Low-grade Cervical Lesions Research Articles

Интраэпителиальные поражения шейки матки низкой степени риска у беременных женщин с впч и нарушениями биоценоза влагалища

Интраэпителиальные поражения шейки матки низкой степени риска у беременных женщин с впч и нарушениями биоценоза влагалища

Factores de riesgo asociados a infecciones vaginales y lesiones escamosas intraepiteliales en estudiantes universitarias de Medellín - Colombia

Objetivo: Explorar factores de riesgo asociados a infecciones vaginales y Lesiones Intraepiteliales Escamosas de Cuello Uterino (LEICU) en estudiantes universitarias de Medellín-Colombia.Materiales y métodos: Estudio observacional analítico transversal con una muestra a conveniencia de 176 estudiantes del área de la salud. Los datos se obtuvieron mediante encuesta anónima que incluyó variables demográficas, clínicas, académicas y de hábitos sexuales. Se realizó toma de muestra por medio de citología cérvico uterina y Gram-Directo de flujo vaginal. Se exploró la asociación estadística de la presencia de IV o LEICU con variables de interés a través de la Odds Ratio (OR) y su intervalo de confianza de 95% (IC95%). Se asumió asociación estadística con p<0,05.Resultados: Se encontró ASCUS en el 9,1%; LEICU de bajo grado en el 4,5%, e infecciones vaginales en el 30.7% de las participantes, siendo la vaginosis bacteriana la infección más común. Además los antecedentes previos de VPH tienen asociación estadística con el ASCUS OR=36,69 IC 95% (3,56-378,15) y con las vaginosis por Gardnerella OR=10,57 IC 95%(1,07-104,64), mientras que las infecciones urinarias tuvieron asociación estadística con la candidiasis OR=4,46 IC 95% (1,21-16,5).Enfermería Global Nº 50 Abril 2018 Página 87Conclusiones: Los hallazgos encontrados pueden servir como información descriptiva acerca de la frecuencia de IV y LEICU en poblaciones universitarias para continuar o mejorar programas de promoción y prevención de la salud sexual y reproductiva, en poblaciones jóvenes. Objective: To explore the risk factors associated with vaginal infections and squamous intraepithelial lesions of the cervix in university students in Medellín, Colombia.Materials and methods: Cross-sectional study; a convenience sample of 176 students from the health care field were included. Data were obtained through an anonymous survey that included demographic, clinical, and academic variables, as well as those pertaining to sexual habits. Cervical cytology and direct gram stain of vaginal fluid were taken. The statistical association for vaginal infections and squamous intraepithelial lesions of the cervix was explored through the odds ratio and the 95% confidence intervals (95% CI). A p value <0.05 was considered statistically significant. Results: Atypical squamous cells of undetermined significance (ASCUS) was found in 9.1% of participants; low-grade cervical squamous intraepithelial lesions in 4.5%, and vaginal infections in 30.7%; bacterial vaginosis was the most common infection. Previous history of HPV has a statistical association with ASCUS OR = 36.69 95% CI (3.56-378.15) and vaginosis by Gardnerella OR = 10.57 CI 95% (1.07-104.64), whereas urinary infections had a statistical association for candidiasis OR = 4.46 CI 95% (1.21-16.5).Conclusions: Our findings can be used as descriptive information regarding the frequency of vaginal infections and squamous intraepithelial lesions of the cervix in university populations, to continue or improve programs for the promotion and prevention of sexual and reproductive health in young populations.

Do Infection Patterns of Human Papillomavirus Affect the Cytologic Detection of High-Grade Cervical Lesions on Papanicolaou Tests?

- Persistent infection with high-risk human papillomavirus (hrHPV) is the major cause of cervical cancer. The effect of HPV infection patterns on cytologic detection of cervical lesions is unknown. - To determine the effect of HPV infection patterns on the sensitivity of cytologic detection of high-grade cervical lesions. - Papanicolaou tests from 257 women with biopsy-confirmed, high-grade squamous intraepithelial lesions were analyzed with respect to HPV infection patterns. - Among 257 biopsy-confirmed, high-grade squamous intraepithelial lesion cases, the preceding cytology showed 20 cases (8%) were benign; 166 cases (65%) were low-grade cervical lesions, including atypical squamous cell of undetermined significance and low-grade squamous intraepithelial lesions; and 71 cases (28%) were high-grade cervical lesions, including atypical squamous cells cannot rule out high-grade squamous intraepithelial lesion (atypical squamous cell-high), atypical glandular cells, and high-grade squamous intraepithelial lesions. In 236 cases tested for HPV, those exhibiting low-grade cervical lesions on cytology were often associated with coinfections of mixed hrHPV genotypes (31 of 40; 78%) or non-16/18 hrHPV (75/103; 73%), compared with single-genotype infections of HPV-16 (33 of 62; 53%) or HPV-18 (2 of 6; 33%) ( P = .001). In contrast, high-grade cervical lesion cytomorphology tended to associate with the single-genotype infection of HPV-16 (20 of 62; 32%) or HPV-18 (3 of 6; 50%), compared with non-16/18 hrHPV (25 of 103; 24%) or multigenotype infection (8 of 40; 20%) ( P = .01). - Our findings suggest that multigenotypic or non-16/18 hrHPV infections often produce deceptive lower-grade cytomorphology, which could result in underdiagnosis and delay of treatment. The HPV infection patterns may offer unrecognized benefit beyond HPV genotyping and should be considered during clinical risk evaluation of women with lower-grade cytology.

Abstract 2217: Detection of circulating immune biomarkers of cervical disease using proteome arrays

Abstract Introduction: More than 260,000 women die of cervical cancer every year. Screening methods have reduced the incidence of cervical cancer in high-income countries, but detection continues to lag in low and middle income countries (LMICs). IgG antibody (Ab) immunity to early (E) HPV antigens (Ags) are potential biomarkers of disease progression. Since HPV16 accounts for only ~50% of invasive cervical cancers, we developed protein microarrays expressing the proteomes of 12 HPV types to detect host IgG Abs to a broad spectrum of viral Ags to detect pre-invasive and invasive cervical disease. Methods: We developed custom HPV protein microarrays displaying the proteomes of two low-risk HPV types (HPV6 and 11) and ten oncogenic high-risk HPV types (HPV16, 18, 31, 33, 35, 39, 45, 51, 52 and 58). Arrays were probed with serum samples obtained from women with invasive cervical cancer (ICC; n=80), no or low-grade cervical disease (CIN 0/I; n=60), and high-grade cervical dysplasia (CIN II/III; n=60). To identify positive serologic responses, arrays were scanned and the signal intensity of each protein spot was quantified and normalized. Visual examination of diffused signal (ring) around each spot was performed and the Ab response to each protein was scored on a scale from 0 to 5. Results: To verify array quality and reproducibility, we confirmed high protein expression levels for 98% (96/98) of the antigens printed and high correlation (R ≥ 0.90) of protein expression signals between different randomly selected arrays. Epitope expression was confirmed using four commercial monoclonal Abs raised against HPV16 Ags. Host Abs to at least one early antigen (E1, E2, E4, E6, or E7) were detected in the sera of 31.2% and 43.3% of ICC and CINII/III patients, respectively, compared with 11.7% of women with CIN 0/I. 73.1% of CINII/III cases detected had Abs only to non-HPV16 Ags. Abs to E1, E2, E4, E6, and E7 Ags were detected in 3.8%, 7.7%, 54%, 19%, and 31% of CINII/III versus 16%, 16%, 32%, 32%, and 40% of ICC cases that were positive. The immunodominant Ags in women with CINII/III and ICC were E4 (54%) and E7 (40%), respectively. These results are consistent with the difference in tissue expression levels of these proteins in these two disease stages. This emphasizes the importance of broadening the scope of serological detection to include non-HPV16 proteomes. Conclusions: This study demonstrates that Abs against high risk HPV16 and non-HPV16 serotypes are potential biomarkers for both the diagnosis of cervical cancer and the differentiation between high grade and low-grade pre-invasive cervical lesions. These results suggest that serology is a potential minimally invasive tool for early detection of cervical cancer. High-throughput immunoprofiling of HPV-associated cancers and pre-invasive lesions may be informative for understanding the diversity of host immunity and viral progression. Citation Format: Radwa Ewaisha, Ian Meshay, Jack Resnik, Tirinder Bharaj, Karen S. Anderson. Detection of circulating immune biomarkers of cervical disease using proteome arrays [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2217. doi:10.1158/1538-7445.AM2017-2217

Prognostic value of human papillomavirus 16/18 genotyping in low-grade cervical lesions preceded by mildly abnormal cytology.

Histological low-grade squamous intraepithelial lesion/cervical intraepithelial neoplasia grade 1 (LSIL/CIN1) preceded by normal or mildly abnormal cytology is recommended for conservative follow-up, with no separated management. In this study, we assessed the triage value of human papillomavirus (HPV) 16/18 genotyping in 273 patients with LSIL/CIN1. HPV16/18 genotyping was performed at baseline and follow-up was at 6-monthly intervals for up to 2 years. At each follow-up, women positive for cytology or high-risk HPV (hrHPV) were referred for colposcopy. Enrollment cytology, HPV16/18 genotyping, and questionnaire-obtained factors were linked to the 2-year cumulative progression rate. Univariate and multivariate analyses were performed taking into account time-to-event with Cox proportional hazard regression. The results showed that 190 cases (69.6%) regressed, 37 (13.6%) persisted, and 46 (16.8%) progressed. HPV16/18 positivity (hazard ratio (HR), 2.708; 95% confidence interval (CI), 1.432-5.121; P=0.002) is significantly associated with higher 2-year cumulative progression rate. Sub-analysis by enrollment cytology and age restricted the positive association among patients preceded by mildly abnormal cytology and aged 30 years or older. Immediate treatment is a rational recommendation for the high-risk subgroup, when good compliance is not assured.

DLG1 polarity protein expression associates with the disease progress of low-grade cervical intraepithelial lesions

Human Discs large tumour suppressor (DLG1) participates in regulating cell polarity and proliferation, suggesting an important connection between epithelial organization and cellular growth control. However, it was demonstrated that DLG1 could acquire oncogenic attributes in some specific contexts. In this work, we evaluated the expression of DLG1 and its contribution to the progress of cervical lesions in order to investigate a potential role of this polarity protein in human oncogenic processes.We analyzed cervical biopsies from women with low-grade squamous intraepithelial lesion (LSIL) diagnosis (n=30), for DLG1 expression by immunohistochemistry. These results were correlated with the clinical monitoring of the patients during a 24-month follow-up period. Our data indicate that while all LSIL patients with a DLG1 staining pattern similar to normal tissues are significantly more likely to regress (n=23, Pattern I), all LSIL biopsy specimens showing a diffuse and intense DLG1 staining likely progress to high-grade lesions (n=4, Pattern II). Finally, all persistent LSIL analyzed showed an undetermined DLG1 staining, with a diffuse distribution without a strong intensity (n=3, Pattern III). We found a significant association between the expression pattern of DLG1 and the evolution of the lesion (p<0.00001).This work contributes to the knowledge of DLG1 biological functions, suggesting that its expression may have an important role in the progression of early dysplastic cervical lesions, giving prognostic information.

Outcome of Persistent Low-Grade Cervical Intraepithelial Neoplasia Treated With Loop Electrosurgical Excision Procedure

The aim of the study was to evaluate the outcome of persistent (≥2 years) low-grade cervical intraepithelial neoplasia (CIN 1) treated with loop electrosurgical excision procedure (LEEP). A study of 252 subjects with persistent biopsy-confirmed CIN 1 diagnosed after low-grade squamous intraepithelial lesions or atypical squamous lesions of undetermined significance on Papanicolaou test and treated with LEEP. Post-LEEP follow-up cytological, colposcopic, and molecular diagnostic examinations were scheduled at 6 months, 1 year, and yearly thereafter. The 252 subjects enrolled had a total number of 1,008 visits per colposcopies (median = 3, range = 1-7) during a median post-LEEP follow-up of 25 months (range = 12-121). The cumulative incidence of CIN 2+ at 2 years and at 3 years of follow-up was 2.3% (4/176) and 5.5% (7/128), respectively, or 1.7 cases (95% CI = 1-2.8) per 100 woman-years. Low-grade cervical lesions during post-LEEP follow-up were diagnosed in 70 subjects (27.8%) or 10 cases (95% CI = 7.9-12.6) per 100 woman-years. Overall, persistent and multiple high-risk HPV infections during follow-up were associated with increased rates of CIN persistence or progression. Women with persistent CIN 1 after atypical squamous lesions of undetermined significance/low-grade squamous intraepithelial lesion treated with LEEP had a low rate of progression to CIN 2+ but remained at a high risk of low-grade cervical abnormalities during follow-up. This information should be taken into account when deciding on the treatment strategy and counseling women with persistent CIN 1.

Immunization with Human Papillomavirus 16 L1+E2 Chimeric Capsomers Elicits Cellular Immune Response and Antitumor Activity in a Mouse Model.

Development of cervical cancer is associated with persistent infections by high-risk human papillomavirus (HPV). Although current HPV L1-based prophylactic vaccines prevent infection, they do not help to eliminate prevalent infections or lesions. Our aims were (i) to generate a vaccine combining prophylactic and therapeutic properties by producing chimeric capsomers after fusion of the L1 protein to different fragments of E2 from HPV 16, and (ii) to evaluate their capacity to generate an antitumoral cellular response, while conserving L1 neutralizing epitopes. Chimeric proteins were produced in Escherichia coli and purified by glutathione S-transferase (GST)-affinity chromatography. Their structure was characterized using size exclusion chromatography, sucrose gradient centrifugation, electron microscopy, and anti-L1 enzyme-linked immunosorbent assay. All chimeric proteins form capsomers and heterogeneous aggregates. One, containing part of the carboxy-terminal domain of E2 and its hinge region (L1Δ+E2H/NC, aa 206-307), conserved the neutralizing epitope H16.V5. We then evaluated the capacity of this chimeric protein to induce a cytotoxic T-cell response against HPV 16 E2. In (51)Cr release cytotoxicity assays, splenocytes from C57BL/6 immunized mice recognized and lysed TC-1/E2 cells, which express and present endogenously processed E2 peptides. Moreover, this E2-specific cytotoxic response inhibited the growth of tumors of TC-1/E2 cells in mice. Finally, we identified an epitope (aa 292-301) of E2 involved in this cytotoxic response. We conclude that the L1Δ+E2H/NC chimeric protein produced in bacteria can be an effective and economically interesting candidate for a combined prophylactic and therapeutic vaccine that could help eliminating HPV16-positive low-grade cervical lesions and persistent viral infections, thus preventing the development of lesions and, at the same time, the establishment of new infections.

Association between folate status and cervical intraepithelial neoplasia.

Background/Objectives:To investigate the effect of folate status on cervical intraepithelial neoplasia (CIN) progression and its relationship with high-risk human papillomavirus (hrHPV).Subjects/Methods:We evaluated 20 000 sexually active women aged <65 years in Yangqu County by using a questionnaire; the subjects were also screened using the ThinPrep cytologic test (TCT). Patients with abnormal TCT results (other than glandular cell abnormalities) who were willing to provide informed consent were further diagnosed using colposcopy and histopathological examination. We investigated 247 cases of low-grade cervical squamous intraepithelial lesions (LSIL), 125 cases of high-grade cervical squamous intraepithelial lesions (HSIL) and 877 controls. A 24-item food frequency questionnaire was filled out by the investigator to estimate the consumption of dietary folate. Positivity for hrHPV from residual exfoliated cervical cells was tested; serum folate was also measured.Results:The hrHPV infection rate in HSIL patients (77.6%) was higher than that in LSIL (33.2%) and control (32.0%) patients. Dietary folate intakes in controls, LSIL and HSIL were 306.9±176.6, 321.8±168.0 and 314.7±193.8 μg/kcal, respectively. The levels of serum folate in controls, LSIL and HSIL were 18.2±7.9, 15.9±7.1 and 14.3±7.5 nmol/l, respectively. Increased CIN correlated with higher rates of hrHPV infection and lower levels of serum folate.Conclusions:Low levels of serum folate may increase the risk of CIN progression. Furthermore, potential synergy may exist between low serum folate levels and hrHPV infection to promote CIN development.

Comparison of Analytical and Clinical Performance of HPV 9G DNA Chip, PANArray HPV Genotyping Chip, and Hybrid-Capture II Assay in Cervicovaginal Swabs

Background:Human papillomavirus (HPV) infection can be detected by using several molecular methods, including Hybrid-Capture II (HC2) assay and variable HPV DNA chip tests, although each method has different sensitivities and specificities.Methods:We performed HPV 9G DNA Chip (9G) and PANArray HPV Genotyping Chip (PANArray) tests on 118 cervicovaginal swabs and compared the results with HC2, cytology, histology, and direct sequencing results.ResultsThe overall and high-risk HPV (HR-HPV) positivity rates were 62.7% and 44.9% using 9G, and 61.0% and 30.5% using PANArray, respectively. The positivity rates for HR-HPV with these two chips were significantly lower than 55.1% when HC2 was used. The sensitivity of overall HPV positivity in detecting histologically confirmed low-grade cervical squamous intraepithelial lesions or higher was 88.7% for all three tests. The specificity was 58.5% for 9G and 61.5% for PANArray, which was significantly lower than the 72.3% for HC2. With the HR-HPV+ genotype threshold, the sensitivity decreased to 75.5% for 9G and 52.8% for PANArray, which was significantly lower than the 88.7% for HC2. Comparison of the two chips showed concordant results in 55.1% of the samples, compatible results in 16.9%, and discordant results in 28.0%, exhibiting poor agreement in detecting certain HPV genotypes. Compared with direct sequencing, 9G yielded no discordant results, whereas PANArray yielded 31 discordant results (26.7%).ConclusionsCompared with HC2, the HPV genotyping tests showed lower sensitivity in histologic correlation. When the two chips were compared, the 9G was more sensitive and accurate for detecting HR-HPV than the PANArray.

Frequency of twelve carcinogenic human papilloma virus types among women from the South Backa region, Vojvodina, Serbia.

The aim of this study was to determine the presence and age distribution of different oncogenic human papilloma virus (HPV) types in women in the South Backa region and its relationship to Pap results. In a group of 1087 women with normal and abnormal cytology, the commercial HR HPV Real-TM kit (Sacace Biotechnologies, Italy) was used. Overall, 50.5% of the women were HPV positive. The presence of HPV types 18, 31, 51, and 58 was significantly influenced by age, while the presence of HPV types 16 and 45 was significantly influenced by cervical cytology. Results of the LSD test show a wide spectrum of high risk HPV among women with normal cytology and women with a low grade cervical lesion rate (atypical squamous cell of undetermined significance (ASCUS) and low grade squamous intraepithelial lesions (LSIL). The most prevalent HPV types found were 16, 31, 51, 18, and 52. In the HSIL group the most prevalent HPV types were 16 and 45. The reported results provide new data on the circulation of oncogenic HPV genotypes and frequency of multiple infections among women in Vojvodina and suggest that a prophylactic vaccine against HPV 16 and 18 has the potential to prevent approximately half of the high-grade lesions.

Prevalence of Human Papillomavirus Genotypes Among Women With High-Grade Cervical Lesions in Beijing, China.

The aim of the study is to investigate the prevalence of high-risk human papillomavirus (hr-HPV) genotypes among Han women with high-grade cervical lesions in Beijing, China.Cervical cell specimens from patients with histopathologically confirmed cervical lesions at 7 hospitals in Beijing were examined with a validated HPV kit for 13 hr-HPV genotypes during the study period. The patients were divided into a low-grade cervical lesions group (cervical intraepithelial neoplasia grade 1, CIN1) and a high-grade cervical lesions group (CIN2+, including cervical intraepithelial neoplasia grade 2, CIN2; cervical intraepithelial neoplasia grade 3, CIN3; squamous cervical cancer, SCC; and adenocarcinoma of the cervix, ACC) based on the histopathology results.A total of 2817 eligible patients were enrolled, including 610 cases identified as CIN1 and 2207 as CIN2+. The hr-HPV positive rates in the CIN1 and CIN2+ groups were 78.2% (477/610) and 93.3% (2060/2207), respectively. The most frequently detected genotypes were HPV16, 58, 52 and18 in the CIN1 group and HPV16, 58, 33, and 52 in the CIN2+ group, in descending order of prevalence. In addition, the prevalence of HPV18 among the patients with ACC was 28.6% (14/49), significantly >7.2% (54/752) prevalence among the SCC patients (P < 0.001). Additionally, significantly more women in the CIN2+ group had multiple infections compared with those in the CIN1 group (38.1% and 24.9%, respectively; P < 0.001). However, as the cervical lesion grade increased, the prevalence of multiple hr-HPV infections gradually deceased to 44.2% in the CIN2 patients, 36.7% in the CIN3 patients, and 35.3% in the cervical cancer (CC) patients, which included SCC and ACC patients. In cases of multiple hr-HPV infections in the CIN2+ group, double infections accounted for ∼76.6%, and HPV16+58, HPV16+52, and HPV16+18 were the most common combinations, in descending order. The most frequent combination for triple infections was HPV16+58+31, with a rate of 4.2%. The highest positive rate occurred in the ≤24 year-old group for all types of cervical lesions.The prevalence of HPV genotypes in the targeted population with high-grade cervical lesions differs from that of other countries. This information could be helpful for the prevention of CC in Beijing, China.

Abstract 5580: Characterization of HPV integration sites in the human genome

Abstract Background. Persistent infection with carcinogenic human papillomaviruses (HPV) is known to be cause of the majority of anogenital cancers as well as some head and neck cancers. With the progression of the disease, the HPV genome is commonly integrated within the host DNA, although the mechanism by which integration may promote the progression of the disease is not well-understood. We performed a systematic review of integration events in HPV-related cancers and analyzed the genomic regions where these events occur. Methods. A PubMed search was performed to collect all published data on integration loci. A total of 29 articles were eligible for the analysis. Determination of genomic integration sites for papers that provided exact nucleotide sequences of integration sites or viral fusion regions was done by using the Basic Local Alignment Search Tool (BLAST, http://blast.ncbi.nlm.nih.gov/Blast.cgi) for the GRCh37/hg19 build. We used the Encyclopedia of DNA Elements (ENCODE) data to investigate if certain genetic features clustered with integrations sites. The Wilcoxon rank sum test was computed to compare the observed and expected distributions of minimal distances from the integration sites with known nucleotide positions to the different genetic features. Results. As of November 2014, approximately 1,200 integration events were reported in the literature from both tumors and cell lines. The majority of integrations sites (53%) were reported in cervical tumors, 9% in head and neck tumors, 4% of oropharyngeal tumors and 23% in cell lines,. Six percent and 2% of the integration sites were found in high-grade and low-grade cervical lesions, respectively. The majority of reported integration events (77%) were related to HPV16. We found that integration sites from tumors were closer to known genes (p &amp;lt;10−14) and CpG islands (p &amp;lt;10−9) than expected by chance. Among repetitive elements, long terminal repeats (LTR) were enriched in integration sites (P = 0.001). Open chromatin based on the FAIRE-seq peaks showed significant enrichment in integration sites (p &amp;lt;10−11). Conclusions. In summary, integration sites occur in genomic regions with enrichment for genetic elements, such as gene-rich regions, CpG sites, repetitive elements and open chromatin regions. This corroborates that accessible regions of the genome are preferentially targeted by integration and suggests that host genes may be functionally altered in development of HPV-related cancers. Citation Format: Clara Bodelon, Micahel Untereiner, Svetlana Vinokurova, Mitchell J. Machiela, Nicolas Wentzensen. Characterization of HPV integration sites in the human genome. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5580. doi:10.1158/1538-7445.AM2015-5580

Utility of HPV genotyping in the management of low-grade squamous intraepithelial cervical lesions (LSIL)

Utility of HPV genotyping in the management of low-grade squamous intraepithelial cervical lesions (LSIL)

Diagnostic performance of HPV E6/E7, hTERT, and Ki67 mRNA RT-qPCR assays on formalin-fixed paraffin-embedded cervical tissue specimens from women with cervical cancer

Human papillomavirus (HPV) is a major cause of cervical cancer, which is the third most common cancer in women. Human telomerase reverse transcriptase (hTERT) and Ki67 are tumor cell markers indicating cancer cell proliferation in cancer patients, and activation of hTERT and Ki67 leads to progressive cervical carcinogenesis. In the present study, we evaluated the CervicGen HPVE6/E7 mRNA RT-qDx assay, which detects 16 HPV high-risk (HR) genotypes (HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68 and 69), and the CervicGen hTERT and Ki67 mRNA RT-qDx assay using 117 formalin-fixed paraffin-embedded (FFPE) cervical cancer tissue samples. The diagnostic validity of the CervicGen HPV RT-qDx assay for detecting histologically proven prevalent squamous cell carcinoma (SCC) was 94% sensitivity, 100% specificity, 77.8% positive predictive value (PPV), and 78.9% negative predictive value (NPV). The most common HPV genotypes detected in FFPE cervical cancer tissue samples were HPV 16 (56%) and HPV 18 (10%).The positivity rate of hTERT and Ki67 mRNA expressions in FFPE cervical cancer tissue samples on RT-qPCR was 65% and 93% respectively. Moreover, the positivity rates were 92% for a combination of HPV E6/E7 and hTERT mRNA expressions, 97% for HPV E6/E7 and Ki67 mRNA expressions, and 99% (99/100) for the combination of HPV E6/E7, hTERT, and Ki67 mRNA expressions. These data showed that SSC FFPE cervical cancer tissue samples correlated more strongly with high Ki67 mRNA expressions than with hTERT mRNA expressions. Notably, hTERT and Ki67 mRNA expression level was increased in high-grade cervical lesions, but was very low in normal samples.Our findings suggest that the combination of HPV E6/E7, hTERT, and Ki67 mRNA expression levels could be used in a complementary manner in diagnosing high-grade cervical lesions. Further studies are required to evaluate these assays as a useful predictive tool for screening low-grade cervical lesions.

P16INK4a immunohistochemical and histopathologic study of Pap test cases interpreted as HSIL without CIN2-3 identification in subsequent cervical specimens.

Tissue biopsy following a pap test diagnosis of high grade squamous intraepithelial lesion (HSIL) sometimes fails to confirm the presence of a corresponding high grade cervical intraepithelial lesion (CIN 2-3), leading to confusion as to how best to manage the patient. It has been shown that these patients are still at higher risk for future detection of CIN 2-3 even if the initial biopsy fails to detect it. It has also been shown that immunohistochemical staining for p16INK4a can be reliably used as a surrogate marker for infection with high risk human papillomavirus in cervical samples, and that it can be used to enhance detection of CIN2-3 in cases where suspicion is high. To evaluate the use of p16INK4a staining in cases of HSIL which were not confirmed on initial biopsy, two pathologists rereviewed Pap and hematoxylin and eosin preparations from all such cases seen within the preceding 3 years. Immunohistochemical study for p16INK4a was performed and graded on representative sections. The results were tabulated and analyzed. Of the identified 596 HSIL Pap cases, 82% had HSIL on initial cervical specimens. Table 1 shows the 56 cases included in the study with graded and stratified p16INK4a results. On review of the p16INK4a slides, only 2 cases could be upgraded to HSIL/CIN2-3 from the original diagnosis. p16INK4a 2-3+ was expressed more frequently in cases initially interpreted on Pap as low-grade cervical lesion as compared with benign (24 of 35 cases). In the younger than 24-yr-old group p16 2-3+ reactivity was more frequent in benign and low-grade cervical lesion/CIN1 groups (benign: 3 of 5 cases, and CIN1: 6 of 8), and p16 negative reactivity was not seen. p16INK4a was graded 0-1+ more frequently in specimens interpreted as benign in the older than 25 yr olds (10 of 16 cases). The study suggests some diagnostic benefit from the use of p16INK4a immunohistochemical study on cervical specimens from women with a HSIL Pap test without HSIL/CIN2-3 on original hematoxylin and eosin review.

Use of hTERT and HPV E6/E7 mRNA RT-qPCR TaqMan assays in combination for diagnosing high-grade cervical lesions and malignant tumors.

Human papillomavirus (HPV) is a major cause of cervical cancer, which is the second most common cancer in women. HPV E6 initiates degradation of cellular tumor suppressor protein p53, induces human telomerase reverse transcriptase (hTERT) activity, and then leads to progressive cervical carcinogenesis. In this study, the CervicGen HPV RT-qDX assay (Optipharm, Osong, Republic of Korea), which detects 16 HPV high-risk subtypes (HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, and 69), and the CervicGen hTERT RT-qDX assay (Optipharm) were evaluated using 545 ThinPrep (Hologic, Bedford, MA) Papanicolaou samples. The positivity for the HPV E6/E7 messenger RNA (mRNA) assay was 94.4%, 95.2%, 82.4%, 46.5%, 25.0%, and 1.1% in squamous cell carcinomas, high-grade squamous intraepithelial lesions (HSILs), atypical squamous cells--cannot exclude HSIL, low-grade squamous intraepithelial lesions, atypical squamous cells of undetermined significance, and normal cytology samples, respectively. Five cervical intraepithelial neoplasia grade 2+ samples were not detected by the HPV E6/E7 mRNA assay, but they exhibited positive signals in the hTERT mRNA assay. Notably, the hTERT mRNA expression level was increased in high-grade cervical lesions but was very low in all 288 normal samples. These data suggest that the combination of HPV E6/E7 and hTERT mRNA expression levels could be used in a complementary manner in diagnosing high-grade cervical lesions and malignant tumors and might be useful as a predictive marker in monitoring low-grade cervical lesions.

Study of 2 years follow-up of referral patients with abnormal Pap smear.

Background:Abnormal Pap smear consists of premalignant or malignant cervical lesions. Many of premalignant cervical lesions will never progress to invasive malignancy, or even may regress over the time. Thus, there is always a risk of overtreatment of patients with an abnormal Pap smear. A long-term follow-up of these patients can reveal final events associated with each subtype of abnormal Pap smear, and, therefore, help us to prevent unnecessary interventions. The aim of our study was to present 2 years follow-up of referral patients with abnormal Pap smear.Materials and Methods:A total of 334 consecutive women aged more than 16 who were referred with an abnormal Pap smear were entered into the study. Patients were followed with biannual Pap smear and annual colposcopy and biopsy for 2 years.Results:At baseline, the majority of patients with abnormal Pap smear were normal on colposcopy and biopsy (68% and 86%, respectively). Six months after first abnormal Pap smear majority of patients in each group showed a significant regress to normal or less invasive lesion (P < 0.001). Twelve patients (4%) had no change in Pap smear, whereas 313 (94%) had at least one stage improvement. Only nine (3%) patients had deteriorated Pap smear after 6 months. All 308 patients who underwent colposcopy and biopsy had normal Pap smear 24 months after the first abnormal Pap smear.Conclusion:Pap smear is associated with a high rate of false-positive results. In addition, the majority of low-grade cervical lesions can spontaneously regress. A long-term follow-up of a patient with abnormal Pap smear can help us to avoid needless interventions.

CD28-, CD45RAnull/dim and natural killer-like CD8+ T cells are increased in peripheral blood of women with low-grade cervical lesions

BackgroundIn response to antigen naive CD8+, T cells differentiate into effector cells, which express Natural killer (NK) receptors, lose CD28 expression, and die by apoptosis. However, in smaller quantities, the cells are retained for subsequent exposure to the same antigen. Knowledge is limited regarding whether the percentages of CD28-, Effector memory (EMRAnull/dim), and the CD16+/CD56 + CD8+ T cells of women with low-grade cervical lesions are altered at a systemic level.MethodsWe enrolled in this study women controls and women with Human papilloma virus infection (HPV-I) without associated cellular neoplastic changes and with Cervical Intraepithelial Neoplastic-I (CIN-I). Flow cytometry (FC) was performed for measurement of CD28-, memory subset, and NK-like CD8 + T cells, and IL-17, IFN-gamma, Tumor necrosis factor (TNF)-alpha, Interleukin (IL)-10, IL-6, IL-4, and IL-2. Finally, we genotyped the HPV.ResultsThe CIN-I group increased the CD8 + CD28− and CD16+/56+ T cell percentage compared with that of HPV-I and controls (p <0.01), and CD8 + CCR7-CD45RAnull/dim (EMRAnull/dim) T cells were also increased in the CIN-I group compared with the controls (p <0.01). These two study groups were HPV- genotyped; 49% were HPV18+, and we did not observe differences in cytokine levels among all groups.ConclusionsIncreased levels of CD28-, EMRAnull/dim, and CD16+/CD56 + CD8+ T cells of peripheral blood in women with CIN-I may be associated with persistent HPV infection and could exert an influence on progression to cervical cancer.Electronic supplementary materialThe online version of this article (doi:10.1186/s12935-014-0097-5) contains supplementary material, which is available to authorized users.

Amplification of 3q26 and 5p15 regions in cervical intraepithelial neoplasia.

To analyze different amplification patterns of 3q26 and 5p15 regions in low-grade and high-grade cervical intraepithelial neoplasia. Experimental research. Department of Obstetrics and Gynecology at a medical faculty in Slovakia. A group of 83 patients referred for colposcopic examination. Amplification of 3q26 and 5p15 regions was analyzed on the 100 most atypical cells from a cervical cytology slide by fluorescent in situ hybridization using a multicolor hybridization probe. Chi-squared and Man-Whitney U-tests were used for statistical analysis. Liquid-based cytology samples and biopsy samples obtained during colposcopic examination correlated with high-risk human papillomavirus status and with amplification patterns of selected regions analyzed by fluorescent in situ hybridization. The number of cells with 3q26 and 5p15 gain rises with the severity of the lesion p < 0.01. The sensitivity of 3q26 amplification for CIN2+ lesions was 72.1% (95% confidence interval 56.3-84.7) and specificity was 90.0% (95% confidence interval 76.3-97.1). The sensitivity of 5p15 amplification for CIN2+ lesions was 69.8% (95% confidence interval 53.9-82.8) and specificity was 85.0% (95% confidence interval 70.2-94.3). Evaluation of telomerase components can help in differential diagnosis of low-grade and high-grade cervical lesions and in individualized management of these patients.