Essential role of mitochondrial antiviral signaling, IFN regulatory factor (IRF)3, and IRF7 in Chlamydophila pneumoniae-mediated IFN-beta response and control of bacterial replication in human endothelial cells.

Chlamydophila pneumoniae infection of the vascular wall as well as activation of the transcription factor IFN regulatory factor (IRF)3 have been linked to development of chronic vascular lesions and atherosclerosis. The innate immune system detects invading pathogens by use of pattern recognition receptors, some of which are able to stimulate IRF3/7 activation and subsequent type I IFN production (e. g., IFN-beta). In this study, we show that infection of human endothelial cells with C. pneumoniae-induced production of IFN-beta, a cytokine that so far has been mainly associated with antiviral immunity. Moreover, C. pneumoniae infection led to IRF3 and IRF7 nuclear translocation in HUVECs and RNA interference experiments showed that IRF3 and IRF7 as well as the mitochondrial antiviral signaling (MAVS) were essential for IFN-beta induction. Finally, C. pneumoniae replication was enhanced in endothelial cells in which IRF3, IRF7, or MAVS expression was inhibited by small interfering RNA and attenuated by IFN-beta treatment. In conclusion, C. pneumoniae infection of endothelial cells activates an MAVS-, IRF3-, and IRF7-dependent signaling, which controls bacterial growth and might modulate development of vascular lesions.

Screening for anal dysplasia associated with human papillomavirus.

Anal dysplasia associated with human papillomavirus (HPV) infection occurs in substantial proportions of HIV-infected men and women and poses risk for anal carcinoma. Whether to routinely screen for HPV-associated anal dysplasia in this population, however, remains a debated question. Anal dysplasia is detectable by Pap screening and colposcopic biopsy; as Pap testing results have relatively low reproducibility, 2 baseline tests may be prudent. Screening should also ascertain risk factors for dysplasia, degree of immunosuppression, and history of prior anal disease. Although treatment options for anal dysplasia are limited by morbidity and high recurrence rates, early detection may permit better tolerance of therapy, and current estimates indicate that routine screening for the condition would be cost-effective. In addition, emerging immunologic therapies offer hope of more effective future treatment. This article summarizes a presentation given by Wm. Christopher Mathews, MD, MSPH, at the November 2002 International AIDS Society-USA course in San Diego.

Identification of Chlamydia trachomatis DNA in human papillomavirus (HPV) positive women with normal and abnormal cytology.

Insofar as infection with human papillomavirus (HPV) and Chlamydia trachomatis (CT) are associated with cervical intraepithelial neoplasia (CIN), the aim of this study was to assess the correlation of HPV and CT infection in patients with normal or abnormal cytology. Endocervical samples from patients (n=121; mean age 33.7+7.0) were assessed for HPV and CT DNA by PCR. While there was no statistically significant difference between HPV-positive (n=44) and HPV-negative (n=77) patients to age pregnancies, higher proportion of smokers, patients with multiple male sex partners, or with abnormal cytology was seen in HPV-positive vs. HPV-negative women, respectively. An infection rate of CT of 21/44 was seen in HPV-positive as compared to 11/77 in HPV-negative patients. Within HPV-positive patients, there was no significant difference between CT-positive and CT-negative patients with regards to the risk factors studied. Collectively, this suggests that a causal relationship between HPV and CT infection in the development of CIN disease.

Chlamydia trachomatis and human papillomavirus infection in Indian women with sexually transmitted diseases and cervical precancerous and cancerous lesions.

OBJECTIVES: Sexually transmitted diseases (STDs) and anogenital cancers are the major health problems in Indian women but no reliable estimate of the prevalence of either genital chlamydial infection or human papillomavirus (HPV) infection in STD patients is available. The aim of this study was to detect the frequency of Chlamydia trachomatis and the most prevalent high-risk HPV type 16 (HPV 16) infection in Indian women, with STDs and precancerous and cancerous lesions of the uterine cervix by polymerase chain reaction (PCR), and their comparison with those of conventional serology and antigen tests used for C. trachomatis detection. METHODS: Endocervical swabs or scrapes were collected from 50 women with STDs and 30 normal healthy women attending the STD clinics of Smt. Sucheta Kripalani Hospital, New Delhi. Scraped cervical cell specimens were also collected from 50 women with precancerous and cancerous lesions of the uterine cervix. Detection of C. trachomatis and HPV was carried out by PCR using chlamydia and HPV genome-specific oligonucleotide primers. The detection of chlamydial antigen and IgG-specific antibodies was carried out by enzyme immunoassay (EIA) and serological enzyme-linked immunosorbent assay (ELISA), respectively. RESULTS: A chlamydia plasmid-based PCR assay detected 50% (25 of 50) positivity of C. trachomatis in STD patients and HPV 16 DNA was found in 30% (15 of 50) of these cases which are significantly higher than those found in healthy controls. The PCR estimate of chlamydia was found to be higher than its reported frequency by tissue culture. The EIA could detect chlamydial antigen in only 13 cases (26%) while serological ELISA revealed evidence of chlamydia IgG-specific antibodies in 26 (52%) cases. Interestingly, in women with precancerous and cancerous lesions, the rate of HPV 16 infection was very high (52% and 72%, respectively), whereas the frequency of chlamydia infection was found to be 12-22% only. Occurrence of other sexually transmitted agents was also evaluated in the women. CONCLUSIONS: This is the first PCR estimate of genital chlamydial (50%) and HPV 16 (30%) infection in STD patients and women with precancerous and cancerous lesions of the uterine cervix in India. The PCR method seems to be a good alternative to tissue culture.

Human papilloma virus DNA detection in oral lesions in the Greek population.

HPVs (Human Papilloma Viruses) are small DNA "epitheliotropic" viruses, implicated in cervical carcinogenesis, particularly the "high-oncogenic-risk" types HPV-16 and HPV-18. Data concerning oral carcinogenesis are however, contradictory. We examined the presence of HPV and subsequently HPV-16 and HPV-18 in 102 specimens of paraffin-embedded oral tissue blocks--81 oral squamous cell carcinomas and 21 oral hyperplasias--using PCR technique followed by agarose gel electrophoresis. Our results demonstrated that 49% (50/102) of the samples were HPV positive. Subsequent analysis of HPV positive lesions revealed 22% positivity for HPV-16 and 44% for HPV-18. HPV-18 was detected only in carcinomas, while HPV-16 was more abundant in papillomatous hyperplasias and in a small percentage of carcinomas. These findings may probably indicate a contributing role for HPV-18 as a potent co-carcinogen in oral epithelial carcinogenesis in the Greek population.

Primary intestinal non-Hodgkin's lymphoma and Epstein-Barr virus: high frequency of EBV-infection in T-cell lymphomas of Mexican origin.

Epstein-Barr virus is universally associated with endemic Burkitt's lymphoma (BL) and undifferentiated nasopharyngeal carcinoma and can be detected in a significant proportion of cases of Hodgkin's disease (HD) and peripheral T-cell lymphoma, but only rarely in sporadic B-NHL. The frequency of EBV-positivity in certain neoplasms shows important geographic variations. Both HD and sporadic BL from Latin America have shown higher rates of EBV-association than cases from Western countries. In T-NHL, the frequency of EBV-positivity is influenced by the site of the primary tumor and the phenotype of the neoplastic cells. Nasal and nasal-type T-NHL, which show a T/NK-cell phenotype with expression of CD56 are virtually always EBV-associated, whereas only a proportion of nodal, gastrointestinal and pulmonary T-NHL are EBV-infected. A recent investigation of primary intestinal lymphomas of Mexican origin demonstrated EBV-positivity in all examined cases of T-NHL and BL and a proportion of other B-NHLs. The presence of EBV was independent of the presence or absence of enteropathy. Two of 6 cases studied showed CD56 expression. The high rate of EBV-positivity independent of histologic subtype is in contrast to the low to intermediate rates of EBV-positivity found in cases of intestinal T-NHL from Western countries and indicates that geographic differences in the frequency of EBV-association of lymphoid neoplasms might also extend to a fraction of peripheral T-cell lymphomas.

Human papillomavirus DNA after treatment of cervical dysplasia: low prevalence in normal cytologic smears.

BACKGROUND: The presence of human papillomavirus (HPV) DNA in relation to cervical cytology was evaluated after treatment of cervical dysplasia. METHODS: Forty patients, 22 with normal and 18 with abnormal cytology (mild or moderate dyskaryosis), with a history of cervical dysplasia were selected. Only patients with HPV DNA positive biopsies obtained before treatment were included. The presence of HPV was assessed in cervical smears at least 1 year after treatment of cervical dysplasia by using a polymerase chain reaction (PCR) with consensus primers (CPI/IIG). HPV typing was done by direct sequence analysis of the CPI/IIG PCR generated amplimers. RESULTS: Smears from 3 of the 22 patients with normal cytology after treatment were positive for HPV DNA (14%). HPV DNA positive smears were found in 13 of the 18 patients with abnormal cytology after treatment (72%) (relative risk: 5.3; 95% confidence interval: 1.78-15.75). In 11 of the 16 HPV DNA positive smears (69%), the HPV type was different from that before treatment. In 35 of 40 patients, the HPV type before treatment could not be detected after treatment (88%). CONCLUSIONS: A minority of the patients with normal cytology after treatment of cervical dysplasia had detectable HPV DNA. In contrast, a high prevalence of HPV DNA was found in cervical smears of patients with abnormal cytology after treatment of cervical dysplasia. After treatment, none of the patients with abnormal cytology but HPV DNA negative smears had recurrence of cervical intraepithelial neoplasia. This suggests the value of supplementary HPV DNA testing during follow-up of patients treated for cervical dysplasia.

[Precancerous and cancerous involvement of the uterine cervix. Results of a survey conducted by the "Genital Cancers" group of Ile-de-France, May 1990-May 1992, based on 8,805 biopsies]. / Pathologie précancéreuse et cancéreuse du col de l'utérus. Résultats d'une enquête conduite par le groupe "cancers génitaux" de PETRI, dans la région Ile-de-France, de mai 1990 à mai 1992, et portant sur 8,805 biopsies.

Results of a study conducted by the "cervix cancer group" of PETRI, in Ile-de-France, from May 1990 to May 1992, and based on 8,805 biopsy specimens. In the absence of Cancer Registry in the Ile de France, no reliable data on invasive and preinvasive neoplasia of the cervix are available concerning this area. The aim of this survey, performed between May 15th, 1990 and May 15th, 1992 in 62 laboratories of pathology under the aegis of the Petri Association (Prévention et épidémiologie des tumeurs en Ile-de-France), was to obtain a better knowledge of this pathology, in which one of the major risk factors is the infection of the cervical epithelium by specific types of human papillomavirus. Over the course of these two years, 8,805 biopsy specimens, taken from neoplastic lesions of the cervix, were analyzed. Intra epithelial neoplasia represented more than 90% of the registered lesions. The average age at the time of the diagnosis was 32.4 years for the cases of condyloma, 32.7 years for CIN I, 33.8 years for CIN II, 36.3 years for CIN III, 45.7 years for micro-infiltrative carcinoma and 50.8 years for infiltrative squamous cell carcinoma. The breakdown of the different histological types of lesions is presented for three characteristic age groups (20-25, 30-35, and 60-70 years old). Differences observed in the eight departments belonging to the Ile-de-France are discussed.

Common Epstein-Barr virus (EBV) type-1 variant strains in both malignant and benign EBV-associated disorders.

In the present study, Epstein-Barr virus (EBV) isolates from 18 malignant tumors (angioimmunoblastic lymphadenopathy [AILD], n = 4; Hodgkin's disease [HD], n = 3; pleomorphic T-cell non-Hodgkin's lymphoma [T-NHL], n = 1; B-cell non-Hodgkin's lymphoma [B-NHL], n = 8; gastric carcinoma, n = 2) as well as from 10 tonsils of EBV-seropositive children and from peripheral blood mononuclear cells of 12 children with uncomplicated infectious mononucleosis (IM) and of a boy with severe chronic active EBV infection were genotyped in the EBV nuclear antigen-2 (EBNA-2) gene. A total of 40 of 41 isolates harbored EBV type 1; in 1 specimen (tonsil), only EBV type 2 was found. Further molecular characterization of EBV type-1 wild-type isolates in the EBNA-2 gene and in the 40-kb distant EBV-encoded small RNAs (EBER) region showed that different groups of stable EBV type-1 variant strains exist in vivo both in benign and malignant lymphatic tissue. Group 1 is composed of EBV type-1 isolates (B-NHL, n = 3; T-NHL, n = 1; HD, n = 1; IM, n = 4) that showed a B95-8-like DNA sequence pattern in both viral genes. Group 2 isolates (HD, n = 1; AILD, n = B-NHL, n = 1; tonsils of EBV-seropositive children, n = 9; IM, n = 20 showed a nucleotide change at position 49095 in the EBNA-2 gene, leading to an amino acid substitution (Pro-->Ser), and EBV type-2 sequences in the EBER region. EBV type-1 isolates that fall into group 3 (AILD, n = 3; HD, n = 1; B-NHL, n = 4; gastric carcinoma, n = 2; IM, n = 6; severe chronic active EBV infection, n = 1) were characterized by typical nucleotide changes and a 3-bp insertion (CTC; extra Leu residue) in the EBNA-2 gene and an EBV type-2-specific sequence pattern in the EBER region. These EBV type-1 variant strains may represent the most prevalent circulating EBV type-1 strains in the exposed population and seem not to be restricted to a certain EBV-associated disease or tumor type. However, analysis of more EBV isolates from benign and malignant lesions must show whether more EBV type-1 substrains exist in vivo.

High expression of latent membrane protein 1 of Epstein-Barr virus and BCL-2 oncoprotein in acquired immunodeficiency syndrome-related primary brain lymphomas.

Nearly all primary brain lymphomas in acquired immunodeficiency syndrome (AIDS) patients are associated withEpstein-Barr virus (EBV). The role of EBV in lymphomagenesis is not totally elucidated. One possible mechanism is the overexpression of the BCL-2 oncoprotein, because the latent membrane protein 1 (LMP1) has been reported to transactivate the bcl-2 gene in vitro. To study the interrelationship beetween LMP1 and BCL-2 in vivo, we have analyzed and compared their expression in 11 AIDS-related primary brain lymphomas and 57 AIDS-related systemic lymphomas by immunoperoxidase technique on frozen sections. In AIDS-related primary brain lymphoma, LMP1 and BCL-2 were expressed in all cases but 1. All positive cases exhibited morphologic immunoblastic features. In contrast, the only negative case was histologically close to Burkitt's lymphoma. In systemic lymphomas, LMP1 was expressed in 21 cases, whereas BCL-2 was positive in only 3 cases, all of which were extranodal. These results indicate that, in addition to the histologic type, the role of EBV genes and BCL-2 expression in lymphomatous cells differ as a function of their localization. In AIDS-related primary brain lymphomas, this correlation between LMP1 and BCL-2 overexpression may have a major implication in lymphomagenesis.

[The correction of the vaginal microflora in patients with a papillomavirus infection associated with cervical intraepithelial neoplasia by using the new bacterial preparation Zhlemik]. / Korrektsiia vaginal'noi mikroflory u bol'nykh s papillomavirusnoi infektsiei v assotsiatsii s tservikal'noi intraépitelial'noi neoplaziei s pomoshch'iu novogo bakteriinogo preparata "Zhlemik".

The microbiological study of vaginal microflora in 40 patients of the reproductive age (20-30 years) with papilloma virus infection in association with cervical intraepithelial neoplasia of the uterine neck revealed dysbiotic disturbances in vaginal microflora, manifested by a decrease in the isolation rate and amount of lacto- and bifidobacteria and by excessive growth of opportunistic microorganisms. The 10-day course of corrective therapy with the new bacterial preparation "Zhlemik" was carried out. Group 1 (81 women) received the preparation intravaginally on a tampon, group 2 (19 patients) received the preparation in the form of vaginal suppositories. The results of this treatment were indicative of a high sanative effect of the preparation irrespective of the method of its application. This was demonstrated by the results of the bacteriological study made after bacterial correction: the amount of the Lactobacillus was restored, and they could be isolated from all patients; the level of contamination of the cervicovaginal niche with opportunistic microbial strains considerably decreased. The positive clinical effect after bacterial therapy with "Zhlemik" was observed in 93-95% of cases, depending on the form of its application.

[The genital tract microflora in patients with a papillomavirus infection]. / Mikroflora genital'nogo trakta u bol'nykh s papillomavirusnoi infektsiei.

In 70 patients of reproductive age (20-30 years) with the papilloma virus infection of the uterine neck the microflora of vaginal contents was studied. The study revealed the specific diversity of bacteria colonizing the vagina and the uterine neck. High occurrence of Chlamydia and Gardnerella was established. The detected dysbiotic disturbances in patients with condylomatosis of the uterine neck were manifested by a decrease in the isolation rate of lactobacteria and bifidobacteria and by an increase in the isolation rate of opportunistic bacteria. The most pronounced dysbiosis in the microflora of the vagina and the uterine neck was characteristic of patients with papilloma virus infection in association with cervical intraepithelial neoplasia of the III degree.

Determination of nonendemic nasopharyngeal carcinoma by in situ hybridization for Epstein-Barr virus EBER1 RNA: sensitivity and specificity in cervical node metastases.

After time-consuming and costly investigations, patients with neck metastases from an occult primary often receive unnecessarily large radiation volumes to treat a possible origin in the nasopharynx. In this study a colorimetric antisense Epstein-Barr early ribonucleoprotein 1 (EBER1) oligonucleotide probe specific for Epstein-Barr virus RNA was hybridized in situ to metastatic tissue obtained from 18 nasopharyngeal, 54 oral and pharyngeal, and 12 occult carcinomas derived from an unselected population. All 16 nonkeratinizing nasopharyngeal carcinomas (NPCs) were positive for EBER1. Both cases of keratinizing NPC and all 54 other metastases were negative. A single positive case of occult carcinoma indicated its origin from NPC. In retrospect, 7 patients with occult carcinoma had received unnecessary treatment with irradiation to the nasopharynx. Nasopharyngeal carcinoma appears to be a less common origin of occult carcinoma than previously considered. In the proper clinicopathologic context EBER1 in situ hybridization (EBER1-ISH) allows exclusion of NPC with a high degree of accuracy. Thus unnecessarily large radiation volumes and their adverse sequelae may be reduced in the treatment of occult carcinoma. Conversely, a positive result of ISH allows exclusion of further extensive diagnostic procedures.

Identification of potential hot spots in the carboxy-terminal part of the Epstein-Barr virus (EBV) BNLF-1 gene in both malignant and benign EBV-associated diseases: high frequency of a 30-bp deletion in Malaysian and Danish peripheral T-cell lymphomas.

In this study, we have sequenced the C-terminal part of the Epstein-Barr virus (EBV)-BNLF-1 gene encoding for the latent membrane protein-1 from tissues of EBV-positive Danish Hodgkin's disease (HD) and of Danish and Malaysian peripheral T-cell lymphomas (PTLs) and from tonsils of Danish infectious mononucleosis (IM). Our study showed that some of the 7 single-base mutations and the 30-bp deletion previously detected between codons of amino acid 322 and 366 in the BNLF-1 gene of the nasopharyngeal carcinoma cell line CAO were present in all Malaysian PTLs and in 60% of the Danish PTLs. In HD and the IM cases, the mutations were present in about 30%. The 30-bp deletion and the single base mutations occurred independently, and mutations were detectable in the majority of EBV type B-positive cases. These findings suggest that the 30-bp deletion and the 7 single-base mutations in the C-terminal part of the CAO-BNLF-1 gene do not characterize a new EBV type A substrain. Rather, some of the positions of single base mutations and the 30-bp deletion are hot spots that may have mutated independently through the evolution of EBV strains.

Abnormal expression of the p53-binding protein MDM2 in Hodgkin's disease.

The possible involvement of p53 tumor suppressor gene in the pathogenesis of Hodgkin's disease (HD) is suggested by the frequent finding of abnormal accumulation of p53 protein in the nuclei of Reed-Sternberg cells and their variants (H-RS) in a large proportion of cases. This finding, besides being consistent with the presence of p53 gene mutations, might represent a consequence of the inactivating interaction between p53 and p53-binding proteins such as the product of the MDM2 cellular oncogene. We have examined an unselected series of 77 HD cases of different histologic patterns for the expression of p53 and MDM2 proteins, using specific monoclonal antibodies and sensitive immunohistochemical techniques in single- and double-marker combination. In the large majority of cases (66/77), a variable proportion of H-RS cells expressed MDM2 that was strictly confined to the nuclei. Coexpression of both MDM2 and p53 was common in the same cells. The abnormal nuclear expression of p53 and MDM2 did not seem to correlate with the presence of Epstein-Barr virus infection, as shown by the results of LMP-1 antigen expression and EBER in situ hybridization analysis. Our data suggest that the abnormal accumulation of MDM2 and p53 proteins in HD might reflect a derangement of molecular mechanisms that could play a pathogenetic role in this disease.