Відділ репродукції вірусів - ІМВ НАН України 🇺🇦

Head of Department, PhD
Svitlana Zagorodnya
svetazagorodnya@ukr.net
+38 (044) 2946965
Skype: svetazagorodnya

AREAS OF ACTIVITY OF THE LABORATORY:

  • Study of regularities of molecular mechanisms of virus-cell interaction on the model of human adenovirus, Epstein-Barr virus, herpes simplex virus type 1 and 2;
  • Identification of new substances with potential anti-herpetic and anti-adenoviral properties and establishment of the targets of their action, which allows to carry out their structural and functional analysis with subsequent adjustment of the direction of synthesis to create highly effective antiviral agents;
  • Development of a strategy for researching the antiviral activity of substances of various nature in the in vitro and in vivo system; conducting preclinical studies;
  • Development of biotechnological aspects of the creation of specific immunobiological preparations of targeted action and diagnostic tools.
Scientific work

Research of human and animal viruses at the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine was started in 1963 in the department of biophysics of viruses (Y.G. Kishko). In 1974, the department of biophysics of viruses was reorganized into the laboratory of animal viruses, which in 1978 was renamed the department of molecular biology of viruses under the leadership of member-cor. National Academy of Sciences of Ukraine, Doctor of Biological Sciences Dyachenko N.S. Since 2004, the department has been renamed the Virus Reproduction Laboratory, the head of which until 2014 was PhD Nesterova Nadiya Vitalyivna. Since 2016, the virus reproduction laboratory has been reorganized into the department of virus reproduction under the leadership of Dr. Zagorodnaya Svitlana Dmitrivna.
Among the long-term scientific achievements of the department's employees is an in-depth study of the physicochemical properties of human adenovirus type 1 proteins, which made it possible to obtain new fundamental knowledge about the localization of adenovirus proteins in an infected cell and the discovery of several new antigenic determinants of the hexon and adenovirus appendage. The different nature of polypeptides (conformational or linear orientation), which depends on the spatial organization of proteins, was revealed, the localization of some epitopes was determined with the help of chemically synthesized antigenically active peptides imitating them. Essential regularities of the structural and functional organization of the hexon of adenoviruses were revealed on the basis of a comparative analysis of antigenic specificity and the primary structure of adenovirus proteins of different types, taxonomically distant from each other. The concept of immunoinactivation (neutralization of infectivity) of adenoviruses was developed and a mathematical model of this process was created for the first time, which determines the contribution of antibodies to several antigenic determinants of hexon and appendage, as well as interferon and complement. The obtained data were summarized in monographs: 

  • •Дяченко Н.С. Аденовирусы.- Киев: Наук.думка, 1974.- 157 с.
  • •Дяченко Н.С.  Пассивная гемагглютинация и ее применение в вирусологии.- Киев: Наук. Думка, 1979.-148 с. 
  • •Дяченко С.С., Синяк К.М., Дяченко Н.С. Патогенные вирусы человека.- Киев: Здоров’я, 1980.- 448 с. 
  • •Носач Л.Н., Дяченко Н.С. Цитопатология аденовирусной инфекции.- Киев:Наук.думка, 1982.- 124 с. 
  • •Djačenko S.S., Sinjak K.M., Djačenko N.S. Patogenni viry člověka .-Praha:Avicenum, 1983.-382 p. 
  • •Дяченко Н., Нас И., Беренчи Д., Носач Л., Ванцак Н., Тарасишин Л., Адам Е.  Аденовирус, клетка, организм. Киев: Наук. думка, 1988. 232 с. 

Focused search and research of the mechanisms of action of anti-adenoviral substances in the department was started in 1979 together with Dr. Chernetsky V.P. and his students Ph.D. Alekseeva I.V. and Shalamai A.S. (Institute of Molecular Biology and Genetics of the National Academy of Sciences of Ukraine). The influence of the original structural analog of cytidine (2-β-D-ribofuranosyl-5-amino-1,2,4-triazine-3(2H)-OH) on the reproduction of human adenoviruses, 6-azacytidine, was investigated in the most detailed manner. Due to its low cytotoxicity and high anti-adenoviral effect, the drug is patented as an antiviral. It has been established that its antiviral activity is associated with inhibition of the synthesis of viral DNA and proteins. A number of 6-azacytidine derivatives were studied and compounds with lower toxicity and better solubility were identified. Anti-adenoviral activity was found in 6-azauridine, a structural analogue of uridine synthesized by Ukrainian chemists. The studied activity of cytosinarabinoside derivative - cyclocytidine and ribavirin's structural analogue - rimamidil, synthesized by memb.-cor. of Academy of Sciences of Latvia M.Yu. Ladakh. It was shown that ribamidil affects the formation of a functionally complete hexon and the maturation of adenovirion. Antiviral activity of proteolysis inhibitors - amoeba and ε-aminocaproic acid was studied. The target of their action is a violation of the morphogenesis of mature infectious adenovirions, which is caused by a violation of the processing of the precursor of the internal polypeptide (VII). Together with the employees of the Odesa Institute of Physics and Chemistry named after A.V. Bogatsky National Academy of Sciences of Ukraine (V.E. Kuzmin, Ph.D., A.G. Artemenko, Ph.D.) studied the antiviral properties of macrocyclic pyridinophanes using 3D and 4D QSAR modeling, which allows establishing the relationship between the structure and activity of the substance, as well as to predict the effectiveness of compounds against viruses. Using QSAR and simplex representation, active structures were discovered and new nitrogen-containing analogues were synthesized, for which encouraging results of antiviral activity were obtained. The work was carried out within the framework of the INTAS 94-292 grant.
The anti-adenoviral activity of some substances isolated from plants and marine organisms was shown. The most active was hyporamine obtained from sea buckthorn at VYLAR (Russia). Hiporamine blocks the reproduction of the adenovirus at the stage of penetration and undressing of the virus. From the aqueous extract from Artemia cysts, the employees of the Institute of Southern Seas of the National Academy of Sciences of Ukraine (PhD I.I. Rudneva and Ph.D. Shaida V.G.) obtained extracts that were patented as an antiviral agent with anti-adenoviral and antiherpetic activities.

The first domestic immunoenzymatic test system was developed for the diagnosis of diseases caused by the Epstein-Barr virus. The antigen was obtained, its specificity was confirmed, and panels of positive and negative sera were created. The conditions for enzyme-linked immunosorbent assay were optimized and the parameters of specificity, sensitivity and compliance of the IFA-AtVEB-strip test system were characterized.
As part of the INTAS 011-2382 grant, the impact of the Epstein-Barr virus on signal transduction processes in malignant B-cells transformed by the virus was investigated. Experimental virus-cell systems were developed, in which the influence of viral proteins on the process of SD-95-mediated apoptosis, in the conditions of drug-induced apoptosis, was studied. Data was obtained on the different degree of influence of viral infection on the sensitivity of BL-41 and DG-75 cells to etoposide and doxorubicin, which are antitumor agents. This may be due to different mechanisms of their action, which in one way or another limit the replicative mechanism of VEB. 
As part of the All-State (national) space program of Ukraine for 2003-2007, research was conducted on the influence of conditions of simulated microgravity on adenovirus and the cell-virus system. It has been established that an intact human adenovirus is able to maintain infectivity for a long time and reproduce in conditions of simulated gravity (Space Science and Technology, 2003, 2004 and 2007). 
The research carried out in the department in recent years was devoted to the modeling of mixed adeno-herpetic infection on a population of suspension lymphoblastoid and monolayer transplanted human and animal cells, further study of the effect of antiviral drugs in the developed models. The research continues the long-term scientific research of the team and is harmoniously combined with the direction of development of world virology, in particular, in relation to adenoviruses and herpesviruses. For the first time, model systems of co-infected cell cultures were obtained, the reproduction of viruses in them was proven, and a detailed analysis of the structural and functional virus-cell interaction was carried out. The anomalous effect of antiviral substances that we discovered, in the case of co-infection of cells with two viruses, indicates the need to use such methodological techniques in the study of promising antiviral substances. The proposed approach of research of promising antiviral compounds on the model of mixed viral infection can take place when creating new and studying the activity of already existing chemotherapeutic drugs, which may help prevent the development of viral resistance to drugs.
Employees of the department pay great attention to the pressing issues of combining scientific achievements with the practical needs of today, particularly in medicine. Methodological recommendations for preclinical studies of the specific antiviral action of drugs in cell culture on the model of adenoviruses have been developed. The use of the proposed unified methods will contribute not only to the introduction of highly effective drugs into medical practice, but also to the entry of domestic drugs into the world market (A.M. Shcherbinska, N.S. Dyachenko, S.L. Rybalko, L.N. Nosach, S. Dyadyun. T., Vrynchanu N.O. Study of the antiviral effect of potential medicinal products//Methodical recommendations of the Ministry of Health, State Pharmacological Center.- 2000.- 40 p.; Nosach L.M., Povnytsia O.Yu. Preclinical study of specific antiviral action of drugs in cell culture on the adenovirus model. Methodological recommendations // Bulletin of Pharmacology and Pharmacy. - 2007, No. 9. - P. 52-64).
Together with "BIOPHARMA PLAZMA" LLC, the production of specific immunoglobulins against TORCH infections (cytomegalovirus, herpes 1, 2 and 6 types, Epstein-Barr virus and chlamydia) has been established. These drugs are successfully used in the treatment of the above-mentioned infections in medical institutions of Ukraine (in urology, obstetrics, neurological departments). 
For diagnostic purposes, immunosensors were created for the detection of specific antibodies to the Epstein-Barr virus, adenovirus, and herpes simplex virus in the blood sera of patients using the Plasmon-6 optoelectronic device.
Laboratory employees take an active part in international scientific and scientific-practical conferences: International Conference on Antiviral Research; International scientific conference of students and postgraduates "Youth and progress in biology"; International Conference "Biologically Active Substances and Materials"; Polish-Ukrainian Weigl Conference on Microbiology; International conferences "Bioresources and viruses"; Congresses of the Society of Microbiologists of Ukraine; Global Virus Network conference of virologists for the Scandinavia-Baltic-Ukraine region; International conference "Fundamental problems of creation of new substances and materials of chemical production"; 1st, 2nd and 3rd International Electronic Conference on Medicinal Chemistry, Department of Organic Chemistry, University of Mons-UMONS, Belgium.
The scientific achievements of employees have been repeatedly recognized with awards and scholarships, in particular, the Award "Intellectual potential of young scientists - to the city of Kyiv" (Zagorodnya S.D., 2003); Award named after D.K. Zabolotny (N.S. Dyachenko, L.N. Nosach, 1999); Acknowledgement from the mayor of Kyiv for a significant personal contribution to the development of national science and strengthening the scientific and technical potential of the capital (Nesterova N.V., 2007); Award named after D.K. Zabolotny (Nesterova N.V., 2002); Certificate of Honor of the Presidium of the National Academy of Sciences of Ukraine (Nesterova N.V., 2008), Award of the President of Ukraine for Young Scientists (Zagorodnya S.D., 2007), NASU Scholarship for Young Scientists (Golovan A.V., 2008 , Bilyavska L.O., 2015), Winner of the contest "Kiyanka - 2009" in the Holosiivska District State Administration in the city of Kyiv in the nomination "Female Scientist of the Year" (Nesterova N.V., 2009), Award for young scientists and students of higher educational institutions for the best scientific papers in 2015 (Bilyavska L.O., Golovan A.V.); Grant of the President of Ukraine to support scientific research of young scientists for 2017 (Bilyavska L.O.).
The department cooperates with leading scientific institutions of Ukraine and abroad, such as: State University "Institute of Epidemiology and Infectious Diseases named after L.V. Gromashevsky" NAMNU, NSC "Institute of Biology and Medicine" of Kyiv National University named after Taras Shevchenko, Institute of Molecular Biology and Genetics of NASU, Institute of Organic Chemistry of NASU, Institute of Semiconductor Physics named after V.E. Lashkaryova NASU, Institute of Experimental Pathology of Oncology and Radiobiology named after RE. Kavetsky NASU, Institute of Organic Chemistry with Center of Phytochemistry, BAS, Bulgaria, Nazarbayev University Research and Innovation System, Astana, Kazakhstan, Institute of Technology, University of Tartu, Tartu, Estonia.

ONGOING PROJECTS

  • • 2015-2019 "Features of the reproduction of human DNA-containing viruses under the influence of potential antiviral substances."
  • • 2017-2021 "Antimicrobial and antiviral activity of new promising drugs and development of approaches to their controlled delivery" (within the framework of the comprehensive program of the Department of Biochemistry, Physiology and Molecular Biology of the National Academy of Sciences of Ukraine "Molecular-genetic and biochemical mechanisms of regulation of cellular and systemic interactions under physiological and pathological conditions".
  • • 2013-2017 "Experimental operation and metrological support of a diagnostic device based on surface plasmon resonance for the diagnosis of the herpes virus. Development of diagnostic methods". (Comprehensive scientific and technical program of the National Academy of Sciences of Ukraine "Sensor devices for medical-ecological and industrial-technological needs: metrological support and experimental operation").
Publications
  1. K Naumenko, A Golovan, G Baranova, S Zagorognya, Yu Shermolovytch. Antiviral and immunostimulatory potential of fluorine containing triazoles // Bulletin of Taras Shevchenko National University of Kyiv. Series: Biology.-2016.-72, N2.-P. 20-24.
  2. Klochko V.V., Zagorodnya S.D., Reva O.N. Genes Encoding Synthesis of Phenazine-1-carboxylic acid in Pseudomonas batumici // Biotechnologia Acta.-2016.-9, N5.-P. 45-53.
  3. Naumenko K., Zagorodnya S., Golovan A., Kovtyn V. Antiviral and apoptosis modulating effect of methylenebisphosphonic acids // Advances in Biology.-2016.-9, 3.- P. 1896-1902.
  4. Pankivska Yu., Biliavska L., Povnitsa O., Zagorodnya S. Antiadenoviral activity of fluoride-containing nucleosides and bisphonates derivatives // Microbiology and biotechnology.-2016.- 36, №3.- P. 60-70.
  5. Болтовець П.М., Нестерова Н.В. Поверхневий плазмонний резонанс: підходи і перспективи використання для дослідження вірус-специфічних взаємодій. Огляд літератури // Мікробіол. журнал.-2016.-78, №4.- С. 109-124.
  6. Alexeeva I., Nosach L., Palchykovska L., Usenko L, Povnitsa O. Synthesis and Comparative Study of Anti-Adenoviral Activity of 6-Azacytidine and Its Analogues // Nucleosides. Nucleotides. Nucleic Acids. – 2015. – 34, №8. – P. 565 – 578.
  7. Загородняя С.Д., Баранова Г.В., Нестерова Н.В. Влияние доксорубицина и этапозида на процесс СD-95 опосредованого апоптоза в ВЭБ-инфицированных клетках BC41 и DG-75 // Микробиол. журнал.-2015.-77, №4.- С.69-74.
  8. Білявська Л.О., Повниця О.Ю., Загородня С.Д., Нестерова Н.В., Шамара Ю.Б., Науменко К.С. Синтез мажорних вірусних білків за умов змішаного інфікування клітин МДВК // Доповіді НАН України.-2015, №8.- С.115-121.
  9. Затовська Т.В., Нестерова Н.В., Баранова Г.В., Рибалко С.Д., Загородня С.Д. Детекція специфічних антитіл до ВПГ-1 за допомогою оптичного біосенсора // Мікробіологія і біотехнологія.-2015.-3.- С.58-66.
  10. Крутьських Т.В., Нестерова Н.В., Загородня С.Д. Вивчення притигерпетичної активності таблеток Альтабор // Фармацевтичний часопис.-2015.-2.-С. 108-110.
  11. Белявская Л.А., Повница О.Ю., Загородняя С.Д., Войчук С.И., Зеленая Л.Б., Шамара Ю.Б., Нестерова Н.В. Моделирование смешанной адено-герпетической инфекции клеток MDBK, ее характеристика и особенности антивирусного действия веществ // Молодой ученый.-2014.-76, №17.- С.117-125.
  12. Dolashka P., Nesterova N., Zagorodnya S., Dolashka A., Baranova G., Golovan A., Voelter W. Antiviral Activity of Hemocyanin Rapana venosa and Its Isoforms Against Epstein-Barr Virus // Global Journal of Pharmacology. – 2014. – 8. – P.206 – 212.
  13. Biliavska L., Povnitsa O., Nesterova N., Zagorodnya S., Bobko T., Golovan A., Voychuk S. Study of adenovirus reproduction in different lymphoblastoid cell lines // Mikrobiologichny zhurnal. – 2014. –76, №6. – С. 24 – 30.
  14. Серый С.А., Тимошенко В.М., Власенко Ю.Г., Баранова Г.В., Загородня С.Д., Нестерова Н.В. Реакции Пуммерера тиопирановых производных как метод получения трифторметилзамещенных тиоланов, проявляющих антивирусную активность // Химия гетероциклических соединений. – 2014. – 4. – С. 513 – 526.
  15. Белявская Л.А., Повница О.Ю., Шермолович ЮГ., Гудзь А.П., Нестерова Н.В. Исследование антивирусной активности новых фторсодержащих гетероциклических соединений // Микробиология и биотехнология.- 2014.-1.-С.19-26.
  16. Головань А.В., Загородня С.Д., Нестерова Н.В. Вивчення апоптоз стимулюючого впливу похідних ізонікотинової кислоти на моделях латентної і гострої ВЕБ-інфекції // Доповіді НАНУ.- 2014.- 1.- С. 153-159.
  17. Рибалко С.Л., Носач Л.М., Повниця О.Ю., Дядюн С.Т., Максименок О.В., Трохимчук Т.Ю., Сокуренко Л.М., Чайковський Ю.Б. Порівняльне дослідження анти-ВІЛ дії нуклеозидних інгібіторів зворотньої транскриптази зидовудину та ламівудину (ЗАТ ФФ «Дарниця», Україна), їх аналогів ретровіру та епівіру («Glaxo Wellcome», Великобританія) // Ліки України плюс – 2013 . –14, №1. –C.34 – 38.
  18. Канищев А.С., Гудзь А.П., Шермолович Ю.Г., Загородня С.Д., Головань А.В., Курова А.О., Нестерова Н.В. Синтез 2-N-замещенных-4-тозил-5-полифторамил-1,2,3-триазолов и изучение их антивирусной активности по отношению к вирусу Эпшнейна-Барр // Журнал органічної та фармацевтичної хімії. – 2012. –1, №37. – С. 30 – 37.
  19. Нестерова Н.В., Рыбалко С.Л., Загородняя С.Д., Баранова Г.В., Головань А.В. Иммуносенсорная тест-система для выявления антител к вирусу Эпштейна-Барр // Биотехнология.-2012.-5, №1.- С. 100-106.
  20. Рибалко С.Л., Носач Л.М., Повниця О.Ю., Дядюн С.Т., Максименок О.В. Дослідження анти-ВІЛ дії препаратів, виробництва ЗАТ «ФФ»Дарниця»(Україна), «Glaxo Wellcome» (Великобританія), Boehringer ingelheim (Німеччина) // Лабораторна діяаностика.-2012.-61, №3.- 37-42.
  21. Загородняя С.Д., Повница О.Ю, Баранова Г.В., Головань А.В., Курова А.О., Белявская Л.О., Нестерова Н.В. Моделирование адено-герпетической инфекции в лимфобластоидных культурах клеток // Мікробіол. журнал – 2012. –74, №4. – С.64 – 72.
Patents
  • Utility model patent UA No. 93023 "2-(3-chlorotetrahydrofuran-2-yl)-4-tosyl-5-(trifluoromethyl)-2H-1,2,3-triazole" L.O. Bilyavska, O.Yu Povnytsia., Shermolovich Y.G., Gudz H.P., Nesterova N.V. - 2014.
  • Utility model patent UA No. 68403 "2-β-D-ribofurinosyl-5-actino-1,2,4-triazine-3(2H)-OH (6-Azicytidine) as an Epstein-Barr virus inhibitor" Nesterova N.V., Zagorodnya S.D., Golovan A.V., Baranova G.V., Alekseeva I.V., Palchikovska L.I. - 2012.
  • Utility model patent UA No. 51125 "Immunosensor test system based on surface plasmon resonance for detection of antibodies against the Epstein-Barr virus" N.V. Nesterova, S.D. Zagorodnya, G.V. Baranova, A.V. Golovan, Ushenin Yu.V., Khristosenko R.V. - 2010.
  • Utility model patent UA No. 46973 "Immunosensor test system for detection of antibodies against human adenoviruses in blood sera" Nesterova N.V., Nosach L.M., Povnytsia O.Yu., Zagorodnya S.D., Baranova G.V ., Golovan A. V., Ushenin Yu. V., Khristosenko R. V. – 2010.
  • Patent of Ukraine for the invention No. 87251 "Use of aqueous extract from Artemia cysts as an antiviral agent with anti-adenoviral and anti-herpetic activities" Rudneva I.I., Shayda V.G., Nosach L.M., Povnytsia O.Yu. - 2009.
  • Utility model patent UA No. 27400 "4-(N-benzyl)aminocarbonyl-1-methylpyridinium iodide (AMIZON) - Epstein-Barr virus inhibitor" Nesterova N.V., Zagorodnya S.D., Danylenko V.P., Bukhtiarova T.A., Baranova G.V., Bobkova L.S., Rybalko S.L. - 2007.
Employees of the department
Olga Povnytsia

Researcher
ph.: +38 (044) 526-34-79
fax +38 (044) 526-23-79
povnitsa@ukr.net

Research interests
• Study of the interaction of adenovirus and herpes simplex virus with the cell during a productive viral infection and the influence of inhibitors of various nature;
•Development and implementation of schemes and methods of preclinical research of the specific antiviral effect of potential drugs to predict their pharmacological activity
• Study of lymphotropic properties of human adenoviruses;
• Modeling of mixed viral infection in vitro, study of the effect of viral infection on cells, mutual influence of viruses and antiviral activity of substances in these conditions;
• Study of the influence of microgravity conditions on the reproduction of human adenovirus in cell culture.

Tetiana Zatovska

Researcher
tzatovskaya@yandex.ru

Research interests
• Investigation of virus-protein interaction by surface plasmon resonance
• Development of biosensor chips based on surface plasmon resonance for the detection of specific antibodies to HSV-1.

Liubov Biliavska

junior researcher, PhD
bilyavskal@ukr.net

Research interests
• Screening of anti-herpetic and anti-adenoviral substances of various origins
• Study of the mechanisms of action of promising antiviral substances
• Studying the influence of virus reproduction on the morphology and functional properties of cells
•Research of the peculiarities of the interaction between viruses under the conditions of co-infection (analysis of nucleic acids and proteins of viruses) and the effectiveness of official and new anomalous nucleotides and nucleosides

Halyna Baranova

Leading engineer
gbar_imv@ukr.net

Research interests
• Research of the antiviral action of substances of various nature against human viruses;
•Research in the field of sensor systems and technologies
• Development of immunobiological drugs for diagnosis and serotherapy of viral infections.

Hanna Holovan

junior researcher, PhD
golovan26@ukr.net

Research interests
• Screening of antiherpetic agents of various nature
• Research of virus-cell and virus-virus interactions (cell structure, viral and cellular factors regulating the viral replicative cycle, analysis of nucleic acids, proteins and other substances produced by the virus)
• Viral carcinogenesis on the example of EBV and research on anticancer drugs

Krystyna Naumenko

junior researcher
Kristin_Naumenko@gmail.com

Research interests
•Research of virus-cell and virus-virus interactions on the model of the latent and lytic form of EBV infection
• Study of viral proteins and their functions
• Predicting the biological activity of compounds using various in silico methods
• Investigation of the features of the action of antiviral compounds in vitro and in vivo.
• Study of the induction of lytic infection by EBV and apoptosis-stimulating compounds

Iuliia Pankivska

PhD student
pankivska.yulia@gmail.com

Research interests
•Study of the structural organization and immunogenic properties of adenovirion capsid proteins for the creation of immunodiagnostics for adenovirus infection;
• The search for new effective antiviral substances of various origins in relation to human adenoviruses;
• Study of the features of mixed viral infection;
• Development of new diagnostic systems for detection of viral pathogens.
• Investigation of the specifics of the effect of antiviral compounds on productive adenovirus infection
• Molecular and biological studies of virus-cell interactions
• In vivo study of antiviral activity of promising in vitro substances (mice, rabbits)

Nina Tkachenko

Laboratory assistant