Effects of human papillomavirus (HPV) type 16 oncoproteins on the expression of involucrin in human keratinocytes

<p>Abstract</p> <p>Background</p> <p>The human papillomavirus (HPV) life cycle is closely linked to keratinocyte differentiation. Oncogenic HPV infection has been shown to hamper the normal differentiation of keratinocytes; however, the underlying mechanisms responsible for this phenomenon are yet to be clarified. Here, we aimed to study the effects of HPV16 E6 and E7 oncogenes on the expression of involucrin (IVL), an established marker of keratinocyte differentiation, in human foreskin keratinocyte (HFK) cells.</p> <p>Results</p> <p>The differentiation of HFK cells by serum and high calcium significantly increased both the mRNA and the protein levels of IVL. The E6 and E7 oncoproteins of HPV16 together caused strong down-regulation of IVL mRNA and protein both in proliferating and in differentiating HFK cells. To study the effects of HPV oncogenes on the <it>IVL </it>promoter, we made transient transfection assays and luciferase tests and found that HPV 16 E6 but not E7 repressed <it>IVL </it>promoter activity in proliferating HFK cells. The inhibitory effect of HPV 16 E6 on the human <it>IVL </it>promoter could be localised to the proximal regulatory region (PRR) of the gene.</p> <p>Conclusions</p> <p>These results suggest that the down-regulation of <it>IVL </it>promoter activity by HPV 16 E6 significantly contribute to the inhibition of endogenous <it>IVL </it>expression by the HPV 16 oncoproteins. In contrast, the down-regulation of endogenous IVL expression by HPV16 E7 is probably not caused by a direct and specific effect of E7 on the <it>IVL </it>promoter.</p

Real-time monitoring of the effectiveness of six COVID-19 vaccines against laboratory confirmed COVID-19 in Hungary in 2021 using the screening method

Several studies have reported the waning effectiveness of COVID-19 vaccines. This study aims to demonstrate the applicability of the screening method for estimating vaccine effectiveness (VE) in a pandemic. We report VE in Hungary, estimated with the screening method, in 2021, covering a period of Alpha and the Delta variant, including the booster dose roll-out. Hungary is in a unique position to use six different vaccines in the same population. All vaccines provided a high level of protection initially, which declined over time. While the picture is different in each age group, the waning of immunity is apparent for all vaccines, especially in the younger age groups and the Sinopharm, Sputnik-V, and AstraZeneca vaccines, which performed similarly. This is clearly reversed by booster doses, more prominent for those three vaccines, where the decline in protection is more evident. Overall, two vaccines, Pfizer/BioNTech and Moderna, tend to produce the best results in all age groups, even with waning immunity considered. Using the screening method in future pandemic waves is worthwhile, especially in countries struggling with a lack of resources or when there is a need to deliver VE results within a short timeframe due to urgent decision-making

Vaccine effectiveness against COVID-19 hospitalisation in adults (≥ 20 years) during Omicron-dominant circulation: I-MOVE-COVID-19 and VEBIS SARI VE networks, Europe, 2021 to 2022

Introduction: The I-MOVE-COVID-19 and VEBIS hospital networks have been measuring COVID-19 vaccine effectiveness (VE) in participating European countries since early 2021. Aim: We aimed to measure VE against PCR-confirmed SARS-CoV-2 in patients ≥ 20 years hospitalised with severe acute respiratory infection (SARI) from December 2021 to July 2022 (Omicron-dominant period). Methods: In both networks, 46 hospitals (13 countries) follow a similar test-negative case-control protocol. We defined complete primary series vaccination (PSV) and first booster dose vaccination as last dose of either vaccine received ≥ 14 days before symptom onset (stratifying first booster into received < 150 and ≥ 150 days after last PSV dose). We measured VE overall, by vaccine category/product, age group and time since first mRNA booster dose, adjusting by site as a fixed effect, and by swab date, age, sex, and presence/absence of at least one commonly collected chronic condition. Results: We included 2,779 cases and 2,362 controls. The VE of all vaccine products combined against hospitalisation for laboratory-confirmed SARS-CoV-2 was 43% (95% CI: 29-54) for complete PSV (with last dose received ≥ 150 days before onset), while it was 59% (95% CI: 51-66) after addition of one booster dose. The VE was 85% (95% CI: 78-89), 70% (95% CI: 61-77) and 36% (95% CI: 17-51) for those with onset 14-59 days, 60-119 days and 120-179 days after booster vaccination, respectively. Conclusions: Our results suggest that, during the Omicron period, observed VE against SARI hospitalisation improved with first mRNA booster dose, particularly for those having symptom onset < 120 days after first booster dose.Key public health message: 1. What did you want to address in this study? In order to understand how well the COVID-19 vaccine is performing in Europe against hospitalisation during the period when the SARS-CoV-2 Omicron variant was circulating, we investigated vaccine effectiveness using data from a multi-country study of complete and booster-dose COVID-19 vaccination among adults aged 20 years and over. 2. What have we learnt from this study? Between December 2021 and July 2022, vaccine effectiveness against hospitalisation with laboratory-confirmed SARS-CoV-2 was 43% for complete vaccination. With addition of an mRNA booster dose, effectiveness was 59% overall. It was higher when onset of illness was close to the date of the last vaccination, at 85% when last booster dose was 14–59 days before onset, at 70% for 60–119 days, and falling below 40% for 120–179 days. 3. What are the implications of your findings for public health? In European hospital settings in 2022, during the Omicron period, COVID-19 mRNA booster vaccine provided an improved benefit for preventing hospitalisation, particularly if disease onset was within 4 months of receiving the booster dose.info:eu-repo/semantics/publishedVersio

Phylogenetic and functional analysis of human papillomavirus (HPV) type 31 certain genomic region

A humán papillomavírusok (HPV) mintegy harmada az anogenitális traktust fertőzi. A magas rizikófaktorú HPV típusok (16, 18, 31, 33, 35 stb.) tehetőek felelőssé a méhnyakrák kialakulásáért. Az E6/E7 valamint az LCR genomi régiókban bekövetkező nukleotidcserék megváltozott transzkripciós aktivitást és/vagy megváltozott onkogén potenciált eredményezhetnek, mindez pedig egy lehetséges mechanizmus arra, hogy a HPV intraípusos variánsok viselkedésében eltérések mutatkozzanak. Munkánk célja az volt, hogy megvizsgáljuk az eddig kevésbé tanulmányozott magas kockázatú HPV 31 LCR, E6 és E7 régióinak genetikai polimorfizmusát egy magyarországi populációban, illetve hogy a variánsok esetleges funkcionális különbségeit detektáljuk. Munkánk során premalignus illetve malignus elváltozásokból származó klinikai mintákkal dolgoztunk. A filogenetikai vizsgálatok alapján három intratípusba (A, B, C) sorolhatók az izolátumok, mely besorolást mind az LCR, mind az E6/E7 régiók alapján el lehet végezni. Nukleotid polimorfizmus szempontjából az LCR bizonyult variábilisabbnak szemben az E6 és E7 régiókkal, melyek fehérjét kódolnak. Szignifikáns különbségeket találtunk az eltérő intratípusos csoportba tartozó HPV 31 LCR variánsok transzkripciós aktivitásában. Deléciós mutánsokkal történő transzfekciók alapján megállapítottuk, hogy az egyes LCR variánsok aktivitás különbségeiért részben az LCR 5’, részben pedig 3’ szakaszán (minimal promoter region) található nukleotidcserék a felelősek. A különféle E6 és E7 variánsokat expressziós vektorba klónozva vizsgáltuk azok hatását p53 illetve adenovírus E2 promóterekre, valamint teszteltük az E6 és E7 variánsok in vivo p53 illetve pRb degradációját indukáló hatását. Az eltérő intratípusba tartozó HPV 31 E7 variánsok, akárcsak a HPV 16 E7 prototípus, megemelték a pAdE2 promóter aktivitását. Az E6 variánsok esetében különbségeket találtunk a variánsok p53 transzaktivációját gátló hatásában, az A és C intratípusba tartozó variánsok csökkentették a p53 aktivitást, hasonlóan a kontrollként használt HPV 16 E6-hoz, a B csoportba tartozó variánsok hatása a p53 aktivitására mérsékeltebb volt. A különböző HPV 31 E6/E7 intratípusos variánsok in vivo p53/Rb degradációs aktivitásában nem találtunk szignifikáns különbségeket. További érdekesség, hogy a B intratípusba tartozó LCR variánsok nagyobb transzkripciós aktivitást mutattak, mint az A illetve C intratípusos variánsok.About one-third of human papillomavirus (HPV) types infect the anogenital tract. High-risk genital HPV types (such as HPV 16, 18, 31, 33, and 35) are linked causally to the development of cervical cancer. Nucleotid changes in the LCR and/or E6 and E7 oncogenes may leads to different transcriptional activities and/or different oncogenic potential and this may be one molecular mechanism that is responsible for the differences in the behavior of intratypic HPV variants. The purpose of present study was to explore genetic variability and functional differences in the complete LCR and E6, E7 regions of the less studied HPV 31 natural variants isolated from Hungarian women with cervical premalignant lesions. According to the phylogenetical tree of the LCR, E6 and E7 variants three major intratypical lineages (A, B, C) were identified. Comparative phylogenetic analysis of the E6/E7 and the LCR region showed that nucleotide sequence variation was lower in the E6/E7 region, but either region could be used with high confidence to identify the variant lineage of an isolate. We found significant differences between the transcriptional activities of the HPV 31 LCR variants belonging to the different variant lineages. On the bases of the experiments with deletion mutants of HPV 31 LCR variants showes, that nucleotid changes, which cause the functional differences, localized at least partially in the 5’ URR domain, on the other hand int he minimal promoter region. To start the functional analysis of the HPV 31 E6 and E7 variants, we tested their effects on p53 and adenovirus E2 promoters. In addition, we also tested the ability of the E6 and E7 variants to induce the in vivo degradation of p53 and pRB. HPV 31 E7 variants belonging to different variant lineages were able to increase E2F transcriptional activity to the same extent as the prototype HPV 16 E7. The E6 variants had comparable abilities to induce the in vivo degradation of p53, there were differences between them in the ability to inhibit the trans-activation function of p53. Namely, the prototype HPV 31 E6 (belonging to variant lineage A), along with the variant belonging to lineage C was active in this function, while the variant belonging to lineage B showed reduced ability to inhibit the trans-activation function of p53. We found no significant differences in the in vivo p53 and pRb degradation activities of HPV 31 E6 and E7 variants belonging to different variant lineages. On the other hand, we recently found that HPV 31 LCR variants belonging to lineage B display higher transcriptional activities than variants belonging to lineage A or C

A genitális humán papillomavírusok diagnosztikájában alkalmazott consensus PCR módszerek összehasonlító vizsgálata

Vizsgálataim célja az volt, hogy összehasonlítsak két consensus PCR technikát, melyek a humán papillomavírusok diagnosztikájára alkalmasak. Az eddig használt módszer a MY09-11 primerpárossal működik, míg az általunk bevezetni kívánt rendszer a PGMY09-11 primereket használja. Az eredmények bizonyítják, hogy ez utóbbi módszer hatékonyabb, specifikusabb, valamint több HPV típus kimutatható, mint az MY09-11 rendszerrel.Mag

Phylogenetic and functional analysis of sequence variation of human papillomavirus type 31 E6 and E7 oncoproteins

High-risk human papillomaviruses (HPV) are the causative agents of cervical and other anogenital cancers as well as a subset of head and neck cancers. The E6 and E7 oncoproteins of HPV contribute to oncogenesis by associating with the tumour suppressor protein p53 and pRb, respectively. For HPV types 16 and 18, intratypic sequence variation was shown to have biological and clinical significance. The functional significance of sequence variation among HPV 31 variants was studied less intensively. HPV 31 variants belonging to different variant lineages were found to have differences in persistence and in the ability to cause high grade cervical intraepithelial neoplasia. In the present study, we started to explore the functional effects of natural sequence variation of HPV 31 E6 and E7 oncoproteins. The E6 variants were tested for their effects on p53 protein stability and transcriptional activity, while the E7 variants were tested for their effects on pRb protein level and also on the transcriptional activity of E2F transcription factors. HPV 31 E7 variants displayed uniform effects on pRb stability and also on the activity of E2F transcription factors. HPV 31 E6 variants had remarkable differences in the ability to inhibit the trans-activation function of p53 but not in the ability to induce the in vivo degradation of p53. Our results indicate that natural sequence variation of the HPV 31 E6 protein may be involved in the observed differences in the oncogenic potential between HPV 31 variants