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<article article-type="research-article" dtd-version="1.3" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xml:lang="ru"><front><journal-meta><journal-id journal-id-type="publisher-id">mimmun</journal-id><journal-title-group><journal-title xml:lang="ru">Медицинская иммунология</journal-title><trans-title-group xml:lang="en"><trans-title>Medical Immunology (Russia)</trans-title></trans-title-group></journal-title-group><issn pub-type="ppub">1563-0625</issn><issn pub-type="epub">2313-741X</issn><publisher><publisher-name>SPb RAACI</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.15789/1563-0625-2017-5-521-528</article-id><article-id custom-type="elpub" pub-id-type="custom">mimmun-1351</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group><subj-group subj-group-type="section-heading" xml:lang="ru"><subject>ОРИГИНАЛЬНЫЕ СТАТЬИ</subject></subj-group><subj-group subj-group-type="section-heading" xml:lang="en"><subject>ORIGINAL ARTICLES</subject></subj-group></article-categories><title-group><article-title>ВЛИЯНИЕ АРГИНИНДЕИМИНАЗЫ STREPTOCOCCUS PYOGENES НА МИГРАЦИОННУЮ АКТИВНОСТЬ И СТРУКТУРУ ЦИТОСКЕЛЕТА ЭНДОТЕЛИАЛЬНЫХ КЛЕТОК ЧЕЛОВЕКА</article-title><trans-title-group xml:lang="en"><trans-title>EFFECT OF ARGININE DEIMINASE FROM STREPTOCOCCUS PYOGENES ON CYTOSKELETON STRUCTURE AND MIGRATION ACTIVITY OF HUMAN ENDOTHELIAL CELLS</trans-title></trans-title-group></title-group><contrib-group><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Старикова</surname><given-names>Э. А.</given-names></name><name name-style="western" xml:lang="en"><surname>Starikova</surname><given-names>E. A.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Старикова Э.А. – кандидат биологических наук, старший научный сотрудник, отдел иммунологии ФГБНУ «Институт экспериментальной медицины».</p></bio><bio xml:lang="en"><p>Starikova E.A., PhD (Biology), Senior Research Associate, Department of Immunology, Institute of Experimental Medicine.</p><p>St. Petersburg.</p></bio><email xlink:type="simple">Starickova@yandex.ru</email><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Маммедова</surname><given-names>Дж. Т.</given-names></name><name name-style="western" xml:lang="en"><surname>Mammedova</surname><given-names>J. T.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Маммедова Дж.Т. – аспирант ФГБОУ ВО «СанктПетербургский государственный технологический институт (технический университет)».</p><p>СанктПетербург.</p></bio><bio xml:lang="en"><p>Mammedova J.T., Research Fellow, St. Petersburg State Technological University.</p><p>St. Petersburg.</p></bio><xref ref-type="aff" rid="aff-2"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Бурова</surname><given-names>Л. А.</given-names></name><name name-style="western" xml:lang="en"><surname>Burova</surname><given-names>L. A.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Бурова Л.А. – доктор медицинских наук, ведущий научный сотрудник, отдел молекулярной микробиологии ФГБНУ «Институт экспериментальной медицины».</p><p>СанктПетербург.</p></bio><bio xml:lang="en"><p>Burova L.A., PhD, MD (Medicine), Leading Research Associate, Department of Molecular Microbiology, Institute  of Experimental Medicine.</p><p>St. Petersburg.</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Соколов</surname><given-names>А. В.</given-names></name><name name-style="western" xml:lang="en"><surname>Sokolov</surname><given-names>A. V.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Соколов А.В. – кандидат биологических наук, заведующий лабораторией, отдел молекулярной генетики ФГБНУ «Институт экспериментальной медицины»; ФГБОУ ВО «СанктПетербургский государственный университет».</p><p>Санкт-Петербург.</p></bio><bio xml:lang="en"><p>Sokolov A.V., PhD (Biology), Head of Laboratory, Department of Molecular Genetics, Institute of Experimental Medicine;  St. Petersburg State University.</p><p>St. Petersburg.</p></bio><xref ref-type="aff" rid="aff-3"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Васильев</surname><given-names>В. Б.</given-names></name><name name-style="western" xml:lang="en"><surname>Vasilyev</surname><given-names>V. B.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Васильев В.Б. – доктор медицинских наук, заведующий отделом молекулярной генетики ФГБНУ «Институт экспериментальной медицины»; ФГБОУ ВО «СанктПетербургский государственный университет».</p><p>Санкт-Петербург.</p></bio><bio xml:lang="en"><p>Vasilyev V.B., PhD, MD (Medicine), Head, Department  of Molecular Genetics, Institute of Experimental Medicine;  St. Petersburg State University.</p><p>St. Petersburg.</p></bio><xref ref-type="aff" rid="aff-3"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Фрейдлин</surname><given-names>И. С.</given-names></name><name name-style="western" xml:lang="en"><surname>Freidlin</surname><given-names>I. S.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Фрейдлин И.С. – доктор медицинских наук, член.-корр. РАН, главный научный сотрудник, отдел иммунологии ФГБНУ «Институт экспериментальной медицины»; ФГБОУ ВО «Санкт-Петербургский государственный университет»; ФГБОУ ВО «Первый СанктПетербургский государственный медицинский университет им. акад. И.П. Павлова».</p><p>Санкт-Петербург.</p></bio><bio xml:lang="en"><p>Freidlin I.S., PhD, MD (Medicine), Corresponding Member, Russian Academy of Medical Sciences, Main Research Associate, Department of Immunology, Institute  of Experimental Medicine; St. Petersburg State University; Pavlov First St. Petersburg State Medical University.</p><p>  St. Petersburg.</p></bio><xref ref-type="aff" rid="aff-4"/></contrib></contrib-group><aff-alternatives id="aff-1"><aff xml:lang="ru"><institution>ФГБНУ «Институт экспериментальной медицины».</institution><country>Россия</country></aff><aff xml:lang="en"><institution>Institute of Experimental Medicine.</institution><country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-2"><aff xml:lang="ru"><institution>ФГБОУ ВО «Санкт-Петербургский государственный технологический институт (технический университет)».</institution><country>Россия</country></aff><aff xml:lang="en"><institution>St. Petersburg State Technological University.</institution><country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-3"><aff xml:lang="ru"><institution>ФГБНУ «Институт экспериментальной медицины»;  ФГБОУ ВО «Санкт-Петербургский государственный университет».</institution><country>Россия</country></aff><aff xml:lang="en"><institution>Institute of Experimental Medicine; St. Petersburg State University.</institution><country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-4"><aff xml:lang="ru"><institution>ФГБНУ «Институт экспериментальной медицины»;  ФГБОУ ВО «Санкт-Петербургский государственный университет»;  ФГБОУ ВО «Первый Санкт-Петербургский государственный медицинский университет  им. акад. И.П. Павлова».</institution><country>Россия</country></aff><aff xml:lang="en"><institution>Institute of Experimental Medicine; St. Petersburg State University;  Pavlov First St. Petersburg State Medical University.</institution><country>Russian Federation</country></aff></aff-alternatives><pub-date pub-type="collection"><year>2017</year></pub-date><pub-date pub-type="epub"><day>18</day><month>10</month><year>2017</year></pub-date><volume>19</volume><issue>5</issue><fpage>521</fpage><lpage>528</lpage><permissions><copyright-statement>Copyright &amp;#x00A9; Старикова Э.А., Маммедова Д.Т., Бурова Л.А., Соколов А.В., Васильев В.Б., Фрейдлин И.С., 2017</copyright-statement><copyright-year>2017</copyright-year><copyright-holder xml:lang="ru">Старикова Э.А., Маммедова Д.Т., Бурова Л.А., Соколов А.В., Васильев В.Б., Фрейдлин И.С.</copyright-holder><copyright-holder xml:lang="en">Starikova E.A., Mammedova J.T., Burova L.A., Sokolov A.V., Vasilyev V.B., Freidlin I.S.</copyright-holder><license xml:lang="ru" license-type="creative-commons-attribution" xlink:href="https://creativecommons.org/licenses/by/4.0/" xlink:type="simple"><license-p>Данная работа распространяется под лицензией Creative Commons Attribution 4.0.</license-p></license><license xml:lang="en" license-type="creative-commons-attribution" xlink:href="https://creativecommons.org/licenses/by/4.0/" xlink:type="simple"><license-p>This work is licensed under a Creative Commons Attribution 4.0 License.</license-p></license></permissions><self-uri xlink:href="https://www.mimmun.ru/mimmun/article/view/1351">https://www.mimmun.ru/mimmun/article/view/1351</self-uri><abstract><p>В последнее время в литературе накапливается всe больше данных о цитопатическом влиянии бактериальной аргининдеиминазы на эндотелиальные клетки человека, однако точные механизмы эндотелиальной дисфункции, вызванной активностью фермента, остаются слабоизученными. Активность аргининдеиминазы истощает запасы аргинина в микроокружении клеток в различных тканях организма-хозяина. Исходя из данных о том, что аргинилирование белков актинового цитоскелета регулирует их функции, мы предположили, что цитопатическое действие аргининдеиминазы S. pyogenes в отношении эндотелиальных клеток может быть связано с нарушением структуры их цитоскелета. Целью исследования было изучение влияния аргининдеиминазы S. pyogenes на миграционную активность и структуру актинового цитоскелета эндотелиальных клеток линии EA.hy926. В работе использовали супернатанты разрушенных S. pyogenes M49-16 и его изогенного мутанта с делецией гена аргининдеиминазы S. pyogenes М49-16delAD, а также супернатанты разрушенных S. pyogenes M22 и аргининдеиминазу, выделенную из стрептококков этого штамма. Влияние бактериальных компонентов на миграционную активность эндотелиальных клеток изучали в модели «раны» in vitro. Для анализа влияния бактериальных компонентов на структуру актинового цитоскелета проводили окрашивание клеток фаллоидин-родамином. Было показано, что супернатанты разрушенных S. pyogenes, так же как выделенная из супернатанта аргининдеиминаза, вызывали достоверное снижение миграционной активности эндотелиальных клеток и изменения структуры их актинового цитоскелета. Супернатант разрушенных S. pyogenes М49-16delAD с делецией гена аргининдеиминазы отличался достоверно ослабленной способностью подавлять миграцию клеток по сравнению с супернатантом разрушенных S. pyogenes М49-16. Это различие между штаммами не сопровождалось существенными различиями в характере влияния на структуру актиновых филаментов у клеток, культивируемых в присутствии соответствующих супернатантов. Добавление экзогенного аргинина в культуру клеток, содержащую супернатанты разрушенных S. pyogenes, не приводило к восстановлению их миграционной активности и структуры актинового цитоскелета. Однако при добавлении аргинина в культуру клеток, содержащую выделенную из сунернатанта аргининдеиминазу, миграционная активность клеток достоверно усиливалась, а структура актинового цитоскелета эндотелиальных клеток восстанавливалась. Cнижение миграционной активности эндотелиальных клеток под влиянием стрептококковой аргинидеиминазы сопряжено с нарушением структуры их актинового цитоскелета на фоне деплеции аргинина.</p></abstract><trans-abstract xml:lang="en"><p>There is a growing body of data about the cytopathic effect of bacterial arginine deiminase on human endothelial cells, but the precise mechanisms of endothelial dysfunction caused by the activity of the enzyme remain poorly understood. Activity of arginine deiminase causes arginine depletion in the microenvironment of the host organism cells. In view that arginylation of beta-actin regulates actin cytoskeleton structure and cell motility, we proposed that the cytopathic effect of arginine deiminase may be associated with disruption of actin in the cytoskeleton of endothelial cells. The aim of this study was to investigate the effect of arginine deiminase from S. pyogenes on migration and actin cytoskeleton structure of the human endothelial cells, line EA.hy926. The supernatant of sonicated S. pyogenes M49-16, its isogenic mutant with a deletion of the arginine deiminase gene (S. pyogenes M49-16delAD), supernatant of sonicated S. pyogenes M22, and arginine deiminase isolated from the latter strain were used. The effect of bacterial factors on migration activity of endothelial cells was studied in the model of "wound healing" in vitro. To analyze the influence of bacterial factors on the actin cytoskeleton structure, cells were stained with phalloidin-rhodamine. It was shown that supernatants of destroyed S. pyogenes, as well as arginine deiminase significantly reduced the migration activity of endothelial cells and altered the structure of their actin cytoskeleton. The supernatants of destroyed S. pyogenes M49-16delAD with  deleted gene of arginine deiminase showed a significantly reduced ability to suppress cell migration as compared with the supernatant of sonicated S. pyogenes M49-16. No significant differences were revealed in the structure of actin filaments in cells cultured in the presence of supernatants of destroyed S. pyogenes M19-16, and cells cultured in the presence of isogenic mutant S. pyogenes M4916delAD. Adding exogenous arginine to the cells cultured with supernatants of destroyed S. pyogenes did not restore their migratory activity and the structure of their actin cytoskeleton. However, if arginine deficiency caused by the activity of arginine deiminase was compensated, endothelial cells migration activity was restored, and the structure of actin cytoskeleton was recovered. A decrease of migration activity of endothelial cells under the influence of streptococcal arginine deiminase was due to the disruption of actin cytoskeleton structure.</p></trans-abstract><kwd-group xml:lang="ru"><kwd>S. pyogenes</kwd><kwd>аргининдеиминаза</kwd><kwd>эндотелиальные клетки</kwd><kwd>метаболизм аргинина</kwd><kwd>миграция</kwd><kwd>цитоскелет</kwd></kwd-group><kwd-group xml:lang="en"><kwd>S. pyogenes</kwd><kwd>arginine deiminase</kwd><kwd>endothelial cells</kwd><kwd>arginine metabolism</kwd><kwd>cell migration</kwd><kwd>cytoskeleton</kwd></kwd-group></article-meta></front><back><ref-list><title>References</title><ref id="cit1"><label>1</label><citation-alternatives><mixed-citation xml:lang="ru">Старикова Э.А., Лебедева А.М., Бурова Л.А., Фрейдлин И.С. 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