Martina De Sanctis

Background/Objectives: Oral human papillomavirus (HPV) infection is emerging as a key driver of HPV-associated oropharyngeal cancer, especially in high-risk groups such as men who have sex with men (MSM) living with HIV (PLWH). However, evidence on oral HPV persistence and the impact of nine-valent HPV vaccination in adults remains limited. We conducted a prospective longitudinal study including 76 MSM PLWH, of whom 64 were nine-valent HPV-vaccinated and 12 unvaccinated. Methods: Oral rinse samples were collected at baseline (T0) and after 6 months (T6). HPV DNA detection and genotyping were performed using the Allplex™ HPV28 assay. Oral HPV dynamics (persistence, clearance, and incidence) were assessed for high-risk (HR) HPV, low-risk (LR) HPV, and vaccine-type HPV genotypes. Results: Baseline oral HPV prevalence was high (59.2%), with HR HPV detected in 43.4% of participants. HPV16 was the most frequent genotype at both T0 and T6. Among participants HPV-positive at baseline, persistence of HPV DNA was high and similar regardless of vaccination status (77.8%). However, incident vaccine-type oral HPV infection was significantly lower among vaccinated individuals than unvaccinated participants (6.3% vs. 33.3%; OR 0.13, 95% CI: 0.03–0.71; p = 0.0441). Finally, reporting ≥10 sexual partners in the previous year was significantly associated with baseline oral HPV positivity (p = 0.0298). Conclusions: In MSM PLWH, oral HPV infection is highly prevalent and persistent, underscoring that it may represent a reservoir for HPV-related oropharyngeal disease. In our small observational cohort, nine-valent HPV vaccination was associated with lower incident detection of vaccine-type oral HPV, supporting targeted vaccination and oral HPV surveillance in high-risk adult populations, while highlighting the need for larger longitudinal studies to confirm these findings and better define the magnitude and durability of vaccine-associated protection at the oral site.

Chronic hepatitis delta is the most severe form of viral hepatitis and is associated with rapid progression to cirrhosis and hepatocellular carcinoma. Although bulevirtide (BLV) effectively inhibits hepatitis D virus (HDV) entry, immunological biomarkers reflecting treatment response and residual viral activity remain poorly defined. This study investigated the serum profiles of interleukin-37 (IL-37) and IL-36 isoforms (IL-36α, IL-36β, and IL-36γ) in 22 HBV/HDV-coinfected patients receiving BLV monotherapy (2 mg/day). Serum cytokine levels were measured by ELISA at baseline (BL) and after 48 weeks of BLV treatment (TW48) and compared with HBV-monoinfected patients under nucleos(t)ide-analogue therapy and healthy donors. Patients were stratified according to virological, biochemical, and combined responses. At both BL and TW48, serum IL-37, IL-36α, and IL-36β levels were significantly higher in HBV/HDV-coinfected patients than in comparison groups (all p < 0.05), independent of treatment response, indicating a persistent cytokine signature during BLV therapy. IL-36β levels significantly decreased over time, particularly in biochemical non-responders (p = 0.0469), whereas IL-36α remained elevated and differed at TW48 between combined responders and non-responders (p = 0.0400). IL-36γ was detectable only in a small subset of patients. Notably, in a subgroup of patients evaluated at week 96, baseline IL-37 levels were significantly lower in those achieving virological response compared with non-responders (p = 0.0275). Moreover, IL-37 was the only cytokine showing a significant positive correlation with HDV RNA levels at TW48 when quantified by the AltoStar^® assay (p = 0.033; R² = 0.7563). Overall, HBV/HDV-coinfected patients display a distinct IL-37/IL-36 cytokine profile during BLV therapy. The association between IL-37 and residual viremia supports further investigation of this cytokine as a complementary biomarker for monitoring low-level viral activity during treatment.

Persistent infection with high-risk human papillomavirus (HPV), particularly genotype 16, is the main driver of anogenital and oropharyngeal cancers. However, data on HPV16 genetic variability in anal and oral samples from people living with HIV are limited. In this study, we investigated the diversity of HPV16 E6 and E7 genes in anal and oral samples collected from HIV-positive men who have sex with men (MSM) according to different cytological outcomes and anatomical site. Among 53 MSM patients, we obtained 51 E6 and 52 E7 sequences. Lineage A predominated (96.1%), mainly represented by sublineages A1 (84.3%) and A2 (9.8%). The E6 region showed higher variability than E7: 31.4% were identical to the reference (K02718), 51.0% carried the L83V (T350G) mutation, while Q14H, D25N, and D25E were also observed. In E7, only one non-synonymous substitution was detected (N29S). No site-specific variants were detected between anal and oral samples. Overall, these findings confirm the predominance of European A1/A2 variants in Italian MSM and suggest that E6/E7 polymorphisms are not directly linked to lesion grade or anatomical localization.

Hepatitis delta virus (HDV) infection is associated with severe hepatic complications and rapid progression towards liver cirrhosis and hepatocellular carcinoma. Accurate measurement of HDV RNA is critical for monitoring therapeutic responses, especially during treatment with novel therapies such as bulevirtide (BLV). This study compared the analytical performance of two HDV RNA quantification assays, Bosphore (Anatolia) and AltoStar^® (Altona), focusing on their sensitivity, specificity, and potential implications for clinical management. Sixty-one clinical samples from twenty-four patients, including fifteen HDV-infected patients receiving BLV treatment and nine controls, were tested using each assay. Of 30 samples identified as HDV-negative by the Bosphore assay, 17 (56.7%) were HDV-positive with AltoStar^®, demonstrating the superior sensitivity (p < 0.0001) of the latter assay. Quantitative analyses revealed consistently higher viral load measurements with AltoStar^® compared to Bosphore, with a difference of 1.23 Log IU/mL and a moderate correlation (r² = 0.7385) between assays. Each assay demonstrated a high specificity, with no false positives detected among control samples. However, our findings suggest that differences in assay sensitivity could impact the evaluation of virological response, highlighting the risk of false-negative results in chronically HDV-infected patients with low-level viremia. This emphasizes the need for careful assay selection to accurately monitor treatment outcomes.

Background: Hepatitis delta virus (HDV) infection represents the most severe form of viral hepatitis and is a significant global health challenge. Bulevirtide (BLV) is a novel therapeutic treatment that has resulted in variable response rates in HBV/HDV-coinfected patients. We evaluated clinical, virological, and polymorphic factors for the purpose of predicting BLV treatment success. Methods: Thirty HBV/HDV-coinfected patients received BLV monotherapy (2 mg/day) for 24 to 48 weeks. Baseline (BL) serum samples were collected to assess clinical parameters and virological markers (HDV RNA, HBV DNA, HBsAg, HBcrAg, anti-HBc IgG) at treatment weeks 24 (TW24) and 48 (TW48). Additionally, full-genome HDV sequencing and a phylogenetic analysis were performed. Finally, analyses of the HDAg protein sequence and HDV RNA secondary structure were conducted to evaluate potential associations with treatment response. Results: A significant reduction in HDV RNA levels was observed at TW48, with a virological response (HDV RNA undetectable or ≥2 Log decline from BL) achieved by 58% of patients. Median BL levels of anti-HBc IgG were significantly different between virological responders (39.3 COI; interquartile range [IQR] 31.6–47.1) and virological non-responders (244.7 COI; IQR 127.0–299.4) (p = 0.0001). HDV genotype 1e was predominant across the cohort, and no specific HDAg polymorphisms predicted the response. However, secondary structure analysis of HDV RNA revealed that a specific pattern of internal loops in the region 63–100 nucleotides downstream of the editing site may influence treatment response by impacting editing efficacy. Conclusions: This study revealed key factors influencing BLV efficacy in HBV/HDV coinfection. Lower baseline anti-HBc IgG levels strongly correlated with a positive virological response, suggesting that the liver’s inflammatory state affects treatment success. Additionally, the analysis of HDV RNA secondary structure in patients receiving BLV treatment revealed a higher editing efficiency in virological responders, highlighting areas for further research.