Fast-Tracking HPV Clearance in Men: Adjunctive Pidotimod Enhances Response to HPV Vaccination

Fast-Tracking HPV Clearance in Men: Adjunctive Pidotimod Enhances Response to HPV Vaccination | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL This is a preprint and has not been peer reviewed. Data may be preliminary. 9 October 2025 V1 Latest version Share on Fast-Tracking HPV Clearance in Men: Adjunctive Pidotimod Enhances Response to HPV Vaccination Authors : Claudio Ucciferri , Livia Moffa , Jacopo Vecchiet , and Katia Falasca 0000-0002-8795-5410 [email protected] Authors Info & Affiliations https://doi.org/10.22541/au.175998250.02547770/v1 413 views 207 downloads Contents Abstract Supplementary Material Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract Background: Human papillomavirus (HPV) infection is a significant global health concern, and while most infections clear naturally, persistent high-risk infections can lead to cancer. The role of immunomodulators in enhancing viral clearance is an area of growing interest. This study aimed to investigate the efficacy of adjunctive pidotimod therapy combined with the HPV vaccine in promoting HPV clearance in men. Methods: This retrospective study included 23 HIV-negative men with anal and/or genital HPV infection. Patients were divided into two groups: 7 receiving standard HPV vaccine and 16 receiving pidotimod starting before each vaccination dose, in addition to the vaccine. HPV genotyping was performed at baseline and at 12 months using real-time PCR. Results: At 12 months, the rate of HPV negative conversion was significantly higher in the pidotimod + vaccine group compared to the control group (62%vs28%, p < 0.005). The treatment group also showed a significant reduction in the number of HPV genotypes from baseline. Interestingly, the use of pre-exposure prophylaxis was associated with a significantly lower rate of HPV clearance in both groups. Conclusion: Adjunctive therapy with pidotimod significantly enhances HPV clearance when combined with the HPV vaccine in men. This approach represents a promising strategy for managing persistent HPV infection. Introduction Human papillomavirus (HPV) is one of the most prevalent sexually transmitted infections worldwide(1), with its unique oncogenic properties. Genital HPV types are typically categorized according to their epidemiological association with low-risk (LR-HPV) or high-risk (HR-HPV). Although HPV infection has a high transmission rate, up to 90% of new HPV infections, including those with high-risk types, clear spontaneously or become undetectable within 24 months after infection (2). In recent years, the use of immunomodulatory agents to enhance host defense mechanisms against HPV has gained significant attention. Among these agents, pidotimod (PDT), a synthetic dipeptide (3-L-pyroglutamyl-L-thiazolidine-4carboxylic acid), has emerged as a promising therapeutic adjuvant due to its well-characterized immunomodulatory properties(3). Pidotimod has demonstrated its ability to enhance both cellular and humoral immune responses, with immunomodulatory activity affecting both innate and adaptive immunity(4). Recent clinical evidence supports the therapeutic potential of PDT in HPV-related conditions. For instance, the combination of PDT with recombinant human interferon α-2b suppositories has been shown to effectively improve HPV negative conversion, accelerate vaginal microecology recovery, and modulate serum inflammatory responses in high-risk HPV patients following loop electrosurgical excision procedures(5) or reduce recurrence of HPV female genital warts (6, 7). Furthermore, prospective studies have evaluated complementary treatment approaches in HPV-associated lesions, suggesting broader applications for enhancing viral clearance. The immunological basis for pidotimod’s efficacy in viral infections stems from its capacity to enhance both cellular and humoral immune responses. In clinical practice, pidotimod has proven effective in the prevention and treatment of various infectious conditions, including respiratory tract infections, demonstrating its potential utility in enhancing viral immunity(8-10). The usefulness of PDT and its role as immunomodulant has been discussed for several decades, with particular focus on its mechanisms and potential for use in infectious diseases, prevention, and enhancing vaccine efficacy(3, 11) Given the persistent challenge of HPV clearance and the need for innovative therapeutic approaches, the role of immunomodulators in HPV-specific immunotherapy is increasingly recognized, especially in individuals who fail to mount a sufficient immune response. The integration of immunostimulant therapy with conventional treatments may offer a synergistic approach to improving clinical outcomes in HPV-infected patients This study aims to investigate the effect of PDT in male subjects with HPV infection in anal or genital sites who were treated with the nona-valent HPV vaccination . Materials and Methods Study Design and Population This retrospective study included 23 HIV-negative male patients who attended the outpatient sexual transmitted infection ambulatory clinic at SS Annunziata Hospital, University of Chieti, Italy, between July and December 2013. All patients underwent genital and anal swab collection for HPV infection. Swabs in solution were analyzed for HPV-DNA by polymerase-chain-reaction followed by type specific hybridization. HPV detection and typing ware performed using real time PCR TOCE technology (Allplex HPV 28 detection Seegene inc). Analysis was repeated at 12 months after complete dose vaccination. Inclusion criteria were male adults aged between 18 and 65 with a confirmed HPV infection in the anal and/or genital area who had not received a previous HPV vaccination. Patients were excluded if they had incomplete medical records, refused HPV vaccination, or were HIV positive. Data from 87 patients who had been followed up for HPV infection during that period were analyzed. Of these, 41 had not undergone any HPV treatment in the last 6 months; of these 31 patients agreed to participate in the study, 23 completed the follow-up and were enrolled in the study. Patients were divided into two groups: Control Group: Received the standard nona-valent HPV vaccination at months 0, 1, and 6. Treatment Group: Received pidotimod 800 mg twice a day, on an empty stomach, starting one to three days before each scheduled vaccination dose for a total of 10 days, in addition to the nona-valent HPV vaccination at months 0, 1, and 6. Data Collection Data were extracted from electronic medical records, including patient demographics, comorbidities, previous history of warts, and treatment details.The primary outcome was HPV negative conversion at 12 months. Secondary outcomes included a reduction in the number of genotype infections and new genital/anal warts.The study was conducted in accordance with the Declaration of Helsinki and approved by the Internal Institutional Review Board. Informed consent was obtained from all subjects involved Sample and Data Collection All specimens were collected by a trained medical practitioner using flocked swabs. Anal and genital sulcus samples were collected by rotating swabs 3 cm into the anal canal and around the coronal sulcus, respectively. Urethral mucosa specimens were obtained by three rotations at the first 2–3 cm of the urethral epithelium. Swabs in solution were analyzed for HPV-DNA using polymerase chain reaction (PCR) followed by type-specific hybridization. HPV detection and typing were performed using real-time PCR (Allplex HPV 28 detection, Seegene Inc.). This test was used to determine the presence of both low-risk genotypes (6, 11, 40, 42, 43, 44, 54, 61, 70) and high-risk genotypes (16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 69, 73, 82). The analysis was repeated at 12 months after the complete vaccination course. Statistical Analysis Data are expressed as mean ± standard deviation (SD) for continuous variables and were compared using Student’s t-test or analysis of variance, as appropriate, based on pre- and post-intervention data or based on different types of variables. Similarly, categorical variables are expressed as number and percentage and were analyzed using χ² and Fisher’s exact tests, as appropriate. A two-way ANOVA with post hoc tests was used to investigate the main effects and interaction effects of the intervention. The authors complied with the ethical standards of the relevant national and institutional committee on human experimentation and with the Declaration of Helsinki of 1975, as revised in 2013. The study was approved by the institutional local review board of the University ‘G. d’ Annunzio’ of Chieti-Pescara (Chieti, Italy). The data security was ensured by the pseudo-anonymization of identification codes from healthcare databases. No patients were personally identifiable for study. Privacy-by-design (or data protection-by-design) principles were adopted in this study. Results A total of 23 HIV-negative males with anal/genital HPV positivity were enrolled: 16 in the PDT + HPV-vaccination group and 7 in the vaccination-only group. The median age was 40,1 +/-9,5 years (41,5 vs 37 p=ns). Eight patients were using PrEP, with four in each group. HPV was detected in 16 males at the anal site and 14 at the genital site, with 8 males positive at both sites. The most frequent genotypes at baseline were HPV-16 (in 9 males) and HPV-6 (in 6 males). HPV-6 strains were the most frequently persistent strains at 12 months; the second most detected was HPV-58 (in 3 males). No new condylomas were detected shown after 12 months. Clearance rates at 12 months were obtained in 12/23 males (52%) but were significantly higher in patients treated with PDT + vaccination compared to vaccination-only patients (62% vs. 28%, p < 0.005) (table1). Regarding the burden of HPV strains, a significant reduction in the number of HPV types was observed in the pidotimod group from baseline to 12 months (39 strains detected vs. 7 strains detected). The vaccination-only group showed 22 strains detected at baseline vs. 12 at the end of the observation period (figure 1-2). A significant correlation between PDT use and the number of strains detected at 12 months was demonstrated (p < 0.009). Also, a significant reduction in the number of HR-HPV genotypes was observed in the PDT group vs control group from baseline to 12 months (p<0.01) but no significant difference was shown in LR-HPV The use of PrEP negatively correlated with complete clearance at 12 months (p <0.005). All HPV-58 strains detected at 12 months were found in PrEP users. Discussion Our results demonstrate that adjunctive treatment with PDT combined with the HPV vaccine significantly enhances HPV clearance in HIV-negative men with anal and genital infections, compared to vaccination alone. The clearance rate of 62% in the PDT plus vaccine group markedly exceeded the 28% observed in the vaccine-only group, underscoring pidotimod’s promising immunomodulatory adjuvant role. Furthermore, a substantial reduction in HPV genotypes—from 42 to 8 strains—was seen in the PDT group at 12 months, while the vaccination group showed a more modest decrease from 24 to 12 strains. These data suggest that PDT may potentiate immune-driven suppression of HPV types, consistent with previous findings demonstrating enhanced immune activation and diminished HPV persistence following immunostimulatory interventions (5, 12). This finding aligns with emerging supporting immunostimulants’ ability to enhance both innate and adaptive immunity, including dendritic cell maturation and T-helper 1 differentiation, which are critical for effective viral clearance. Such effects have been documented in respiratory (3, 8, 13) as well as HPV-related conditions, where immunomodulation reduces genital wart recurrence and improves post-surgical clearance (5-7). Furthermore, prior studies also indicate that pidotimod can enhance vaccine efficacy and mitigate adverse effects when administered alongside vaccination (14, 15). The observed reduction in the burden of oncogenic HPV genotypes in our treatment group also supports the role of pidotimod in targeting high-risk viral strains, which are strongly implicated in carcinogenesis. This efficacy can be also related to the molecular affinity of pidotimod with imiquimod, a molecule widely used to treat condylomatosis(16, 17). Interestingly, HPV type 6 and 58 persisted more frequently despite treatment, which is consistent with data suggesting that HPV subtypes may exhibit differential clearance dynamics (1) and higher diffusion in MSM population(18-20). ). The negative correlation between PrEP use and HPV clearance, with all HPV-58 detections at 12 months occurring in PrEP users supports recent observations of persistent HPV infections in HIV-negative men on PrEP. This persistence is likely related to behavioral risk factors, such as increased exposure without barrier protection, rather than direct pharmacological effects of PrEP itself(18, 20). The clinical implications of this finding warrant further investigation, especially considering the widespread use of PrEP and its potential influence on HPV epidemiology within this population (18). Current knowledge suggests that, although up to 90% of new HPV infections are resolved naturally within two years, additional immunostimulatory therapies such as pidotimod may accelerate this process and improve long-term outcomes in populations with impaired immune responses or high viral loads(5). Overall, our study highlights the promising therapeutic potential of integrating immunomodulators with prophylactic vaccination to overcome challenges in managing persistent HPV infections, especially in patients mounting suboptimal immune responses to vaccination alone. Limitations of this study include the relatively small sample size, retrospective design, and lack of longer-term follow-up to assess the durability of clearance and clinical outcomes. Future larger randomized controlled trials should validate these findings and explore immunological correlations of response to better elucidate the mechanisms underlying Pidotimod’s effects. . Additionally, deeper exploration of the interplay between PrEP use, HPV persistence, and immune function is warranted Conclusion Pidotimod appears to be a safe and effective immunoadjuvant that potentiates HPV vaccine-induced clearance, representing a novel adjunctive strategy in managing persistent HPV infection in men. These findings contribute to the growing evidence base advocating for immunomodulation as a complementary approach in the prevention and treatment of HPV-associated pathologies. References 1. Dunne EF, Unger ER, Sternberg M, McQuillan G, Swan DC, Patel SS, et al. Prevalence of HPV infection among females in the United States. JAMA. 2007;297(8):813-9.2. Bonde J, Bottari F, Iacobone AD, Cocuzza CE, Sandri MT, Bogliatto F, et al. Human Papillomavirus Same Genotype Persistence and Risk: A Systematic Review. J Low Genit Tract Dis. 2021;25(1):27-37.3. Ucciferri C, Chiappini F, Vecchiet J, Falasca K. From Legacy to Innovation: Pidotimod’s Expanding Therapeutic Horizon. Mediterr J Hematol I. 2025;17(1).4. Puggioni F, Alves-Correia M, Mohamed MF, Stomeo N, Mager R, Marinoni M, et al. Immunostimulants in respiratory diseases: focus on Pidotimod. Multidiscip Respir Med. 2019;14:31.5. Li X, Qiu Y. Pidotimod plus recombinant human interferon alpha-2b suppository boosts HPV clearance in high-risk patients following loop electrosurgical excision procedure. Am J Transl Res. 2025;17(3):2276-82.6. Zervoudis S, Iatrakis G, Peitsidis P, Peitsidou A, Papandonopolos L, Nikolopoulou MK, et al. Complementary treatment with oral pidotimod plus vitamin C after laser vaporization for female genital warts: a prospective study. J Med Life. 2010;3(3):286-8.7. Guerra B, Perino A, Polatti F, Scala M. Pidotimod in the management of vulvar papillomatosis: double-blind clinical trial versus placebo. Am J Ther. 1998;5(3):147-52.8. Ucciferri C, Di Gasbarro A, Borrelli P, Di Nicola M, Vecchiet J, Falasca K. New Therapeutic Options in Mild Moderate COVID-19 Outpatients. Microorganisms. 2022;10(11).9. Xu L, Zhao Y, Wu S, Song Q, Ouyang Z, Zhang X, et al. Effects of adjuvant pidotimod therapy on levels of inflammatory factors and expressions of serum GM-CSF and KL-6 in elderly patients with mycoplasma pneumonia. Am J Transl Res. 2021;13(10):11899-907.10. Marogna M, Ciprandi G. Pidotimod as add-on therapy in patients with pollen-induced allergic rhinitis and asthma and associated respiratory infections. J Biol Regul Homeost Agents. 2021;35(3):1053-8.11. Niu H, Wang R, Jia YT, Cai Y. Pidotimod, an immunostimulant in pediatric recurrent respiratory tract infections: A meta-analysis of randomized controlled trials. Int Immunopharmacol. 2019;67:35-45.12. Smith JA, Gaikwad AA, Mathew L, Rech B, Faro JP, Lucci JA, 3rd, et al. AHCC((R)) Supplementation to Support Immune Function to Clear Persistent Human Papillomavirus Infections. Front Oncol. 2022;12:881902.13. Zhao N, Liu C, Zhu C, Dong X, Liu X. Pidotimod: a review of its pharmacological features and clinical effectiveness in respiratory tract infections. Expert Rev Anti Infect Ther. 2019;17(10):803-18.14. R. C. Pidotimod activity in patients affected by COPD. Minerva Pneumol. 2014;53:21-6.15. Ucciferri C, Vecchiet J, Auricchio A, Falasca K. Improving BNT162b2 mRNA Vaccine Tolerability without Efficacy Loss by Pidotimod Supplementation. Mediterr J Hematol Infect Dis. 2022;14(1):e2022023.16. Wang DY, Cui YY, Zhang WW, Fan MS, Qiu KX, Yan L. Effect of different interventions on the treatment of high-risk human papillomavirus infection: a systematic review and network meta-analysis. Front Med (Lausanne). 2024;11:1274568.17. Gallio N, Preti M, Casetta E, Albuquerque A, Vieira-Baptista P, Borella F, et al. Imiquimod for Anal High Grade Intraepithelial Neoplasia: A Systematic Review. Curr Oncol Rep. 2025;27(7):833-43.18. Ucciferri C, Tamburro M, Falasca K, Sammarco ML, Ripabelli G, Vecchiet J. Prevalence of anal, oral, penile and urethral Human Papillomavirus in HIV infected and HIV uninfected men who have sex with men. J Med Virol. 2018;90(2):358-66.19. Tartaglia E, Falasca K, Vecchiet J, Sabusco GP, Picciano G, Di Marco R, et al. Prevalence of HPV infection among HIV-positive and HIV-negative women in Central/Eastern Italy: Strategies of prevention. Oncol Lett. 2017;14(6):7629-35.20. Sammarco ML, Ucciferri C, Tamburro M, Falasca K, Ripabelli G, Vecchiet J. High prevalence of human papillomavirus type 58 in HIV infected men who have sex with men: A preliminary report in Central Italy. J Med Virol. 2016;88(5):911-4. 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Keywords disease control epidemiology immunity/immunization immunodulators papillomavirus virus classification Authors Affiliations Claudio Ucciferri Universita degli Studi Gabriele d'Annunzio Chieti Pescara Dipartimento di Medicina e Scienze dell'Invecchiamento View all articles by this author Livia Moffa Universita degli Studi Gabriele d'Annunzio Chieti Pescara Dipartimento di Medicina e Scienze dell'Invecchiamento View all articles by this author Jacopo Vecchiet Universita degli Studi Gabriele d'Annunzio Chieti Pescara Dipartimento di Medicina e Scienze dell'Invecchiamento View all articles by this author Katia Falasca 0000-0002-8795-5410 [email protected] Universita degli Studi Gabriele d'Annunzio Chieti Pescara Dipartimento di Medicina e Scienze dell'Invecchiamento View all articles by this author Metrics & Citations Metrics Article Usage 413 views 207 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Claudio Ucciferri, Livia Moffa, Jacopo Vecchiet, et al. 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