Multiplex-Serologie - Simultane Analyse von Antikörper-Antworten gegen humane Papillomviren (HPV) mit Protein-Arrays

Mehr als 100 humane Papillomvirus (HPV)-Typen können den Menschen infizieren und teilweise maligne Erkrankungen auslösen. HPV-Seroepidemiologie erlaubt es, die Infektion und Krankheits-Assoziation von HPV zu untersuchen und Krebs-spezifische Antikörpermuster zu detektieren, wird jedoch durch die Vielfalt der Virustypen und -proteine und die Typspezifität der gegen sie gerichteten Antikörper erschwert. Der bisher zur Detektion von Antikörpern genutzte GST capture ELISA ist methodisch ausgereift und gut validiert, im Hinblick auf die Zahl parallel analysierbarer Antigene jedoch beschränkt. Ziel dieser Arbeit war daher die Entwicklung eines Antikörperscreeningsystems, das eine Hochdurchsatzanalyse von Seren auf Antikörper gegen viele in situ-affinitätsgereinigte Antigene in einem Reaktionsansatz erlaubt. Zwei solche Systeme wurden entwickelt: Ein auf Protein Microarray-Technologie basiertes System (Chip-ISA), für das sich die in situ-Affinitätsreinigung der Antigene nicht realisieren ließ, und ein System namens Multiplex-Serologie, das auf einem Suspensions-Array von farbkodierten Polystyrolbeads beruht, die sich in einem Duchflusszytometer unterscheiden lassen. Nach Etablierung einer Methodik zur Derivatisierung dieser Beads mit Glutathion konnten die als GST-Fusionsproteine exprimierten Antigene direkt aus Bakterienlysat aufgereinigt werden. Multiplex-Serologie wurde vollständig validiert und ist gut reproduzierbar, sehr sensitiv und spezifisch. Die Methode wurde zur Analyse großer Serumzahlen adaptiert und erlaubt jetzt die Untersuchung von bis zu 1000 Seren auf Antikörper gegen bis zu 100 simultan getestete Antigene pro Tag. Über die technische Entwicklung hinaus wurden mit Multiplex-Serologie verschiedene seroepidemiologische Studien analysiert und die Ergebnisse von dreien im Detail vorgestellt. Erstmals wurde die Seroprävalenz gegen die L1-Proteine von 13 HPV-Typen in der deutschen Normalbevölkerung untersucht und auf Alters- und geschlechtsabhängige Dynamik analysiert. In einer zweiten Studie wurde die Assoziation von Antikörpern gegen die L1-Proteine einiger sogenannter EV-HPV-Typen mit nicht melanotischem Hautkrebs untermauert. Ebenfalls zum ersten mal wurde gezeigt, dass typspezifische Antikörper gegen die E6- und E7-Proteine seltener sogenannter Hoch-Risiko-HPV-Typen existieren und mit Zervix-Karzinomen assoziiert sind. Die Entwicklung von Multiplex-Serologie erlaubt erstmals die simultane Analyse von komplexen Antikörperantworten gegen bis zu 100 Proteine und stellt möglicherweise einen Quantensprung für (HPV-)Seroepidemiologie dar

Elimination of HPV-associated oropharyngeal cancers in Nordic countries

Incidence of human papillomavirus (HPV, most notably HPV type 16) associated oropharyngeal squamous cell carcinoma (OPSCC) among middle-aged (50?69 year-old) males has tripled in four high income Nordic countries (Denmark, Finland, Norway and Sweden) over the last 30 years. In Finland and Sweden, this increase was preceded by an HPV16 epidemic in fertile-aged populations in the 1980?s. The recent implementation of school based prophylactic HPV vaccination in early adolescent boys and girls will gradually decrease the incidence, and eventually eliminate the HPV-associated OPSCCs (especially tonsillar and base of tongue carcinomas) in the Nordic countries. However, beyond the adolescent and young adult birth cohorts vaccinated, there are approximately 50 birth cohorts (born in 1995 or before) that would benefit from screening for HPV-associated OPSCC. This article reviews the need, prerequisites, proof-of-concept trial and prospects of preventing HPVassociated OPSCC in the Nordic countries.Peer reviewe

Prospective Study of Human Papillomavirus Seropositivity and Risk of Nonmelanoma Skin Cancer

Cutaneous human papillomaviruses (HPVs) have been associated with squamous cell carcinoma (SCC) in case-control studies, but there are limited data from prospective studies assessing whether virus exposure predicts risk of future cancer development. Two major biobanks, the Southern Sweden Microbiology Biobank (1971-2003) and the Janus Biobank (1973-2003) in Norway, containing samples from 850,000 donors, were searched for incident skin cancer for up to 30 years using registry linkages. Altogether, 2,623 donors with samples taken before diagnosis of SCC or basal cell carcinoma (BCC) of the skin were identified. Prediagnostic samples and samples from 2,623 matched controls were tested for antibodies against 33 types of HPV. Baseline seropositivity to HPV types in genus beta species 2 was associated with SCC risk (odds ratio = 1.3, 95% confidence interval: 1.1, 1.7); this was also the case for samples taken more than 18 years before diagnosis (odds ratio = 1.8, 95% confidence interval: 1.1, 2.8). Type-specific persistent seropositivity entailed elevated point estimates for SCC risk for 29 HPV types and decreased point estimates for only 3 types. After multiple hypothesis adjustment, HPV 76 was significantly associated with SCC risk and HPV 9 with BCC risk. In summary, seropositivity for certain HPV types was associated with an increased risk for future development of SCC and BCC

Squamous cell carcinoma of the nasal cavity:A descriptive analysis of cases from the Head and Neck 5000 study

OBJECTIVES: This paper aims to provide contemporary epidemiological data on squamous cell carcinoma (SCC) of the nasal cavity, which represents a rare type of head and neck cancer.DESIGN, SETTING &amp; PARTICIPANTS: A descriptive analysis of people with nasal cavity SCC treated with curative intent from the Head and Neck 5000 study; a multicentre clinical cohort study of people from the UK with head and neck cancer. People with tumours of the nasopharynx, paranasal sinuses and other sub-sites of the head and neck were excluded.MAIN OUTCOME MEASURES: Demographic data and treatment details are presented for all participants. The main outcomes were overall survival and survival according to categories of characteristics (e.g. smoker vs non-smoker); these were explored using Kaplan-Meier plots.RESULTS: Thirty people with nasal cavity SCC were included in the study, of which most were male (67%) and current or ex-smokers (70%). The majority (70%) presented with early stage (T1/2, N0) tumours. Cervical lymph node metastases at presentation were rare, occurring in only one person. Nine people died during the follow up period (30%). Worse survival outcomes were seen in people with moderate or severe co-morbidities.CONCLUSIONS: This paper provides epidemiological data on nasal cavity SCC in the UK. Patterns of disease and survival outcomes are described, identifying high-risk groups. Further studies should explore whether primary treatment modality alters survival. This article is protected by copyright. All rights reserved.</p

Immunostimulatory Membrane Proteins Potentiate \u3cem\u3eH. pylori-\u3c/em\u3eInduced Carcinogenesis by Enabling CagA Translocation

Infection with Helicobacter pylori is the single greatest risk factor for developing gastric adenocarcinoma. In prospective, population-based studies, seropositivity to the uncharacterized H. pylori proteins Hp0305 and Hp1564 was significantly associated with cancer risk in East Asia. However, the mechanism underlying this observation has not been elucidated. Here, we show that Hp0305 and Hp1564 act in concert with previously ascribed H. pylori virulence mechanisms to orchestrate cellular alterations that promote gastric carcinogenesis. In samples from 546 patients exhibiting premalignant gastric lesions, seropositivity to Hp0305 and Hp1564 was significantly associated with increased gastric atrophy across all stomach conditions. In vitro, depletion of Hp0305 and Hp1564 significantly reduced levels of gastric cell-associated bacteria and markedly impaired the ability of H. pylori to stimulate pro-inflammatory cytokine production. Remarkably, our studies revealed that Hp1564 is required for translocation of the oncoprotein CagA into gastric epithelial cells. Our data provide experimental insight into the molecular mechanisms governing novel H. pylori pathogenicity factors that are strongly associated with gastric disease and highlight the potential of Hp0305 and Hp1564 as robust molecular tools that can improve identification of individuals that are highly susceptible to gastric cancer. We demonstrate that Hp0305 and Hp1564 augment H. pylori-mediated inflammation and gastric cancer risk by promoting key bacteria-gastric cell interactions that facilitate delivery of oncogenic microbial cargo to target cells. Thus, therapeutically targeting microbial interactions driven by Hp0305/Hp1564 may enable focused H. pylori eradication strategies to prevent development of gastric malignancies in high-risk populations

Amino acid variation in HLA class II proteins is a major determinant of humoral response to common viruses

The magnitude of the human antibody response to viral antigens is highly variable. To explore the human genetic contribution to this variability, we performed genome-wide association studies of the immunoglobulin G response to 14 pathogenic viruses in 2,363 immunocompetent adults. Significant associations were observed in the major histocompatibility complex region on chromosome 6 for influenza A virus, Epstein-Barr virus, JC polyomavirus, and Merkel cell polyomavirus. Using local imputation and fine mapping, we identified specific amino acid residues in human leucocyte antigen (HLA) class II proteins as the most probable causal variants underlying these association signals. Common HLA-DRβ1 haplotypes showed virus-specific patterns of humoral-response regulation. We observed an overlap between variants affecting the humoral response to influenza A and EBV and variants previously associated with autoimmune diseases related to these viruses. The results of this study emphasize the central and pathogen-specific role of HLA class II variation in the modulation of humoral immune response to viral antigens in humans

SP-0489: HPV-transformation in the cervix and at non-cervical sites

Pla general d'un dels panells horitzontals sobre espais verds de Barcelona a l'exposició Ciutat. Barcelona projecta a l'Edifici Fòrum. Exposició sobre la planificació urbanística i arquitectònica de Barcelon

Disease trajectories, place and mode of death in people with head and neck cancer: findings from the ‘Head and Neck 5000’ population-based prospective clinical cohort study

Background: Few large studies describe initial disease trajectories and subsequent mortality in people with head and neck cancer. This is a necessary first step to identify the need for palliative care and associated services. Aim: To analyse data from the Head and Neck 5000 study to present mortality, place and mode of death within 12 months of diagnosis. Design: Prospective cohort study. Participants: In total, 5402 people with a new diagnosis of head and neck cancer were recruited from 76 cancer centres in the United Kingdom between April 2011 and December 2014. Results: Initially, 161/5402 (3%) and 5241/5402 (97%) of participants were treated with ‘non-curative’ and ‘curative’ intent respectively. Within 12 months, 109/161 (68%) in the ‘non-curative’ group died compared with 482/5241 (9%) in the ‘curative’ group. Catastrophic bleed was the terminal event for 10.4% and 9.8% of people in ‘non-curative’ and ‘curative’ groups respectively; terminal airway obstruction was recorded for 7.5% and 6.3% of people in the same corresponding groups. Similar proportions of people in both groups died in a hospice (22.9% ‘non-curative’; 23.5% ‘curative’) and 45.7% of the ‘curative’ group died in hospital. Conclusions: In addition to those with incurable head and neck cancer, there is a small but significant ‘curative’ subgroup of people who may have palliative needs shortly following diagnosis. Given the high mortality, risk of acute catastrophic event and frequent hospital death, clarifying the level and timing of palliative care services engagement would help provide assurance as to whether palliative care needs are being met