Low-grade Squamous Intraepithelial Lesion Cases Research Articles

Risk of Cervical Precancer and Cancer in Women Aged 30 Years and Older With an HPV-Negative Low-Grade Squamous Intraepithelial Lesion Screening Result

To characterize the 6- and 18-month cumulative risk of cervical intraepithelial neoplasia grade 2 or worse (CIN 2+) and grade 3 or worse (CIN 3+) in women aged 30 years and older after a low-grade squamous intraepithelial lesion (LSIL) cytology and high-risk human papillomavirus (HPV)-negative screening result in routine clinical practice. Comprehensive quality assurance databases of screening test and biopsy results from the Regional Laboratory of the Kaiser Permanente Northern California Health Plan were reviewed. All women aged 30 years and older with LSIL cytology were sorted by high-risk HPV status. Associated biopsy results were tabulated, and the corresponding risks of CIN 2+ and CIN 3+ diagnosed within 18 months after LSIL cytology were calculated overall and by decade of age. During the 6-year period, from 2003 to 2008, 4,113 LSIL cases were interpreted in women aged 30 years and older for which corresponding high-risk HPV and biopsy results were available. The proportion of women with LSIL testing positive for HPV declined with age, from 89% in the group aged 30 to 39 years to 76% in women older than 50 years (p < .001). Of 622 women with HPV-negative LSIL cytology, there was no case of cancer detected at colposcopy occurring within 6 months of the screening test. The 18-month risks of CIN 2+ and CIN 3+ were 3.5% and 1.4%, respectively. The risk of CIN 3+ is sufficiently low in women aged 30 years and older with high-risk HPV-negative LSIL that 1 year follow-up rather than immediate colposcopy should be considered when it occurs in routine clinical practice.

Micronucleus in Cervical Intraepithelial Lesions and Carcinoma

Aims and Objectives: To score and compare micronucleus (MN) in the whole spectrum of cervical lesions including normal, inflammatory, abnormal squamous cell of undetermined significance (ASC-US), low-grade squamous intraepithelial lesion (LSIL), high-grade squamous intraepithelial lesion (HSIL) and invasive cancer (IC) and to evaluate the role of MN as a biomarker in different pre-neoplastic and neoplastic lesions. Materials and Methods: A total of 224 slides, comprised of normal (40), inflammatory (40), ASC-US (30), LSIL (38), HSIL (30) and IC (46), were studied. All the cases of HSIL, IC and ASC-US had histopathology. The LSIL, normal and inflammatory smears were again reviewed by 2 experienced cytopathologists independently. Two observers separately and independently counted the number of micronucleated cells per 1,000 of epithelial cells in oil immersion magnification (×100 objective) which was expressed as MN score per 1,000 cells. Results: The mean MN scores ± SD in normal, inflammatory, ASC-US, LSIL, HSIL and IC cases of cervical lesions were 1.02 ± 1.59, 0.4250 ± 0.71208, 2.87 ± 2.21, 4.7368 ± 5.62179, 21.30 ± 17.18 and 18.50 ± 9.54, respectively. MN scores of IC and HSIL were significantly high compared to the normal (p < 0.000), the inflammatory (p < 0.000), the ASC-US (p < 0.000) and to the LSIL (p < 0.000) group (analysis of variance test). LSIL showed significant difference with the normal (p = 0.043), the inflammatory (p = 0.019), the HSIL (p < 0.000) and the IC (p < 0.000) group but not with the ASC-US (p = 0.342) group. Conclusions: MN scoring on the epithelial cells of cervix could be used as a biomarker in cancer screening. This is an easy, simple, reliable, reproducible and objective test which can be performed on routinely stained smears.

Significance of High-Risk Human Papillomavirus DNA Detection in Women 50 Years and Older With Squamous Cell Papanicolaou Test Abnormalities

Data on cytologic screening and follow-up disproportionately reflect findings from frequently screened younger women, and data from screened women 50 years and older using newer screening technologies remain limited. To better understand the utility of adjunctive high-risk human papillomavirus (hrHPV) testing for disease risk stratification in women 50 years and older with a range of liquid-based cytology, abnormal, squamous cell Papanicolaou test results. Liquid-based cytology cases interpreted as high-grade squamous intraepithelial lesion (HSIL); low-grade squamous intraepithelial lesion (LSIL); atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion (ASC-H); and atypical squamous cells of undetermined significance (ASC-US) at Magee-Womens Hospital (Pittsburgh, Pennsylvania) were retrospectively identified for a 36-month period between July 1, 2005, and June 30, 2008, from women 50 years and older who also had hrHPV DNA test results. Histopathologic follow-up diagnoses were analyzed. During the study period, 4855 women 50 years and older had HSIL, LSIL, ASC-H, or ASC-US Papanicolaou test results and hrHPV testing. In 89.3% of HSIL cases, 71.0% of LSIL cases, 38.9% of ASC-H cases, and 14.2% of ASC-US cases, hrHPV test results were positive. The positive predictive value of a positive hrHPV test for histopathologic cervical intraepithelial neoplasia 2/3(+) detection was 90.5% with HSIL, 15% with ASC-H, 9.8% with LSIL, and 3.2% with ASC-US. A negative hrHPV test result had a 100% negative predictive value for histopathologic cervical intraepithelial neoplasia 2/3(+) in both LSIL and ASC-H cases. In women 50 years and older, a positive hrHPV test result significantly increased the likelihood of follow-up histopathologic diagnoses of cervical intraepithelial neoplasia 2 /3(+) in patients with HSIL, LSIL, and ASC-H Papanicolaou test results compared with women with negative hrHPV test results. No cervical intraepithelial neoplasia 2/3(+) diagnoses were documented in women 50 years and older with LSIL or ASC-H Papanicolaou test results and negative hrHPV test results.

Atypical squamous cells and low-grade squamous intraepithelial lesion in cervical cytology: cytohistological correlation and implication for management in a low-resource setting

To perform an audit of all cervical smears reported as atypical squamous cells (ASC) and low-grade squamous intraepithelial lesion (LSIL) as in the Bethesda system (TBS) 2001, and determine their histological follow-up and outcome when available, in order to define the threshold for colposcopic referral. A total of 25,203 cervical smears were screened over a period of 3 years (January 2006 - December 2008) and all ASC and LSIL smears were reviewed with the corresponding histological follow-up. All cervical intraepithelial neoplasia (CIN) grade 2 lesions and above (CIN2+) were considered as clinically significant lesions for analysis. Out of 25,203 cervical smears, 424 (1.7%) were reported as ASC and 113 (0.4%) as LSIL. Additionally, three were reported as atypical cells, not otherwise specified. The ASC : SIL ratio was 2.18 : 1. Follow-up histology was available in 153 (36.8%) of the ASC cases and revealed CIN2+ lesions in 22 (14.4%). Follow-up histology was available in 50 (44.2%) of LSIL cases and revealed clinically significant abnormalities in five (10%), all of which were CIN2. CIN3 and invasive squamous carcinomas were seen in 5.9% and 1.4%, respectively, of cases of ASC, and not seen in LSIL. Reclassification of ASC smears into ASC-US (ASC-undetermined significance) and ASC-H (ASC- high grade SIL not excluded) revealed ASC-H in 2.6% of all ASC smears, with a clinically significant outcome in 45.4%. In a low-resource setting where human papillomavirus testing is unaffordable, the threshold for colposcopic referral and follow-up histology should be ASC rather than SIL.

SERVİKOVAGİNAL SMEARLERDE SİTOLOJİK BULGULARIN BETHESDA SİSTEMİNE GÖRE DEĞERLENDİRİLMESİ “SON BEŞ YILDAKİ (2005-2010) SONUÇLARIMIZ”

Amac: invaziv kansere donusebilecek oncu lezyonlarla seyreden serviks kanseri onlenebilir bir hastaliktir. Erken tanida servikovajinal sitolojiler onemlidir. Amacimiz, 2001 Bethesda Sistemine gore serviko-vaginal smear olgularimizin sitopatolojik bulgularini guncel bilgiler esliginde yeniden degerlendirerek sunmaktir. Gerec ve yontem: Calismamizda 2005 – 2010 arasinda Mustafa Kemal Universitesi Tip Fakultesi Patoloji laboratuvarinda Bethesda sistemine gore tani almis 11946 olgunun raporlari tekrar gozden gecirildi. Bulgular: 11946 servikovaginal smearda ASC-US (Atipical Squamous Cells-Undetermined Significance) %0.92, ASC-H (Atipical Squamous Cells-Cannot Exclude High Grade Squamous Intraepithelial Lesions) %0.1, LSIL(Low Grade Squamous Intraepithelial Lesion): %0.08, HSIL(High Grade Squamous Intraepithelial Lesion) %0.07 ve malign hucre %0.02 olarak bulundu. ASH:SIL (Atipical Squamous Cells: Squamous Intraepithelial Lesion) orani 144:19 olarak belirlendi. Sonuc: ASC-US, ASC-H, LSIL, HSIL ve skuamoz hucreli karsinom olgularimiz az sayidadir. Laboratuvardaki kalite kontrolunu belirlemede ASH:SIL orani onemlidir. Fakat daha iyi yorumlama yapilabilmesi icin genis diziler iceren calismalar yapilmalidir. ABSTRACT Aim: Servical cancer is a preventable disease, which manifests as prior lesions that can turn into invasive cancers. Servico-vaginal sitologies are important in early diagnosis. We aim to offer the sitopathologic findings of servico-vaginal smear according to 2001 Bethesda System in view of updated knowledge. Materials methods: In our study, the 5961 case reports diagnosed according to Bethesda System in Mustafa Kemal University Laboratuary of Pathology between 2005-2007 have been reviewed. Results : Of 11946 cases, the findings in servico-vaginal smear were as fallows: ASC-US (Atipical Squamous Cells-Undetermined Significance) 0.92%, ASC-H (Atipical Squamous Cells-Cannot Exclude High Grade Squamous Intraepithelial Lesions) 0.1%, LSIL(Low Grade Squamous Intraepithelial Lesion) 0.08%, HSIL (High Grade Squamous Intraepithelial Lesion) 0.07%, and malign cells 0.02%. The ratio of ASH to SIL (ASH:SIL) has been determined as 144:19. Conclusion: The cases of ASC-US, ASC-H, LSIL, HSIL and squamous cell carcinoma are relatively few. In determining the quality control in the laboratuary the ASH:SIL ratio is important. However, for a better evaluation, studies with wider series should be carried out.

Age-specific prevalence of HPV genotypes in cervical cytology samples with equivocal or low-grade lesions

Background:To define the spectrum of human papillomavirus (HPV) types and establish an age limit for triage HPV testing in atypical squamous cells of undetermined significance (ASCUS) and low-grade squamous intraepithelial lesion (LSIL).Materials and methods:343 liquid-based cytological samples from the population-based screening programme with minor abnormalities were subjected to HPV genotyping (Linear Array, Roche, Basel, Switzerland).Results:High-risk human papillomavirus (HR-HPV) was found in 71% of LSIL and 49% of ASCUS cases (P<0.001). High-risk human papillomavirus prevalence was age-dependent in LSIL (P=0.01), with decreasing prevalence until the age of 50 years, followed by a slight increase. Human papillomavirus type 16 was the most common HR-HPV, found in 23% of HPV-positive women. Human papillomavirus type 18 was the sixth most common, found in 9.9% (P<0.001). An age-dependent quadratic trend was observed for multiple infections (P=0.01) with a trough at about 42 years. The most common HR-HPV types to show a coinfection with HPV16 (clade 9) were HPV39 (28%), 45 (38%), and 59 (46%), belonging to HPV18 clade 7. The frequency of low-risk (LR) vs probable HR and HR-HPV also followed an age-dependent quadratic trend.Conclusions:After the age of 25 years, HR-HPV prevalence is similar in LSIL and ASCUS cases, motivating a low age limit for triage HPV testing. Multiple infections and LR/HR-HPV dominance are age-dependent. Genotyping in longitudinal design is needed to elucidate the importance of multiple infections in cancer progression and in cross-protection from vaccination.

3q26 ( hTERC) gain studied by fluorescence in situ hybridization as a persistence-progression indicator in low-grade squamous intraepithelial lesion cases

Gains of 3q26 chromosome region, where the human telomerase RNA gene (hTERC) is located, have been previously documented in cervical carcinomas and preneoplastic lesions. The aim of our study was to define the value of 3q26 gains related to persistence-progression in cervical specimens with cytologic diagnosis for low-grade squamous intraepithelial lesions, using liquid-based cytology (ThinPrep; Hologic, Marlborough, MA) and fluorescence in situ hybridization. For these purposes, 55 patients were included in the study: 25 cases with a negative cytologic diagnosis for squamous intraepithelial lesion or malignancy (20 premenopausal and 5 postmenopausal women, used as control negative cases) and 30 low-grade squamous intraepithelial lesion cases. The follow-up was performed using cytology at 6, 12, and 24 months after the low-grade squamous intraepithelial lesion diagnosis. When the cytology result showed a high-grade lesion, colposcopy and biopsy were performed. Fluorescence in situ hybridization technique with a 3q26 2-color commercial probe was performed to determine the number of hTERC copies. There were no differences between premenopausal and postmenopausal normal cases. Low-grade squamous intraepithelial lesion cases with regression in the follow-up at 6, 12, and 24 months showed a percentage of cells with 3q26 gains similar to the control cases and lower than low-grade squamous intraepithelial lesion cases with persistence or progression (P < .05). Fluorescence in situ hybridization results were similar in preserved and frozen samples. However, in frozen samples, the number of cells suitable to be evaluated by fluorescence in situ hybridization was lower than in preserved (nonfrozen) cases. In conclusion, the determination by fluorescence in situ hybridization of 3q26 gains in low-grade squamous intraepithelial lesion cases could be useful to predict the persistence-progression of such cervical lesions using both preserved and frozen cervical material.

여성에 있어 DNA 칩검사에 의한 인유두종바이러스 감염률의 조사

우리는 549명의 여성을 대상으로 HPV DNA Chip을 이용하여 자궁경부암의 주요 원인인 HPV 감염률을 조사하고 Pap 도말 염색법을 이용한 세포학적 검사를 실시하였다. 전체 대상여성 549명 중 237명 HPV DNA Chip 검사에서 양성 이었다(43.17%). 237명 중 203명이 고위험군 HPV 아형에 감염되었고(88.61%, 고위험군), 17명이 저위험군 HPV 아형에 감염되었고(7.17%, 저위험군), 나머지 17명은 고위험군 아형, 저위험군 아형, 미확인 아형에 감염 되었다(7.17%, 혼합형). 연령별 감염률은 20대가 1.26%, 30대 15.61%,40대 31.65%, 50대 23.21%, 60대 이상이 13.92%으로 확인되었다. 저위험군과 혼합군에서 고위험군보다 HPV 감염의 빈도가 더 낮게 나타났다. 세포학적 진단결과(224명의 여성)와 HPV chip 양성(237명의 여성)여성 간의 비교에서 고위험군의 경우 194명중 132명(68.04%)이 ASCUS (atypical squamous cells of undetermined significance, 7.22%), LSIL (low grade squamous intraepithelial lesion, 15.98%), HSIL (high grade SIL, 23.20%), 자궁경부암(21.65%) 등과 같은 자궁경부질환이 있었다. 저위험군(224 여성 중 14명)의 경우 ASCUS 1예와 LSIL 6예였는데 비해 혼합군(224 여성 중 4명)의 경우에는 단지 2예의 ASCUS 만이 있었다. 고위험 HPV 아형16 및 18에 감염된 여성은 각각 26예 및 7예의 자궁경부암이 있었으나 저위험군 HPV 아형 및 기타 아형의 경우 자궁경부암 발생이 매우 낮거나 없었다. 결론적으로 본 연구에서 43.17%의 HPV 유병률을 보였고 고위험 HPV 아형16이 감염된 여성들에 있어 전암 병변 또는 자궁경부암의 원인이 되는 주요인자이며 HPV DNA chip 검사는 HPV 감염 유무를 진단하는 정밀하고 유용한 방법임을 시사하고 있다. We determined the prevalence of human papillomavirus (HPV) by DNA chip test in 549 women and cytologic diagnosis. 237 of 549 women (43.17%) subjected with HPV DNA Chip examination were found positive for HPV. 210 (88.60%, High group) were infected with high-risk HPV types. 17 (7.17%, Low group) were infected with low-risk HPV types (6, 11, 40, 44, 70) and 17 (7.17%, Mixed group) were infected with mixed types. According to age, in their twenties, thirties, forties, fifties and over sixties, the prevalence of infection with high-risk HPV types were 1.26% (3/237), 15.61% (37/237), 31.65% (75/237), 23.21% (55/237), and 13.92% (33/237), respectively. In the Low and Mixed group, percentages of infection with HPV were significantly lower than that of the High group. On the comparison of cytologic diagnosis (224 women) by Pap smear and DNA chip positive (237 women) for HPV, 132 out of 194 cases in the High group (68.04%) suffered cervical lesions with ASCUS (atypical squamous cells of undetermined significance, 7.22%), LSIL (low grade squamous intraepithelial lesion, 15.98%), HSIL (high grade SIL, 23.20%) and ICC (invasive cervical cancer, 21.65%). The Low group (14/224 women) showed 1 case of ASCUS and 6 cases of LSIL, whereas the Mixed group (4/224 women) had only 2 cases of ASCUS. According to the HPV subtypes, the high-risk types 16 and 18 induced 26 and 7 cases of ICC, respectively, whereas other HPV subtypes induced lower or no ICC incidence. In conclusion, the present data imply that the prevalence of HPV was 43.17%, high-risk HPV type 16 is a major factor, which causes precancerous and/or cervical cancer in woman and that HPV DNA chip is an accurate and useful tool for detecting HPV.

Staining characteristics of p16INK4a: Is there a correlation with lesion grade or high‐risk human papilloma virus positivity?

The aims of this study were to evaluate the efficiency of p16INK4a in showing cervical lesions and to determine any relationship between lesion grade and high-risk human papilloma virus (HR-HPV) infection and p16INK4a staining characteristics. Immunohistochemical analysis of p16INK4a was performed on 13 low-grade squamous intraepithelial lesions (LSIL), 22 high-grade squamous intraepithelial lesions (HSIL), 23 squamous cell carcinoma (SCC) and 25 normal tissue samples. The distribution, staining pattern and intensity of p16INK4a expression were assessed and correlated with HR-HPV positivity determined by real-time polymerase chain reaction. All HSIL and SCC cases, but only 46.2% of LSIL cases, were positive for p16INK4a. Although positive staining of p16INK4a in showing HR-HPV-positive lesions was statistically significant (P=0.000), we could not find a significant correlation for distribution (P=0.319), staining pattern (P=0.057) or intensity (P=0.057) of p16INK4a in showing HR-HPV in cervical epithelium. These parameters were correlated only with the increasing grade of the lesion (P=0.000). p16INK4a is a highly sensitive marker of cervical intraepithelial neoplasia and cervical cancer. There is a good correlation between p16INK4a expression and cervical lesion grade and HR-HPV positivity. The distribution, staining pattern and intensity of this marker are significantly correlated with the increasing grade of cervical lesions, suggesting that diffuse distribution, full thickness staining pattern and strong intensity of this marker are highly supportive of HSIL and cervical cancer, while focal distribution, scattered/basal staining pattern and weak intensity suggest LSIL. However, these parameters are not correlated with HR-HPV status in cervical lesions.

Human papillomavirus genotype distribution in low-grade squamous intraepithelial lesions in France and comparison with CIN2/3 and invasive cervical cancer: The EDiTH III study

Objectives In the present study (EDiTH III study), the genotype-specific prevalence of HPV in low-grade squamous intraepithelial lesions (LSIL) was estimated to predict the potential benefit of HPV vaccination in France. This prevalence was compared to that previously reported in France in high-grade cervical intraepithelial neoplasia (CIN2/3, EDiTH II study) and squamous cell carcinoma (SCC, EDiTH I study) to identify the genotypes preferentially associated with a progression to malignancy. Methods 397 smears with LSIL diagnosis (Preservcyt®) were retrospectively collected in different centres in France and genotyped using the INNO-LiPA assay allowing the detection of 24 HPV genotypes. Results HPV was found in 98% of cases. The most prevalent genotypes in LSIL in France were HPV 66 (25%), HPV 16 (21%), HPV 53 (18%), 51 (17%) and 52 (14%). HPV 16 and/or 18 were present in 28% and HPV 6, 11, 16 and/or 18 in 33% of LSIL. The highest SCC/LSIL prevalence ratios were shown for HPV 16, 33 and 18. Conclusions With a 95% vaccine efficacy on CIN1 and theoretical vaccine coverage of 100%, HPV vaccination might prevent 27% (with a 16, 18 bivalent vaccine) and up to 32% (with a 6, 11, 16, 18 quadrivalent vaccine) of LSIL cases in France. In this study, LSIL related to HPV 16, 18 or 33 are at highest risk of progression to malignancy and thus could require a stringent surveillance. Conversely, anxiety and over-treatment could be avoided in women with low risk of progression.

Human Papillomavirus Infection and Cervical Disease in Human Immunodeficiency Virus-1–Infected Women

To report on the natural history of high-risk human papillomavirus (HPV) infection and cervical disease in human immunodeficiency virus (HIV)-1-infected women living in Cape Town, South Africa. This was a prospective study of 400 untreated, HIV-1-infected women who underwent high-risk HPV DNA testing, cytology, colposcopy, histology, and CD4 count testing every 6 months for 36 months. Human immunodeficiency virus viral loads and HPV type distribution were determined at entry and after 18 months. Sixty-eight percent of the women were high-risk HPV DNA positive at entry, 35% had a cytologic diagnosis of low-grade squamous intraepithelial lesion (LSIL), and 13% had high-grade squamous intraepithelial lesion (HSIL). There were no cancers. Abnormal cytology and high-risk HPV positivity were strongly correlated with low CD4 counts and high HIV viral loads. The most prevalent types of HPV were HPV-16, -52, -53, -35, and -18. Incident high-risk HPV infection occurred in 22%, and of those infected with high-risk HPV, 94% of infections persisted over an 18-month period, and 6% cleared their infections. Cytologic progression to SIL from normal/atypical squamous cells of undetermined significance cytology occurred in 17% of cases, but only 4% of cases of LSIL progressed to HSIL. There is a high level of high-risk HPV infection in HIV-1 infected women, but progression to HSIL over 36 months occurred in the minority of cases. We recommend an initial colposcopy for an abnormal test, and if no high-grade lesion is identified, triennial screening would be appropriate. Human papillomavirus type 16 was the commonest, and HPV-18 was the fifth commonest, suggesting that vaccination against these two types would have a significant effect. II.

Low-Grade Squamous Intraepithelial Lesions of the Cervix With Marked Cytological Atypia-Clinical Follow-Up and Human Papillomavirus Genotyping

The majority of low-grade squamous intraepithelial lesions (LSILs) of the cervix regress spontaneously; however, a small proportion of LSILs progress to high-grade squamous intraepithelial lesion (HSIL) if the lesion is not excised. The guidelines for which patients with LSIL should be treated and which may be followed safely are not well established. The goal of this study was to identify a subgroup of patients with LSIL who may require surgical treatment. We hypothesized that patients with LSILs with marked cytological atypia (LSIL-MA) may be at higher risk for subsequent HSIL. In addition, we were interested in whether LSIL-MA was associated with specific human papillomavirus genotypes. Consecutive patients with biopsy diagnosis of LSIL (n = 30) and LSIL-MA (n = 30) were identified. Marked atypia was defined as 5 or more cells with nuclear enlargement of at least 5 times the size of an intermediate cell nucleus or multinucleation with 5 or more nuclei. Patient follow-up was recorded for up to 24 months. Human papillomavirus genotyping was performed using SPF10 PCR and line probe assay. High-grade squamous intraepithelial lesion on follow-up was significantly more common in patients with LSIL-MA (36%) than in patients with LSIL (7%), and negative follow-up was significantly more common in patients with LSIL (50%) than LSIL-MA (23%). Cases of LSIL and LSIL-MA showed similar spectrum of human papillomavirus genotypes. Marked cytological atypia in LSILs identifies a subset of patients with a high rate of HSIL on follow-up. In such patients, an excisional cone biopsy should be strongly considered.

Evaluation of p16INK4a, minichromosome maintenance protein 2, DNA topoisomerase II α, ProE X C, and p16INK4a/ProE X C in cervical squamous intraepithelial lesions

p16INK4a has been shown to be overexpressed in nearly all high-grade squamous intraepithelial lesions (HSILs). Other cell-cycle regulators, such as minichromosome maintenance protein 2 (MCM2), DNA topoisomerase IIalpha (TOP IIA), and ProE(X) C (a cocktail of MCM2 and TOP IIA), have also demonstrated some value in identifying squamous intraepithelial lesions. Data on direct comparison of those cell regulatory proteins in the detection of squamous intraepithelial lesions, with a focus on low-grade squamous intraepithelial lesions (LSILs), are limited. We immunohistochemically evaluated the diagnostic value of p16, MCM2, TOP IIA, ProE(X) C, and a cocktail of p16 and ProE(X) C in 62 cervical biopsy specimens, including 14 cases of benign squamous mucosa (group 1), 34 cases of LSILs (group 2), and 14 cases of HSILs (group 3). The staining intensity and distribution were recorded. The results demonstrated that positive staining for p16 and the p16/ProE(X) C was observed in 100% of cases in group 3, whereas 79%, 86%, and 79% of cases were positive for CM2, TOP IIA, and ProE(X) C, respectively. ProE(X) C and the p16/ProE(X) C showed positive staining in 94% and 100% of cases in group 2, respectively. In contrast, immunoreactivity for p16, MCM2, and TOP IIA was detected in only 76% of cases in group 2. Importantly, all 8 p16-negative cases in group 2 were positive for p16/ProE(X) C (P = .003). Our data indicate that (1) p16 is a more sensitive and specific marker for identifying HSILs; (2) ProE(X) C is a better marker for the detection of LSILs; and (3) p16/ProE(X) C provides the highest diagnostic value for the detection of both HSILs and LSILs.

Accuracy of ThinPrep Imaging System in Detecting Low-Grade Squamous Intraepithelial Lesions

The ThinPrep Imaging System (Imager) for cervical cytology is used in many US laboratories, but the ability of the system to identify classic changes of low-grade squamous intraepithelial lesion (LSIL) has not been independently reported. To evaluate the accuracy of this system in detecting classic LSIL cells. A total of 114 imaged ThinPrep LSIL cases from April to June 2005 were reviewed to determine whether the most diagnostically relevant cells were present in the 22 fields selected by Imager. Those LSIL specimens from January to June 2005 that were initially screened by the Imager and classified as "negative" in the 22 fields were also reviewed. The average number of classic koilocytes per slide was compared among cases that had koilocytes within 22 fields with cases upgraded during full review. Twelve (10.5%) of 114 LSIL cases from April to June 2005 did not show diagnostic LSIL cells within the initial 22 fields. Full manual rescreening was performed on 1025 cases from January to June (20.4% of negative cases). Nine cases that were initially negative in the 22 fields were revised to LSIL (2.4% of all 381 LSIL cases reported from January to June 2005). An average of 10 to 11 koilocytes were present in these 2 groups, which was significantly lower than the average of 75 koilocytes in cases in which the 22 Imager-selected fields showed LSIL. Although the ThinPrep Imager finds abnormal cells in most LSIL cases, the system may have limitations in detecting koilocytes in the 22 Imager-selected fields. When 10% quality control rescreening is performed as required by federal regulations, full manual rescreening will provide the most accurate results.

The Distribution of Low and High-risk HPV Types in Vulvar and Vaginal Intraepithelial Neoplasia (VIN and VaIN)

It has been proposed that low-grade vulvar and vaginal lesions (VIN 1 and VaIN 1) are flat condylomas and should be designated as such. Moreover, their relationship to high-grade lesions (VIN 3 and VaIN 3) is unclear. Accordingly, this study was undertaken to address these issues by comparing the distribution of human papillomavirus (HPV) types in vulvar and vaginal intraepithelial lesions. We identified 33 cases of VIN 1, 34 cases of VIN 3, 17 cases of VaIN 1, and 16 cases of VaIN 3. In addition, 36 cases of low-grade squamous intraepithelial lesion (LSIL) in the cervix and 116 cases of cervical high-grade squamous intraepithelial lesion were used for comparison. Polymerase chain reaction analysis was performed using both the Roche PGMY and DDL SPF 10 systems. In cases where HPV was detected, the majority of low-grade and high-grade lesions contained a single HPV type. However, a minority of cases were found to have multiple HPV types. Of the VIN 1 cases, a low-risk virus was seen in 22 (67%), with HPV 6 or 11 accounting for 14 (42%). A high-risk virus was detected in 14 (42%) of cases of which 2 (6%) contained HPV 16. Of the VIN 3 cases, all had high-risk HPV of which 31 (91%) were found to have HPV 16. Of the VaIN 1 cases, 6 (35%) were found to have low-risk HPV types. HPV 6 or 11 were not found in these cases. High-risk virus was seen in 13 (76%) VaIN 1 cases, with 1 (6%) containing HPV 16. HPV was detected in 15 of 16 (94%) VaIN 3 lesions, all of which had high-risk types. HPV 16 was found in 8 (50%). In contrast, 2 (6%) of cervical LSIL had low-risk HPV (HPV 6 and 11), whereas 34 (94%) of LSIL cases had high-risk HPVs. Of the cervical high-grade squamous intraepithelial lesion cases, 100% had high-risk HPVs of which 87 (75%) were found to have HPV 16. The findings demonstrate that a significant number of low-grade vulvar and vaginal lesions contain high-risk HPV types, supporting their designation as low-grade intraepithelial lesions rather than flat condylomas. The low frequency of HPV 16 in VIN 1 compared with VIN 3 suggests they are distinct lesions or that HPV 16 is critical in the progression to VIN 3. Finally, comparison of the distribution of HPV in the vagina and vulva suggests that VaIN is more closely related to cervical intraepithelial neoplasia than to VIN.

Human papillomaviral load changes in low-grade squamous intraepithelial lesions of the uterine cervix

To better predict risk of progression of low-grade squamous intraepithelial lesions (LSILs) of the uterine cervix in women with human papillomavirus (HPV) infections, 294 baseline cervical specimens from women with LSILs were evaluated. Specimens were tested for HPV DNA using hybrid capture 2 (HC2) and PCR-reverse line blotting. 65 LSILs with HPV DNA types 16, 18, 52, or 58 were examined for physical status, E2/E6 ratio and viral load at two time points, along with patient age. Women with LSILs whose viral loads increased between baseline and 6 month follow-up had a 45% risk of developing HSIL (OR=7.6, 95% CI=1.9–29.4, P<0.01), as evaluated by real-time PCR and a 44% risk (OR=6.1, 95% CI=1.6–22.7, P<0.01), as evaluated by HC2. The two viral load measures correlated well (Person's coefficient, r=0.687, P<0.001). Such evaluations of viral load changes (increased or not increased) through repeat HPV DNA testing could predict progression of disease in LSIL cases of HPV types 16, 18, 52, and 58, which correlates to clinical implications.

Testing automated liquid-based cytology samples with a manual liquid-based cytology method using residual cell suspensions from 500 ThinPrep cases

We report a technical improvement upon a previously disclosed manual liquid-based cytology (MLBC) method; and, we use the improved method to prepare slides from residual ThinPrep specimens in order to see how often ThinPrep diagnoses correspond to diagnoses derived from exhaustive examination of their parent sample suspensions. Residual cell suspensions from 500 ThinPrep cases comprising (1) 20 low-grade squamous intraepithelial lesions (LSILs); (2) 200 high risk (HR) negatives and 20 ASC-US; and (3) 260 screening cytology specimens were studied. Institutional review committee guidelines allowed us to know diagnoses by groups of specimens, but did not allow us to know individual patient diagnoses, so we could not perform case-by-case matched outcome-comparisons. Cells were concentrated by conventional centrifugation and sedimented into a polymer gel that was then vortex-mixed and converted into a viscous cell-rich suspension. The cell suspension was smeared between two clean glass slides, which were air-dried and stained with the Papanicolaou stain. Two study-sets were created, comprising one slide from each case. Each of the two study sets was examined by two cytopathologists, and discordant diagnoses were adjudicated. Because of the ambiguity involved in the "atypical" (ASC-US, ASC-H, AGC) diagnosis categories, only outcomes at the level of LSIL or greater were recorded. All MLBC SILs were digitally imaged and abnormal slides plus digital images were sent to the laboratory that provided the residual automated liquid-based cytology (ALBC) suspensions. The final diagnoses were confirmed by the laboratory that provided the residual ALBC specimens. MLBC slides of the 20 LSIL cases afforded 2 high-grade squamous intraepithelial lesions (HSILs) and 18 LSILs. Those of the 200 HR-Negatives showed 3 HSILs and 30 LSILs; and those of the 20 HR-ASC-US showed 3 HSILs and 9 LSILs. MLBC slides of the 260 screening cytology specimens showed 1 Carcinoma, 3 HSILs and 20 LSILs; affording 3 HSILs and 14 LSILs more than originally diagnosed. The MLBC method of this report is useful for preparing cell suspensions for cytological examination. Our analytical method was exhaustive and used nearly all of the cell material that was provided to us for analysis; therefore, we conclude that this approach is useful for determining how well ALBC instruments represent their parent sample suspensions. It appears that "rare events" may be overlooked when limited sample aliquots are analyzed by ALBC instruments, and this probably accounts for our increased discovery of SILs by the MLBC method. Also, SILs often present as aggregates of cohesive cells which, if overlooked or ineffectively transferred to ALBC slides, would not be diagnosed.

Papanicolaou Tests With Mixed High-Grade and Low-Grade Squamous Intraepithelial Lesion Features: Distinct Performance in the College of American Pathologists Interlaboratory Comparison Program in Cervicovaginal Cytopathology

Previous studies have shown that in gynecologic cytology, cases of low-grade squamous intraepithelial lesion (LSIL) and high-grade squamous intraepithelial lesion (HSIL) perform differently on interpretive review. The performance of cases with mixed LSIL and HSIL features is unknown. To compare the performance of gynecologic cytology cases of "pure" LSIL and HSIL with cases showing mixed LSIL and HSIL features. We compiled performance data from the College of American Pathologists Interlaboratory Comparison Program in Cervicovaginal Cytopathology from the years 2003 and 2004, and compared the performance of slides showing relatively pure LSIL and HSIL (< or = 10% misclassification as HSIL and LSIL, respectively) with slides showing mixed LSIL or HSIL features (cases misclassified as LSIL or HSIL > 10% of the time). Interpretations from a total of 4508 cases (2452 HSIL and 2056 LSIL) were analyzed. Overall, the sensitivity of participants on slides with a reference diagnosis of HSIL was 97.3%, and of LSIL was 95.9%. Performance trends for pure versus mixed cases varied by slide type and reference diagnosis. For conventional slides, participant sensitivity on pure HSIL cases was greatest (98.0%) and on pure LSIL cases was least (95.2%), while participant performance on cases with mixed features was intermediate (97.0% for mixed HSIL and 96.7% for mixed LSIL). In contrast, participant performance on ThinPrep slides showed the greatest sensitivity for mixed LSIL slides (97.9%), while performance on mixed HSIL slides showed the lowest sensitivity (95.7%); slides with pure features had intermediate sensitivity levels (96.3% for both HSIL and LSIL). Further evaluation demonstrated that conventional pure HSIL slides performed significantly better than mixed HSIL slides (P = .006), whereas mixed LSIL slides performed better than pure LSIL slides (P = .01). For ThinPrep slides, pure HSIL cases performed similarly to mixed HSIL cases (P = .43), while mixed LSIL cases performed better than pure LSIL cases (P = .04). Slides with mixed LSIL and HSIL features have measurably distinct performance characteristics in comparison to slides with pure LSIL or HSIL features. Participant performance on conventional mixed cases is distinctly different from performance on ThinPrep mixed cases.

Immunocytochemistry of p16INK4a in liquid-based cervicovaginal specimens with modified Papanicolaou counterstaining

Aim: To evaluate the feasibility and value of a modified Papanicolaou counterstain for p16INK4a immunostaining in liquid-based cervicovaginal samples. Methods: Immunocytochemical analyses were carried out with p16INK4a and modified Papanicolaou...

Quality assurance in cervical screening of a high risk population: A study of 65,753 reviewed cases in Parana screening program, Brazil

Epidemiological data from the Quality Assurance Unit (QAU) of the Cervical Cancer Screening Program of Paraná is discussed. Slides (65,753) reviewed by the QAU, from October 1997 to July 1999, were analyzed. Relations between agreement rate, diagnostic categories, and age groups were tested. The overall agreement rate was 97.04% (kappa +/- 95%, confidence interval = 0.888 +/- 0.003). Low-grade squamous intraepithelial lesion (LSIL) showed its peak prevalence in patients < or = 19 yr (8.3%). High-grade squamous intraepithelial lesion (HSIL) was more common in women between 40 and 49 yr (5.0%). Squamous-cell carcinoma (SCC) was almost exclusively seen in women > or = 40 yr. The lowest agreement rates were seen in adenocarcinoma (54.5%; kappa = 0.667) and LSIL cases (76.3%, kappa = 0.822), and in women < or = 19 yr (95.8%). It was concluded that constant monitoring to assess specific errors allows directed measures. The 10% rescreening model of quality assurance proved to be effective in a low resources setting with a positive overall financial impact.