Topoisomerase II inhibitors induce DNA damage-dependent interferon responses circumventing Ebola virus immune evasion

Ebola virus (EBOV) protein VP35 inhibits production of interferon alpha/beta (IFN) by blocking RIG-I-like receptor signaling pathways, thereby promoting virus replication and pathogenesis. A high-throughput screening assay, developed to identify compounds that either inhibit or bypass VP35 IFN-antagonist function, identified five DNA intercalators as reproducible hits from a library of bioactive compounds. Four, including doxorubicin and daunorubicin, are anthracycline antibiotics that inhibit topoisomerase II and are used clinically as chemotherapeutic drugs. These compounds were demonstrated to induce IFN responses in an ATM kinase-dependent manner and to also trigger the DNA-sensing cGAS-STING pathway of IFN induction. These compounds also suppress EBOV replication in vitro and induce IFN in the presence of IFN-antagonist proteins from multiple negative-sense RNA viruses. These findings provide new insights into signaling pathways activated by important chemotherapy drugs and identify a novel therapeutic approach for IFN induction that may be exploited to inhibit RNA virus replication

Ebolavirus species-specific interferon antagonism mediated by VP24

Members of the Ebolavirus genus demonstrate a marked differences in pathogenicity in humans with Ebola (EBOV) being the most pathogenic, Bundibugyo (BDBV) less pathogenic, and Reston (RESTV) is not known to cause a disease in humans. The VP24 protein encoded by members of the Ebolavirus genus blocks type I interferon (IFN-I) signaling through interaction with host karyopherin alpha nuclear transporters, potentially contributing to virulence. Previously, we demonstrated that BDBV VP24 (bVP24) binds with lower affinities to karyopherin alpha proteins relative to EBOV VP24 (eVP24), and this correlated with a reduced inhibition in IFN-I signaling. We hypothesized that modification of eVP24-karyopherin alpha interface to make it similar to bVP24 would attenuate the ability to antagonize IFN-I response. We generated a panel of recombinant EBOVs containing single or combinations of point mutations in the eVP24-karyopherin alpha interface. Most of the viruses appeared to be attenuated in both IFN-I-competent 769-P and IFN-I-deficient Vero-E6 cells in the presence of IFNs. However, the R140A mutant grew at reduced levels even in the absence of IFNs in both cell lines, as well as in U3A STAT1 knockout cells. Both the R140A mutation and its combination with the N135A mutation greatly reduced the amounts of viral genomic RNA and mRNA suggesting that these mutations attenuate the virus in an IFN-I-independent attenuation. Additionally, we found that unlike eVP24, bVP24 does not inhibit interferon lambda 1 (IFN-λ1), interferon beta (IFN-β), and ISG15, which potentially explains the lower pathogenicity of BDBV relative to EBOV. Thus, the VP24 residues binding karyopherin alpha attenuates the virus by IFN-I-dependent and independent mechanisms

Filovirus refseq entries: Evaluation and selection of filovirus type variants, Type sequences, And names

Sequence determination of complete or coding-complete genomes of viruses is becoming common practice for supporting the work of epidemiologists, ecologists, virologists, and taxonomists. Sequencing duration and costs are rapidly decreasing, sequencing hardware is under modification for use by non-experts, and software is constantly being improved to simplify sequence data management and analysis. Thus, analysis of virus disease outbreaks on the molecular level is now feasible, including characterization of the evolution of individual virus populations in single patients over time. The increasing accumulation of sequencing data creates a management problem for the curators of commonly used sequence databases and an entry retrieval problem for end users. Therefore, utilizing the data to their fullest potential will require setting nomenclature and annotation standards for virus isolates and associated genomic sequences. The National Center for Biotechnology Information's (NCBI's) RefSeq is a non-redundant, curated database for reference (or type) nucleotide sequence records that supplies source data to numerous other databases. Building on recently proposed templates for filovirus variant naming [<virus name> (<strain>)/<isolation host-suffix>/<country of sampling>/<year of sampling>/<genetic variant designation>-<isolate designation>], we report consensus decisions from a majority of past and currently active filovirus experts on the eight filovirus type variants and isolates to be represented in RefSeq, their final designations, and their associated sequences.Other co-authors: Ralf G. Dietzgen, Norman A. Doggett, Olga Dolnik, John M. Dye, Sven Enterlein, Paul W. Fenimore, Pierre Formenty, Alexander N. Freiberg, Robert F. Garry, Nicole L. Garza, Stephen K. Gire, Jean-Paul Gonzalez, Anthony Griffiths, Christian T. Happi, Lisa E. Hensley, Andrew S. Herbert, Michael C. Hevey, Thomas Hoenen, Anna N. Honko, Georgy M. Ignatyev, Peter B. Jahrling, Joshua C. Johnson, Karl M. Johnson, Jason Kindrachuk, Hans-Dieter Klenk, Gary Kobinger, Tadeusz J. Kochel, Matthew G. Lackemeyer, Daniel F. Lackner, Eric M. Leroy, Mark S. Lever, Elke Mühlberger, Sergey V. Netesov, Gene G. Olinger, Sunday A. Omilabu, Gustavo Palacios, Rekha G. Panchal, Daniel J. Park, Jean L. Patterson, Janusz T. Paweska, Clarence J. Peters, James Pettitt, Louise Pitt, Sheli R. Radoshitzky, Elena I. Ryabchikova, Erica Ollmann Saphire, Pardis C. Sabeti, Rachel Sealfon, Aleksandr M. Shestopalov, Sophie J. Smither, Nancy J. Sullivan, Robert Swanepoel, Ayato Takada, Jonathan S. Towner, Guido van der Groen, Viktor E. Volchkov, Valentina A. Volchkova, Victoria Wahl-Jensen, Travis K. Warren, Kelly L. Warfield, and Stuart T. Nichol Output Type: Lette

Infection of Ciliated Cells by Human Parainfluenza Virus Type 3 in an In Vitro Model of Human Airway Epithelium

We constructed a human recombinant parainfluenza virus type 3 (rPIV3) that expresses enhanced green fluorescent protein (GFP) and used this virus, rgPIV3, to characterize PIV3 infection of an established in vitro model of human pseudostratified mucociliary airway epithelium (HAE). The apical surface of HAE was highly susceptible to rgPIV3 infection, whereas only occasional cells were infected when virus was applied to the basolateral surface. Infection involved exclusively ciliated epithelial cells. There was little evidence of virus-mediated cytopathology and no spread of the virus beyond the ciliated cell types. Infection of ciliated cells by rgPIV3 was sensitive to a neuraminidase specific for α2-6-linked sialic acid residues, but not to a neuraminidase that cleaves α2-3- and α2-8-linked sialic acid residues. This provided evidence that rgPIV3 utilizes α2-6-linked sialic acid residues for initiating infection, a specificity also described for human influenza viruses. The PIV3 fusion (F) glycoprotein was trafficked exclusively to the apical surface of ciliated cells, which also was the site of release of progeny virus. F glycoprotein localized predominately to the membranes of the cilial shafts, suggesting that progeny viruses may bud from cilia per se. The polarized trafficking of F glycoprotein to the apical surface also likely restricts its interaction with neighboring cells and could account for the observed lack of cell-cell fusion. HAE derived from cystic fibrosis patients was not more susceptible to rgPIV3 infection but did exhibit limited spread of virus due to impaired movement of lumenal secretions due to compromised function of the cilia

Convergence of a common solution to broad ebolavirus neutralization by glycan cap directed human antibodies

Antibodies that target the glycan cap epitope on ebolavirus glycoprotein (GP) are common in the adaptive response of survivors. A subset is known to be broadly neutralizing, but the details of their epitopes and basis for neutralization is not well-understood. Here we present cryo-electron microscopy (cryo-EM) structures of several glycan cap antibodies that variably synergize with GP base-binding antibodies. These structures describe a conserved site of vulnerability that anchors the mucin-like domains (MLD) to the glycan cap, which we name the MLD-anchor and cradle. Antibodies that bind to the MLD-cradle share common features, including the use of IGHV1-69 and IGHJ6 germline genes, which exploit hydrophobic residues and form beta-hairpin structures to mimic the MLD-anchor, disrupt MLD attachment, destabilize GP quaternary structure and block cleavage events required for receptor binding. Our results collectively provide a molecular basis for ebolavirus neutralization by broadly reactive glycan cap antibodies

Emerging Infectious Diseases: a 10-Year Perspective from the National Institute of Allergy and Infectious Diseases

Advances in infectious disease research over the past 10 years have allowed breakthroughs in th

Organization of Integrated Formation Model with Applying of Dynamic Programming Principles

Шовкопляс А. Ш. Постановка моделі інтегрованого формування з використанням принципів динамічного програмування / А. Ш. Шовкопляс, С. А. Букреєв, Ю. О. Банніков // Вісник Приазовського державного технічного університету : зб. наукових праць / ПДТУ. – Маріуполь, 2014. – Вип. 28. – С. 105-110. – (Серія : Економічні науки).Зроблено аналіз підходів і методів організації партнерства сільськогосподарських товаровиробників і переробних підприємств. Розглянуто одна з варіантів реалізації спільних інтересів у вигляді математичної моделі взаємовигідних стосунків членів інтегрованого формування – асоціації. Наведені перевагами цієї моделі у встановленні партнерських стосунків.Підкреслено ряд актуальних положень, які не знайшли віддзеркалення в моделі і вимагають подальшої розробки та врахування при інтеграції суб'єктів господарської діяльності. Ліквідувати ці недоліки пропонується з використанням принципів динамічного програмування при проектуванні зв’язків в рамках інтегрованого об’єднання. Наведені завдання моделювання інтегрованого формування, які полягають у відображенні результативності сумісних дій підприємств у формуванні умов ефективного виробництва та створення фінансово-економічного результату. Запропонована модель представлена в рамках апробації з метою визначення оптимальних параметрів виробничої діяльності підприємств ТОВ "Колос", ТОВ ”Племзавод імені Літвінова” та ЗАТ ”Слов’яносербський молокозавод”, об’єднаних в інтегроване формування з метою формування виробничо-збутового ланцюга з виробництва молочної продукції кінцевогоспоживання. Розглянуто альтернативні варіанти виробництва кормової продукції в ТОВ "Колос", варіанти виробництва молока в ТОВ ”Племзавод імені Літвінова” та виробництва асортименту молочної продукції в ЗАТ ”Слов’яносербський молокозавод”. Основним стимулом до створення інтегрованих формувань є формування можливостей поліпшення процесу ресурсного та фінансово-економічного забезпечення підприємства, а також ліквідувати недолік підпорядкування економіки підприємств друг другу.Сделан анализ подходов и методов организации партнерства сельскохозяйственных товаропроизводителей и перерабатывающих предприятий. Рассмотрен один из вариантов реализации общих интересов в виде математической модели взаимовыгодных отношений членов интегрированного формирования - ассоциации. Приведены преимущества этой модели в установлении партнерских отношений. Подчеркнуто ряд актуальных положений, которые не нашли отражения в модели и требуют последующей разработки и учета при интеграции субъектов хозяйственной деятельности. Ликвидировать эти недостатки предлагается с использованием принципов динамического программирования при проектировании связей в рамках интегрированного объединения. Приведены задания моделирования интегрированного формирования, которые заключаются в отображении результативности совместимых действий предприятий в формировании условий эффективного производства и создания финансово- экономического результата. Предложена модель, представленная в рамках апробации с целью определения оптимальных параметров производственной деятельности предприятий ООО "Колос", ООО ”Племзавод имени Литвинова” и ЗАО ”Славяносербский молокозавод”, объединенных в интегрированное формирование с целью формирования производственно-сбытовой цепи по производству молочной продукции конечного потребления. Рассмотрены альтернативные варианты производства кормовой продукции в ООО "Колос", варианты производства молока в ООО ”Племзавод имени Литвинова” и производства ассортимента молочной продукции в ЗАО ”Славяносербский молокозавод”. Основным стимулом для создания интегрированных формирований является формирование возможностей улучшения процесса ресурсного и финансово- экономического обеспечения предприятия, а также ликвидировать недостаток подчинения экономики предприятий друг другу.The analysis of the partnership of agricultural producers and processing enterprises organization approaches and methods is done. One of the variants of common interests realization in the way of mathematical model of mutually advantageous relations of integrated formation – members - association - is considered. The advantages this model organizing partnership relations are given. A set of actual statements which weren’t discrebed in the model and require further research and control in integration of economic activity entites is analized. It is proposed to eliminate these disadvantages using dynamic programming principles in planning relations within integrated association. The tasks of integrated formation modelling which are found in the demonstration of enterprises joint actions in forming the effective production conditions and in financial and economic result are given. The proposed model is demonstrated as an approbation with the aim of the most optimal parameters determining the productivity such of enterprises as Ltd. “Kolos”, Ltd. “Breeding plant by Litvinov” and closed joint stock company “Slov’yanoserbs’k milk processing plant”, united in the integrated formation with the aim of production and sales chain based on milk production of the final consumption. Alternative variants of fodder production in Ltd. “Kolos”, variants of milk production in Ltd. “Breeding plant by Litvinov” and production of the milk products assortment in closed joint stock company “Slov’yanoserbs’k milk processing plant” are considered. The main incentive for integrated formations creation is forming the possibilities of resource and financial - economic enterprises providing process improvement and also elimination of the lack of enterprises economy subordination to each other

Mucosal Immunization of Cynomolgus Macaques with the VSVΔG/ZEBOVGP Vaccine Stimulates Strong Ebola GP-Specific Immune Responses

(ZEBOV) produces a lethal viral hemorrhagic fever in humans and non-human primates.We demonstrate that the VSVΔG/ZEBOVGP vaccine given 28 days pre-challenge either intranasally (IN), orally (OR), or intramuscularly (IM) protects non-human primates against a lethal systemic challenge of ZEBOV, and induces cellular and humoral immune responses. We demonstrated that ZEBOVGP-specific T-cell and humoral responses induced in the IN and OR groups, following an immunization and challenge, produced the most IFN-γ and IL-2 secreting cells, and long term memory responses.We have shown conclusively that mucosal immunization can protect from systemic ZEBOV challenge and that mucosal delivery, particularly IN immunization, seems to be more potent than IM injection in the immune parameters we have tested. Mucosal immunization would be a huge benefit in any emergency mass vaccination campaign during a natural outbreak, or following intentional release, or for mucosal immunization of great apes in the wild