In vitro expression of HPV16 E7 linked to HMGB1 immunoadjuvant in mammalian cells.

BACKGROUND: E7 is the major transforming protein of human papillomavirus (HPV) that plays important role in maintaining the proliferative state in HPV-infected cells. Furthermore, high mobility group 1 protein (HMGB1) is a highly conserved component of chromatin that can be secreted by macrophages and activated monocytes and thus functions as an inflammation mediator. METHODS: In the current study, cloning of HMGB1 gene and also HPV16E7-HMGB1 was performed in pEGFP-N1 eukaryotic expression vector in order to evaluate their expression in mammalian cells. For this purpose, the HEK-293T cells were transfected by pEGFP-E7, pEGFP-HMGB1 and pEGFP-E7-HMGB1 using TurboFect delivery system. The levels of protein expression were assessed by flow cytometry and fluorescent microscopy at 48 hr after transfection, as well as by western blot analysis using anti-GFP polyclonal antibody. RESULTS: Our data showed a clear band of ~ 684 bp and ~ 981 bp related to HMGB1 and E7-HMGB1 genes in agarose gel, respectively. The expression of HMGB1-GFP and E7-HMGB1-GFP proteins was confirmed for the bands of ~ 53 kDa and ~ 64 kDa in the transfected cells using western blot analysis, respectively. The linkage of HMGB1 gene to E7 could likely neutralize the negative charges of E7, thus a clear band of 64 kDa was detected instead of 76 kDa in western blot analysis. Moreover, the percentage of expression for E7-GFP, HMGB1-GFP and E7-HMGB1-GFP was 76 %, 55 %, and 52 %, in comparison with pEGFP-N1 (~82 %) as a positive control. Indeed, HMGB1 linked to HPV16 E7 gene decreased transfection efficiency of E7 DNA in HEK-293T cells. CONCLUSION: Generally, the electrophoretic mobility of HPV16 E7 was changed due to the linkage of HMGB1 gene. Furthermore, the fusion protein could be efficiently expressed in mammalian cells for the next use in immunotherapy (Fig. 3, Ref. 51).

Estimation of influenza and severe acute respiratory illness incidence (burden) in three provinces of the Islamic Republic of Iran, 2012 and 2013.

There are few estimates of influenza burden in the WHO Region for the Eastern Mediterranean. In this study we estimated the burden of severe acute respiratory infection (SARI) and influenza-associated SARI (F-SARI) in selected provinces of Islamic Republic of Iran, the trends of SARI and confirmed cases of influenza (F-SARI) over 12 months (seasonality), and the age groups most at risk. Using the electronic Iranian influenza surveillance system and data of cases in sentinel hospitals of 3 selected provinces, we estimated the monthly trend (seasonality) of incidence for SARI and F-SARI, overall incidence of SARI and F-SARI and their disaggregation by age with the aid using the Monte Carlo technique. The age groups most at-risk were children aged under 2 years and adults older than 50 years.

Estimation of influenza and severe acute respiratory illness incidence [burden] in three provinces of the Islamic Republic of Iran, 2012 and 2013

There are few estimates of influenza burden in the WHO Region for the Eastern Mediterranean. In this study we estimated the burden of severe acute respiratory infection [SARI] and influenza-associated SARI [F-SARI] in selected provinces of Islamic Republic of Iran, the trends of SARI and confirmed cases of influenza [F-SARI] over 12 months [seasonality], and the age groups most at risk. Using the electronic Iranian influenza surveillance system and data of cases in sentinel hospitals of 3 selected provinces, we estimated the monthly trend [seasonality] of incidence for SARI and F-SARI, overall incidence of SARI and F-SARI and their disaggregation by age with the aid using the Monte Carlo technique. The age groups most at-risk were children aged under 2 years and adults older than 50 years

Peu d'estimations sont disponibles sur la charge de la grippe dans la Région OMS de la Méditerranée orientale. Dans la présente étude, nous avons estimé la charge de morbidité due aux infections respiratoires aiguës sévères [IRAS] et aux IRAS associés à la grippe dans certaines provinces de la République islamique d'Iran, ainsi que les tendances des IRAS et des cas de grippe confirmés sur 12 mois [saisonnalité], et les groupes d'âge les plus exposés. En utilisant le système électronique iranien de surveillance de la grippe et les données relatives aux cas des hôpitaux sentinelles de trois provinces sélectionnées, nous avons estimé la tendance mensuelle [saisonnalité] de l'incidence des IRAS et des IRAS dus à la grippe, l'incidence globale des IRAS et des IRAS dues à la grippe et leur ventilation par âge grâce à la méthode de Monte Carlo. Les groupes d'âge les plus exposés au risque étaient les enfants de moins de 2 ans et les adultes de plus de 50 ans

Measles vaccination coverage and seroprevalence of anti-measles antibody in south-east Islamic Republic of Iran

كثيراً ما توجد تباينات بين تغطية التطعيم المسجلة وبين المستوى الحقيقي للمناعة في المجتمع. ولتقدير تغطية التلقيح ضد الحصبة في جنوب شرق جمهورية إيران الإسلامية تم إجراء دراسة مقطعية في 3 مناطق خلال صيف عام 2011.فتم اختيار عينة عنقودية من 1368 طفلاً تتراوح أعمارهم ما بين 30-54 شهراً باستخدام الاحتمال المتناسب مع الحجم. وتم فحص عينات مصلية من 663 منهم - ممن تلقوا حقنتين من اللقاح الثلاثي ضد الحصبة والنكاف والحصبة الألمانية [MMR]- للتأكد من وجود الجلوبيولين المناعي G [IgG]المضاد للحصبة. لقد بلغت تغطية التطعيم بالجرعة الثانية من لقاح MMR 93.7%.وكان انتشار الجلوبيولين المناعي G المضاد للحصبة لدى أولئك الذين تلقوا جرعتين على الأقل من لقاح MMR 94.6%.وكان هناك ارتباط ذو دلالة إحصائية بين النتائج السيرولوجية وبين المتغيرات التي تعكس ضعف إمكانية الوصول إلى الخدمات الصحية. وبجمع النتائج السيرولوجية مع بيانات التغطية قُدِّرت نسبة الجمهور المحمي ضد الحصبة بـ 88.6%، والتي كانت أقل من الحدود المعينة لأهداف التخلص من الحصبة

Discrepancies often exist between recorded immunization coverage and the real immunity level in a community. To estimate the vaccination coverage against measles in south-east Islamic Republic of Iran, a crosssectional study was conducted in 3 districts during summer 2011. Using probability proportional to size cluster sampling, 1368 children aged 30–54 months were selected. Serum samples of 663 who had received 2 injections of mumpsmeasles-rubella (MMR) vaccine were checked for anti-measles IgG. Vaccination coverage for the second dose of MMRvaccine was 93.7%. The prevalence of anti-measles IgG in those who had received at least 2 MMR vaccine doses was 94.6%. There was a statistically significant association between the serological results and variables that reflected pooraccessibility to health services. Combining serological results with coverage data, the proportion of the community protected against measles was estimated as 88.6%, which was below the limits defined for the measles elimination goals.

Il existe souvent des écarts entre la couverture vaccinale enregistrée et le niveau réel d'immunité d'une communauté donnée. Afin d'estimer la couverture vaccinale antirougeoleuse dans le sud-est de la République islamique d'Iran, une étude transversale a été menée dans trois districts durant l'été 2011. En appliquantl'échantillonnage en grappes avec probabilité proportionnelle à la taille, 1368 enfants âgés de 30 à 54 mois ont été sélectionnés. Des prélèvements de sérum de 663 enfants de l'échantillonnage ayant reçu deux injections du vaccincontre la rougeole, les oreillons et la rubéole (ROR) ont été analysés à la recherche d'anticorps IgG antirougeoleux. La couverture vaccinale pour la deuxième dose de vaccin ROR était de 93,7 %. La prévalence des anticorps IgG antirougeoleux chez les enfants ayant reçu au moins deux doses du vaccin ROR était de 94,6 %. Il existait une association statistiquement significative entre les résultats sérologiques et les variables qui reflétaient l'accès insuffisant aux services de santé. En combinant les résultats sérologiques et les données de couverture, la proportion de la communauté protégée contre la rougeole a été estimée à 88,6 %, soit un taux inférieur aux seuils définis pour les objectifs d'élimination de la rougeole.

Sequence and phylogenetic analyses of neuraminidase gene of Iranian seasonal influenza H1N1 viruses from 2005-2009 and corresponding vaccine strains.

Prophylaxis of influenza A virus infections is based on the vaccines inducing antibodies to the major viral antigens, hemagglutinin (HA) and neuraminidase (NA). Since these antigens continuously change during virus replication in various hosts, only the currently circulating strains should be used in the vaccines. Besides, monitoring of the naturally occurring changes in HA, NA, and respective genes, especially those associated with resistance to the NA inhibitors is necessary. The NA genes of 30 Iranian isolates of influenza H1N1 virus from the seasons 2005-2009 were sequenced and subjected to the sequence and phylogenetic analyses. The seasonal isolates turned out to be closely related to the corresponding vaccine strains, except for the 2007-2008 isolates, which also displayed a higher nucleotide variation. A resistance to the NA inhibitors was found in the 2008-2009 isolates only. The average nucleotide identities of the isolates with corresponding vaccine strains for the years 2005-2009 were 98.83%, 98.55%, 98.7%, 97.55%, and 98.76%, respectively.

Direct identification of non-polio enteroviruses in residual paralysis cases by analysis of VP1 sequences.

BACKGROUND: The 66 serotypes of human enteroviruses (EVs) are classified into four species A-D, based on phylogenetic relationships in multiple genome regions. Partial VP(1) amplification and sequence analysis are reliable methods for identifying non-polio enterovirus serotypes, especially in negative cell culture specimens from patients with residual paralysis. OBJECTIVES: In Iran during the years 2000-2002, there were 29 residual paralysis cases with negative cell (RD, HEp(2) and L(20)B) culture results. STUDY DESIGN: The genomic RNA was extracted from stool specimens from cases of residual paralysis and detected by amplification of the 5'-nontranslated region using RT-PCR with Pan-EV primers. Partial VP(1) amplification by semi-nested RT-PCR (snRT-PCR) and sequence analysis were done. RESULTS: Specimens from the 29 culture-negative cases contained echoviruses of six different serotypes. CONCLUSIONS: The global eradication of wild polioviruses is near and study of non-polio enteroviruses, which can cause poliomyelitis, is increasingly important to understand their pathogenesis. The VP(1) sequences, derived from the snRT-PCR products, allowed rapid molecular analysis of these non-polio strains.

Characterization of mutations in the VP(1) region of Sabin strain type 1 polioviruses isolated from vaccine-associated paralytic poliomyelitis cases in Iran.

BACKGROUND: The live-attenuated oral polio vaccine used to interrupt poliovirus transmission is genetically unstable. Reversion of some attenuating mutations, which normally occurs during vaccine strain replication in some recipients, and can rarely cause vaccine-associated paralytic poliomyelitis (VAPP). The poliovirus eradication program designed by the World Health Organization (WHO) includes immunization with OPV in addition to careful surveillance of all acute-flaccid paralysis (AFP) cases. OBJECTIVES: In Iran we last isolated imported wild poliovirus in 2000 and the immunization coverage was 100% in 2002. During 2001, there were three AFP cases with residual paralysis from which Sabin-like type 1 polioviruses were isolated in our national polio laboratory. STUDY DESIGN: The complete VP(1) region of the three isolates was sequenced and amino acid substitutions associated with these neurovirulent isolates were recorded. RESULTS: These isolates had either 4, 2 or 1 nucleotide substitution(s) in the VP(1) region, corresponding to amino acid change in the VP(1) of isolate 1 of either (H-[149]->Y), (T-[106]->A) or (I-[90]->L), respectively. CONCLUSIONS: Surveillance of the VAPP cases in countries where endemic transmission has recently ceased increases our understanding of the important neurovirulent mutations in vaccine-strain isolates and assists in planning the next step in the eradication program in these countries.

Prevalence of human papillomavirus in Mazandaran Province, Islamic Republic of Iran.

We report the prevalence of human papillomavirus (HPV) types in 100 cervical biopsy specimens in Mazandaran province. HPV DNA was detected in 78.6% of cervical carcinoma cases, 64.3% of dys/ metaplasia and 9% of normal cases. Significant correlation was found between the presence of HPV DNA and development of cervical carcinoma. HPV types 16 and 18 were detected in 60.6% of HPV-positive cervical carcinoma cases, whereas HPV31 and 33 were found in 21.2%, and HPV6 and 11 in 18.2%. Among HPV-positive dys/metaplasia cases, 55.6% were positive for HPV16 and 18, 22.3% for HPV6 and 11, and 11.1% for HPV31 and 33. Only HPV6 and 11 were detected in 4 (100%) normal biopsy specimens.

Prevalence of human papillomavirus in Mazandaran Province, Islamic Republic of Iran

We report the prevalence of human papillomavirus [HPV] types in 100 cervical biopsy specimens in Mazandaran province. HPV DNA was detected in 78.6% of cervical carcinoma cases, 64.3% of dys/ metaplasia and 9% of normal cases. Significant correlation was found between the presence of HPV DNA and development of cervical carcinoma. HPV types 16 and 18 were detected in 60.6% of HPV-positive cervical carcinoma cases, whereas HPV31 and 33 were found in 21.2%, and HPV6 and 11 in 18.2%. Among HPV-positive dys/metaplasia cases, 55.6% were positive for HPV16 and 18, 22.3% for HPV6 and 11, and 11.1% for HPV31 and 33. Only HPV6 and 11 were detected in 4 [100%] normal biopsy specimens