High-risk Human Papillomavirus Load Research Articles

Analysis of related factors of cervical intraepithelial neoplasia complicated with vaginal intraepithelial neoplasia.

To explore the underlying risk factors and to prevent misdiagnosis of cervical intraepithelial neoplasia (CIN) coexisted with vaginal intraepithelial neoplasia (VaIN). Clinical data of patients pathologically diagnosed with CIN were collected from January 2017 to December 2018. A total of 446 cases were analyzed, including 406 cases of single lesions ('CIN single' group) and 40 cases complicated with VAIN ('VAIN concurrent' group). The median age of the VAIN concurrent group was 53years (46.25-59years), and the median age of the CIN single group was 44years (36-50years). Regarding menopausal status, there were 28 cases (70.0%) in the VAIN concurrent group and 89 cases (21.9%) in the CIN single group (P < 0.005). The median load of high-risk human papillomavirus (Hr-HPV) in the VAIN concurrent and CIN single group was 923.4 relative light units/cutoff (RLU/CO) (145-2172.2 RLU/CO) and 229.155 RLU/CO (18.615-638.1275 RLU/CO), respectively (P = 0.037). The results revealed that the menopausal status was an independent risk factor for VAIN occurrence in CIN patients. The risk of VAIN in menopausal patients was higher than that in non-menopausal CIN patients (OR = 8.311, 95% CI 4.062-17.005). Age and HPV load were also related to the concurrence of VAIN and CIN. Examinations regarding vaginal screening are of great importance in the diagnosis of perimenopausal and postmenopausal CIN patients, especially patients with Hr-HPV load. Colposcopy and tissue biopsy should also be performed, when necessary, to avoid misdiagnosis and the appearance of vaginal lesions.

Preliminary study of the use of E6/E7mRNA detection in screening and triage management of HR-HPV infection during pregnancy.

BackgroundDetection of E6 and E7 mRNA load of high-risk human papillomavirus (HR-HPV) infection during pregnancy was compared with conventional cytopathology and DNA detection by pathological examination as colposcopy to evaluate the application of E6 and E7 mRNA detection in the diagnosis and management in HR-HPV infection for high -grade cervical lesions during pregnancy.MethodsFrom January 2014 to June 2019, 1,058 pregnant women of childbearing age who were filed for regular obstetrics in the Department of Obstetrics and Gynecology, Xuanwu Hospital of Capital Medical University, were separately assessed using cervical liquid-based cytology and HPV DNA detection. If the results were abnormal, colposcopy was performed as a follow-up. The presence of HR-HPV E6/E7 mRNA fragments was detected through the HR-HPV E6/E7 mRNA test, and monitored at the same time as colposcopy. The diagnostic efficacy of the HR-HPV DNA test versus the HR-HPV E6/E7 mRNA test for high-grade cervical lesions during pregnancy was compared.ResultsThe positive rate of HR-HPV E6/E7 mRNA detection in the overall cervical intraepithelial neoplasia (CIN) and above during pregnancy was lower than that with HR-HPV DNA detection, and there was a significant statistical difference between the two methods. In CIN I and normal or inflammatory results, the positive rate of HR-HPV E6/E7 mRNA detection was lower than that of HR-HPV DNA detection, while in the results of CIN II and CIN III, the positive rate of the two was not significantly different. HR-HPV E6/E7 mRNA detection is the same as HR-HPV DNA detection, both of which increased with the severity of cervical lesions, and the positive rate increased. In cases of maintenance or progression of cervical lesions, the positive rate of HR-HPV E6/E7 mRNA detection during pregnancy can reach 81.8%. High-grade cervical lesions during pregnancy had a higher rate of reversal to a lower level after delivery.ConclusionsThe results suggested that the use of HR-HPV E6/E7 mRNA detection in cases of positive HR-HPV DNA detection can significantly improve the diagnostic specificity of CIN II and above high-grade cervical lesions.

Patterns of multiple human papillomavirus clearance during 5-aminolevulinic acid-based photodynamic therapy in patients with genital warts

BackgroundMultiple human papillomavirus infections are commonly encountered in genital warts. Infection can be eliminated using 5-aminolevulinic acid-based photodynamic therapy. The aim of this study was to identify the preponderant genotypes and patterns of human papillomavirus co-infection clearance during photodynamic therapy in patients with genital warts. MethodsWe conducted a retrospective study between January 2020 and February 2021 at two hospitals in Guangzhou, China. Human papillomavirus typing and quantitative detection were performed before starting treatment and after each session of photodynamic therapy. ResultsA total of 97 patients participated in the study. Co-infections with low-risk and high-risk types were common in genital warts. Types 6 was the most common type detected, followed by types 52, 11, 58, 51 and 56. Patients with multiple infections were more likely to have high-risk human papillomavirus infection. The viral load of high-risk human papillomavirus before treatment was significantly lower than that of low-risk human papillomavirus, and decreased faster during therapy. In addition, high-risk types were cleared more readily than low-risk types, and 51.4% of high-risk types were eliminated after three rounds of therapy. A transient increase in viral load, especially low-risk types, was observed after two sessions of therapy. ConclusionsLow-risk human papillomavirus was preponderant in patients co-infected with low-risk and high-risk types. Photodynamic therapy effectively eliminated multiple human papillomavirus infections. The response to photodynamic therapy was mainly determined by the low-risk types in patients infected with combinations of low-risk and high-risk types.

Increased High-Risk Human Papillomavirus Viral Load Is Associated With Immunosuppressed Microenvironment and Predicts a Worse Long-Term Survival in Cervical Cancer Patients.

To evaluate the correlation between tumor-infiltrating lymphocytes (TILs) and the viral load of high-risk human papillomavirus (HR-HPV) in cervical cancer patients. A total of 62 cervical cancer patients were recruited during 1993-1994 and assigned into four groups treated with radiotherapy alone or radiotherapy combined with chemotherapy and/or thermotherapy. Ki67+ tumor cells, CD4+, CD8+, FoxP3+, OX40+ and granzyme B+ TILs were detected by immunohistochemistry. The viral load of HR-HPV in biopsy tissues before therapy was detected by in situ hybridization. The patients with high HPV viral load showed a significantly lower 15-year survival rate and an advanced International Federation of Gynecology and Obstetrics (FIGO) stage and increased recurrence rate. The distribution of Ki67+ tumor cells, FoxP3+ TILs, and CD8+/FoxP3+ ratio was obviously different between low and high HPV viral load groups. A worse clinical outcome was also implicated with increased HPV viral load tested by Cox regression analysis. Patients with increased HR-HPV viral load tend to be resistant to therapy with decreased immune surveillance in the immune microenvironment. Thus, HR-HPV viral load would influence the local immune microenvironment, and then further affect the survival of cervical cancer patients.

Viral Load in the Natural History of Human Papillomavirus Infection Among Men in Rural China: A Population-based Prospective Study.

High-risk human papillomavirus (HR-HPV) load is predictive of HR-HPV persistence and subsequent carcinogenesis in women. However, in men, data on genital HPV load and its effect on the natural history of HPV infection are limited. The subjects included 1532 men aged 25-65 years with up to 7 biannual visits for evaluation of genital HPV load in rural China during 2009-2013 who were positive for ≥1 of the 18 selected HPV types (including 10 HR-HPV types) detected by general primer-mediated polymerase chain reaction (PCR) and sequencing. Type-specific HPV load was quantified with real-time PCR and dichotomized based on median values. Men with multiple lifetime sex partners were more likely to have higher overall levels of HR-HPV load across visits (adjusted odds ratio, 2.42; 95% confidence interval [CI], 1.12-5.24; 2 partners vs 0-1 partner). Higher levels of HR-HPV load at the time of the first HPV diagnosis conferred an increased probability of the subject remaining type-specific HPV-positive up to 12 months and an increased probability of persistent/intermittent infection (virus detected repeatedly with or without a period of intercurrent negativity) versus transient infections (single-time positive). Higher overall HR-HPV levels were predictive of reduced HR-HPV clearance rates (adjusted hazard ratio, 0.47; 95% CI, .27-.83). Having multiple lifetime sex partners is associated with increased male genital HR-HPV load. Higher HR-HPV load predicts persistence of HR-HPV in men from rural China.

High-Risk Human Papillomavirus Infection in Latvian Male Kidney Transplant Recipients / Augsta Riska Cilvēka Papilomas Vīrusa Infekcija Vīriešu Dzimuma Nieru Transplantāta Saņēmējiem Latvijā

Abstract Kidney transplant recipients have higher incidence of human papillomavirus (HPV)-related malignancies, but studies on the natural history of HPV infection are insufficient, especially regarding in male recipients. The aim of this study was to evaluate the course of high-risk HPV (HR-HPV) infection after kidney allograft transplantation in male recipients: to estimate frequency and activity of HR-HPV infection under immune system suppression. Twenty male renal recipients (age 20 - 68) were enrolled in this investigation and examined in dynamics. Peripheral EDTA-blood samples and urine samples were collected from each patient 2 weeks, 6 months and 12 months after transplantation. Polymerase chain reaction (PCR) with consensus primers was used for initial detection of high range HPV types, a commercial qPCR kit for detection of HR-HPV load in urine samples and ELISA for detection of serum IgG class antibodies to HR-HPV L1-capsid protein. Overall, combining molecular (HR-HPV genomic sequences detected by real-time PCR) and serological studies (IgG class antibodies to HR-HPV L1-capsids’ protein), high frequency of HRHPV infection among male kidney transplant recipients (9/20; 45%) was showed. However, the majority of HR-HPV positive recipients (7/9; 78%) showed signs of infection clearance. It means that, despite the applied immune suppressive therapy, the host’s immune system is capable of dealing with HR-HPV infection up to the 12th month after transplantation. However, the sample size should be increased to enable through statistical analysis before final conclusions are made.

High-risk human papillomavirus load and biomarkers in cervical intraepithelial neoplasia and cancer

The purpose of this study was to examine the implication of high-risk human papillomavirus (HPV) load in cervical intraepithelial neoplasia (CIN) and cancer, and to detect biomarkers in cervical disease. We conducted high-risk HPV DNA load and cervical cytology tests in 343 women, cervical tissue biopsy in 143 women, and immunohistochemistry for p16(INK4A), cyclin D1, p53, cyclooxygenase-2, Ki-67, GLUT1, hPygopus2, and beta-catenin. As a result, HPV load [relative light units (RLU) value] was correlated with the histological severity of cervical disease (p < 0.05). In the 'atypical squamous cells of undetermined significance' cytology group, 2.385 of HPV load seemed to be the cut-off value at which 'benign' or CIN I can be differentiated from 'CIN II or more severe' (AUC = 0.712), but not statistically significant. The relative risk (odds ratio) of p16(INK4A) and GLUT1 overexpression increased gradually according to the histological severity of cervical disease. The p16(INK4A) showed statistically significant odds ratios in CIN II, CIN III, and cancer; GLUT1, in CIN II and CIN III; hPygopus2, in CIN III; and beta-catenin, in CIN III and cancer. Conclusively, HPV load, p16(INK4A) , and GLUT1 can be instrumental in predicting the severity of HPV-related cervical disease. The beta-catenin/hPygopus2 signaling may be involved in proceeding to CIN III.

P300 Expression is Related to High-risk Human Papillomavirus Infections and Severity of Cervical Intraepithelial Neoplasia But Not to Viral or Disease Outcomes in a Longitudinal Setting

To evaluate the role of the expression of the transcription factor p300 as an independent predictor of high-risk human papillomavirus (HR-HPV) infections and outcome of the cervical disease.Cervical biopsy samples taken at enrolment from 225 women of the Latin American Screening study cohort were analyzed for p300 using immunohistochemistry to assess its value as predictor of (a) cervical intraepithelial neoplasia (CIN) grade, and (b) HR-HPV at baseline, as well as (c) outcomes of HR-HPV infections, and (d) development of incident CIN as surrogate endpoints of progressive disease.There was a significant linear trend in increasing upregulation (=pattern shift) of p300 (P=0.0001) in parallel with increasing grade of CIN. When dichotomized (normal/moderately increase vs. strong-intense), upregulated p300 expression predicted CIN3+ with odds ratio=4.16 (95% confidence interval: 1.95-8.86) (P=0.0001) and CIN2+ with odds ratio=3.48 (95% confidence interval: 1.86-6.48) (P=0.0001). p300 was upregulated more often in HR-HPV+ lesions than in those remaining negative. Semiquantitative viral loads were also directly related to upregulation of p300 (P=0.036), but p300 was not a significant predictor of disease progression to either CIN1+ or CIN2+.p300 expression was upregulated in CIN lesions and related to detection and viral load of HR-HPV but not to their outcome or to incident CIN.

High‐risk human papillomavirus testing for monitoring patients treated for high‐grade cervical intraepithelial neoplasia

The aim of the present study was to examine the accuracy of high-risk human papillomavirus (HR-HPV) DNA detection as a predictor of residual or recurrent cervical intraepithelial neoplasia (CIN) after treatment of high-grade CIN. Ninety-five women treated by conization and loop electrosurgical excision procedure (LEEP) for high-grade CIN were followed-up with HR-HPV DNA testing (Hybrid Capture II test, Digene Diagnostics, Gaithersburg, MD, USA) and cytology at 3, 6, 12, 18 and 24 months after treatment. The outcome of our study was to detect the presence of CIN using colposcopy-directed biopsy within 24 months after treatment. Women with recurrent or residual disease did not differ from women who were cured within clinicopathologic parameters at treatment. Pre-treatment HR-HPV testing was positive in all cases. In the recurrent pre-treatment group HR-HPV loads were significantly higher than in the group with no recurrence (1065.5 +/- 852.3 vs 527.7 +/- 669.6, P = 0.003). Residual or recurrent disease was identified in 17 patients (17.9%) during a 24-month period. A Pap smear significantly predicted disease recurrence at the first follow-up visit only. At the 6-month visit, HR-HPV testing showed 100% sensitivity and negative predictive value (NPV). When both a Pap smear and a HR-HPV test were used together, 82% sensitivity, 76% specificity and 95% NPV was noted. Performance of resection margins (odds ratio (OR) 9.8; 95% confidence interval (CI), 3.0-32.7) and post-treatment HR-HPV load >100 RLU (OR 9.3; 95% CI, 2.2-38.2) were also significant. HR-HPV testing in conjunction with Pap smear offers clear advantages over single cytology when monitoring women treated for high-grade CIN.

Association between high-risk human papillomavirus DNA load and different histological grades of cervical neoplasia

To determine the association between viral load of high risk human papillomavirus (HR-HPV) and cervical intraepithelial neoplasia (CIN). Cervical exfoliated cells were collected from 18 186 women aged 17 -59 from six urban areas and eight rural areas when they were screened in the cross-sectional population-based studies from 1999 to 2008. HR-HPV was detected by the Hybrid Capture 2 (hc2) system, and viral load was measured by the ratio of relative light units to standard positive control (RLU/PC). RLU/PC was categorized for analysis into four groups: negative [0, 1.00), low viral load [1.0, 10.00), moderate viral load [10.00, 100.00), and high viral load > or = 100.00. Cervical lesions were diagnosed by biopsies as normal, CIN 1, CIN 2, CIN 3 and squamous cervical cancer (SCC). Association between HR-HPV viral load and CIN was evaluated by unconditional multinomial logistic regression. The HR-HPV infection rate of the population was 14.51% (2515/17334). 100.00% (29/29) of SCC, 97.63% (206/211) of CIN 3, 93.43% (199/213) of CIN 2, 75.04% (421/561) of CIN 1 and 10.17% (1660/16320) of normal women were positive for HR-HPV DNA. The median RLUs for the HR-HPV positive women with SCC, CIN 3, CIN 2, CIN 1 and normal were 320.85, 158.05, 143.70, 125.34 and 9.64, respectively. There were significant differences among the distributions of viral loads in each lesion (chi2 = 6190.40, P < 0.01). The severity of CIN increased with the viral load (chi2 = 5493.35, P <0.01). Compared with the risks of CINs in HR-HPV negative population, the risks of CINs in low, moderate and high viral loads were increased gradually [OR(95% CI) : CIN 1 : 9.01(6.31 - 12.87), 24.96(18.23 - 34.17) and 68.42(51.40 - 91.08); CIN 2 : 26.44(12.07 - 57.95), 98.53(49.54 - 195.98) and 322.88(168.62 - 618.27); CIN 3+ : 72.89(24.02-221.18); 343.58(121.81-969.09) and >999.99(473.38 - >999.99)], and there were obvious dose-response relationships (chi2trend was 3115.05, 2413.95 and 3098.57, respectively. P< 0.01). In each age group of the HR-HPV positive population,the risks of CIN 2 + in the women with moderate or high viral load were higher than the one with low viral load [OR(95% CI): <35 : 4.71(1.23 - 18.09) and 15.06(4.40 - 51.49); 35 -: 4.01 (1.62 -9.90) and 14.09(6.15 -32.28); 40 - : 3.06(1.52 -6.16) and 7.78(4.05 -14.95); > or =45: 3.50(1.36 -9. 01) and 7.57(3.13 - 18. 30)], and there was a positive correlation between the risk of CIN 2+ and the viral load (chi2trend was 51.33, 66.28, 53.64 and 51.00, respectively. P <0.01). The risk of CIN 2 + was highest among the women aged 40 - with high viral load [OR (95% CI) : 2.02 (1.15 - 3.52)]. There is strong correlation between the HR-HPV viral load and the severity of CIN, and so is the correlation between the HR-HPV viral load and the risk of CIN 2 +. A moderate to high viral load of HR-HPV should be the major risk factor for the cervical cancer and CIN 2 and CIN 3, and there is a higher risk in the women aged 35 or older than the younger ones. Considering both the age and viral load could help the doctors to manage the screening women more effectively.

P16INK4a Immunostaining Identifies Occult CIN Lesions in HPV-positive Women

To evaluate whether p16 staining could help to recognize underestimated cervical intraepithelial neoplasia (CIN) in women positive for high-risk human papillomavirus (HR-HPV) with negative biopsy. Out of 1,259 women undergoing a histologic study and a simultaneous HR-HPV detection using the Hybrid Capture 2 test, we selected all patients testing positive for HR-HPV and having a negative biopsy (n=139), as well as all women testing negative for HR-HPV with a biopsy of either CIN 1 (26 cases) or CIN 2 to 3 (11 cases). Of the remaining 1,083 women, we randomly selected for the purpose of controls, 50 cases negative for HR-HPV with negative biopsy and 100 cases positive for HR-HPV and with biopsy of CIN (50 CIN 1, 50 CIN 2-3). In all cases, immunohistochemical staining for p16 and a second evaluation of the initial biopsy was carried out. Thirty-four out of 139 biopsies (24.5%) testing positive for HR-HPV but having a negative biopsy were positive for p16. Thirty of these cases (21.6%) were classified as harboring a CIN (11 CIN 1, 19 CIN 2/3) after reevaluation. Both the number of cases reclassified as CIN of any grade, or as CIN 2/3, were significantly higher for cases with HR-HPV load above 100 relative light unit (P<0.005). Particular attention should be paid to biopsies from patients having positive Hybrid Capture 2. The risk of harboring undetected CIN of any type or CIN 2/3 is significantly higher for patients with high HR-HPV load. Immunostaining with p16 should be considered as a highly desirable addition to the histologic evaluation of cervical biopsy specimens in HR-HPV-positive women.

The association of pre-conization high-risk HPV load and the persistence of HPV infection and persistence/recurrence of cervical intraepithelial neoplasia after conization

Objective. The aim of this study was to evaluate whether the pre-conization high-risk human papilloma virus (HR-HPV) load is predictive for the persistence of HR-HPV infection and the persistence/recurrence of cervical intraepithelial neoplasia (CIN) after conization of the cervix. Materials and methods. A retrospective review was performed on 236 women who underwent conization due to CIN at the Center for Uterine Cancer, National Cancer Center, Korea, between March 2001 and March 2006. The samples for pre-conization HR-HPV test were obtained at least within 3 weeks before conization. All patients underwent HR-HPV testing and cytology between 3 and 6 months after conization, and subsequent follow-up of 3- to 6-month interval was performed thereafter. The persistence of HR-HPV infection and persistence/recurrence of histologic abnormality after conization were analyzed by age, parity, menopausal status, method of conization, glandular extension, margin status, severity of CIN, and pre-cone HR-HPV load in univariate and multivariate analysis. Results. In univariate analysis, high pre-cone HR-HPV load was the only risk factor for the persistence of HR-HPV infection after conization (persistent HR-HPV infection; 19.8% [23/116] of patients with an HR-HPV load ≥ 100 RLU/PC vs. 10.0% [12/120] of patients with a load < 100 RLU/PC, P = 0.034). Multivariate analysis showed that an HR-HPV load ≥ 100 RLU/PC was a risk factor for persistence/recurrence of histological abnormalities after conization ( P = 0.040, OR = 5.748, 95% CI = 1.082–30.526). Conclusion. Patients with a pre-conization HR-HPV load ≥ 100 RLU/PC had a higher rate of persistent HR-HPV infection and a higher rate of persistent/recurrent histological abnormalities after conization for CIN compared to patients with a load < 100 RLU/PC.

The expression of p16 (INK4a) and Ki-67 in relation to high-risk human papilloma viral load and residual disease after conization with positive margins

The purpose of this study was to investigate the correlations between high-risk human papillomavirus (HPV) load and p16 (INK4a) or Ki-67, and to identify biomarkers that may predict residual disease after conization with positive margins. The following samples were analyzed: 49 paraffin-embedded specimens from patients with cervical intraepithelial neoplasia (CIN), including 12 CIN 2 conization specimens and 37 CIN 3 conization specimens. Immunohistochemical analysis was performed with antibodies to p16 (INK4a) and Ki-67. Hybrid Capture II testing was used to detect high-risk HPV DNA. The mean HPV loads within each of the p16 (INK4a)-staining cases were 9.5 (relative light units/positive control) RLU/PC for negative staining, 531.8 RLU/PC for 1+ staining, 140.2 RLU/PC for 2+ staining, and 545.1 RLU/PC for 3+ staining. HPV loads differed significantly according to p16 (INK4a) expression (P= 0.0021). The mean HPV loads within Ki-67 staining cases were 28.2 RLU/PC for 1+ staining, 189.6 RLU/PC for 2+ staining, and 563.3 RLU/PC for 3+ staining. HPV loads differed significantly according to Ki-67 expression (P= 0.0259). The expression of p16 (INK4a) (P= 0.0012) and Ki-67 (P= 0.0006) were significantly associated with the CIN grade. In univariate and multiple logistic regression analysis, age, parity, cytology, lesion grade in the cone, high-risk HPV load, and the expression of p16 (INK4a) and Ki-67 were not significantly associated with residual lesions after conization with positive margins (P&gt; 0.05). In conclusion, high-risk HPV load showed significant differences according to the expression of p16 (INK4a) and Ki-67, while none of the prognostic factors were significantly associated with residual disease after conization with positive margins.

Risk factors predicting residual disease in subsequent hysterectomy following conization for cervical intraepithelial neoplasia (CIN) III and microinvasive cervical cancer

Objective. To determine factors predicting post-cone residual disease in cervical intraepithelial neoplasia (CIN) III and microinvasive cervical cancer. Methods. Of 337 patients who underwent conization due to CIN III and microinvasive cervical cancer between November 2001 and March 2006, 77 underwent hysterectomy within 6 months of conization. We analyzed their demographic features, pathologic parameters and pre-cone high-risk human papilloma virus (HR-HPV) load measured by Digene Hybrid Capture II. Results. In univariate analysis, age ≥ 50 years ( P = 0.048, relative risk [RR] = 2.74, 95% confidence interval [CI] = 1.0–7.4), positive resection margin ( P = 0.004, RR = 4.35, 95% CI = 1.5–12.3), and pre-cone HR-HPV load ≥ 300 relative light units (RLU)/positive control (PC) ( P = 0.009, RR = 3.41, 95% CI = 1.3–8.7) were significant factors associated with residual disease. While postmenopausal status showed borderline significance ( P = 0.065, RR = 2.65, 95% CI = 0.9–7.5), parity ≥ 3, severity of disease (CIN III vs. microinvasive cancer), conization method (large loop excision of transformation zone vs. cold knife conization), and glandular extension were not significant. In multivariate analysis only positive margin ( P = 0.023, RR = 3.56, 95% CI = 1.2–10.7) and pre-cone HR-HPV load ≥ 300 RLU/PC ( P = 0.034, RR = 2.96, 95% CI = 1.1–8.1) were significant factors associated with residual disease. Conclusion. Positive margin and pre-cone HR-HPV load ≥ 300 RLU/PC were the only significant factors predicting post-cone residual disease in multivariate analysis. Appropriate application of these predictive factors may avoid post-cone hysterectomy.

P16ink4aexpression 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

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. 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. 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. 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. 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.

The expression of p16 (INK4a) and Ki-67 in relation to high-risk human papilloma viral load and residual disease after conization with positive margins

The purpose of this study was to investigate the correlations between high-risk human papillomavirus (HPV) load and p16 (INK4a) or Ki-67, and to identify biomarkers that may predict residual disease after conization with positive margins. The following samples were analyzed: 49 paraffin-embedded specimens from patients with cervical intraepithelial neoplasia (CIN), including 12 CIN 2 conization specimens and 37 CIN 3 conization specimens. Immunohistochemical analysis was performed with antibodies to p16 (INK4a) and Ki-67. Hybrid Capture II testing was used to detect high-risk HPV DNA. The mean HPV loads within each of the p16 (INK4a)-staining cases were 9.5 (relative light units/positive control) RLU/PC for negative staining, 531.8 RLU/PC for 1+ staining, 140.2 RLU/PC for 2+ staining, and 545.1 RLU/PC for 3+ staining. HPV loads differed significantly according to p16 (INK4a) expression (P = 0.0021). The mean HPV loads within Ki-67 staining cases were 28.2 RLU/PC for 1+ staining, 189.6 RLU/PC for 2+ staining, and 563.3 RLU/PC for 3+ staining. HPV loads differed significantly according to Ki-67 expression (P = 0.0259). The expression of p16 (INK4a) (P = 0.0012) and Ki-67 (P = 0.0006) were significantly associated with the CIN grade. In univariate and multiple logistic regression analysis, age, parity, cytology, lesion grade in the cone, high-risk HPV load, and the expression of p16 (INK4a) and Ki-67 were not significantly associated with residual lesions after conization with positive margins (P > 0.05). In conclusion, high-risk HPV load showed significant differences according to the expression of p16 (INK4a) and Ki-67, while none of the prognostic factors were significantly associated with residual disease after conization with positive margins.

Pre- and post-conization high-risk HPV testing predicts residual/recurrent disease in patients treated for CIN 2–3

Objective. To evaluate whether high-risk human papillomavirus (HR-HPV) detection and viral load prior to treatment and status of cone margins can predict residual/recurrent disease as well as the ability of current diagnostic tools to identify residual/recurrent disease during follow-up of high-grade cervical intraepithelial neoplasia (CIN) treated by conization using loop electrosurgical procedure (LEEP). Methods. Two hundred and three women (mean age 38.6 ± 9.7; range 22–83) with CIN2–3 treated by LEEP conization and confirmed in the surgical specimen, attending follow-up visits were included. Age, HR-HPV detection and viral load determined by HybridCapture 2, and cone margins were evaluated as possible predictors of residual/recurrent disease. Value of single and repeated cytology as well as HR-HPV detection and viral load during follow-up were analyzed as screening tools of recurrence. Results. Residual/recurrent disease was demonstrated by colposcopy guided biopsy in 36 patients (17.7%). High HR-HPV load (>1000 RLU) prior to LEEP and positive cone margins were significantly associated with higher risk of recurrence (31.8% vs. 9.4%, P = 0.005; and 36.4% vs. 11.9%, P < 0.001 respectively). HR-HPV detection at 6–12 m after LEEP showed higher sensitivity than a single or repeated cytology (97.2% vs. 83.3% and 94.4% respectively) although it showed less specificity (81.4% vs. 92.2% and 82.6%). The combination of HR-HPV detection and the first cytology during follow-up detected all patients with residual/recurrent disease (sensitivity 100%, negative predictive value 100%) with an acceptable specificity (76.6%). Conclusion. The inclusion of HR-HPV testing with cytology in follow-up of patients treated for CIN2–3 would allow for fewer post-treatment visits and avoid unnecessary cytologies. High HR-HPV load prior to LEEP or positive margins should be considered as risk factors for developing residual/recurrent disease.

Factores de riesgo de cáncer cervicouterino invasor en mujeres mexicanas

To evaluate the association between invasive Cervical Cancer (CC) and high risk Human PapillomaVirus (HR-HPV) (viral load and type 16), along with other gynecological and socioeconomic factors. Individually matched case-control study (215 women with invasive CC and 420 controls). The study population was recruited between 2000 and 2001. A set of variables traditionally linked with CC (gynecological and socioeconomic factors) and two variables related to HPV infection (viral load and type 16) were assessed. Hybrid Capture II was used to detect HR-HPV DNA. Viral load was measured by light measurements expressed as relative light units (RLU) and they were categorized for analysis into four groups: negative (< 1 RLU), low viral load (1-49 RLU), middle load (50-499 RLU) and high load (>499 RLU). The analysis included univariate, bivariate and multivariate techniques being the final step the estimation of Odds Ratios (OR) by means of conditional logistic regression models. Results. The probability of having invasive CC was 78 times higher in patients with infection of HR-HPV. Risk increases with HPV type 16 (OR = 429.7) as compared with other types of HR-HPV (OR = 64.1). An important trend was observed with the increase of the viral load (from 46.6 with low viral load; to 250.7 with intermediate and 612.9 with high load). The findings also indicate significant diferences in the viral load between cases and controls according to age groups and HR-HPV types (16 versus others high risk types). Finally, the partner-demographic and obstetrical variables related to the disease increased the risk of invasive CC. No association between CC and smoking was observed in this population. This study helps in identifying women at higher risk of developing invasive CC as a subset of those patients infected with HR-HPV. The findings point strongly to the importance of the viral load in HR-HPV as a co-factor in the development of this disease. This biomarker contributes to improving the prevention and early detection of this disease and also to identify women at higher risk who carry a high viral load of HR-HPV.

Acquisition of high-risk human papillomavirus infections and pap smear abnormalities among women in the New Independent States of the Former Soviet Union.

The rates of acquisition and the times of incident high-risk (HR) human papillomavirus (HPV) infections and Pap smear abnormalities and their predictive factors were analyzed in women participating in a multicenter screening study in three countries of the New Independent States of the former Soviet Union. The 423 patients were prospectively monitored for a mean of 21.6 months. At the baseline, 118 women were HR HPV DNA negative (Hybrid Capture II assay) and Pap smear negative (group 1), 184 were HPV DNA positive and Pap smear negative (group 2), and 121 were HPV DNA negative and Pap smear positive (group 3). The time to the acquisition of an incident abnormal Pap smear (19.4 months) was significantly longer in group 1 than in group 2 (9.2 months) (P = 0.0001). The times of acquisition of incident HR HPV infection were 16.6 and 11.0 months in group 1 and group 3, respectively (P = 0.006). The monthly rates of acquisition of incident HR HPV infections were very similar in group 1 (1.0%) and group 3 (0.8%), whereas the rate of acquisition of an abnormal Pap smear was significantly higher in group 2 (3.1%) than in group 1 (1.5%) (P = 0.0001). The acquisition of HR HPV infection (but not a positive Pap smear result) was significantly (P = 0.0001) age dependent. The only significant independent (P = 0.001) predictor of the incidence of an abnormal Pap smear result was a high HR HPV load of >20 relative light units/control value (CO) (rate ratio, 2.050; 95% confidence interval, 1.343 to 3.129). Independent predictors of incident HR HPV infection were patient category (a sexually transmitted disease) and ever having been pregnant. The time of acquisition of HR HPV infection was 3 months shorter than that of an abnormal Pap smear. At the baseline the high load of a particular HR HPV type is the single most important predictor of an incident Pap smear abnormality, whereas young age and having a sexually transmitted disease predict incident HR HPV infections.

Abstracts of the 2nd joint meeting of the British Division of the International Academy of Pathology and the Pathological Society of Great Britain and Ireland, and the 185th meeting of the Pathological Society of Great Britain and Ireland. 1-4 July 2003, Bristol, United Kingdom.

Given the fact that infection with high-risk human papillomavirus (HPV) is causally involved in cervical cancer, addition of high-risk HPV testing to a cervical smear may improve the efficacy of cervical cancer screening programmes, the triage of women with equivocal or borderline Pap smears, and the monitoring of women who have been treated for cervical intraepithelial neoplasia grade 3 (CIN 3). Compared to a cervical smear HPV tests revealed a superior sensitivity (ie clinical sensitivity) for lesions >/= CIN 3, and a negative predictive value approaching 100%. However, a potential complication is the availability of several HPV testing methods, all displaying a different sensitivity and specificity to detect HPV-positive women (ie analytical sensitivity and specificity). There is now compelling evidence that the clinical sensitivity and specificity of HPV tests are not simply synonymous to their analytical sensitivity and specificity, respectively. In fact, a distinction between so-called clinically relevant and irrelevant high-risk HPV infections should be made when considering HPV tests for primary screening, triage policies, or post-treatment monitoring. Here, we discuss the potential importance of HPV load in the context of currently widely applied HPV detection methods, to distinguish clinically relevant from irrelevant HPV infections. From this it can be concluded that it is of utmost importance to define criteria, involving viral load threshold and the type of HPV detection method that should be fulfilled by an HPV test before implementation of such a test in clinical practice and population-based cervical cancer screening programmes.