Evidence for an association between Chlamydia trachomatis and cervical intraepithelial neoplasia

Summary. Attempts were made to isolate Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum from women with cervical intraepithelial neoplasia (CIN), from those with microinvasive or invasive cervical cancer, and from a control group of similar women with other gynaecological problems. C. trachomatis was found in a significantly greater proportion of women with CIN (8%) or cervical cancer (18%) than in the control group (1%), whereas M. hominis and U. urealyticum were approximately as common in each group. Histological examination of the cervix in cone biopsies or hysterectomy specimens from 26 women with CIN and from 39 women of similar age with no evidence of CIN showed lymphoid follicles, previously reported to be associated with chlamydial infection, in nine of the specimens with CIN, but in none of the specimens without CIN.     

Detection of HPV DNA in the uterine cervical lesions by polymerase chain reaction and in situ hybridization]

Human papillomavirus (HPV) is found in close association with carcinogenesis of the uterine cervix. We applied a new in vitro gene amplification technology, the polymerase chain reaction (PCR) to detect HPV 16 and 18 in cervical exfoliated cells. HPV infections were detected in 5 (16%) of 31 women with no pathological lesions of the uterine cervix (normal), 16 (24%) of 67 with cervical intraepithelial neoplasia (CIN) and 6 (38%) of 16 with invasive cervical cancer. Moreover, 10% formalin-fixed and paraffin-embedded tissue sections were prepared from the uterine cervix of these 27 women with PCR-proven HPV infection and were examined for the histological localization of HPV-DNA by in situ hybridization with biotin-labeled DNA probes of HPV types 6/11, 16/18 and 31/33/35. HPV-DNA type 16/18 was detected in 3 of 5 normal women, 2 of 4 CINs I, 2 of 3 CINs II, 6 of 9 CINs III and 6 of 6 invasive cervical cancers. HPV-DNA type 6/11 was detected in 6 of 6 condylomas. Viral DNA sequence was detected in the superficial cells of CIN I and II, and it was distributed through entire thickness layer of undifferentiated cells derived from CIN III and squamous cell carcinoma. In addition, the staining intensity became weak as the lesion progressed. These differences between lesions might be due to the difference in the viral form in the nuclei, ie whether an episomal or integrated form. Thus, an in situ hybridization technique with a biotin-labeled DNA probe as well as the PCR method is useful for the detection of HPV in clinical samples.     

Study on the prevalence of human papillomavirus in the anal canal of women with cervical intraepithelial neoplasia grade III

Objective

The objective was to evaluate the prevalence of human papillomavirus (HPV) in the anal canal of women with cervical intraepithelial neoplasia (CIN) grade III.

Study design

Two groups were compared. In group I (study group), 40 women who had undergone cervical biopsy with a histopathological result indicating CIN III were evaluated. Group II (control) consisted of 40 women with normal results from colposcopic examination and colpocytological tests. The women in group I who presented high-grade neoplasia in colpocytological tests underwent collection of material from the uterine cervix and anal canal for investigating HPV DNA using the Hybrid Capture II^® technique. Colposcopy and cervical biopsy were then performed. If CIN III was confirmed, HPV DNA was investigated in the material collected. In group II, colpocytological tests and colposcopy were performed and, if normal, the procedure was similar to that followed for group I, except that no biopsy was performed.

Results

In group I, 39 women (97.5%) were positive for HPV in the uterine cervix and 14 women (35%) in the anal canal. In group II, only four women (10%) had a positive HPV test, for both the uterine cervix and the anal canal.

Conclusions

The prevalence of HPV in the anal canal of the women with CIN III was greater than in the women without CIN III.     

Chlamydia trachomatis modulates expression of tumor suppressor gene caveolin-1 and oncogene C-myc in the transformation zone of non-neoplastic cervical tissue

OBJECTIVES: The obligate intracellular bacterium Chlamydia trachomatis is frequently found in association with benign proliferative, pre-neoplastic and malignant changes in cervical epithelium. The present study addresses the possible role of C. trachomatis infection of the uterine cervix in modulating human cancer gene expression. METHODS: RNA was extracted from both C. trachomatis infected and non-infected human fibroblast cultures treated with ITFgamma. The extracted RNA was used for cDNA microarrays carrying 33,000 human genes to detect abnormal gene expression induced by Chlamydia. Forty specimens of cervix dissected from the transformation zone had previously tested negative for HPV and positive for C. trachomatis by standard DNA PCR (20). These samples were subjected to RT-PCR to detect the expression of the abnormal genes induced by Chlamydia infection. RESULTS: The ITFgamma-induced, non-replicative Chlamydia-infected fibroblast cultures showed significant modulation of gene expression. The cultures showed a 2-fold decrease in the expression of the gene coding for the tumor suppressor caveolin-1, and increased expression of the oncogene C-myc, a promoter of cervical carcinogenesis. In tissues from the Chlamydia-infected cervical transformation zone, real-time RT-PCR demonstrated a highly significant average 4.7-fold reduction of caveolin-1 mRNA (P < or = 0.0001) and an average 2.1-fold increase in C-myc (P < 0.05). CONCLUSIONS: Human ITFgamma-treated fibroblasts as well as non-neoplastic cervical tissues responded to C. trachomatis with a strong down-regulation of caveolin-1 mRNA and a light up-regulation of C-myc mRNA. These changes were independent of the HPV high-risk types. This study reveals possible mechanisms by which C. trachomatis infection may contribute to neoplastic changes in the transformation of uterine cervix. These possible mechanisms require further evaluation.     

The relation between the type of immunoreactive cells found in human papillomavirus (HPV) lesions of the uterine cervix and the subsequent behaviour of these lesions

Human papillomavirus (HPV) lesions in the skin are rejected by immunological mechanisms, but very little is known about such mechanisms in HPV lesions of the uterine cervix. We studied the mononuclear cell infiltrates in cervical biopsies from 65 women with an HPV lesion (flat, inverted or papillomatous condyloma) who were followed for at least twelve months. We examined cryostat sections using the ANAE (acid alpha-naphthylacetate esterase) technique to demonstrate B cell, T cells, and mononuclear phagocytes (MPs), and monoclonal antibodies (OKT3, OKT4, OKT6, and OKT8) for the T cell subsets. The percentages of B-, T-, and MPs in HPV lesions with concomitant cervical intraepithelial neoplasia (CIN) were similar to those lesions without CIN. Similarly, no significant differences in cell type percentages were found in HPV lesions which regressed (n = 24), remained stationary (n = 24) or progressed (n = 3) during the follow-up period. The ratio OKT4+/OKT8+ (T helper/T suppressor, cytotoxic) cells was 1.44 and 0.81 in HPV lesions with CIN and those without CIN respectively (p less than 0.025). No statistically significant differences in that ratio were found between the lesions which remained stationary, regressed or progressed. The tendency to progress was directly related to the degree of CIN associated with HPV.     

Chlamydia trachomatis infection in cervical intraepithelial neoplasia and invasive carcinoma

We analyzed 149 women (81 with cervical intraepithelial neoplasia and with invasive carcinoma of the cervix and 68--as a control group). The influence of Chlamydia trachomatis (Cht) infection into expression of EGFR, TGF-alpha, Ki 67, HPV 16 and 18 was examined. IS-PCR was used to measure the level of antibodies in the serum. We detected that chlamydial infection may cause cervical hypertrophy in women with and without cervical intraepithelial neoplasia and invasive carcinoma. Infections of both Cht and HPV correlate with high expession of Ki 67 in epithelium. Cht infection also increased the expression of HPV16 in CIN I. These results suggest that Cht infection modifies the activity of viruses. In our research we have confimed that Cht infection increases the expression of EGFR and TGF-alpha. These facts may explain variants other than the HPV-mechanism of cervical carcinogenesis.     

Analysis of transforming gene regions of human papillomavirus type 16 in normal cervical smears

Summary Exfoliated cells from the uterine cervix of 102 Hungarian women with no cytological abnormality were screened using the polymerase chain reaction (PCR) for human papillomavirus (HPV) type 16 infection. Twenty-nine patients with histologically confirmed cervical intraepithelial neoplasia (CIN) served as reference cases. PCR was performed with 2 different HPV 16 specific oligonucleotide primer pairs flanking a 300 and a 200 base-pair fragment from the early 6 (E6) and early 7 (E7) genes, position 215–514 and 605–805. The specimens exhibited the same proportions of type 16 sequences specific for the tested regions. 8.8% (9/102) of normal samples showed amplification for HPV type 16 E6 and E7 regions, while 48.3% (14/29) of CIN biopsies were positive for the same gene sequences.     

In situ hybridization analysis of HPV DNA in cervical precancer and cervical cancers from China

Summary A series of 103 cervical biopsies derived from 103 women during July 1958 to September 1963 from Beijing, China were investigated with in situ hybridization for the presence of HPV6, 11, 16, 18, 31 and 33 DNA. The mean age of the patients was 46.1 + 10.6 years with a range of 24–74 years. Morphological features of HPV infection were found in 80 (77.7%) biopsies. Invasive cervical cancer was diagnosed in 43 biopsies and cervical intraepithelial neoplasia CIN I, CIN II and CIN III in 9, 9, and 27 cases, respectively. A total of 63.1% (65/103) of the lesions had morphological features of HPV infections associated with CIN or invasive carcinomas. Altogether, 31.1% (32/103) of the biopsies were shown to contain HPV DNA. Of the cases showing HPV morphology, 43.1% were HPV DNA positive. HPV16 (30/32) was the most frequent type, followed by HPV11 and 18, whereas no lesions with HPV6, 31 or 33 were found. A total of 19/43 (44.2%) of the invasive carcinomas contained HPV DNA. HPV DNA positivity and the grade of CIN showed a statistically significant correlation (P=0.0011). Our study demonstrated the presence of HPV in cervical lesions among Chinese women in the late 1950's and early 1960's when a single sexual partner was the rule and also supports the concept that HPV has as an important etiological role in cervical cancer, the highest risk being associated with HPV type 16. The applicability of in situ hybridization in retrospective assessment is emphasized.     

Prostitution and other cofactors in preinvasive and invasive lesions of the cervix

OBJECTIVE: To examine risk factors for preinvasive and invasive lesions of the cervix in Venezuelan female sex workers (FSW). METHODS: A total of 438 FSW were analysed. Each FSW had their clinical history recorded, a gynaecological examination, a Pap smear and a colposcopic examination of the cervix. A cervical biopsy was taken under colposcopic guidance when there was an abnormal epithelium. The statistical methods used were Student's t-test, chi(2) test and logistic regression. RESULTS: The sex workers' mean age was 32.1 +/- 7.9 years old (mean +/- standard deviation). The age of the first sexual intercourse activity for the FSW was 15.9 +/- 2.1 years. Sixty-four of the 84 women (76.2%) who had cervical biopsies had preinvasive and invasive lesions; 41 (9.4%) had cervical intra-epithelial neoplasia (CIN) 1, 12 (2.7%) had CIN 2, eight (1.8%) had CIN 3, two (0.5%) had microinvasive carcinoma, and one (0.2%) had invasive carcinoma. Forty of these 64 women (62.5%) with preinvasive or invasive lesions had a concurrent histological diagnosis of human papilloma virus (HPV) infection (P < 0.0001). The FSW with cervical pathology were younger (P < 0.05) and had their first sexual intercourse earlier (P < 0.02) than FSW with no pathology. Two hundred and seventy-six (63%) of the FSW were current smokers, and 47 of these women (17.6%) had cervical pathology (P < 0.03). CONCLUSIONS: First sexual experience before 20 years of age, cigarette smoking and HPV infection were high-risk factors for preinvasive and invasive lesions of the cervix in Venezuelan FSW.     

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.     

Cervical biopsy specimens and human papilloma virus positivity in patients with external genital warts

Cervical biopsy specimens were taken from 16 women with external genital warts in order to diagnose concomitant cervical human papillomavirus (HPV) infection and cervical intraepithelial neoplasia (CIN) according to pathology. Positive HPV infection was detected 56% (9/16) of the patients. Among those patients 66% (6/9) had concurrent subclinical low-grade CIN lesions. One of those patients had a high-grade CIN lesion. It is concluded that cervical HPV infection was rather common in women with external genital warts and it is valuable to examine cervical HPV infection by biopsy to detect concomitant CIN.     

Factors associated with progression of cervical human papillomavirus (HPV) infections into carcinoma in situ during a long-term prospective follow-up

Summary. In the course of a prospective study of 508 women with papillomavirus (HPV) lesions of the uterine cervix, 66 lesions that progressed into carcinoma in situ (CIS) were identified and treated by conization during a mean follow-up period of 35 months. The lesions were investigated with light microscopy and with in-situ DNA hybridization using ³⁵S-labelled probes for HPV 6,11,16,18, 31 and 33. After radical cone treatment, 11 of the 66 women (16-7%) have presented with a recurrent HPV infection. The recurrence rate increased with the duration of the follow-up period from <10% at the mean follow-up of 25 months to 16.7% at the most recent follow-up at 35 months. Most of these 66 HPV lesions (89%) presented with concomitant CIN in the first punch biopsy, but it is noteworthy that the other 11 % presented without concomitant CIN. HPV DNA of at least one of the six types examined was found in 73% of the first biopsies and it is noteworthy that the so-called 'low-risk' types, HPV 6 and 11, were found as frequently as the 'high-risk' types, HPV 16 and 18 (18% and 17%, respectively). This would suggest a similarity in the biological behaviour of these two HPV groups. Although the concept of the 'high-risk' and 'low-risk' HPV types may remain at least partially valid, it is imperative to realize that infection by HPV 6 and 11 by no means excludes the possibility for clinical progession into CIS and eventually to an invasive carcinoma.     

Herpes simplex virus type 2 and Chlamydia trachomatis in adenocarcinoma of the uterine cervix

INTRODUCTION: Adenocarcinoma of the uterine cervix (AC) occurs in 15-20% of primary cervical neoplasias. Although some etiologic factors for squamous cell carcinoma are well defined, and its relationship with sexually transmitted disease as human papillomavirus (HPV) is established, we still do not know about the causative factors of most of AC besides HPV infection. OBJECTIVES: To determine the presence of herpes simplex virus type 2 (HSV-2) and Chlamydia trachomatis (CT) DNA in AC specimens, and its correlation with HPV infection. METHODS: 206 paraffin-embedded cases of AC were selected to DNA extraction. The specimens and the DNA were isolated. Samples were first screened for beta-globin DNA sequences, and 67 cases were considered adequate to further analysis. In a previous analysis, DNA of HPV was identified in 79.4% of specimens included in this series (51% HPV 18 and 34% HPV 16).The local ethical committee approved the study. RESULTS: All samples were negative for HSV-2 DNA and CT DNA. CONCLUSIONS: In our series HSV-2 DNA and CT DNA were not found to be integrated to the genome of adenocarcinoma of the uterine cervix and do not seem to be a co-factor for HPV on the etiology of this histologic subtype.     

宫颈鳞癌和上皮内瘤变组织中TRF1、TRF2及HPV16、HPV18的检测及其意义

目的研究端粒结合蛋白TRF1、TRF2在宫颈鳞癌发生发展中的作用并分析HPV16、HPV18感染与TRF1、TRF2蛋白表达的关系。方法随机选择南华大学附属第一医院病理科2005年9月至2006年10月期间的组织石蜡块标本共86例,采用原位杂交方法检测HPV16、HPV18在15例正常宫颈上皮、36例宫颈上皮内瘤变(CIN)和35例宫颈鳞癌组织中的感染情况;采用免疫组化方法检测所有组织标本中TRF1、TRF2蛋白的表达。结果(1)HPV16、HPV18阳性感染率CIN组[63.9%(23/36)]和宫颈鳞癌组[97.1%(34/35)]显著高于正常组[20.0%(3/15)](χ2=30.639,P<0.01)。(2)TRF1阳性表达率宫颈鳞癌组[40.0%(14/35)]显著低于CIN组[63.9%(23/36)]和正常组[86.7%(13/15)](χ2=10.237,P<0.01);CINⅢ组[42.9%(6/14)]显著低于CINⅠ组[90.0(9/10)](χ2=5.531,P<0.01)。TRF2阳性表达率宫颈鳞癌组[80.0%(28/35)]显著高于CIN组[52.8%(19/36)]和正常组[...     

检出高级别子宫颈病变的阴道镜活检程序的优化

目的:探讨阴道镜下子宫颈活检对高级别子宫颈病变［即子宫颈上皮内瘤变Ⅱ级及以上病变（CIN Ⅱ +）］检出的价值,并通过基于不同风险条件组合的阴道镜检查实践,进一步优化阴道镜活检程序。 方法:前瞻性研究2017年4月至2019年12月在南京医科大学第一附属医院因子宫颈癌筛查结果异常转诊阴道镜检查的3...     

Natural history of cervical human papillomavirus (HPV) infections based on prospective follow-up

Summary. To assess the natural history of human papillomavirus (HPV) lesions in the uterine cervix, a prospective follow-up of untreated lesions has been conducted since late 1981. The present report summarizes the data on 343 women with cervical HPV lesions currently followed-up for a mean of 18.7 (SD 15.2) months by colposcopy and PAP smears (group R) or by additional punch biopsy (group A). Initially these tw7o groups were classified on the first PAP smears, presenting with HPV-induced cytopathic changes, and either with (group A) or without (group B) concomitant changes suggestive of cervical intraepithelial neoplasia (CIN). The clinical course of the HPV lesions could not be predicted adequately from the findings of the first PAP smears, as evidenced by the higher progression rate (154%) in the 214 women initially classified in group B, compared with 116% in the 129 women classified in group A. Furthermore, the number progressing to careinomia in situ requiring conization was equal (seven patients) in the two groups. This necessitated a more flexible approach to follow-up, permitting transfer of patients between groups, which resulted in a final allocation of 261 women to group A, and 82 to group B. To date, 25% of the total of 343 HPV lesions have regressed. 61% have persisted, and 14% have progressed. Of the latter, a total of 14 (4.1%) have been coned due to progression to carcinoma in situ . The rate of regression seems to be inversely related, and progression directly related. to the degree of HPV-associated CIN. The results are discussed in the light of the well-established natural history of classical CIN. Generally recognized as premalignant tesions. The present results, demonstrating that the natural history of cervical HPV lesions is very similar to that o f CIN, further substantiate the concept of HPV lesions as precursors of cervical cancer. With the potential to progress to carcinoma in situ when left untreated.     

Detection of Chlamydia trachomatis by several methods in the uterine cervix of pregnant women

Most pregnant women who have Chlamydia trachomatis (C. trachomatis) in the uterine cervix are asymptomatic. Several ways of detecting C.trachomatis were tested on 331 pregnant women, as well as 146 female patients attending our STD clinic as a control. 1) The detection rates for C.trachomatis in the cervix of pregnant women were 5.1% using the cell culture method, 2.4% with Micro Trak, and 2.2% employing Chlamydiazyme. These rates were higher in those patients visiting the STD clinic. 2) In pregnant women, the positive rate of Chlamydiazyme was 66.7% in the cell culture-positive cervical specimens, whereas Micro Trak was positive in 33.3%. 3) The antibody-positive rate was 84.6% in cases with PID caused by C.trachomatis. The antibody was found in only 17.7% of the pregnant women. Additionally, no significant correlation was noted between the antibody titer and C.trachomatis colonization in specimens obtained from the cervix of pregnant women. Although Micro Trak, Chlamydiazyme and possibly the microplate immunofluorescence antibody technique can be substituted for a cell culture method for detecting C.trachomatis in cases of symptomatic infection, these tests are not considered to be useful for screening Chlamydia-positive pregnant women.     

Human papillomavirus (HPV) infections involved in the neoplastic process of the uterine cervix as established by prospective follow-up of 513 women for two years

A total of 513 women with cervical HPV infections have been followed-up since 1981 (mean 25.6 months) to establish the biological potential of HPV in cervical carcinogenesis. On each attendance, the patients were subjected to colposcopy accompanied by Papanicolaou (PAP) smears and/or punch biopsies. The latter were analysed for HPV particles on TEM, for the cytopathic changes of HPV, as well as for HPV structural proteins. The local immunocompetent cell (ICC) infiltrates are enumerated using ANAE-technique to define B-, MPS- and T cells, and monoclonal antibodies for T cell subsets, NK and K cells and Langerhans cells. HPV DNA typing was accomplished by Southern blot and spot hybridization using the DNA probes for HPV 6, 11, 16 and 18. Antibody titres for HSV were measured, and Chlamydia trachomatis isolations completed in cervical swabs. No correlation with the clinical course, e.g. regression (RE), persistence (PE), progression (PR) or recurrence (RC) of the HPV lesions could be established for the following factors; expression of HPV antigens, presence of HPV particles on TEM, Chlamydia in cervical swabs, the levels of HSV antibodies, and the levels of the ICCs. The OKT4+/OKT8+ cell ratio, however, was inversely correlated with PR, being most markedly reduced in recurrent lesions. Of the 513 lesions, 24.8% regressed, 59.8% remained persistent, and 14.1% progressed, 11.9% having been coned due to progression into CIS. So far, 1.1% of lesions have recurred after such a treatment. The progression rate was highest (45.5%) in HPV 16 lesions, followed by that (27.3%) in HPV 18 lesions, as contrasted to 0% and 13.3% for HPV 6 and 11, respectively. The results clearly confirm that cervical HPV infections are capable of progressing into CIS and thus show a natural history equivalent to that of classical CIN. The inherent potential of HPV 16 and HPV 18 lesions for clinical progression was clearly established, supporting the concept on HPV 16 and HPV 18 as the high risk HPV types in cervical carcinogenesis.     

Prevalence of human papillomavirus type 16 DNA sequences in cervical intraepithelial neoplasia and invasive carcinoma of the cervix

Summary. The frequency of human papillomavirus (HPV) type 16 in premalignant and malignant lesions of the cervix was investigated and compared with the detection of HPV type 6. In cervical intraepithelial neoplasia (CIN) grades I-III HPV 6 was detected in 28% and HPV 16 in 62% of patients whereas 90% of malignant lesions contained HPV 16 only. In the CIN lesions there was an increase in HPV 16 detection as the severity of disease increased while the level of detection of HPV 6 decreased. Only three (18%) of the cervices that were colposcnpically and histologically normal contained HPV genomes: although two of these three women had either a history of genital warts or a sexual partner with penile warts.     

Prevalence of human papillomavirus type 16 DNA sequences in cervical intraepithelial neoplasia and invasive carcinoma of the cervix

The frequency of human papillomavirus (HPV) type 16 in premalignant and malignant lesions of the cervix was investigated and compared with the detection of HPV type 6. In cervical intraepithelial neoplasia (CIN) grades I-III HPV 6 was detected in 28% and HPV 16 in 62% of patients whereas 90% of malignant lesions contained HPV 16 only. In the CIN lesions there was an increase in HPV 16 detection as the severity of disease increased while the level of detection of HPV 6 decreased. Only three (18%) of the cervices that were colposcopically and histologically normal contained HPV genomes; although two of these three women had either a history of genital warts or a sexual partner with penile warts.