioxidante e anti-inflamatória.

Recent studies suggest that some cases of prenatal depression may be associated with reactivation of latent infections of the herpesvirus family. The possible relationships among stress, prenatal depression, and herpes viral reactivation in pregnancy are understudied and the molecular pathways such as the neuroimmune biogenic amine pathway are unidentified. Chronic stress shifts the T helper-1 cell (Th1) cytokine profile to a Th2 profile, which favors virus induced pathogenesis and survival. Pregnancy is also associated with a similar Th2 dominance. In non-pregnant individuals, exposure to psychological or physical stress may be associated with latent herpes viral reactivation and could result in behavioral deficits and depression. Normally, type-1 cytokines such as Interferon-gamma (IFN -gamma) and inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) induce indoleamine-2, 3-dioxygenase (IDO) activation which inhibits herpes virus replication and reactivation, decreases tryptophan production, and alters phenylalanine /tyrosine metabolism. Thus it is possible that prenatal depression may occur from tryptophan stealing through the IDO pathway which results in decreased serotonin as well as increased risk for latent herpes viral reactivation. The purpose of this study is to analyze the relationships among stress, herpes viral titers, depression, and metabolites of IDO activation, which involves tryptophan and guanosine-triphosphate-cyclohydrolase-1(GTP-CH1) pathways. This study builds on Influence of Lactation on Postpartum Stress and Immunity (Grant number: R01-NR05000) which investigated perinatal immune, endocrine, and inflammatory changes in pregnancy and the postpartum. A secondary data analysis was conducted on baseline data from women collected at 16 to 25 gestational weeks. This data set included some herpes viral titers, and additional ones were measured in stored plasma samples. The aim of this study is to examine relationships among stress, herpes viral reactivation, depression, and the IDO activation pathway. The results of this study provide information about the possible role of further relationships of prenatal stress, latent herpes viral reactivation, and depression mechanisms. The results will be important in health promotion and disease prevention during pregnancy.

Objective: The objective of this study was to describe the prevalence of sexually transmitted co-infections and assess factors associated with a single infection and co-infections. Methods: A total of 388 women were included in this study. At enrollment of the EVRI trial women were tested for five STIs: Human papilloma virus (HPV), Chlamydia trachomatis, Neisseria gonorrhoeae, Herpes simplex virus-2 (HSV-2) and Treponema pallidum. Prevalence of STI infections and proportion of women with different combinations of co-infections were calculated. Factors associated with single infection and STI co-infection were assessed using a polytomous logistic regression model and odds ratio (OR) and 95% confidence intervals (95% CI) were reported as the measure of association. Results: Prevalence of a single STI and concurrent STI co-infections were high. Prevalence of chlamydia was 33%, syphilis 6%, HSV-2 46% and HPV 71%. The most common STI co-infection pattern was HPV-HSV (32%), followed by HPV-chlamydia (17%). The odds of single STI compared to no STIs was significantly lower for women who had education level of grade 7 compared to women who had some college or technical education (OR 0.16, 95% CI: 0.03-0.79).The odds of a single STI compared to no STIs were significantly higher (OR 3.02, 95% CI: 1.05-8.64) and the odds of concurrent STIs compared to no infection were significantly higher (OR 3.86, 95% CI: 1.42-10.48) for women with three or more lifetime partners compared to one life time partner. Conclusions: STIs, single and multiple concurrent infections, are common among this cohort of South African women. These results strengthen the recommendation that STI screening and treatment needs to be a component of multiple intervention strategies among high-risk women residing in communities with high STI prevalence.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第23467号 / 医博第4774号 / 新制||医||1053(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 森信 暁雄, 教授 上野 英樹, 教授 小柳 義夫 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Chlamydia trachomatis/HSV-2 vaginal co-infections are seen clinically, suggesting that these sexually transmitted pathogens may interact. We previously established an intravaginal Chlamydia muridarum/HSV-2 super-infection model and observed that chlamydial pre-infection protects mice from a subsequent lethal HSV-2 challenge. However, the mechanism of protection remains unknown. The type I interferon, IFN-β, binds to the type I interferon receptor (IFNR), elicits a host cellular antiviral response and inhibits HSV replication in vitro and in vivo. Previous studies have demonstrated that C. muridarum infection stimulates genital tract (GT) IFN-β production; therefore, we hypothesized that chlamydial pre-infection protects mice from HSV-2 challenge via the IFN-β/IFNR-induced antiviral response. To test this prediction, we quantified IFN-β levels in vaginal swab samples. Detection of IFN-β in C. muridarum singly infected, but not in mock-infected animals, prompted the use of the super-infection model in IFNR knockout (IFNR-/-) mice. We observed that C. muridarum pre-infection reduces HSV-2-induced mortality by 40% in wild-type mice and by 60% IFNR-/-mice. Severity of HSV-2 disease symptoms and viral shedding was also similarly reduced by C. muridarum pre-infection. These data indicate that, while chlamydial infection induces GT production of IFN-β, type I IFN-induced antiviral responses are likely not required for the observed protective effect.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal. / Le cancer du col utérin est la troisième néoplasie en importance chez la femme mondialement. Plusieurs études cliniques et épidémiologiques démontrent que le virus du papillome humain (HP V) est un agent étiologique impliqué, nécessaire au développement des néoplasies du col. Par contre, de plus en plus d'évidences suggèrent une interaction de multiples cofacteurs dans la progression vers le cancer. Nous avons démontré précédemment que des cellules cotransfectées par HPV-16 et la séquence Xho-2 du virus herpès simplex type 2 (HSV-2) induisaient des tumeurs dans des souris athymiques, contrairement à des cellules transfectées par HPV-16 ou HSV-2 seuls. Nous nous proposons d'étudier dans cette étude transversale le rôle de HSV-2, ainsi que de Chlamydîa trachomatis (CT) et du virus herpès humain type 6 (HHV-6) comme cofacteurs aux HPV dans la progression des lésions néoplasiques du col. Une cohorte de femmes a été recrutée consécutivement dans une clinique de colposcopie. Des cellules exfoliées du col ont été soumises à des tests PCR standardisés pour la présence de CT et de HHV-6. 27 génotypes de HPV ont été identifiés avec le Line Blot assay. La région transformante Bgl IIC de HSV-2 a été détectée par PCR. La région Xho-2 de HSV-2 a été détectée par une technique de détection par PCR déjà bien établie (PCR Xho-2) auparavant. Parmi les 439 femmes recrutées, 244 étaient normales, 80 avaient des lésions du col de bas grade (LSIL), 65 avaient des lésions de haut grade (HSIL) et 50 avaient im cancer du col. Des HP V à haut risque étaient détectées dans 93.3% des femmes avec HSIL et 20% des femmes normales. HPV-16 était le plus fréquemment détectés parmi les génotypes de HPV. Tous les spécimens étaient négatifs pour les régions transformantes Xho-2 et Bgl IIC de HSV-2. HHV-6 a été détecté dans 10 spécimens toutes lésions confondues. 3 spécimens de femmes avec un cancer étaient positifs pour CT, contrairement à aucun des spécimens de femmes sans lésions (p<0.031). En conclusion, notre étude a démontré que même si la séquence Xho-2 de HSV-2 transformes des cellules in vitro, elle n'a pas été détectée chez des femmes avec des lésions de haut grade du col. Cependant, C. trachomatis et HHV-6 ont été associés significativement avec les lésions de haut grade et cancer du col et pourraient être des cofacteurs aux HPV.

Most epidemiologic studies of sexually transmitted diseases (STD) are based on patients seeking help or advice at various health care settings. Because many STD:s are subclinical, epidemiologic surveys can be strengthened by a population-based approach. The aims of the present study were to investigate the prevalence and incidence of STDs in a population of young women, and to assess associations between STDs and social background, education, previous genital infections, sexual behaviour, contraceptive use and reproductive experience. All women belonging to the 19-, 21-, 23- and 25-year age cohorts and living in the catchment area of a community health center, were invited by mail to participate in the study. In the presence of the investigator, participants answered a structured questionaire regarding their social background, education, previous genital infections, sexual behaviour, contraceptive use and reproductive experience. A gynecologic examination was performed. Cervical scrapes for human papillomavirus (HPV) DNA, as well as samples for wet smear, cervical pap smear, and Chlamydia trachomatis (CT) culture were taken. The presence of genital warts was noted, and a colposcopy was performed 2-5 minutes after application of 5% acetic acid on the cervix and vulva. Acetowhite changes were then assessed. A serologic test for CT and herpes simplex virus type 2 (HSV-2) antibodies were performed. Of the 816 women available, 611 (75%) participated in the study. One out of four women reported symptoms from the lower genital tract. The most common were itching, followed by discharge and soreness. There was a significant correlation between the womens" complaint of vaginal discharge, and previous CT infection, lack of lactobacilli and the presence of leucocytosis in wet smear. Twenty-two percent of the women were HPV DNA positive and acetowhitening at the cervix was observed in 16% of the women. The sensitivity of detection of HPV infection by acetowhitening of the cervix was 22% (95%CI 18%, 26%), and the specificity was 90% (95% Cl 87%, 93%). C.trachomatis culture positivity was found in 2.7% of the women and the seroprevalence of CT was 24.7 %. Atypical cytology was found in 3.4% of the women and 6.6% was HSV-2 seropositiv. Of the women studied 23.6% reported having had at least one STD previously and the laboratory analysis showed 45.4% to have had at least one STD. Multivariate logistic regression analysis showed that the number of sexual partners, age at first coitus, history of therapeutic abortion, and previous pelvic inflammatory disease (PID) was independently correlated with CT seropositivity. Lifetime number of sexual partners was the only independent risk factor for HPV. Multivariate analysis showed that increasing age, early sexual debut, and a history of spontaneous abortion were independently related to the presence of HSV-2 antibodies. The lifetime number of sexual partners and coitus on first date were independently associated with a previous STD. Conclusion, We found that one out of four women had some kind of lower genital tract complaint, almost every other women had at sometime in their life an STD, and STDs were often asymptomatic. Acetowhitening of the cervix and vulva has low sensitivity, to low to warrant its use as a predictor of subclinical HPV infection. The pattern of risk factors differed between STDs. / <p>Härtill 6 uppsatser</p> / digitalisering@umu

Clinical and experimental studies have shown that estradiol (E2) can enhance protection against sexually transmitted infections such as HSV-2 and HIV-1. Antigen presenting cells (APCs) such as Dendritic cells (DCs) are critical for generating immune responses against these infections, and it is unclear whether unique factors present in the genital mucosa can influence immune responses by directly modulating the phenotype and function of local APCs. To address this, I hypothesized that sex hormones, such as E2 and innate factors in the local microenvironment can regulate the phenotype and function of vaginal APCs. The work summarized in this thesis addressed this central hypothesis. In the first section of the thesis, I examined whether vaginal APCs were distinct in their phenotype and function compared to those in other mucosal tissues or spleen. The results show that the vagina was enriched in CD11c+ CD11b+ MHCII− DCs. Functionally, vaginal tissue cells (TC) and CD11c+ DCs were more potent inducers of Th17 responses in co-cultures with CD4+ T cells, compared to lung, small intestine or spleen APCs. E2 was critical for the conditioning of vaginal DCs to prime these Th17 responses through an IL-1-dependent pathway, indicating that sex hormones such as E2 can directly influence the function of vaginal APCs. In the next section, I determined whether other co-factors in the genital microenvironment such as microflora and innate lymphocytes could also influence vaginal APC functions. We found that while microflora was not essential, IL-17 produced by innate lymphocytes was critical for the induction of IL-1 from DCs, and consequently for potentiating Th17 responses. Finally, I attempted to develop an in vivo mouse model where the effect of E2 on vaginal APCs could be examined in the context of genital HSV-2 infection. I tested a 7-day injectable E2 and a 21-day E2 pellet delivery model, and found that both regimes had limitations for examining E2-effects on anti-viral responses. Yet, subsequent to the work done in this thesis, we were able to confirm our observations of E2-conditioned Th17 responses in vivo in an intranasal immunization model utilizing E2 pellet delivery, and thereby addressed the mechanism underlying enhanced anti-viral protection following E2-treatment. In conclusion, this is the first study to show the effect of E2 on genital tract APCs and their ability to prime Th17 responses. It provides future avenues to examine whether modulation of this microenvironment can help optimize vaccine-induced immune responses against STIs. On a more fundamental level, it highlights the need to consider the inherent distinctions in APC populations among different mucosal tissues. / Dissertation / Doctor of Philosophy (PhD)

<p>Sexually transmitted infections (STIs), especially HIV/AIDS and HSV-2, continue to be a devastating burden on societies around the world. The close link between HSV-2 and HIV-1, the role of inflammation in driving these infections, and the limited success and availability of prophylactic and therapeutic measures underscore the need for continued search of alternative means of protection. Characterization of endogenous antimicrobials, especially those local to the female genital tract and actively regulating inflammatory and antiviral responses, could be beneficial for microbicidal trials. Although regulators of mucosal immunity, such as serine antiproteases, trappin-2 and elafin (Tr/E), have been associated with resistance to HIV-1, their antiviral activity remains poorly understood. Thus, the research presented in this thesis centers on characterization of antiviral properties of Tr and E individually and their potential mechanisms in defense against HSV-2 and HIV-1 in the female genital mucosa. Chapter 2 examines Tr/E contribution to antiviral host defense responses elicited by a synthetic mimic of viral dsRNA, polyI:C. Chapter 3 documents the presence and characteristics, including potential mechanisms, of antiviral activity of Tr/E against in vitro and in vivo HSV-2 infection. Chapters 4 and 5 determine the contribution of Tr/E to the natural anti-HIV-1 protection of CVL and structural characteristics, mode(s) of action, and cellular distribution/localization of antiviral Tr/E proteins. Therein, we present novel properties of each Tr/E by demonstrating their inhibitory and multiple effects against both HSV-2 and HIV-1. These effects appear to be mediated either through virus or cells and be associated with altered viral attachment/entry, transcytosis and infection, innate viral recognition, modulated inflammation and increased antiviral protection of cells. Reported antiviral activity of Tr/E was also contextual and exerted, at least in HEC-1A cells, via autocrine/paracrine mode and depended on elafin’s nuclear localization and its unmodified N-terminus. Tr/E may represent viable candidates for further studies in the field of STIs.</p> / Doctor of Philosophy (PhD)

Background: Herpes Simplex Virus Type 2 (HSV 2) is one of the most common sexually transmitted infections (STIs) worldwide. HSV 2 infection is a risk factor for the acquisition and transmission of other STIs. Aim: The aim of this study is to determine prevalence and predictors of HSV 2 infection in Durban, South Africa by using available data that has not been previously analysed for the purpose of adding scientific evidence to the existing body of knowledge relating to HSV 2. Method: The study involves secondary analyses of data collected as a prospective study which enrolled women who participated in a clinical trial. A total of 3472 sexually active women were screened in the primary study from two clinics in Durban. All consenting participants were tested for HIV, HSV 2, Trichomonas vaginalis (TV), Neisseria gonorrhoea (NG) and Chlamydia trachomatis (CT) infection. Results: There were 2532 women who had HSV 2 giving a prevalence of 73%. Of these, 53% also tested positive for HIV infection. In univariate analysis, co-infection with HIV was strongly associated with HSV2 (Odds Ratio (OR): 7.4, 95% Confidence Interval (CI): 6.0, 9.1, p<0.001). There was also an association between other STIs, such as CT, NG and syphilis and HSV 2, although only NG was significantly associated with prevalent of HSV 2 (OR: 2.3, 95% CI: 1.3, 4.1, p=0.005). Women older than 25 years of age more likely to have HSV 2 (OR: 2.4, 95% CI: 2.0,2.8, p<0.001). A risk of being infected with HSV 2 increased with the number of reported lifetime sexual partners Those with two and three or more were 2,5 and 4.6 times more likely to have HSV2 respectively (OR: 2.5, 95% CI: 2.1,3.1, p<0.001 and OR: 4.6, 95% CI: 3.8, 5.6, p<0.001 respectively). Women who had less than high school education were also found to have higher risk for HSV 2. Conclusion: The secondary analysis showed a high prevalence of HSV 2 infection and a strong association of HSV 2 and HIV. A significant association of HSV 2 was noted in women having more than two sex partners and lower high school education. Therefore, it is recommended that screening for HSV 2 among high risk populations be incorporated into the STI screening and treatment packages. / Prevalence and risk factors associated with HSV 2 / Herpes Simplex Virus Type 2 / HSV 2 / Department of Health Studies / M. (Public health)