Imiquimod leads to a decrease of human papillomavirus DNA and to a sustained clearance of anal intraepithelial neoplasia in HIV-infected men

Anal intraepithelial neoplasia (AIN), a human papillomavirus (HPV)-associated precursor lesion of anal carcinoma, is highly prevalent in HIV-infected men having sex with men (MSM). This prospective follow-up study evaluated the long-term results of imiquimod treatment of AIN in 19 HIV-infected MSM. Standardized follow-up examinations included high-resolution anoscopy, anal cytology/histology, HPV typing, and DNA load determination for HPV types 16, 18, 31, and 33. Mean follow-up time was 30.3 months. A total of 74% (14/19) of the patients remained free of AIN at the previously treated site. Five patients (26%) had recurrent high-grade AIN after a mean time of 24.6 months. At the end of follow-up, the numbers of HPV types as well as high-risk HPV-DNA loads were significantly lower than before therapy. During follow-up, 58% of all patients (11/19) developed new anal cytological abnormalities in previously normal, untreated anal regions. 55% of these new AIN lesions were high-grade lesions and most of them were located intra-anally and associated with high-risk HPV types not detectable before therapy. These results demonstrate that imiquimod leads to a high rate of long-term clearance of AIN in HIV-positive men together with a prolonged decrease of high-risk HPV-DNA load. However, new AIN lesions associated with previously undetected HPV types frequently occur in untreated areas.     

Anal carcinoma in human immunodeficiency virus‐positive men: results of a prospective study from Germany

Background Anal intraepithelial neoplasia (AIN), a human papillomavirus (HPV)‐associated potential precursor lesion of anal cancer, is frequent among human immunodeficiency virus (HIV)‐positive men who have sex with men (MSM). There is a paucity of data published on the progression of high‐grade AIN to invasive cancer as well as on clinical and virological characteristics comparing anal margin and anal canal carcinoma. Objectives To search for anal carcinoma and AIN in a large series of HIV‐positive MSM, to assess treatment response of anal carcinoma, and to analyse lesional HPV spectrum of anal cancers. Methods Detection of anal carcinoma and AIN was performed using cytology, high‐resolution anoscopy, and histology in case of abnormal findings. Additionally, HPV analyses for 36 high‐ and low‐risk α‐HPV types were performed in patients with anal carcinoma. Results In total, 446 German HIV‐positive MSM were examined within an observation period of 5 years and 10 months. Of these, 116 (26·0%) patients had normal findings, 163 (36·5%) had low‐grade AIN, 156 (35·0%) had high‐grade AIN, and 11 (2·5%) had anal carcinoma as evidenced by the highest grade of cytology/histology. Five patients with anal cancer, who had refused treatment of their precancerous lesions, had progressed from high‐grade AIN to invasive cancer within a median time of 8·6 months. All anal cancers carried high‐risk α‐HPV types. All five squamous cell carcinomas (SCCs) of the anal canal were HPV16 positive. In contrast, only one of the four anal margin SCCs were HPV16 positive (HPV31, HPV33 and HPV33 + HPV68 were found in the other three anal margin SCCs). HPV59 was found in two adenocarcinomas, one of which additionally carried HPV33. In contrast to the cancer biopsies, a broad spectrum of surface high‐ and low‐risk HPV types was found in anal swabs of the patients. Surgical excision resulted in long‐term disease control of all anal margin carcinomas, whereas combined chemoradiotherapy in carcinomas of the anal canal was associated with high recurrence rates, high toxicity, and high mortality. Conclusions Anal carcinoma and AIN are frequent in HIV‐positive men, even in patients participating in anal cancer prevention programmes. High‐grade dysplasia in these patients can progress to invasive cancer within a short period of time. Anal margin carcinoma and anal canal carcinoma differ substantially in their lesional HPV spectrum, prognosis and treatment response.     

p16ink4a expression decreases during imiquimod treatment of anal intraepithelial neoplasia in human immunodeficiency virus-infected men and correlates with the decline of lesional high-risk human papillomavirus DNA load

BACKGROUND: Human papillomavirus (HPV)-associated anogenital cancers and their precursor lesions occur in excess in human immunodeficiency virus (HIV)-infected patients despite the initiation of highly active antiretroviral therapy. In this context, a drastically increased relative risk for anal intraepithelial neoplasia (AIN) exists in HIV-infected men having sex with men (MSM). In a pilot study, imiquimod, a topical immune response modifier, has been reported to be beneficial in the treatment of AIN. OBJECTIVES: To investigate the role of several biomarkers as potential adjuncts in the course of imiquimod treatment for AIN, and to determine whether these markers correlate with the course of high-risk HPV DNA load during imiquimod therapy. METHODS: Immunohistochemical staining was performed for p16(ink4a), minichromosome maintenance protein (MCM), Ki67, proliferating cell nuclear antigen (PCNA) and p21(waf1) expression before and after 16 weeks of imiquimod treatment for AIN. High-risk HPV DNA load determinations were performed by real-time polymerase chain reaction with type-specific primers and probes for HPV types 16, 18, 31 and 33. RESULTS: Histopathological and virological analyses were performed in 21 HIV-infected MSM with histologically confirmed AIN. Eighteen (86%) patients had a complete histological clearance of AIN after imiquimod therapy. As previously shown, lesional high-risk HPV DNA load significantly decreased during imiquimod therapy. Moreover, a significant decline of p16(ink4a), Ki67, MCM and PCNA expression after treatment was observed, while p21(waf1) expression changed nonsignificantly after imiquimod therapy. A significant correlation between the course of high-risk HPV DNA load and p16(ink4a) expression was observed during imiquimod treatment of AIN, whereas the decline of high-risk HPV DNA load did not significantly correlate with MCM, Ki67, PCNA or p21(waf1) expression. CONCLUSIONS: The significant decrease in p16(ink4a) expression in correlation with the drop of lesional high-risk HPV load suggests that p16(ink4a) may be a useful adjunct for the evaluation of treatment response in HPV-associated malignancies and their precursor lesions.     

Topical 5‐fluorouracil treatment of anal intraepithelial neoplasia in human immunodeficiency virus‐positive men

Background Anal intraepithelial neoplasia (AIN), a human papillomavirus (HPV)‐induced potential precursor lesion of anal cancer, is frequent among human immunodeficiency virus (HIV)‐positive men who have sex with men (MSM). So far, only a few prospective studies have been performed on the topical treatment of AIN, especially at the intra‐anal location. Objectives To evaluate the efficacy and safety of self‐administered topical 5‐fluorouracil (5‐FU) treatment of AIN in HIV‐positive MSM. Methods High‐resolution anoscopy (HRA) was performed and patients with AIN (grade 1–3) were treated with 5‐FU twice weekly for a total of 16 weeks. HRA‐guided lesional biopsies were repeated after 5‐FU treatment for histopathological evaluation. Lesional swabs were obtained before and after treatment for HPV typing and HPV‐DNA load determination of the high‐risk types HPV16, 18, 31 and 33. Responding patients returned 6 months after treatment for follow‐up. Results A total of 46 patients with AIN were included in this open prospective pilot study; 76% had multifocal disease and 74% had high‐grade lesions (AIN 2 or 3). In an intention‐to‐treat analysis, 26 of 46 patients (57%) responded to 5‐FU treatment. Eighteen patients (39%) had a complete clearance of AIN and eight patients (17%) had a partial response. Seventeen patients (37%) did not respond (unchanged grade of AIN in 16 patients and progression from low‐ to high‐grade AIN in one patient). 5‐FU treatment led to a significant decrease of HPV16‐DNA load and cumulative high‐risk HPV‐DNA load in both responding and nonresponding patients. Thirty‐nine patients (85%) experienced side‐effects during therapy, but only two discontinued 5‐FU treatment. One patient was lost to follow‐up. Six months later, 50% of the complete responders had a recurrence. Conclusions A substantial proportion of HIV‐positive MSM with AIN completely cleared their lesions with topical 5‐FU treatment. In those with partial response, pretreatment with topical 5‐FU might facilitate subsequent ablative therapy.     

Expression of human beta-defensin-2 and -3 in verrucae vulgares and condylomata acuminata

The human beta defensins (hBDs) 2 and 3 contribute to the inducible antimicrobial peptides in human skin. Besides an important role in fighting bacteria, they may also exert antiviral function. Verrucae vulgares and condylomata acuminata are typical cutanous viral diseases caused by different subtypes of the human papillomavirus. Their tendency for spontaneous regression could be caused by antiviral proteins like defensins. In a retoperspective study, we investigated lesions of verrucae vulgares and condylomata acuminata for the presence of hBD-2 and hBD-3 by immunohistochemistry. All of the specimens of verrucae vulgares ( n  = 20) were positive for hBD-2 and hBD-3. The specimens of condylomata acuminata ( n  = 15) exhibited expression for hBD-2 and hBD-3, with the exception of three and two samples, respectively. The expression levels in condyloma acuminata were lower for both defensins compared with verrucae vulgares. Normal skin tissue exhibited no staining of both hBD-2 and hBD-3 with the exception of two cases. Taken together, this is the first report of an increased expression of the hBD-2 and hBD-3 in cutaneous papillomavirus infections.     

Imiquimod treatment of anal intraepithelial neoplasia in HIV-positive men

OBJECTIVE: To evaluate the treatment of anal intraepithelial neoplasia (AIN) with the local immune response modifier imiquimod in human immunodeficiency virus (HIV)-positive men who have sex with men (MSM). DESIGN: Prospective, nonrandomized, open-label pilot study, with a mean follow-up time of 9(1/2) months. SETTING: Dermatology department of a university hospital. Patients Twenty-eight consecutive HIV-positive MSM with histologically confirmed perianal (n = 23) or intra-anal (n = 5) AIN. Intervention Overnight treatment with self-applied imiquimod cream (perianal AIN) or suppositories (intra-anal AIN) 3 times a week for 16 weeks. MAIN OUTCOME MEASURES: Response to treatment was documented using clinical, cytologic, and histologic criteria. Human papillomavirus (HPV) typing and HPV DNA load determination for the high-risk HPV types 16, 18, 31, and 33 were performed. RESULTS: Seventeen (61%) of all 28 patients included in the study and 17 (77%) of the 22 patients with AIN, who applied imiquimod as instructed, showed clinical and histologic clearance at the end of therapy. Four patients had residual AIN and 1 patient did not improve. Clinical response was accompanied by a sharp decline in HPV DNA loads and by a reduction in the number of HPV types, but long-term HPV clearance was rarely achieved. In the follow-up period, AIN cleared in 3 patients with residual AIN. Fourteen (78%) of 18 imiquimod responders with at least 5 five months of follow-up had a normal cytologic and clinical picture at the end of the follow-up period. Three primary responders developed a recurrence. In 6 noncompliant patients, there was no clinical or morphological improvement and the HPV DNA loads remained high. CONCLUSIONS: Imiquimod appears to be a safe and effective treatment option for AIN in HIV-positive MSM. Clinical response is accompanied by a significant decrease in high-risk HPV DNA load. These results should encourage controlled randomized studies of imiquimod treatment of AIN.     

Penile intraepithelial neoplasia is frequent in HIV-positive men with anal dysplasia

Anogenital human papillomavirus (HPV)-infection is common in HIV-infected men who have sex with men (HIV+MSM). These patients have a strongly increased risk of HPV-induced anal cancer and its precursor lesion, anal intraepithelial neoplasia (AIN), and a moderately increased risk for penile cancer. Only limited data exist on penile intraepithelial neoplasia (PIN) in HIV+MSM. We determined the prevalence and evaluated the virologic characteristics of PIN and AIN in 263 HIV+MSM. In case of histologically confirmed PIN (and AIN), HPV-typing, HPV-DNA load determination, and immunohistochemical staining for p16(INK4a) were performed. PIN was detected in 11 (4.2%) and AIN in 156 (59.3%) patients. Ten PIN patients also had AIN within the observation period. Four clinical types of PINs could be distinguished. High-risk-alpha-HPV-DNA was found in 10 PIN lesions, with HPV16 being the most frequent type. Infections with multiple HPV-types were common. All high-grade lesions had high-risk-HPV-DNA-loads > or = 1 HPV-copy/beta-globin-gene-copy. Cutaneous beta-HPVs were found in PIN and AIN, but beta-HPV-DNA loads were very low, irrespective of the histological grade. p16(INK4a) Expression was detectable in all PIN lesions and correlated both with the histological grade and with high-risk HPV-DNA loads. In view of the PIN prevalence found in our study, all HIV+MSM should be screened for PIN in addition to AIN screening.     

Differential suppression of epidermal antimicrobial protein expression in atopic dermatitis and in EFAD mice by pimecrolimus compared to corticosteroids

It has been suggested that the increased rate of bacterial infection in atopic dermatitis (AD) may be caused by reduced antimicrobial protein (AMP) expression. We were interested whether common treatments in AD affect antimicrobial defense. We investigated the effects of topically applied corticosteroids betamethasone valerate (BV) and triamacinolone acetonide (TA) and those of the calcineurin inhibitor pimecrolimus for 3 weeks on AMP expression in AD. BV and TA treatment in AD led to a significant reduction in AMP expression; protein expression of human beta-defensins (hBD)-2 and hBD-3, psoriasin, RNase 7 and cathelicidin LL-37 was below the level in skin of healthy controls. After pimecrolimus treatment, AMP expression was also reduced but less compared to BV and TA; the expression levels of hBD-2, psoriasin and RNase 7 still remained above the control levels. In essential fatty acid-deficient (EFAD) mice, a model of chronic skin barrier disease with inflammation, expression of the mouse beta-defensins mBD-1, mBD-3 and mBD-14 (orthologues for hBD-1, hBD-2 and hBD-3, respectively), was reduced by both treatments, again more pronounced by BV compared to pimecrolimus. In summary, we found that treatment for AD with corticosteroids in human skin and EFAD mice caused a strong reduction in AMPs; reduction was less with pimecrolimus. This result may explain the clinical observation that prolonged treatment with topical corticosteroids sometimes leads to bacterial infection.     

Human papillomavirus infection in men who have sex with men participating in a Dutch gay-cohort study

BACKGROUND: To develop strategies for prevention and early treatment of human papillomavirus (HPV) anal and penile cancer, a better understanding of related sexual behavior risk factors is needed. GOAL: The goal of this study was to establish the prevalence of anal and coronal sulcus HPV in a group of men who have sex with men participating in a Dutch gay-cohort study, to identify risk factors associated with HPV infection in this group, and to investigate the presence of identical HPV types in couples with stable relationships. STUDY DESIGN: A cross-sectional study of 241 HIV-negative and 17 HIV-positive men who have sex with men visiting the sexually transmitted disease clinic of the Erasmus MC for a regular and scheduled examination. Participants underwent a routine venereological examination including HIV serologic analysis, and swabs were taken from the coronal sulcus and anus for HPV DNA testing. All subjects were asked to complete a questionnaire on sexual risk behavior. RESULTS: HPV DNA was detected at the coronal sulcus in 23.5% of the HIV-positive men and in 15.8% of the HIV-negative men (P=0.492). In anal specimens, HPV DNA was detected in 64.7% of the HIV-positive men and 32.8% of the HIV-negative men (P=0.015). High-risk HPV types (P=0.007) and 2 or more different HPV genotypes (P=0.006) were seen more often in anal specimens of HIV-positive persons than in specimens of HIV-negative persons. A factor possibly associated with the presence of anal HPV infection was a concomitant anal infection with Chlamydia trachomatis, gonococci, or herpes simplex virus (P=0.059). In only 16.7% of HPV-positive steady couples, both companions showed the presence of one or more identical HPV genotypes. CONCLUSION: In this study, anal HPV DNA was detected more often than HPV DNA at the coronal sulcus. HIV positivity was associated with a higher prevalence of high-risk, but not with low-risk HPV types, at the anus. No association was found between HIV positivity and presence of high-risk HPV at the coronal sulcus. No sexual behavioral determinants for the presence of HPV could be identified. Concomitant anal infection with C trachomatis, gonococci, or herpes simplex virus may be associated with HPV infection. In the majority of steady couples, partners were infected with different HPV types.     

Anal verrucous carcinoma and penile condylomata acuminata

A 50-year-old man presented with recurrent tumorous lesions on the penis and the anal region. The anal lesion was histologically diagnosed as verrucous carcinoma (VC) and the penile lesions were in line with condylomata acuminata. Samples taken from tumors of both sites were human papillomavirus (HPV) DNA positive. Two of them taken from the penis and the perianal region scored HPV DNA 6 positive by using polymerase chain reaction and the Southern blot method. Treatment of both VC and condylomata acuminata consisted in surgery and adjuvant immune therapy. Neither tumor recurrence nor metastases occurred up until 6 months after therapy.     

Anal intraepithelial neoplasia in HIV infection

Human papillomavirus (HPV) infections belong to the most common sexually transmitted infections worldwide. While the immune system eliminates most HPV infections over time in immunocompetent individuals, HPV infections tend to persist in immunodeficient individuals. In HIV‐infected men who have sex with men (MSM), anal HPV prevalence is more than 90% and infections with multiple HPV types are common. Consequently, HPV‐associated anogenital malignancies occur with high frequency in patients with HIV infection. Anal intraepithelial neoplasia (AIN) is a potential precursor lesion of squamous cell carcinoma of the anus. Like its cervical counterpart, cervical intraepithelial neoplasia (CIN), AIN is causally linked to persistent infections with high‐risk HPV types such as HPV16 or HPV18. As AIN and CIN share distinct biological similar‐ities, AIN screenings analogous to Pap smear programs for CIN have been recommended in high‐risk populations to reduce the incidence of anal carcinoma. These screenings include cytological analysis followed by high resolution anoscopy in case of anal dysplasia. Treatment guidelines for AIN are not yet available. Therapeutic strategies can be divided into topical (e.g. trichloroacetic acid, podophyllotoxin, imiquimod, photodynamic therapy) and ablative (e. g. surgical excision, laser ablation, infrared coagulation, electrocautery) measures. However, controlled studies on AIN treatment have not been performed. The impact of HPV vaccination on AIN development will also need to be assessed. Long‐term follow‐up of these patients is essential to gain more insight into the natural history of anogenital HPV infection in HIV‐positive MSM.     

Human papillomavirus (HPV) viral load and HPV type in the clinical outcome of HIV-positive patients treated with imiquimod for anogenital warts and anal intraepithelial neoplasia

OBJECTIVE: To evaluate the efficacy of 5% imiquimod in HIV-positive male patients with anogenital warts or anal intraepithelial neoplasia (AIN), and to elucidate whether human papillomavirus (HPV) type and viral load were important for clinical outcome and recurrences. METHODS: Thirty-seven patients with histologically proven anogenital warts or AIN were enrolled. Topical 5% imiquimod was applied three times per week for more than 8 h overnight for 16 weeks, although patients were allowed to continue therapy for 4 more weeks if they did not have complete clearance of lesions. RESULTS: Mean age was 34 years. The perianal area was the main lesion location. Thirty-three patients had CD4 counts of < 500 cells/mm(3). Eighteen patients had a histopathological diagnosis of AIN-1. Main HPV types detected corresponded to low-risk HPV types. At 20 weeks of therapy, 46% patients achieved total clearance whereas 14 patients had > 50% clearance. Recurrence was observed in 5 of 17 patients who cleared. Clearance was not influenced by patients' CD4 counts, wart location, HIV viral load or HPV viral load. CONCLUSIONS: The assumption that visible perianal warts are benign lesions in HIV-positive patients has to be reevaluated since an important number of such lesions could correspond to low-grade anal disease, which in turn could progress to high-grade anal disease or cancer. In addition, our results in this preliminary study indicate that imiquimod appears to be effective in treating AIN in HIV-positive patients. Further studies are needed to document its utility to prevent high-grade dysplasia and/or anal cancer.     

Anale intraepitheliale Neoplasie und Analkarzinom

Anal dysplasia is common in HIV patients, especially in HIV-positive men having sex with men (MSM). High-grade anal dysplasia can progress to invasive anal cancer. As in cervical carcinoma, there is a cause and effect relationship between anal cancer and human papillomavirus (HPV) infection, especially with high-risk types such as HPV16. Several experts have recommended screening programs for anal cancer, including anal cytology along the lines of the Pap smear in women. Such screenings should only be performed if pathological findings result in further diagnostic steps and, if necessary, appropriate treatment. Clinical inspection, lesion biopsy, and treatment of anal dysplasia are performed under high-resolution anoscopy. Anal cancer is divided into cancer of the anal margin and cancer of the anal canal. This classification is important because of the difference in treatment regimens. Early cancer of the anal margin is excised akin to squamous cell cancer of the exposed skin, whereas cancer of the anal canal is treated by radiochemotherapy. HIV-positive and HIV-negative patients have similar response rates to combined radiochemotherapy. However, side effects, especially acute post-irradiation skin toxicity, early local recurrences, and abdominoperineal rectal excision are more common in HIV-positive patients. Physicians working in the field of HIV/AIDS should regularly screen their patients for the presence of anal dysplasia and anal cancer. Basic diagnostic workup includes clinical inspection of the perianal area, digital rectal examination, and anal cytology.     

No clinical predictors of intraepithelial neoplasia in HIV-positive patients with external condilomata acuminata

To identify clinical parameters in association with human papilloma virus (HPV) genotypes and histopathology diagnosis in HIV-positive patients with external condylomata acuminata (ECA), 400 Chilean HIV-positive patients were included in the study. Forty-seven patients presented ECA. Clinical parameters and socio demographic data were recorded. Histopathology study and HPV linear array genotyping assay were performed. Intraepithelial neoplasia (IEN) grade 2 or 3 was found in 8.5% of patients, associated to HPV-16. Patients were mainly single, MSM, with history of sexually transmitted disease (STD), multiple sexual partners, receiving antiretroviral therapy and with recurrent lesions. All ECA were mainly perianal, grey or pink colored, exophytic with less than two years evolution. No clinical parameter could predict the development of high grade IEN in HIV patients with ECA. It seems necessary to perform biopsy and genotype all HIV positive patients with ECA.     

A comparison between cytology and histology to detect anal intraepithelial neoplasia.

INTRODUCTION--Anal intraepithelial neoplasia (AIN), which may be a precursor of anal carcinoma, has been identified on histology following minor anal surgical procedures, in particular the removal of perianal condylomata, in increasing numbers of homosexual and bisexual men. Anal cytology has recently been proposed as a useful method of identifying AIN lesions. OBJECTIVE--To compare anal cytology with histology as a method of detecting AIN. METHODS--215 homosexual and bisexual men attending a central London sexually transmitted diseases clinic had an anal cytological smear performed under standard conditions. The perianal area and anal canal were then examined using a colposcope, and areas macroscopically suggestive of intraepithelial neoplasia were biopsied. RESULTS--176 of the 215 patients were biopsied of whom 76 had AIN on histology. 154 of the 215 patients had an adequate anal smear of whom 46 and 85 had cytological features of both HPV and AIN, or HPV alone respectively. Including features of HPV alone as an abnormal smear, anal cytology, when compared with anoscopy and histology as the gold standard for diagnosing AIN, resulted in a sensitivity of 87.5%, a specificity of 16.3%, a positive predictive value of 37.4% and a negative predictive value of 69.6%. Restricting abnormal smears to those with features of both HPV and AIN resulted in a sensitivity of 33.9%, a specificity of 72.5%, a positive predictive value of 41.3% and a negative predictive value of 65.7%. CONCLUSION--Anal cytology is a sensitive but nonspecific method of identifying patients with biopsy proven AIN if cytological features of HPV alone are included as abnormal smears. Specificity is improved by restricting abnormal smears to those with features of both HPV and AIN but this markedly lowers the sensitivity of the test. At present, anoscopy and histology are required in addition to anal cytology to differentiate between patients who simply have anal condylomata and those who also have AIN.     

寻常性银屑病患者皮损中抗菌肽LL-37和人β防御素-2 mRNA的表达

目的探讨寻常性银屑病患者皮损中抗菌肽LL-37与人β防御素-2(hBD-2)mRNA的表达.方法应用逆转录(RT)-PCR方法检测31例寻常性银屑病患者皮损和16例正常人皮肤组织中LL-37和hBD-2 mRNA的表达.结果与正常人皮肤组织相比,寻常性银屑病患者皮损中LL-37和hBD-2 mRNA的表达水平明显升高(P＜0.001).结论寻常性银屑病患者皮损中LL-37和hBD-2 mRNA表达水平的上调可能与银屑病患者很少发生皮肤感染等有关.