Low-grade Anal Intraepithelial Neoplasia Research Articles

DNA Methylation Analysis to predict Regression of high-grade anal Intraepithelial Neoplasia in HIV+ men (MARINE): a cohort study protocol

IntroductionAnal cancer precursors, or high-grade anal intraepithelial neoplasia (HGAIN), are highly prevalent in HIV-seropositive (HIV+) men who have sex with men (MSM). Around 30% of lesions regress within 1 year,...

Cytology-based Screening for Anal Intraepithelial Neoplasia in Immunocompetent Brazilian Women with a History of High-Grade Cervical Intraepithelial Neoplasia or Cancer.

To determine the prevalence and possible variables associated with anal intraepithelial neoplasia and anal cancer in immunocompetent women with high-grade cervical intraepithelial neoplasia. A cross-sectional study involving immunocompetent women with a histological diagnosis of high-grade cervical intraepithelial neoplasia and cervical cancer, conducted between January 2016 and September 2020. All women underwent anal cytology and answered a questionnaire on characterization and potential risk factors. Women with altered cytology were submitted to anoscopy and biopsy. A total of 69 women were included in the study. Of these, 7 (10.1%) had abnormal anal cytology results: (high-grade lesion, atypical squamous cells of undetermined significance, and atypical squamous cells, cannot exclude high-grade lesions: 28,5% each; low grade lesion: 14,3%). Of the anoscopies, 3 (42.8%) showed alterations. Of the 2 (28,5% of all abnormal cytology results) biopsies performed, only 1 showed low-grade anal intraepithelial neoplasia. The average number of pregnancies, vaginal deliveries, and abortions was associated with abnormal anal cytology. However, the highest mean regarding the cesarean sections was associated with normal cytology. The prevalence of anal intraepithelial neoplasia was compatible with data from recent studies, especially those conducted in Brazil. Opportunistic screening for anal intraepithelial neoplasia in this high-risk population should be considered. Anal cytology is suitable for this purpose, due to its low cost and feasibility in public health services.

Cytology-based screening for anal intraepithelial neoplasia in women with a history of cervical intraepithelial neoplasia or cancer.

High-risk human papillomavirus (HPV) has been identified in the pathogenesis of anal cancer. The purpose of this study was to assess the prevalence of abnormal anal cytology and HPV in women aged ≥40 years who have a history of high-grade cervical squamous intraepithelial lesion (SIL) or cancer and to estimate the prevalence of anal intraepithelial neoplasia (AIN) using cytology as the primary screening modality. Women who had a history of high-grade cervical SIL or cancer and were ≥40 years of age were included in this prospective study. Anal cytology with HPV-DNA testing was performed. All patients with abnormal anal cytology were referred for high-resolution anoscopy (HRA), and abnormal lesions were biopsied and treated if pathologically confirmed. Abnormal anal cytology correlated with HPV status, HRA findings, and clinical and demographic characteristics. A total of 317 women completed the study. Of these, 96 (30.3%) had abnormal anal cytology (high-grade SIL, 12.5%; low-grade SIL, 19.8%; atypical squamous cells, cannot exclude high-grade SIL, 6.3%; atypical squamous cells of undetermined significance, 61.5%) and 101 (31.9%) were HPV-DNA-positive. There was a significant association between abnormal cytology results and the presence of high-risk HPV. Of the 96 patients with abnormal cytology, 30 (31.3%) had biopsy-proven AIN on HRA, representing 9.5% of the total patient cohort; of these, 10 (33.3%) had low-grade AIN and 20 (66.7%) had high-grade AIN. Older age and smoking were significant risk factors for abnormal anal cytology. Women aged ≥40 years with a history of high-grade cervical SIL or cancer have a high rate of AIN. Screening for anal cancer may therefore be considered in this patient population. The optimal screening approach should be addressed in future studies.

Utility of Stathmin-1 as a Novel Marker in Evaluating Anal Intraepithelial Neoplasia (AIN).

The aim of this study is to determine whether immunohistochemistry for Stathmin-1 enhances diagnostic accuracy of anal dysplasia. The study included 40 biopsies with diagnosis of benign anal transitional zone (n=10), low-grade anal intraepithelial neoplasia (AIN) (AIN1, n=10), and high-grade AIN (AIN2, n=10, AIN3, n=10). The cases were selected to represent classic features. Immunohistochemistry for Stathmin-1, p16, and Ki-67 was performed and assessed for distribution within epithelial thickness. Stathmin-1 was expressed only in the basal layer of benign anal epithelium. Similar pattern of distribution was seen in all low-grade AIN cases (100%). In total, 40% of AIN2 showed Stathmin-1 staining pattern similar to AIN1. The other 60% of cases showed staining extending into the middle third of the epithelial thickness. Of AIN3 cases, 20% showed staining confined to the lower third epithelium, 20% showed staining extending to the middle third, and 60% showed staining extending into the upper third epithelium. The pattern of stain distribution suggested that staining extending above the lower one-third of the epithelial thickness discriminates between low-grade and high-grade AIN. With this cutoff, the sensitivity for the diagnosis of high-grade AIN was 70%, specificity was 100%, positive predictive value equaled 100%, and negative predictive value equaled 77%. P16INK4a showed 100% sensitivity for AIN2 and AIN3, whereas Ki-67 had 100% sensitivity for any AIN grade. In conclusion, Stathmin-1 has excellent specificity for the diagnosis of high-grade AIN; however, Stathmin-1 alone may not be sufficiently sensitive. Use in conjunction with other sensitive markers, such as p16 or Ki-67 may be considered.

18 - Identification of productive and transforming anal intraepithelial neoplasia using immunohistochemical markers p16INK4a and HPV E4

Background Immunohistochemical (IHC) biomarkers enable standardised, reproducible grading of anal intraepithelial neoplasia (AIN) based on progression of HPV infection from transient productive infection to complete neoplastic transformation. We investigated the expression of IHC biomarker HPV E4, a marker of productive HPV infection, in different grades of AIN caused by low-risk (lr) and high-risk (hr) HPV infection. Methods A reference grading of AIN was established using expert, consensus, subjective HE diagnoses supported by p16 expression for 183 anal biopsies: normal squamous epithelium 37, AIN1 67, AIN2 43, AIN3 36. Causative HPV genotype of the worst lesion was identified using laser capture microdissection and SPF10-PCR. Results Most AIN1 lesions were caused by lrHPV (72%) whereas most AIN2 (84%) and AIN3 (92%) was caused by hrHPV. None of the normal biopsies showed E4 positivity and 49% (33/67) of AIN1, 56% (24/43) of AIN2 and 3% (1/36) of AIN3 showed E4 positivity. While the large majority of E4 positive AIN1 (26/33) and AIN2 (16/24) lesions showed extensive positivity, the E4 positive AIN3 lesion showed only focal, superficial E4 positivity. HPV E4 positivity was seen in lesions caused by lr and hr HPV. Conclusions Our results show that when using HPV E4 in addition to p16, productive infections among both low-grade and high-grade AIN can be identified. Studies that further explore progression and regression in transforming (E4 negative) and productive (E4 positive) high-grade AIN, are necessary.

Incidence of Anal Dysplasia in a Population of High-Risk Women: Observations at a Cervical Pathology Unit.

This study aimed to evaluate the incidence of anal dysplasia in women at high risk of developing those lesions and to assess the relationships between positive anal cytology and different risk factors. We performed an observational cross-sectional study involving a patient survey and chart review. The study was conducted at the cervical pathology unit at the Hospital Universitario 12 de Octubre, Madrid, Spain, from 2011 to 2015. Patients were 215 women aged 18 to 65 years old with risk factors for anal dysplasia, for whom anal evaluation was indicated. Anal cytology was performed in all patients. High-resolution anoscopy and anal biopsy were used to investigate abnormalities. All patients completed a survey regarding sexual practices. The patients' demographic and clinical data were collected by using a retrospective chart review. The main outcome measure was the incidence of anal dysplasia in this population. Of the 215 patients, 45 (21.0%) presented with cytological abnormalities (atypical squamous cells of undetermined significance, 13.5%; low-grade squamous intraepithelial lesion, 5.6%; high-grade squamous intraepithelial lesion, 1.9%). Anoscopy was performed in 31 patients (14.4%) and 2 patients (0.9%) had abnormal findings. One case (0.5%) of low-grade anal intraepithelial neoplasia was identified. Only immunosuppression (p = .01) and smoking status (p = .02) were significantly correlated with positive anal cytology results. Limitations of the study include the small single-center sample, a lack of controls, the retrospective design, potential survey response biases, and the nonstandardized survey. The incidence of abnormal cytological findings was 21.0%, whereas low-grade anal intraepithelial was confirmed in 0.5% of the patients. There is a higher incidence of cytological abnormalities among immunosuppressed women and smokers.

Methylation of HPV and a tumor suppressor gene reveals anal cancer and precursor lesions.

We studied DNA methylation patterns of human papillomavirus (HPV) and tumor suppressor gene EPB41L3 in 148 anal and perianal biopsies to determine whether high levels of methylation would be associated with anal intraepithelial neoplasia (AIN). The most prevalent HPV type was HPV16, detected in 54% of the 30 benign biopsies, 33% of the 43 low-grade AIN (lgAIN), 82% of the 59 high grade AIN (hgAIN) and 4 of the 5 anal cancers. A methylation score was developed (0.561*HPV16me+0.439*EPB41L3) which had increasing values with severity of disease: the mean was 8.1% in benign, 13.2% in lgAIN, 22.3% in hgAIN and 49.3% in cancers (p < 0.0001). The methylation score as a triage classifier at a cut-off of 8.8 gave a sensitivity of 90.6% (95% CI: 82.8, 96.9), specificity of 50.7% (95% CI: 39.7, 61.6) and area under the curve of 0.82 (95% CI: 0.75–0.89) for separating hgAIN and cancer from benign and lgAIN biopsies. We conclude that methylation of HPV16 and EPB41L3 show highly significant association with increasing severity of AIN and cancer and may be useful as biomarkers in anal disease.

Anal study in immunocompetent women with human papillomavirus related lower genital tract pathology

ObjetiveTo estimate the prevalence of anal dysplasia in immunocompetent women with cervical intraepithelial dysplasia. Study designWe did a prospective cohort study, in which we enrolled 166 women with gynecological pathology related to human papilloma virus (HPV) infection. All patients underwent an anal cytology and HPV detection. Statistical analysis with a 95% confidence interval was used for prevalence calculations. A Χ2 test and Fisher’s exact one were used to determine differences between groups of qualitative variables. Differences between normally distributed and non-normally distributed groups in quantitative variables were accounted for using Student’s t-test or Mann-Whitney’s U test, respectively. ResultsOut of the 166 patients studied, high risk HPV in the anal canal was detected in 107 (64.46%) cases. The most prevalent genotype observed was non 16/18 high risk HPV, present in 54 (50.47%) patients. There was no a significant association with smoking, use of condom, anal intercourse, or anal benign pathology. However, a significant correlation between the presence of high risk HPV in the anal canal and the antecedent of condylomas was observed (p=0.047) (CI95%: 1.00%–12.58%). Women with cervical intraepithelial neoplasia (CIN) grade 1 had a significantly increased presence of high risk HPV in the anal canal (p=0.044). Out of the 166 women, 6 (3.61%) had abnormal anal cytology results, and were referred to high-resolution anoscopy. Anal biopsy was performed in these six cases. In 2 patients the biopsy reported low-grade Anal Intraepithelial Neoplasia: 1.20% (0.15%–4.28%). ConclusionsWomen with cervical intraepithelial dysplasia have 1.20% prevalence of anal intraepithelial neoplasia, so that it does not seem necessary to screen this population.

Colorectal Disorders in Acute Human Immunodeficiency Virus Infection: A Case Series.

Background. The gastrointestinal (GI) tract is important in the pathogenesis of human immunodeficiency virus (HIV) infection. We report a case series of lower GI endoscopic and histopathologic findings of HIV-infected individuals after presentation with acute infection. Methods. We performed a retrospective case review of individuals infected with HIV who enrolled between August 2010 and April 2013 in a primary infection treatment trial. All participants started the trial during acute infection and underwent colonoscopy with biopsies at baseline and after the start of antiretroviral treatment. Results. Twenty acutely infected individuals were included in the study (mean age, 33 years; range, 20-54 years). All participants were male who reported having receptive anal sex as an HIV risk factor. Nine individuals (45%) had at least 1 finding by colorectal pathology; 1 person had 2 diagnoses (diverticulosis and focal active proctitis). The histopathological findings revealed anal dysplasia in 3 cases: 2 had high-grade anal intraepithelial neoplasia (AIN) and 1 had low-grade AIN. Two persons had a colorectal polyp, 1 hyperplastic and 1 adenomatous. Three persons were diagnosed with diverticulosis, and 2 persons were diagnosed with proctitis, including 1 with focal active proctitis and 1 with cytomegalovirus proctitis. Conclusions. To our knowledge, this is the first case series report of lower GI disorders in acute HIV-infected individuals. Although the causal relationship remains uncertain, we describe the endoscopic findings that were observed during acute HIV infection among men who have sex with men. Understanding the prevalence of these pathologies may likely shed light on how acute HIV infection damages the lower GI tract.

Endoscopic submucosal dissection for anal intraepithelial neoplasia.

A 68-year-old woman was found to have a 20-mm superficial lesion at the dentate line (Fig. 1). Narrow band imaging (NBI) revealed a well-demarcated area with irregular intrapapillary capillary loops, similar to the findings seen in esophageal superficial squamous cell carcinoma [1]. Endoscopic biopsy revealed a high-grade squamous intraepithelial lesion (HSIL). After delineating the lesion under magnifying NBI, endoscopic submucosal dissection (ESD) was performed using an endo-surgical knife (Flushknife BT 1.5 mm, DK2618JB-15, Fujifilm, Tokyo, Japan). A transparent hood (D-201-12704, Olympus Medical Systems, Co. Ltd., Tokyo) was fitted on the tip of the colonoscope. For submucosal injection, 0.5% procaine hydrochrolide was used in the anal canal and 0.4% sodium hyaluronate (Mucoup, Johnson & Johnson Medical Company, Tokyo, Japan) at the rectal side. The lesion was completely removed without any complication (Fig. 2). Histological findings of the resected specimen confirmed HSIL with clear resection margins. Immunohistochemical staining showed p16 protein expression, suggesting infection by the human papilloma virus (HPV). Two months later, the patient was asymptomatic and repeat endoscopy showed a well-healed scar. Anal intraepithelial neoplasia (AIN) is a precursor of invasive anal cancer. Although low-grade AIN can be followed up conservatively, HSIL is recommended to be treated [2]. The traditional treatment for HSIL has been local surgical excision with mapping biopsy, though in some cases local recurrence and complications, i.e. anal stenosis or fecal incontinence, may occur [3]. In this case, magnifying NBI contributed accurately to the diagnosis of AIN, and ESD enabled complete removal of the lesion.

The effectiveness of electrocautery ablation for the treatment of high-grade anal intraepithelial neoplasia in HIV-infected men who have sex with men.

Electrocautery is one of the main treatment options for high-grade anal intraepithelial neoplasia (HGAIN). However, data regarding its efficacy are scarce. The aim of the study was to evaluate the effectiveness of electrocautery for the treatment of HGAIN. An observational study of HIV-infected men who have sex with men (MSM) who underwent screening for anal dysplasia was carried out. The on-treatment effectiveness of electrocautery was evaluated (according to biopsy findings measured 6-8weeks after treatment) in patients with HGAIN. A complete response was defined as resolution of anal intraepithelial neoplasia (AIN), a partial response as regression to low-grade AIN and recurrence as biopsy-proven HGAIN during follow-up. From May 2009 to November 2014, 21.9% (126 of 576) of patients screened were found to have HGAIN. Electrocautery effectiveness was evaluated in 83 patients. A complete response was observed in 27 patients [32.5%; 95% confidence interval (CI) 23.4-53.2%], a partial response in 28 patients (33.7%; 95% CI 24.5-44.4%) and persistence in 28 patients (33.7%; 95% CI 24.5-44.4%). The patients with the most successful results (81.8%) required two to four sessions of electrocautery. After a mean follow-up of 12.1months, 14 of 55 patients with a response (25.4%; 95% CI 15.8-38.3%) developed recurrent HGAIN within a mean time of 29.9months (95% CI 22-37.7months). No patient progressed to invasive cancer during the study or developed serious adverse events after treatment. No factors associated with poor response or recurrences were observed. Although electrocautery is the standard treatment for anal dysplasia, almost 50% of patients with HGAIN in our study did not respond or relapsed. New treatment strategies are necessary to optimize the management of anal dysplasia.

Which lesions should be biopsied during high-resolution anoscopy? Prospective descriptive study of simple morphological criteria.

High-resolution anoscopy (HRA) is a useful screening tool for anal intraepithelial neoplasia (AIN), although reputedly challenging for interpretation of suspected lesions. All consecutive patients who underwent biopsies for at least 1 lesion suspicious for AIN during HRA from November 2010 to March 2012 were prospectively included. The characteristics (borders, staining with acetic acid and Lugol iodine solution, and patterns of epithelium and vascularization) of the lesions detected during HRA were compared with histology. A total of 168 suspicious anal lesions were biopsied and analyzed in 103 patients (68% men, mean age ± standard deviation = 49.8 ± 9 y, 57% positive status on human immunodeficiency virus infection). According to histology, 41.7% of the lesions were high grade, 34.5% were low grade, and 23.8% were nondysplastic. Lesions with irregular epithelial pattern (or irregular vascularization) were twice as likely to be high grade compared with lesions with regular epithelial pattern (or regular vascularization). The incidences of acetic acid-induced whitening were 91.4%, 94.8%, and 70% among the high-grade AIN, low-grade AIN, and nondysplastic samples, respectively. Among the high-grade AIN, 62.9% were not stained by Lugol solution (vs 31% of the low-grade AIN). The positive predictive value of a combination of these simple morphological criteria was 68.6%. Several simple morphological criteria are significantly associated with high-grade AIN and are found less often in low-grade AIN. A combination of these morphological criteria provides sufficient positive predictive value to guide biopsy placement during HRA.

Clinical effectiveness and cost-effectiveness of quadrivalent human papillomavirus vaccination in HIV-negative men who have sex with men to prevent recurrent high-grade anal intraepithelial neoplasia

We examined the long-term clinical and economic benefits of quadrivalent human papillomavirus (qHPV) vaccine as a secondary/adjunct prevention strategy in the prevention of recurrent high-grade intraepithelial neoplasia (HGAIN) in HIV-negative men who have sex with men (MSM) and are 27 years or older. We constructed a Markov model to evaluate the clinical effectiveness and cost-effectiveness of two strategies: (1) no qHPV vaccine after treatment for HGAIN versus (2) qHPV vaccine after treatment for HGAIN. Model parameters, including natural history of anal cancer, vaccine efficacy measured in terms of hazard ratio (HR) (decrease in the risk of recurrent HGAIN), HGAIN treatment efficacy, utilities, and costs, were obtained from the literature. The outcomes were measured in terms of lifetime risk of anal cancer, lifetime cost, quality-adjusted life years, and incremental cost-effectiveness ratios (ICERs). Sensitivity analysis was conducted on all model parameters. We found that vaccinating HIV-negative MSM reduced the lifetime risk of anal cancer by 60.77% at an ICER of US$87,240 per quality-adjusted life-year. The results were highly sensitive to vaccine efficacy, transition of HGAIN to anal cancer, cost of treatment for HGAIN, vaccine degree of protection over time, and the vaccine duration of protection and less sensitive to HPV clearance, cost of qHPV vaccine, and the transitions from normal to low-grade anal intraepithelial neoplasia (LGAIN) and normal to HGAIN. With an HR of 0.3, the ICER was well below a $50,000 willingness-to-pay threshold; with an HR of 0.5, the ICER was still below a threshold of $100,000. The most critical disease-related factor influencing the cost-effectiveness was the progression of HGAIN to anal cancer. At an annual transition probability below 0.001, the ICER was below $50,000. Vaccinating HIV-negative MSM treated for HGAIN decreases the lifetime risk of anal cancer and is likely to be a cost-effective intervention.

High-Risk HPV DNA Test for Screening Anal Lesions in HIV-Positive and Negative Patients

The high incidence of anal dysplasia is related to many factors. The infections caused by HPV and HIV seem to be determinants of the following anal Pap smear changes: atypical squamous cells of undetermined significance (ASCUS), low-grade anal intraepithelial neoplasia (LAIN) and high-grade anal intraepithelial neoplasia (HAIN). Highrisk HPV infection is confirmed by the HPV DNA test, using hybrid capture technology. Screening for anal lesions should be complemented with the anal Pap smear protocol and high-resolution anoscopy. The aim of this study was to evaluate the influence of the high-risk HPV DNA test on screening for anal lesions. We evaluated 70 HIV-positive and negative patients who had previously had anal intercourse, at CRT/AIDS Sao Paulo from January 2013 to December 2013. All of the patients underwent the high-risk HPV DNA test, anal Pap smear protocol and highresolution anoscopy. The anal lesions were treated with 90% trichloroacetic acid (TCA) and 5% imiquimod for 12 weeks. The statistical analysis was performed using hypothesis tests on proportions and the significance level was set at less than 5%. 31 HIV-positive male patients, 19 HIV-negative male patients and 20 HIV-negative female patients were evaluated. 28 HIV-positive patients were positive for the high-risk HPV DNA test and 29 patients were positive for the high-risk HPV DNA test and had anal dysplasia. Our conclusion is that it was more common for HIVpositive patients to be infected with oncogenic HPV and the oncogenic HPV was more frequent in patients with anal dysplasia.

Anal cytology as a predictor of anal intraepithelial neoplasia in HIV‐positive men and women

Human immunodeficiency virus (HIV)-infected individuals have increased risk of anal intraepithelial neoplasia (AIN) and squamous cell carcinoma (SCC). Cytologic screening is invaluable in the detection of cervical neoplasia, therefore many clinicians have adopted anal cytology as part of anal cancer screening in patients at high-risk for anal neoplasia. The purpose of this study is to determine whether anal cytology is a valuable screening test for identifying AIN in HIV+ patients. The cohort included 228 HIV+ patients who underwent anal cancer screening with collection of 318 anal cytology specimens between January 2006 and December 2009. Of this group, 74 (32.5%) patients had associated anal biopsies within a 6-month period, with a total of 89 comparison cases. The anal cytology samples were classified using the 2001 Bethesda System terminology. The sensitivity of anal cytology in detecting ASC-US, AIN 1-3 or SCC was 93%. Cytology was 88% sensitive for detecting low-grade AIN (AIN 1), but only 20% sensitive for detecting high-grade AIN (AIN 2-3) or SCC. Atypical squamous cells of undetermined significance cases were distributed evenly between low- and high-grade AIN, with two cases having normal histology. Only six cases had negative cytology, all of which were associated with AIN on biopsy, for a false negative rate of 7%. Anal cytology is a good predictor of AIN, as confirmed by the high degree of sensitivity. However, there is poor correlation between the cytological and histological grade of AIN. Cytology underestimates the grade of dysplasia compared to the corresponding biopsy.

9. Cellular immune response to anal dysplasia in HIV-positive men on highly active antiretroviral therapy

Background The role of the cellular immune response to anal dysplasia progression is poorly understood. Extrapolated from the cervical model, CD4+ (killed by HIV) and CD8+ cells may have crucial anti-tumour activity. We report pilot data on key immune responses in HIV+ patients on HAART with anal dysplasia. Methods: High-resolution anoscopy and biopsies were performed. Ten tissue biopsies with high-grade anal intraepithelial neoplasia (HGAIN) were stained with immunohistochemistry for CD4+, CD8+, CXCL12+ and FOXP3+, and compared with 10 samples with low-grade anal intraepithelial neoplasia (LGAIN). Serum CD4+ and CD8+ counts were available. Tissue photographs were obtained and analysed utilising MetaXpress software. Results: In all dysplastic tissue, we found a higher number of CD4+, CD8+ and FOXP3+ cells in the subepithelium than epithelium. In HGAIN, the number of FOXP3+, intraepithelial CD4+ cells, and the ratio of intraepithelial to serum CD4+ were higher than in LGAIN. CXCL12+ staining was more abundant in HGAIN. CD4+ cells outnumbered CD8+ cells in all dysplastic tissue. Conclusions: The higher CD4+ to CD8+ ratio in HGAIN may be explained by the excess demand for antitumour activity, which is known to be a function of CD4+ cells against HPV-dysplasia in the analogous cervical dysplasia model. Cytotoxic CD8 cells, although known to have an important anti-tumour role, were less abundant in our study. The pro-tumour FOXP3+ cells, being more abundant in HGAIN, may have a key role in the progression of anal dysplasia. Strategies to block CXCL12+ may be useful in the treatment of HPV-related dysplasia.

Ano-genital human papillomavirus type 97 infection is detected in Canadian men but not women at risk or infected with the human immunodeficiency virus

BackgroundHuman papillomavirus type 97 (HPV97) DNA was detected in nearly 5% of anal samples collected from HIV-seropositive men living in Montreal, Canada. The rate of detection of HPV97 in the genital tract of Canadian women is unknown. Whether HPV97 is a local epidemic in HIV-seropositive men living in Montreal is also unknown. The prevalence of human papillomavirus type 97 (HPV97) was assessed in cervicovaginal cells from women living in Canada and in anal samples from HIV-seropositive men living in Toronto.FindingsCervicovaginal lavages collected from 904 women (678 HIV-seropositive, 226 HIV-seronegative) women living in Canada and anal cells collected from 123 HIV-seropositive men living in Toronto were tested for the presence of HPV97 with PCR. HPV97-positive samples were further tested by PCR-sequencing for molecular variant analysis to assess if all HPV97-positive men were infected with the same strain. All cervicovaginal samples were negative for HPV97. HPV97 was detected in anal samples from 6 HIV-seropositive men (4.9%, 95% confidence interval 2.0-10.5%), of whom five had high-grade and one had low-grade anal intraepithelial neoplasia, in addition to 2 to 8 HPV genital genotypes per sample. Four HPV97 variants were defined by four variation sites in the viral control region.ConclusionThese findings indicate that HPV97 infects in the anal canal of HIV-seropositive men but is not detected in the genital tract of women.

Anal Intraepithelial Lesions in Women With Human Papillomavirus–Related Disease

This study aimed to determine the prevalence of anal intraepithelial lesions in women with histologic diagnosis of intraepithelial lesions of the lower genital tract. This was a cross-sectional study conducted at the Lower Genital Tract and Colposcopy Unit of Hospital de Clínicas "José de San Martín," University of Buenos Aires, Argentina. A total of 481 women with histologically confirmed low-grade and high-grade cervical, vaginal, or vulvar intraepithelial lesions were evaluated between 2005 and 2011. They were referred for cytologic samples and examination with high-resolution anoscopy. We obtained biopsy specimens of any suspicious colposcopic images. Of a total of 481 patients, 404 (84%) were immunocompetent, 31 (6.4%) were HIV+, and 46 (9.6%) had other causes of immunosuppression. Moreover, of the 481 patients, 134 (27.86%) had anal intraepithelial neoplasia (AIN); 28 (5.82%) had high-grade AIN and 106 (22%) had low-grade AIN. Women with high-grade cervical intraepithelial neoplasia (CIN 2, 3) had 2 times the odds of developing AIN compared with women with low-grade CIN (CIN 1) (odds ratio = 1.91, 95% confidence interval = 1.1-3.6). Regarding localization, we found statistically significant difference between the frequency of vulvar and anal lesions. Women with vulvar condylomata and vulvar intraepithelial neoplasia (VIN) may be more likely to develop AIN. Immunocompetent women with CIN, vaginal intraepithelial neoplasia, or VIN may also present high-grade or low-grade anal intraepithelial lesions so we should consider AIN as part of multicentric disease of the lower genital tract. Cervical intraepithelial neoplasia, VIN, condyloma accuminatta, and vaginal intraepithelial neoplasia could be warning signs of anal intraepithelial lesions.

The reporting of anal cytology and histology samples: establishing terminology and criteria

Our understanding of the human papillomavirus (HPV) related cytomorphology and histopathology of the anal canal is underpinned by our knowledge of HPV infection in the cervix. In this review, we utilise cervical reporting of cytological and histological specimens as a foundation for the development of standardised and evidence-based terminology and criteria for reporting of anal specimens. We advocate use of the Australian Modified Bethesda System 2004 for reporting anal cytology. We propose the use of a two-tiered histological reporting system for noninvasive disease - low-grade and high-grade anal intraepithelial neoplasia. These classification systems reflect current understanding of the biology of HPV and enhance diagnostic reproducibility. Biomarkers such as p16(INK4A) may prove useful in further improving diagnostic accuracy. Standardisation is important because it will increase the value of the data collected as Australian centres develop programs for screening for anal neoplasia.

Abstract LB-177: HPV16 CpG and de novo-cytosine methylation is differentially associated with low-grade versus high-grade anal intraepithelial neoplasia in HIV-infected men

Abstract Objective: To determine if cytosine methylation in CpG and de novo sites is differentially associated with high- or low-grade anal intraepithelial neoplasias (HG-, LG-AIN), HPV16 DNA from182 clinical specimens. Background: Intra-anal cancer (IAC) is an emerging health crisis for gay, bisexual, transgender and other men who have sex with men (MSM) and rates have risen sharply among HIV-infected MSM despite introduction of HAART. High-risk HPVs are a causal risk factor for IAC and are especially common where MSM show HIV-coinfection. Further, cytosine methylation has been posited as an epigenetic transcription regulator that is poorly described in intra-anal dysplasias and cancers. Methods: HPV16 DNA was extracted from anal swab specimens and tested using bisulfite modification, PCR, cloning and sequencing of ∼10 clones/sample. Cervical cancer cell lines, CaSki and SiHa, and paraffin-embedded anal cancer specimens were similarly characterized as controls. Genomic sequences were evaluated using CLUSTAL and BiQ Sequence Alignment analysis software. Descriptive, tabular and multivariate analyses were performed using SAS (Version 9.2). Graphical representations were compiled using SIGMAPLOT (Version 9.0). Findings: Overall, methylation was relatively low in clinical AIN specimens. For 25,085 cytosines, the mean prevalence of methylation was 2.7% (SEM=0.4%) across 3′ L1 and LCR; the prevalence me-CpG, -CpA, -CpT, and -CpC was 3.2% (0.2%), 2.6% (0.1%), 2.5% (0.1%), and 2.7% (0.1%), respectively. However, some variation in HPV16 genomes was observed across specific functional gene sequences for HG- and LG-AIN specimens. Specifically, LG-AINs showed 1.30–2.62 times greater mean prevalence of methylation when compared to HG-AINs, across 24 of the 31 cytosines between nt7428–7564 (p-values&amp;lt;0.05) in the 5′ LCR and enhancer. In total, across the 148 cytosine positions, 37 sites showed statistically significantly greater prevalence of meC in LG-AINs and none showed higher methylation in HG-AINs, yielding a false discovery rate of 0.2 (7.4/37). The nt7428–7564 contains binding sites for E2–1, TEF-1, NF-1, YY1, OCT-1, GRE and potential deamination targets for ApoBec3G. Analyses are ongoing, nonetheless, multivariate analyses show risk for HG-AIN decreases by 27% for each cumulative increase of 100% cytosine-methylation across nt7220–7558, even after we controlled for age, CD4+ T-lymphocyte count, and repeated measurements. Conclusions: Data suggest the HPV16 5′LCR/enhancer region contains a large concentration of cytosine-containing transcription regulatory elements where binding interference by methylation might alter expression of the p97 promoter. Methylation in the HPV16 5′LCR/enhancer may decrease risk for HG-AIN. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-177. doi:10.1158/1538-7445.AM2011-LB-177