K. Heim

Spontaneous regression of CIN and delayed-type hypersensitivity to HPV-16 oncoprotein E7

We investigated delayed-type hypersensitivity to human papillomavirus (HPV) in women with cervical dysplasia or cancer. Women were challenged by skin tests with synthetic HPV-16 E7 oncoprotein peptides. 11 women were regressors (cleared disease without treatment) and 37 were progressors (required surgery). Antibodies to early antigens (markers for progression) were detectable in a higher proportion of cancer patients than all other patients, particularly progressors with cervical intraepithelial neoplasia (CIN). By contrast, cellular immunity to HPV-16 E7, measured by skin test, was significantly (p=0.0001) associated with clinical and cytological resolution of HPV-induced CIN, indicating that E7-specific T-helper cells have a role in control of HPV.

Clinical Relevance of Dominant-Negative p73 Isoforms for Responsiveness to Chemotherapy and Survival in Ovarian Cancer: Evidence for a Crucial p53-p73 Cross-talk In vivo

Purpose: We aimed to determine the clinical role of the p53 family members p53 and p73 in the responsiveness to platinum-based chemotherapy and survival in ovarian cancer, considering their cross-talk and the p53 polymorphism at codon 72. Experimental Design: A detailed analysis of p53 and p73 in a series of 122 ovarian cancers was done. We used a functional yeast-based assay to determine the p53 mutational status. Red yeast colonies, indicating mutant p53, were subsequently sequenced to determine the specific p53 alteration. p53 mutations were divided into two groups according to their previous characterization in the literature: those that efficiently inhibit transcriptionally active TAp73 function and those that do not. A p53 polymorphism at codon 72 was determined in corresponding normal tissue or blood of ovarian cancer patients. Isoform-specific p73 expression analysis using real-time reverse transcription-PCR has previously been done in the majority of ovarian cancers included in this study. In a retrospective chart review, responsiveness to chemotherapy was assessed, and survival data with long follow-up times were collected. Results: Eighty of 122 (65.6%) of ovarian cancers harbored p53 mutations. p53 mutational status was an important determinant of responsiveness to platinum-based chemotherapy in all patients with a residual tumor of <2 cm in diameter after initial surgery (wild-type versus mutant, P = 0.029). In addition, p53 mutational status was a strong prognosticator for recurrence-free and overall survival (P < 0.0001 and P = 0.003, respectively) in univariate analyses. High expression levels of dominant-negative p73 isoforms (ΔNp73 and ΔN′p73) significantly correlated with chemotherapeutic failure (P = 0.048) and with worse recurrence-free and overall survival in patients with p53 mutant cancers (P = 0.048 and P = 0.005, respectively). Eight p53 mutations, present in 19 cases, were found that efficiently inhibit TAp73 (i.e., 175H, 220C, 245S, 245D, 248W, 248Q, 266E, and 273H). Patients with p53 mutations that efficiently inhibit TAp73 function had a significantly shorter overall survival than patients with p53 mutations of unknown effect on TAp73 (P = 0.044). The p53 polymorphism at codon 72 had no influence on responsiveness to chemotherapy or survival. Conclusion: We provide the first clinical evidence that dominant-negative p73 isoforms contribute to drug resistance in vivo, underscoring the importance of a p53-p73 cross-talk. NH2-terminally truncated p73 isoforms were of significant clinical effect by providing an additional unfavorable factor for response to platinum-based chemotherapy and survival in p53 mutant ovarian cancers.

Specific serum IgG, IgM and IgA antibodies to human papillomavirus types 6, 11, 16, 18 and 31 virus-like particles in human immunodeficiency virus-seropositive women.

To evaluate the humoral immune response to human papillomavirus (HPV) in women infected with human immunodeficiency virus (HIV), serum samples of 83 HIV-positive individuals were analysed by ELISA for specific antibodies of the isotypes IgG, IgA and IgM recognizing HPV-6, -11, -16, -18 and -31 L1 virus-like particles (VLPs). Papillomavirus-related lesions were present in 30 of 83 HIV-positive women. Twenty-one women (25%) presented with high-/intermediate-grade anogenital squamous intraepithelial lesions. PCR analysis and sequencing for HPV typing was done from biopsy specimens of 18 women; PCR-positive results were obtained in 90% of cases. In addition, HPV DNA hybrid capture assays were performed from cervical swabs of 58 HIV-positive women, 53% of whom had a positive result for high-risk HPV. Overall, positive IgG reactivity to HPV-6/-11 and HPV-16/-18/-31 was seen in 19%/31% and 49%/30%/24% of HIV-positive women, respectively. HPV-seropositivity was even higher than in 48 HIV-negative cervical intraepithelial neoplasia/cancer patients with percentages as follows: 8%/2% and 31%/15%/15%. This difference was significant for HPV-16 (P=0.046). IgA responses were comparable to IgG. IgM responses were low. The extraordinarily high rate of antibodies to the capsid protein L1 of high-risk HPVs (HPV-16, -18 and/or -31) in 58% of HIV-positive women compared to 19% (P=0.00001) of 102 healthy HIV-negative control women suggests a high lifetime cumulative exposure to HPV and increased expression of capsid proteins due to cellular immunodeficiency in HIV-infected women.

Childbirth as a Biological Model for Stress

Objective: The aims of this investigation were to measure corticotropin-releasing hormone (CRH), corticotropin (ACTH) and cortisol before, during and after delivery searching for an endocrine intercorrelation of the hypothalamic-pituitary-adrenal (HPA) axis and to correlate these findings with obstetrical variables. Methods: Blood was sampled from 50 women with singleton pregnancies at term without uterine contractions, during delivery (after full cervical dilatation) and on the 4th postnatal day. Hormones were measured by radioimmunoassay (RIA). The correlation between obstetric variables, sociodemographic and endocrine data were evaluated using the Spearman rank coefficient. Group comparisons for continuous variables were calculated using the Mann-Whitney U test and Kruskal-Wallis test. Results: Maternal plasma ACTH and cortisol increased significantly during labor, declining toward the 4th postnatal day (p < 0.001) and showing a significant intercorrelation (p < 0.01). Compared to women without uterine contractions CRH rose during labor (p < 0.05) and decreased rapidly to the 4th postnatal day (p < 0.001). No correlations between CRH and ACTH or cortisol were observed. None of the obstetrical variables (parity, newborn’s weight, duration of delivery) revealed any significant correlation with ACTH. Analgetic medication (pethidine hydrochloride) was not able to influence the endocrine response to labor stress. Conclusions: Stressful experience during childbirth has an impact on endocrine response. However, this is not fully evident along the HPA axis in a simple biological model with monocausal dependencies. This ‘biological stress model’ is not sensitive enough to detect different childbirth conditions and the hormones in the maternal compartment have partially fetal (placental) origin.

Anale HPV-Infektionen

Anogenitale Infektionen mit humanen Papillomaviren (HPV) verzeichnen in den letzten Jahren gerade in jüngeren Generationen durch frühe sexuelle Aktivität eine steigende Inzidenz. Neue Erkenntnisse der ätiologischen, epidemiologischen und pathophysiologischen Zusammenhänge erfordern eine Überarbeitung der proktologischen Position HPV-assoziierter Erkrankungen der Analregion. Dies ist auch aufgrund der ansteigenden Inzidenz präkanzeröser und invasiver Neubildungen nötig. In Ermangelung großer, kontrollierter Studien stellt dieser Konsensusbericht über weite Strecken die Expertenmeinung der auf dem Gebiet der HPVInfektion erfahrenen Autoren aus den Gebieten Proktologie, Dermatologie, Gynäkologie, Radiologie, Strahlentherapie und Pathologie dar. Anale HPV-Infektionen1

An lnterleukin-6 Gene Promoter Polymorphism and Unexplained Late Intrauterine Fetal Death: A Multicenter Study

Objective: Interleukin-6 (IL-6)-mediated inflammatory processes have been proposed to be involved in the pathogenesis of pregnancy-associated complications such as late unexplained intrauterine fetal death (IUFD). Therefore we determined whether a common guanine/cytosine polymorphism at position-174 of the promoter of the IL-6 gene (1L6) known to affect in vivo protein activity can serve as candidate genefor this condition. Methods: In a multicenter case-control study, we evaluated the IL6 promoter polymorphism by pyrosequencing in 92 women with IUFD. Ninety-four healthy women with at least one uncomplicated full-term pregnancy and no history of IUFD served as the control group. Results: No significant association was found between the presence of at least one mutant allele of the IL6 promoter polymorphism (P = .2; odds ratio = 1.5 [95% confidence interval, 0.8-2.7]) and the incidence of IUFD. In women with IUFD, the presence of at least one mutant allele of the IL6 promoter polymorphism did not influence timing offetal death (33.9 [5. 1]gestational weeks vs 34. 1 [4.9]gestational weeks, P = .8) or birth weight (2055 [1119] g vs 1963 [992] g, P = .7). Conclusion: To our knowledge, we are thefirst to report on a common polymorphism of the IL6 promoter gene in women with late IUFD. The investigated IL6 promoter polymorphism can not be seen as candidate gene for IUFD in Caucasian women.

Spezifische serologische Untersuchungen mit einem neuartigen authentischen HPV-Antigen (Virus-like Particles) auf HPV-6 Antikörper bei gynäkologischen Patientenkollektiven

OBJECTIVE A serological assay for genital HPV infection would provide important additional information to HPV DNA diagnostic methods, since it would evaluate prior exposure to the viruses, detect significant systemic immunologic response to virus infection, and could be performed in most clinical laboratories. METHODS Serum samples from three groups of patients attending a gynecology clinic were analysed by direct ELISA for specific IgG antibodies to baculovirus-expressed HPV-6 and BPV-1-L1-VLPs. RESULTS Positive IgG reactivity to HPV-6-L1-VLPs were 4/72 (6%) in a control group, 28/73 (38%) in a condyloma group and 17/62 (17%) in cervical intraepithelial neoplasia patients. Individual IgG ELISA values of condyloma and CIN patients for HPV-6-L1-VLPs demonstrated no correlation to results with BPV-1-L1-VLPs. CONCLUSIONS These data show that HPV-6-L1-VLPs are effective antigens for serological studies and can detect species specific antibodies with important implications for diagnosis, epidemiology, insights to natural course of disease, prognosis and evaluation of vaccination.

Immunologische Aspekte bei Infektionen mit Papillomviren Aussichten auf einen Impfstoff

ZusammenfassungHumane Papillomviren (HPV) sind Ursache häufiger epithelialer Proliferationen und an der Entstehung von anogenitalen Karzinomen wesentlich beteiligt. Bei der Entwicklung eines Impfstoffes gegen krebsassoziierte HPV wird infolge zweier grundsätzlicher Ziele sowohl in Richtung der Prophylaxe als auch einer therapeutischen Anwendung gearbeitet. Eine Vakzine zur Verhinderung einer Infektion mit HPV scheint in naher Zukunft herstellbar. Antikörper gegen dreidimensionale Kapsidstrukturen schützen im Tiermodell vor dem Eindringen der Papillomviren in die Wirtszelle, und diese wichtigen neutralisierenden Antikörper können durch DNA-freie virusähnliche Kapsidpartikel aus spontan rearrangierten Strukturproteinen induziert werden. Solche Partikel gelten deshalb als sicherer, prophylaktisch wirksamer Impfstoff. Eine ideale HPV-Vakzine sollte zusätzlich die durch zelluläre Immunmechanismen vermittelte Rückbildung von bereits bestehenden präkanzerösen und auch von malignen Läsionen vermitteln. Die transformierenden Frühproteine der Papillomviren, die in Basalzellen der Epidermis als ein hypothetisches Antigen spezifischer T-Zellen exprimiert sind, könnten in Form von synthetischen Peptiden für einen solchen Impfstoff eingesetzt werden.SummaryHuman papillomaviruses induce benign or malignant epithelial proliferations in both skin or mucosa and are involved in the development of anogenital cancer. One can consider two approaches towards vaccine development: prevention of infection by prophylactic induction of virus neutralizing antibodies or therapeutic vaccination which would lead to regression of already existing lesions. Neutralizing antibodies are directed against 3-dimensional structures on intact virions. Such antibodies which are protective in model systems can be induced safely by DNA-free “virus-like-particles”. These constructs are candidates for a prophylactic vaccine. A further approach to optimize the vaccination strategy concentrates on immunotherapy of precancerous or malignant lesions by induction of a specific cell-mediated immune response. Hypothetically the transforming papillomavirus proteins which are expressed in basal layers of the epidermis could be used as a therapeutic vaccine in the form of synthetic peptides.

Determination of light-chain myosin in pregnancy and under tocolysis

Cardiac toxicity of tocolysis containing hexoprenaline (0.3 microgram/min) and the beta 1-blocking agent metoprolol (0.01 mg/min) was investigated using a recently developed monoclonal antibody arised against light-chain myosin in 15 patients and compared with the results in 51 control subjects. There was an increase in the median and 80th percentile values 2, 6, 12 and 24 hours after the beginning of tocolysis but no statistically significant difference between these values and the normal ones could be shown.