Aldo Di Carlo

Prevalence and determinants of anti‐lytic and anti‐latent antibodies to human herpesvirus‐8 among Italian individuals at risk of sexually and parenterally transmitted infections

Three hundred seventy‐nine individuals [137 non‐injecting drug using (non‐IDU) heterosexuals, 130 homosexual men and 112 IDU] attending the human immunodeficiency virus (HIV) testing program of a sexually transmitted disease (STD) clinic in Rome were studied to estimate the prevalence and to identify the modalities of transmission of human herpesvirus‐8 (HHV‐8) infection. Serological analysis was performed by using an immunofluorescence assay able to detect anti‐latent and anti‐lytic HHV‐8 antibodies. Twelve acquired immunodeficiency syndrome (AIDS)‐Kaposis sarcoma (KS) patients and 94 blood donors were tested as reference population groups. Anti‐lytic antibodies were detected in 185 (48.8%) individuals; 52 of them (13.7%) also had anti‐latent antibodies. Both anti‐lytic and anti‐latent antibody prevalence were higher among homosexual men (66.9% and 27.7%, respectively) than among IDU (49.1% and 8.0%, respectively) and non‐IDU heterosexuals (31.4% and 5.1%, respectively), and tended to increase with age. Anti‐lytic HHV‐8 antibodies were associated with syphilis [odds ratio (OR) = 3.81] but not with hepatitis C virus (HCV) seropositivity. HIV‐infected homosexual men were more likely to have HHV‐8 antibodies than those who were HIV‐negative. When using anti‐latent antibodies the direction of the OR remained the same, although the associations did not often reach statistical significance. Among AIDS‐KS patients, 83.3% had anti‐lytic and 66.6% had anti‐latent antibodies. Among blood donors, 28% had anti‐lytic antibodies and 2 of them (2.1%) also had anti‐latent antibodies. Our data indicate that HHV‐8 seroprevalence increases with age and is higher among homosexual men, particularly those infected with HIV. This is consistent with sexual transmission of HHV‐8 infection. In addition, the presence of HHV‐8 antibodies in HIV‐negative non‐IDU heterosexual contacts and in healthy blood donors is consistent with the high incidence of classic KS in Italy. Int. J. Cancer. 77:361–365, 1998.

Phase I therapeutic trial of the HIV-1 Tat protein and long term follow-up

A randomized, double blind, placebo-controlled phase I vaccine trial based on the native Tat protein was conducted in HIV-infected asymptomatic individuals. The vaccine was administered five times subcute with alum or intradermally without adjuvant at 7.5microg, 15microg or 30microg doses, respectively. The Tat vaccine was well tolerated both locally and systemically and induced and/or maintained Tat-specific T helper (Th)-1 T-cell responses and Th-2 responses in all subjects with a wide spectrum of functional anti-Tat antibodies, rarely seen in HIV-infected subjects. The data indicate the achievement of both the primary (safety) and secondary (immunogenicity) endpoints of the study.

Prevalence, genotype diversity and determinants of anal HPV infection in HIV-uninfected men having sex with men.

BACKGROUND Anal human papillomavirus (HPV) infection is very common in men having sex with men (MSM), but the available data on its burden and characteristics mainly concern HIV-infected individuals. OBJECTIVES This study aimed to assess the prevalence, spectrum of genotypes, and determinants of the anal HPV infection in metropolitan HIV-1 uninfected MSM. STUDY DESIGN A cohort of 258 MSM (median age 32 years, IQR 26-39) enrolled at an STI Clinic was screened for anal HPV infection using a highly sensitive PCR-based genotyping method. Medical history and behavioral data were collected. RESULTS Overall, 74.8% of the MSM were HPV-positive, with 56.2% of the participants being infected by high-risk (HR) types. A multiple infection was detected in 65.3% of the HPV-positive MSM, with up to 10 different HPV types detected in the same sample. A broad spectrum of infecting HPV types was observed, with 36 different types found overall and HPV16 representing the most common type (17.8%). The lifetime and recent number of sexual partners as well as having receptive anal sex were significantly associated with the anal HPV infection, confirming the role of sexual behavior in risk of HPV infection. However, neither younger age at first intercourse nor inconsistent use of condom was significantly associated with the infection. CONCLUSIONS The present findings highlight the need to create a more significant awareness about the risk of anal HPV infection among HIV-uninfected MSM and warrant the investigation of possible anal intraepithelial lesions, particularly in view of the increasing anal cancer incidence in high-risk populations.

The preventive phase I trial with the HIV-1 Tat-based vaccine.

The native HIV-1 Tat protein was chosen as vaccine candidate for phase I clinical trials based on its role in the natural infection and AIDS pathogenesis, on the association of Tat-specific immune response with the asymptomatic stage as well as on its sequence conservation among HIV clades. A randomized, double blind, placebo-controlled phase I study (ISS P-001) was conducted in healthy adult volunteers without identifiable risk of HIV infection. Tat was administered 5 times monthly, subcute in alum or intradermic alone at 7.5 microg, 15 microg or 30 microg, respectively (ClinicalTrials.gov identifier: NCT00529698). Vaccination with Tat resulted to be safe and well tolerated (primary endpoint) both locally and systemically. In addition, Tat induced both Th1 and Th2 type specific immune responses in all subjects (secondary endpoint) with a wide spectrum of functional antibodies that are rarely seen in natural infection, providing key information for further clinical development of the Tat vaccine candidate.

The therapeutic phase I trial of the recombinant native HIV-1 Tat protein.

The native HIV-1 Tat protein was chosen as a vaccine candidate based on its key role in the virus life cycle and on the correlation of Tat-specific immune responses with the asymptomatic stage and lower disease progression rates, but also due to its sequence conservation amongst the various HIV clades as well as the adjuvant effects on dendritic cells. Safety, immunogenicity and efficacy data in monkeys support the development of this vaccine concept.

Increased HIV incidence among men who have sex with men in Rome.

Among 976 men who have sex with men (MSM) who had undergone repeat HIV testing between 1984 and 2003 at a sexually transmitted infection clinic in Rome, Italy, we observed a dramatic increase in HIV incidence in 2002 and 2003, with the cumulative incidence for 2000–2003 being twice as high as that for 1984–1995, and significantly higher than that for 1996–1999. This trend suggests the need for interventions aimed at encouraging behavioural changes among MSM.

Incidence of Human Herpesvirus 8 (HHV-8) infection among HIV-uninfected individuals at high risk for sexually transmitted infections

BackgroundThe occurrence of, and risk factors for, HHV-8 infection have yet to be definitively determined, particularly among heterosexual individuals with at-risk behavior for sexually transmitted infections (STI). The objective of this study was to estimate the incidence and determinants of HHV-8 infection among HIV-uninfected individuals repeatedly attending an urban STI clinic.MethodsSera from consecutive HIV-uninfected individuals repeatedly tested for HIV-1 antibodies were additionally tested for HHV-8 antibodies using an immunofluorescence assay. To identify determinants of HHV-8 infection, a nested case-control study and multivariate logistic regression analysis were performed.ResultsSera from 456 HIV-uninfected individuals (224 multiple-partner heterosexuals and 232 men who have sex with men (MSM]) were identified for inclusion in the study. The HHV-8 seroprevalence at enrollment was 9.4% (21/224; 95% C.I.: 6.0–14.2%) among heterosexuals with multiple partners and 22.0% (51/232; 95% C.I.: 16.9–28.0%) among MSM. Among the 203 multiple-partner heterosexuals and 181 MSM who were initially HHV-8-negative, 17 (IR = 3.0/100 p-y, 95% C.I.: 1.9 – 4.8) and 21 (IR = 3.3/100 p-y, 95% C.I:.2.1 – 5.1) seroconversions occurred, respectively. HHV-8 seroconversion tended to be associated with a high number of sexual partners during the follow-up among MSM (> 10 partners: AOR = 3.32 95% CI:0.89–12.46) and among the multiple-partner heterosexuals (> 10 partner; AOR = 3.46, 95% CI:0.42–28.2). Moreover, among MSM, HHV-8 seroconversion tended to be associated with STI (AOR = 1.80 95%CI: 0.52–7.96).During the study period the HIV-1 incidence was lower than that of HHV-8 among both groups (0.89/100 p-y among MSM and 0.95/100 p-y among multiple-partner heterosexuals).ConclusionThe large difference between the incidence of HHV-8 and the incidence of HIV-1 and other STIs may suggest that the circulation of HHV-8 is sustained by practices other than classical at-risk sexual behavior.

Comparative evaluation of different DNA extraction methods for HPV genotyping by Linear Array and INNO-LiPA

In order to investigate the influence of DNA extraction on two PCR‐based HPV genotyping tests (Linear Array, Roche and INNO‐LiPA Extra, Innogenetics), three different procedures were used to purify DNA from 28 cervico‐vaginal samples tested previously by the Hybrid Capture 2: the AmpliLute Liquid Media Extraction kit (Roche), the QIAamp DNA Blood mini kit (QIAGEN), and the NucliSENS EasyMAG automated platform (bioMérieux). All HC2‐positive samples were found positive by both assays, independently of the extract used. Type‐specific concordance (i.e., identical HPV type‐specific profile in all the extracts of the same sample) was observed in 55% and 75% of the cases testing samples by the Linear Array and the INNO‐LiPA, respectively. Using the DNA extracted with the two manual methods the results were concordant in 75% of the cases both for the Linear Array and the INNO‐LiPA. When comparing the Linear Array results obtained on either of the two manual extracts with those obtained following automated extraction, 65% of the samples showed type‐specific concordance in both cases. The INNO‐LiPA results were concordant in 80% of the cases comparing the AmpliLute versus the automated extract, while concordant results were observed in 90% of the cases when comparing the QIAGEN versus the automated extract. In conclusion, the Linear Array and INNO‐LiPA results are affected by the method of DNA extraction. Consequently, different HPV type‐specific profiles may be observed using different extracts of the same sample. The use of consistent protocols for DNA purification is a priority to guarantee intra‐assay reproducibility over time. J. Med. Virol. 83:1042–1047, 2011.

Anal cytological abnormalities and epidemiological correlates among men who have sex with men at risk for HIV-1 infection

BackgroundThe incidence of anal cancer, a Human Papillomavirus (HPV)-related neoplasia, has been increasing in recent decades, mainly in men who have sex with men (MSM). Cytological changes of the anal epithelium induced by HPV can be detected through an anal pap smear. This study aimed to evaluate the prevalence and epidemiological correlates of anal cytological abnormalities among relatively young MSM at risk for HIV-1 infection, to help clarify whether or not this population deserves further investigation to assess the presence of anal cancer precursor lesions.MethodsMSM were recruited among attendees of a large STI clinic for a HIV-1 screening program. Anal samples, collected with a Dracon swab in PreservCyt, were used both for liquid-based cytology and HPV testing by the Linear Array HPV Genotyping Test. Data regarding socio-demographic characteristics and sexual behavior were collected in face-to-face interviews.ResultsA total of 346 MSM were recruited (median age 32 years). Overall, 72.5% of the individuals had an anal HPV infection, with 56.1% of them being infected by oncogenic HPV genotypes. Anal cytological abnormalities were found in 29.8% of the cases (16.7% ASC-US and 13.1% L-SIL). Presence of ASC-US+ was strongly associated with infection by any HPV type (OR=4.21, 95% CI: 1.97-9.23), and particularly by HPV 16 and/or 18 (OR=5.62, 95% CI: 2.33-13.81). A higher proportion of ASC-US+ was found in older MSM, in those with a higher number of lifetime partners and in those with a history of ano-genital warts. However, none of these variables or the others analyzed showed any significant association with abnormal cytological findings.ConclusionsThe presence of anal cytological abnormalities in about one third of the recruited MSM and their strong association with HPV infection, in particular that caused by HPV 16 and/or 18, might provide a further complement to the data that now support the introduction of HPV vaccination among MSM to protect them from the development of HPV-associated diseases. Additional studies are needed to determine whether and how screening for anal cancer precursor lesions should be performed in younger MSM.

AIDS-Kaposi Sarcoma and Classic Kaposi Sarcoma: are different ultrasound patterns related to different variants?

BackgroundKaposi Sarcoma (KS) is a malignancy of endothelial skin cells with multifocal localization on the skin, lymph nodes and visceral organs. Although all clinical variants are associated with HHV-8 infection, specific differences in the clinical onset and in the natural history of AIDS-KS and Classic-KS have been described. The present randomised prospective-observational study aimed to investigate whether the ultrasound pattern and color Doppler flow imaging of vascularisation of skin lesions of patients with Classic KS (CKS) or AIDS-KS could provide useful information to the evaluation of clinical activity of the disease.MethodsCutaneous lesions of 24 patients with histologically confirmed KS were investigated using very high frequency ultrasound probes; 16 patients had CKS and 8 had AIDS-KS. HHV-8 infection was confirmed in all patients by investigating the specific humoral response to viral antigens. Immunological and virological parameters were also assessed to monitor HIV or HHV-8 viral infection. For each patient, a target skin lesion was selected on the basis of size (diameter from 0.4 to 2 cm). Each lesion was analyzed in terms of size, depth and color Doppler pattern.ResultsThe B-mode ultrasound patterns of skin lesions did not differ when comparing CKS patients to AIDS-KS patients, whereas the color Doppler signal, which is associated with vascular activity, was detected in the KS lesions of 6/8 AIDS-KS patients (75.0%) and in 2/16 CKS (16,7%); the latter two patients showed a clinically progressive and extensive disease stage (IV B).ConclusionsOur preliminary results suggest that small cutaneous KS lesions - in both CKS and AIDS-KS patients- display similar B-mode ultrasound patterns ( hypoechoic, well defined, superficial lesions). However, the color Doppler signal, which is associated with endothelial activity and angiogenesis, which play a substantial role in KS progression, could constitute a useful tool for evaluating disease activity.