Diagnosis of cervical intraepithelial neoplasia and human papillomavirus infection: punch biopsy versus cervical smear

Summary In 102 patients referred to our colposcopy clinic because of one to three Papanicolaou smears indicating cervical intraepithelial neoplasia (CIN) and/or abnormal colposcopy, routine smears and colposcopically directed punch biopsies were taken simultaneously. For detection and typing of human papillomavirus (HPV)-DNA in situ hybridization was performed in all biopsies and in 46 of the cervical smears. In cases of dysplastic lesions the number of HPV 16/18 (40.5%) and 31/33 (42.9%) was markedly higher than HPV 6/11 (16.6%) infection rate. In cases where simultaneous in situ hybridization in biopsy specimen and cervical smears was performed 21.7% showed a HPV negative smear and a positive biopsy, in 6.5% the results were the other way round. In 34.9% of cases with CIN I and 9.5% of cases with CIN II verified by punch biopsy the cytological smear did not indicate dysplasia. Our data show that mild and moderate CIN lesions of the cervix as well as HPV infection are detected more frequently by a combination of cervical smear and colposcopically directed punch biopsy than by cervical smear alone.     

Human papillomavirus-type persistence patterns predict the clinical outcome of cervical intraepithelial neoplasia

BACKGROUND: Persistent infection with high-risk (HR) human papillomavirus (HPV) genotypes is required for the development of cervical carcinoma, and integration of HPV testing into cervical screening programs is under investigation. For the clinical value of HPV testing to be fully established, genotyping studies are needed to identify HR HPV persistence in samples of known cytology and histology, and to determine the relationship with clinical outcome. To date, methods for genotyping have been research-based, and subject to variation. The availability of the Roche prototype line blot assay (LBA) offers a PCR-based, reproducible genotyping method, with a 37-type target spectrum and many potential applications. METHODS: We applied the LBA to determine persistence of HR HPV in 54 women with low-grade histology. Median interval between genotyping was 12.5 months (range 5-48). RESULTS: All 15 lesions that progressed to CIN3 (PD) were associated with HR HPV persistence. Regression of lesions (REM) was observed in 31 HPV+ women, of whom nine had clearance of existing HPV infections, with one patient then acquiring additional types. Eight HPV+ patients had no change in lesions observed (NC). Persistence of HPV type 16 was more common in the PD group (60%), compared with the REM group (27%) and the NC group (38%). CONCLUSION: Our results show that the LBA is a useful tool to identify HPV persistence patterns under anonymized conditions, with potential for research and clinical studies.     

Serologic response to human papillomavirus 16 among Australian women with high-grade cervical intraepithelial neoplasia

This study evaluated the detection of human papillomavirus (HPV) 16 antibody in HPV 16-associated cervical intraepithelial neoplasia (CIN) in Australian women. Seroreactivity to HPV 16 L1 virus-like particles was assessed in patients with CIN 2 (n= 169) and CIN 3 (n= 229) lesions previously tested for the presence of HPV DNA. Seropositivity was significantly commoner in women with HPV 16 DNA-positive lesions (98/184) than in women with no HPV DNA in the lesion (15/47) or with HPV of types other than 16 in the lesion (43/167) (P= 0.0004). In addition, seropositivity was observed in 33% (55/169) of women with CIN 2 and 46% (106/229) of women with CIN 3, in keeping with the lower fraction of CIN 2 (57/169) than CIN 3 (127/229) biopsies positive for HPV 16 DNA. HPV 16 seropositivity is most common in women with HPV 16-associated CIN, but many patients with HPV-associated CIN 3 are seronegative, and HPV 16 seropositivity is common in women with CIN associated with other HPV types. Overall, HPV 16 serology is a poor predictor of presence of HPV 16-associated CIN 3 in patient population studied.     

Risk factors for cervical intraepithelial neoplasia recurrence after conization: a 10-year study

Objective

To evaluate the risk factors potentially involved in the development of cervical intraepithelial neoplasia (CIN) recurrence after cervical conization in a long-term follow-up period.

Study design

Consecutive patients with histologically proven CIN who had undergone either cold knife conization or a loop electrosurgical excision procedure were enrolled and scheduled for serial follow-up examinations over a 10-year period. Data were stored in a digital database. Multivariate analysis was performed to identify factors for recurrence.

Results

Between January 1999 and December 2009, 282 patients fulfilled the inclusion criteria and were included in the final statistical analysis. After a median follow-up of 26.7 months (range 6–100), 64 (22.7%) women developed histologically confirmed recurrence. The 2-year recurrence-free survival was 83.7% and 66.7% for women with negative and positive margins, respectively (p = 0.008). The 5-year recurrence-free survival was 75.4% and 50.3% for patients with negative and positive margins, respectively (p = 0.0004). Positive surgical margin was the most important independent predictor of recurrence [HR 2.5 (95%CI 1.5–4.5), p = 0.0007; Wald 11.338]. After multinomial logistic regression the indication for conization based on persistent CIN1 was the only independent predictor for negative margin [OR 0.3 (95%CI 0.1–0.7), p = 0.008].

Conclusions

Our study demonstrated that the surgical margin status represents the most important predictor for CIN recurrence after conization. After excisional therapy, close follow-up is mandatory for the early detection of recurrent disease. The identification of risk factors for recurrence may guide clinical decision-making on expectant management versus re-intervention.     

Incidence of ano-genital and head and neck malignancies in women with a previous diagnosis of cervical intraepithelial neoplasia

Objective

The objective of this study was to determine if women with a history of Cervical Intraepithelial Neoplasia grades 2 and 3 (CIN2 and CIN3) are at increased long-term risk for developing non-cervix HPV-related malignancies.

Material and methods

Women diagnosed with CIN2 or CIN3 between 1980 and 2005 were identified from the British Columbia (BC) Cancer Agency Cervical Cancer Screening Program's database. These patients' records were then cross-referenced with the BC Cancer Registry for diagnosis of vulvar, vaginal, anal or head and neck (HN) cancers during the period subsequent to their diagnosis of CIN. Standardized incidence ratios (SIR) were generated according to expected rates of each cancer.

Results

54,320 women with a diagnosis of CIN2 or CIN3 were identified between 1985 and 2005. The crude incidence rate for non-cervix HPV-related cancers was 35.4 per 100,000 person-years (8.6 for vagina, 17.6 for vulva, 3.7 for anal canal and 5.5 for HN). The SIR was 1.9 (95% CI 1.3–2.7) for all non-cervix cancers, 6.7 (95% CI: 3.0–12.8) for vagina, 2.9 (95% CI: 1.7–4.6) for vulva, 1.8 (95% CI: 0.4–4.7) for anal canal, and 0.6 (95% CI: 0.2–1.4) for HN. There were statistically significant increases in anal cancers for years 5–9 and in HN cancers for years 0.5–5.

Conclusion

BC women with a history of CIN2 or CIN3 are at relatively high risk of developing non-cervical HPV-related malignancies. The findings of this study suggest that interventions such as vaccination against high-risk HPV or long-term screening for these other cancers should be evaluated.     

744例宫颈上皮内瘤样病变的诊断分析

目的　分析宫颈上皮内瘤样病变 (CIN) 10年来发病情况 ,并对其筛查诊断方法进行回顾性分析。方法　回顾总结 1993年 1月～ 2 0 0 2年 12月经阴道镜检查并行宫颈活组织检查的 2 10 7病人 ,检出 74 4例CIN ,分析CIN检出的情况及宫颈涂片细胞学和阴道镜检查对其诊断的意义。结果　CIN的检出率逐年升高 ,宫颈涂片细胞学人工阅片筛查CIN的敏感度为 4 3 9% ,电脑辅助细胞学分析系统 (CCT)筛查CIN的敏感度为72 7% ,阴道镜诊断CIN的敏感度为 88 0 %。 35岁以下未明确诊断意义的不典型鳞状上皮细胞 (ASCUS)患者中CIN检出率明显高于 35岁以上者 ,差异有显著性。结论　CCT对CIN的检出有明显提高 ,优于人工阅片 ,阴道镜指导下宫颈活组织检查是诊断CIN的敏感方法。 35岁以下ASCUS患者的处理更应重视     

Alterations in human papillomavirus-related biomarkers after treatment of cervical intraepithelial neoplasia

Objective

This study aims to assess the alterations in various HPV-related biomarkers 6 months post-treatment and how these relate to various risk factors and individual characteristics; their role for the prediction of treatment failure was also evaluated.

Material and methods

Design: Prospective observational study.Population: Women planning to undergo treatment for cervical intraepithelial neoplasia.Intervention: A liquid-based cytology sample was taken pre-operatively. This was tested for HPV genotyping, Nucleic Acid Sequence Based Amplification, flow cytometric evaluation and p16 immunostaining. A repeat LBC sample was obtained 6 months post-treatment and was tested for the same biomarkers.Outcomes: The alterations of the biomarkers 6 months post-treatment were recorded. Their relation to individual characteristics and risk factors (age, smoking, sexual history, use of condom, CIN grade, excision margin status, crypt involvement) as well as their role for the prediction of residual/recurrent disease were assessed.Analysis: The accuracy parameters (sensitivity, specificity, positive and negative predictive value and the likelihood ratios) of each biomarker for the prediction of recurrent/residual CIN were calculated.

Results

A total of 190 women were recruited. All biomarkers had significantly higher negativity rates post-treatment compared to pre-treatment ones. Multivariate analysis demonstrated that consistent condom use post-treatment significantly reduces the high-risk HPV positivity rates in comparison to no use (OR = 0.18; 95% CI: 0.09-0.38). Sensitivity and specificity for all high risk HPV DNA testing were 0.5/0.62, respectively; the relevant values for only type 16 or 18 DNA typing were 0.5/0.92, for NASBA 0.5/0.94, for flow 0.5/0.85 and for p16 0.25/0.93.

Conclusion

CIN treatment reduces positivity for all HPV-related biomarkers. Consistent condom use significantly reduces high-risk HPV positivity rates. More cases of treatment failures are required in order to specify whether different combinations of HPV-related biomarkers could enhance the accuracy of follow up, possibly in the form of a Scoring System that could allow tailored post-treatment surveillance.     

Demographic variables routinely collected at colposcopic examination do not predict who will default from conservative management of cervical intraepithelial neoplasia I

OBJECTIVE: As a result of the low incidence of progression from low grade epithelial abnormalities to cervical intraepithelial neoplasia (CIN) 3 or cervical cancer, a conservative approach to management is supported, especially in young women. Loss to follow-up is a recognised problem with a conservative approach however, with women defaulting known to experience higher rates of cancer. AIM: To determine if any routinely collected demographic variables could predict which Australian women would subsequently default from care having initially elected to have conservative management of CIN 1 lesions. METHODS: Prospectively collected data was audited on 279 women with a colposcopically directed biopsy diagnosis of CIN 1, confirmed on external review, who were enroled by their own choice into a conservative management program and monitored until a definitive lesion outcome was determined. Women who defaulted from follow-up and were lost to care providers despite follow-up appointments and reminder letters were compared to women who completed follow-up with either lesion resolution or progression requiring treatment, to establish if there were any demographic variables to predict default from care. RESULTS: Fifty-two (18.5%) women subsequently defaulted from follow-up. There were no significant differences in age, parity, proportion of women who were pregnant at diagnosis, smoking status, immunosuppressed or had a 'human papillomavirus (HPV) effect' reported on Pap-smear or colposcopic examination. CONCLUSION: We cannot easily identify a subgroup of women who are more likely to default from follow-up of CIN 1 using routinely collected demographic data. Default from follow-up is a major risk with conservative approaches and further research to reduce default rates are required.     

Immunohistochemical expression of laminin-5 in cervical intraepithelial neoplasia

OBJECTIVES: Laminin-5 is an attachment protein for epithelial cells. Several studies of a variety of cancers have reported increased expression of laminin-5 in carcinoma in situ and invasive cancer. This study was designed to investigate the correlation between the grade of cervical intraepithelial neoplasia and the immunohistochemical expression of laminin-5 in the cytoplasm and in the basement membrane underlining dysplastic squamous cells. METHODS: We used immunohistochemical methods to stain paraffin-embedded sections of cervical cone biopsies with a monoclonal antibody specifically targeting the 2-chain of human laminin-5 protein. The study sample included 175 slides: 7 normal cervical epithelium, 36 lesions of mild dysplasia, 50 lesions of moderate dysplasia, 81 lesions of severe dysplasia, and 1 invasive squamous cell carcinoma. RESULTS: We found a statistically significant correlation between the grade of cervical intraepithelial neoplasia and laminin-5 immunoreactivity in the cytoplasm (P < 0.01) and in the basement membrane (P = 0.03) by use of the Wilcoxon rank-sum test. CONCLUSIONS: According to previously published reports we confirmed with a higher number of cases a correlation of laminin-5 expression in the cytoplasm and/or basement membrane and grade of dysplastic lesion in the cervical epithelium. This study warrants further investigations with special interest to follow-up to investigate whether laminin-5 is a marker to predict the risk of progression of cervical intraepithelial neoplasia lesions.     

Characterization of high- and low-grade cervical intraepithelial neoplasia

A need exists to characterize the various grades of cervical intrapithelial neoplasia (CIN), and attempt to differentiate between high- and low-grade lesions, that may have different behavioral and progressive potentials. The identification of patients with high- or low-grade CIN is useful, as it may allow identification of those patients that have true cancer precursors. Fifty patients referred for colposcopy with abnormal cytology were studied. Univariate analysis identified three factors as important predictors of histologic grade; the colposcopic opinion, lesion surface area and the index cytology ( P < 0.005). Colposcopic opinion was associated with the index cytology ( P < 0.01) and the lesion surface area ( P < 0.005). Only the colposcopic opinion and the index cytologic smear appeared in the final model using a stepwise logistic regression analysis, indicating their independent prognostic importance in prediction of grade of abnormality in cervical intraepithelial neoplasia. The study demonstrates the value of colposcopic training and experience being necessary prior to utilizing excisional treatment methods if overtreatment is to be avoided.     

Duration of HPV-associated risk for high-grade cervical intraepithelial neoplasia

OBJECTIVE: To evaluate the duration of high-risk HPV-associated cancer risk. STUDY DESIGN: Patients who had had a routine diagnostic Hybrid Capture Tube Test (HCT) due to squamous cell abnormalities of the uterine cervix were followed-up until the endpoint of histologically diagnosed cervical intraepithelial neoplasia (CIN). RESULTS: Six hundred and thirty-eight women were followed during a cumulative follow-up of 16,423 patient months. The adjusted relative risk associated with the positive HR-HCT test for high-grade CIN/52.0 (20.9-19.2)/ proved to be higher than that of the cytological atypia/5.44 (2.52-11.77)/. At the end of the 30 months of follow-up the crude and adjusted risks for CIN2+ were 214.3 (28.4-1615.7) and 196.7 (25.4-1525.2), respectively in the HPV 16/18 group, and after 30 months, the crude and adjusted RR decreased to 57.6 (10.4-318.9) and 29.2 (5.02-170.0). In the groups of other high-risk types and possibly high-risk types the general tendency was the same. However, new CIN2+ cases were not detected after the 30th month of follow-up in these later groups. CONCLUSIONS: HPV16/18 associated relative risk is nearly 200 times higher than that of the HPV negative population and an outstanding risk persists with duration of about 30 months. The risk is manifested in progression to high-grade CIN lesions mainly within a 2 years interval after the first detection of HPV 16/18 infection.     

Cervical intraepithelial neoplasia treated by perilesional injection of interferon

Twelve patients with cervical intraepithelial neoplasia (CIN) were studied by colposcopy and cervical smears to elucidate the local response to interferon alpha and beta given by perilesional injections and to assess long-term disease remission. In all patients, local inflammatory response was seen and persisted throughout the treatment period. Selective toxic effect was evident by exfoliation of increasing proportion of degenerate to viable dyskaryotic cells, while the normal epithelial cells appeared unaffected, and by the progressive regression of the lesion. Interferon alpha induced complete remission in six of seven (85.7%) patients but three had recurrence 12 to 24 months later, while interferon beta induced complete remission in only two of five (40%) patients.     

HPV-related vulvar intraepithelial neoplasia: outcome of different management modalities.

OBJECTIVE: To evaluate the outcome of various management schemes for HPV-related vulvar intraepithelial neoplasia (VIN, usual type). METHODS: Retrospective chart review of patients with histologically diagnosed grade 2/3-VIN who had at least one year of follow-up. The variables that were collected included patient characteristics, management modalities, and clinical outcome. RESULTS: Fifty patients with a median age of 45 years old were evaluated. The median duration of follow-up was 43.5 months (12-186). Complete response (CR) and partial response occurred in 28 (56%) and 4 (8%), respectively. Nineteen of 28 patients with CR recurred with VIN. Surgical excision yielded higher CR (77%) than did either ablational techniques (21-33%) or topical immunotherapy (33%). CONCLUSION: In this experience, surgical excision for VIN, usual type, resulted in better therapeutic success rates than other treatment modalities. Management schemes should be individualized based on extent of disease and patient compliance.     

Age distribution of HPV genotypes in cervical intraepithelial neoplasia

Objective

Recent data showed that HPV16 infections in young women can lead to CIN3 formation very quickly and questioned the common assumption that invasive cervical cancer develops through slowly progressing pre-cancer lesions, CIN1, CIN2 and CIN3. The aim of the study is to compare the age distribution of HPV 16/18 related and HPV16/18 not related CIN.

Methods

We used the data generated from the clinical use of HPV genotyping (LINEAR ARRAY, Roche Diagnostics). Patients were grouped on the basis of histology, CIN1 vs. CIN2+ and on HR-HPV genotype status.

Results

The probability to develop a CIN2+ seemed to decrease with age in patients infected with HR-HPV genotype 16/18 while the inverse effect was observed in CIN2+ patients who were HR-HPV positive but HPV16/18 negative (Chi-square test, p^trend = 0.01). Only in HR-HPV positive but HPV 16/18 negative patients, a relative reduction of CIN1 vs. CIN2+ was observed with increasing age (Cochran-Armitage test, p^trend = 0.01); finally, in HR-HPV non-16/18 infected patients only a statistically significant difference in mean age between CIN1 and CIN2+ patients below age 35 was observed.

Conclusions

Besides the limitations of the present cross-sectional analysis, these data suggest a genotype specific natural history of cervical cancer precursors development: one type, more frequent, HPV16/18 related, which develops quick and early in life; another one, non-16/18 HR-HPV related, which develops later, slowly, through low- to high-grade lesions. If confirmed, this hypothesis could influence screening policies, especially in the vaccinated population.     

Accuracy of cervical smears in predicting the grades of cervical intraepithelial neoplasia

This is a retrospective study carried out to assess the correlation between the cytology and histology of cervical intraepithelial neoplasia in 1325 women. A poor correlation between the cytologic and histologic diagnosis of the various grades of CIN was shown. Forty-one percent of smears with repeated borderline change and 50% of those predicting CIN1 showed a higher grade of CIN on histology. The overall apparent false negative rate of cervical smears for high grade CIN (CIN2 and CIN3) was 19% and for CIN3 alone was only 3%. It is therefore concluded that there is a consistent tendency for cervical cytology to underestimate the severity of histologic lesions and it is therefore important that the clinicians ensure adequate follow-up of patients whose smears show a lesser degree of abnormality.     

Heat shock fusion protein-based immunotherapy for treatment of cervical intraepithelial neoplasia III

OBJECTIVES: SGN-00101 (HspE7, Nventa, San Diego, CA) is a novel therapeutic vaccine consisting of a fusion protein containing an M. bovis BCG heat shock protein (Hsp65) covalently linked to the entire sequence of HPV 16 E7. This trial was designed to evaluate the efficacy and toxicities of HspE7 in women with CIN III. METHODS: HIV (-) women with biopsy-proven CIN III were eligible. Two cohorts were accrued; one cohort to establish efficacy and a second cohort with a longer follow-up period to improve the precision of the trial to estimate response rates. Each patient underwent 3 monthly subcutaneous vaccinations with 500 microg of HspE7 followed by monthly colposcopic follow-up for 1 month in cohort 1 and an extended observation period (2 months) in cohort 2. All patients then underwent a LEEP or cone biopsy of the cervix. A complete pathologic response (pCR) was defined as no evidence of CIN or CIN I (only HPV changes). A partial response (PR) was defined as colposcopic lesion regression of >50% in size. Cervicovaginal lavage samples were obtained at each visit for HPV typing using MY09/ MY11 HPV PCR. RESULTS: Seventy-two patients were registered and screened, of whom 64 were eligible. Fifty-eight patients completed the trial and were evaluable (31 in cohort 1, 27 in cohort 2). There were no significant epidemiologic or HPV type differences between the 2 cohorts so responses were combined for analysis. Of the 58 evaluable patients, 13 (22.5%) had a pCR; 32 (55%) had a PR and 11 (19%) had stable disease. Two (3.5%) patients in cohort 2 had microinvasive disease and were defined as progressive disease. Thirty-three of 58 (57%) of the patients were infected with HPV 16 prior to vaccination or in subsequent visits. There was no significant difference in regression in women infected with HPV 16 compared to those without HPV 16 infection (88% vs. 70%; p=0.12). Women who had a previous LEEP or ablation for CIN were 2.7 times more likely to have a complete response compared to patients without previous treatment, although the difference was not statistically significant (95% CI for rate ratio: 0.95-6.19, p=0.10). At a cellular level, there was a significant association between local inflammation and response; lower grade of lesional inflammation correlated with a response to HspE7 (p=0.04 using Wilcoxon rank sum test). CONCLUSIONS: HspE7 appeared to demonstrate activity in women with CIN III and met a priori assumptions for efficacy; however, it is unclear whether this response was due to natural regression rather than treatment effect. HspE7, which targets the HPV 16 E7 oncoprotein, had efficacy in patients infected with HPV types other than 16, suggesting cross-reactivity. A larger randomized, controlled trial is needed to better define efficacy and to identify subsets of women most likely to benefit from immunotherapy.     

CD1a和E-cadherin在宫颈上皮内瘤变中的表达

目的:探讨CD1a和E-cadherin与宫颈癌发生发展的关系及作为早期癌变生物学指标的可能性。方法:采用免疫组化SP法检测CD1a和E-cadherin在同期56例宫颈上皮内瘤变、56例宫颈鳞癌和15例正常宫颈组织中的分布及表达。结果:(1)CD1a+朗格汉斯细胞在正常宫颈、CIN和宫颈癌各组中数量逐渐减少,两两比较有显著差异(P0.01);CIN中该细胞数量随病变严重程度减少,CINⅠ和CINⅡ、CINⅠ和CINⅢ组之间两两比较有显著差异(P0.05),而CINⅡ与CINⅢ组间无显著差异(P0.05);(2)E-cadher-in在正常宫颈、CIN和宫颈癌各组中的阳性表达率及强度逐渐下降,两两比较有显著差异(P0.05);在CIN中阳性表达率及表达强度随病变严重程度呈下降趋势,CINⅠ和CINⅡ、CINⅠ和CINⅢ之间两两比较有显著差异(P0.05),而CINⅡ和CINⅢ之间无显著差异(P0.05);(3)宫颈组织中CD1a+朗格汉斯细胞的细胞数与E-cadherin的阳性表达率及强度呈正相关(r=0.912,P0.05)。结论:CD1a+朗格汉斯细胞与E-cadherin可能在宫颈癌的发生、发展过程中起重要作用。     

Cervical intraepithelial neoplasia and condyloma: an analysis of diagnostic accuracy of posttreatment follow-up methods

The appropriate follow-up method of patients treated for cervical intraepithelial neoplasia and condyloma is controversial. One school of thought favors cytologic testing alone, whereas the other favors cytologic testing combined with colposcopy. We have analyzed the records of 750 patients treated for cervical intraepithelial neoplasia or condyloma or both in our Colposcopy Clinic. Cytologic testing, colposcopy, and histologic examination were done routinely at 3 to 4, 6 to 8, and 12 to 15 months after treatment of the above conditions in 750 patients. Totals of 95% and 5% of all 128 persistent lesions were detected by histologic examination at the first (3 to 4 months) and second (6 to 8 months) posttreatment visits, respectively. The combined false negative rates for both visits were 23% for cytologic testing, 8% for colposcopy, and 5% for histologic examination. False negative rates at first visits were 1.5% for colposcopy combined with cytologic testing. These observations suggest that colposcopy and, if appropriate, histologic examination significantly contribute to cytologic testing for diagnosing the majority of persistent disease during the first posttreatment visit. Cytologic testing alone seems to be sufficient for the subsequent follow-up of patients treated for cervical intraepithelial neoplasia and/or condyloma.