Josè Ramon Fiore

In Vitro and In Vivo Human Herpesvirus 8 Infection of Placenta

Herpesvirus infection of placenta may be harmful in pregnancy leading to disorders in fetal growth, premature delivery, miscarriage, or major congenital abnormalities. Although a correlation between human herpesvirus 8 (HHV-8) infection and abortion or low birth weight in children has been suggested, and rare cases of in utero or perinatal HHV-8 transmission have been documented, no direct evidence of HHV-8 infection of placenta has yet been reported. The aim of this study was to evaluate the in vitro and in vivo susceptibility of placental cells to HHV-8 infection. Short-term infection assays were performed on placental chorionic villi isolated from term placentae. Qualitative and quantitative HHV-8 detection were performed by PCR and real-time PCR, and HHV-8 proteins were analyzed by immunohistochemistry. Term placenta samples from HHV-8-seropositive women were analyzed for the presence of HHV-8 DNA and antigens. In vitro infected histocultures showed increasing amounts of HHV-8 DNA in tissues and supernatants; cyto- and syncitiotrophoblasts, as well as endothelial cells, expressed latent and lytic viral antigens. Increased apoptotic phenomena were visualized by the terminal deoxynucleotidyl transferase-mediated deoxyuridine nick end-labeling method in infected histocultures. Ex vivo, HHV-8 DNA and a latent viral antigen were detected in placenta samples from HHV-8-seropositive women. These findings demonstrate that HHV-8, like other human herpesviruses, may infect placental cells in vitro and in vivo, thus providing evidence that this phenomenon might influence vertical transmission and pregnancy outcome in HHV-8-infected women.

Changes in at-risk behavior for HIV infection among HIV-positive persons in Italy.

Many HIV-positive persons reportedly continue to engage in at-risk behavior. We compared the sexual and drug-using practices of HIV-positive persons before and after the diagnosis of HIV infection to determine whether their behavior had changed. To this end, in 2006, we conducted a cross-sectional study involving clinical centers in five Italian cities. Each center was asked to enroll 100 persons aged 18 years or older who had a diagnosis of HIV infection that dated back at least 2 years. Data were collected with a specifically designed questionnaire, administered during a structured interview. The McNemar chi2 test was used to compare the data before and after the diagnosis. A total of 497 persons participated (65.5% males; median age of 40 years; age range, 34-45 years). The most common exposure categories were: heterosexual contact (43.4%), homosexual contact (27.2%), and injecting drug use (20.6%). Although the percentage of drug users significantly decreased after diagnosis, 32.4% of injectors continued to use drugs, and approximately half of them exchanged syringes. Regarding sexual behavior, after diagnosis there was a significant decrease in the number of sexual partners and in stable relationships and an increase in condom use, both for persons with stable partners and those with occasional partners, although the percentage varied according to the specific sexual practice. These results indicate that though at-risk behavior seems to decrease after the diagnosis of HIV infection, seropositive persons continue to engage in at-risk practices, indicating the need for interventions specifically geared toward HIV-positive persons.

Immunological markers in HIV-infected pregnant and non-pregnant women.

OBJECTIVEnTo assess the influence of pregnancy on the course of HIV infection by comparing the behaviour of total lymphocyte counts and lymphocyte subsets (CD4(+) and CD8(+) and their ratio) in a cohort of infected pregnant women.nnnSETTINGnTertiary referral centre for high risk obstetrics and infectious diseases in pregnancy.nnnPATIENTS AND METHODSnA prospective study was designed, HIV infected women being enrolled at the beginning of pregnancy and sampled each trimester and in the puerperium. As controls, a group of non-pregnant HIV-infected women, cross-matched for age, risk factors and stage of disease were included and similarly evaluated in the same period.nnnRESULTSnAll the parameters, when longitudinally evaluated, were stable during gestation. Compared with non-pregnant subjects, patients had higher CD4(+) counts at the beginning and increased values of total lymphocytes count and subsets during the puerperium. Antepartum and postpartum risk factors such as drug abuse, smoking, antiretroviral therapy, length of gestation, maternal complications and HIV status of the neonate were not influential on the total lymphocytes counts and subsets.nnnDISCUSSIONnAccording to this data, pregnancy per se seems to have a negligible influence over the course of HIV infection, at least as far as immune parameters are concerned.

HTLV-I and HTLV-II infections in subjects at risk for HIV-1 infection from southeastern Italy (Apulia region)

To assess the prevalence of HTLV‐I and HTLV‐II infections in different groups at risk for HIV‐I infection, a study on 867 subjects was carried out by means of serological and PCR analyses. Serum specimens were collected from 268 intravenous drug users (IVDU), 66 homosexual men, 248 subjects with sexually transmitted diseases (STD), 105 thalassemics and 180 hemophiliacs. Sera from 3 IVDU and a sample from an STD patient were confirmed as HTLV‐II seropositive; a thalassemic patient was seropositive for HTLV‐I; a homosexual man, though confirmed as HTLV‐I/II‐seroreactive, could not be typed by serological methods. No hemophiliac was found to be HTLV‐I/II‐reactive. All 3 HTLV‐II‐seroreactive IVDU and the HTLV‐I‐infected thalassemic were confirmed by PCR; an additional sample from an IVDU, indeterminate by Western blot, was confirmed to be positive for HTLV‐II by PCR. Subtyping of HTLV‐II samples indicated the presence of II/b subtype in all 4 cases. Up to now, the reservoir for HTLV‐II infection in southeastern Italy is mainly represented by IVDU, while HTLV‐I infection seems to be sporadic.

Long-term efficacy and safety of switching from lamivudine + adefovir to tenofovir disoproxil fumarate in virologically suppressed patients

BACKGROUND AND AIMnTenofovir disoproxil fumarate (TDF) is recommended as first-line monotherapy for nucleos(t)ide (NA)-naïve chronic hepatitis B (CHB) patients and as a second-line rescue therapy for NA-experienced patients with a previous treatment failure. However, data regarding the efficacy of TDF monotherapy in patients with lamivudine resistance (LAM-R) successfully treated with LAM+adefovir (ADV) are limited. Herein, the efficacy and safety of switching from LAM+ADV to TDF monotherapy in clinical practice have been evaluated.nnnMETHODSnSixty LAM-R HBeAg-negative CHB patients treated with ADV add-on therapy and stable viral suppression, were switched to TDF monotherapy and prospectively evaluated for virological response, liver and renal function, and bone mineral density.nnnRESULTSnDuring a median period of 57 months of TDF monotherapy, all patients maintained a virological response, four of whom cleared HBsAg (6.6%) and discontinued treatment. Monitoring of renal function showed no case of the Fanconi syndrome, no significant alterations of median serum creatinine, eGFR and phosphate levels, although a reduction of TDF dosage was required in five patients (8.3%). Despite the stable virological suppression, five cirrhotic patients and one CHB patient developed hepatocellular carcinoma.nnnCONCLUSIONSnOur results demonstrate the efficacy of switching to TDF monotherapy in virologically suppressed CHB patients receiving long-term LAM+ADV therapy, with a low rate of adverse events.

Serum IgG Antibodies from Pregnant Women Reacting to Mimotopes of Simian Virus 40 Large T Antigen, the Viral Oncoprotein

Simian virus 40 (SV40) large T antigen (LT) coding sequences were revealed in different human samples, whereas SV40 antibodies (Ab) were detected in human sera of cancer patients and healthy individuals, although with a lower prevalence. Previous studies carried out by the neutralization assay gave a SV40 seroprevalence, in the general population, up to 8%, although higher rates, 12%, were detected in kidney transplant children, in a group of HIV-positive patients, and in healthy females. In this study, serum samples from pregnant women, together with those from non-pregnant women, were analyzed to check the prevalence of IgG Ab reacting to SV40 LT antigens. Serum samples were collected from pregnant and non-pregnant women, with the same mean age. Women were in the range of 15–48u2009years old. Samples were assayed by an indirect ELISA employing specific SV40 LT mimotopes as antigens, whereas functional analysis was performed by neutralization of the viral infectivity in cell cultures. As a control, sera were analyzed for Ab against BK polyomavirus (BKPyV), which is a human polyomavirus homologous to SV40. Statistical analyses employed chi-square with Yates’ correction, and Student’s t tests. Indirect ELISAs indicated that pregnant women tested SV40 LT-positive with a prevalence of 17% (23/134), whereas non-pregnant women had a prevalence of 20% (36/180) (Pu2009>u20090.05). Ab against BKPyV were detected with a prevalence of 80% in pregnant women and with a prevalence of 78% in non-pregnant women. These data indicate that SV40 infects at a low prevalence pregnant women. We may speculate that SV40, or a close human polyomavirus still undetected, could be transmitted from mother to fetus.

Correlation between knowledge on transmission and prevention of HIV/STI and proficiency in condom use among male migrants from Africa and Middle East evaluated by a Condom Use Skills score using a wooden penile model

BackgroundMigrants in Italy are prevalently young adults, with a higher risk of sexual transmitted infections (STI) and HIV infection. Promoting consistent as well as correct use of condoms could reduce failure rate due to their improper use. The aim of our study was to evaluate Condom Use Skills among a migrant population recently landed in Italy, hosted in a government center for asylum seekers.MethodsThe study sample was composed of 80 male migrants. Sanitary trained interviewers submitted a questionnaire to participants to investigate age, provenience, marital status, educational level and knowledge about transmission and prevention of HIV/STI. Then, we assessed participants’ level of condom use skill with the Condom Use Skills (CUS) measure by using a wooden penile model. The interviewer filled in a checklist and assigned 1 point for correct demonstration of each behavior that may prevent condom failure during sex.ResultsParticipants’ median age was 26xa0years and the sample was composed of 54 migrants from sub-Saharan Africa and 26 from Middle East. Most of them were married, with a lower middle level of education, up to 8 or 5xa0years. Half of the sample achieved the highest score in the questionnaire and our CUS showed a large number of people with middle high score classes. The Spearman’s rho was 0.30, therefore answers to the questionnaire and CUS score appeared correlated (pxa0<xa00.05). In the multivariate model, to have a higher CUS score resulted to be associated to be older than 26xa0years (pxa0<xa00.05), with a higher level of education (pxa0=xa00.001), and a higher score in the questionnaire (pxa0<xa00.05). There were no significant differences in the level of CUS between single or married men and between African and Middle Asian migrants of the sample.ConclusionsOur study shows that educational level influences the quality of knowledge and awareness about STI/AIDS and contribute to correct condom use. Since the half of participants had a low educational level and linguistic problems, the risk of missing campaigns messages or misunderstanding informative materials increases. Direct observation of condom-application on penile model may offer realistic assessment of application skills in these individuals.

Human herpesvirus 8 (HHV-8) “in vitro” infection of human placental histocultures

Most human Herpesvirus infect placental cells and may be harmful in pregnancy, leading to obstetrical and/or neonatal complications. Although a correlation between human herpesvirus 8 (HHV-8) infection and abortion or low birth weight in children has been reported [1,2] presently no information has been published regarding HHV-8 tropism for placenta.

In vitro Zidovudine-Induced Apoptosis Demonstrated by a Model of Placental Histoculture Useful for the Study of the Apoptotic Effects of Antiretroviral Drugs

This study aims to evaluate the use of a placenta histoculture system based on placental villi culture on collagen sponge gels at the liquid/air interface, for the detection of zidovudine-induced apoptosis. Explant cultures from full term placentas were exposed to different concentrations of zidovudine (1, 10, 100 uM) up to 7 days and samples were analysed for apoptosis with different methods after 3, 24, 48 hours and 7 days of exposure. Apoptotic phenomena were demonstrated at 10 and 100 uM zidovudine, more delayed at the lowest concentration: this demonstrates that zidovudine-induced placenta apoptosis in vitro is both time- and dose-dependent and that also concentrations near to those achievable in vivo might cause placenta apoptosis. This has to be taken into account when considering possible consequences of antiretroviral treatments during pregnancy. Placental histocultures on collagen sponge cells are a reliable tool for the study of antiretroviral-induced toxicity in placenta: in fact, explants cultured for more prolonged periods (up to 7–14 days) in a “physiological milieu” allow the detection of biological effects otherwise not evident in the commonly used, short-term, placental cultures.