vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJ17.289vavilov-1193Research ArticleМОЛЕКУЛЯРНАЯ ГЕНЕТИКА И КЛЕТОЧНАЯ БИОЛОГИЯMOLECULAR GENETICS AND CELL BIOLOGYЭкспрессия генов цитокинов, антигенов дифференцировки и факторов транскрипции в дендритных клетках человека, активированных двуцепочечной ДНКExpression of genes of cytokines, transcription factors and differentiation antigens in human dendritic cells activated by double-stranded DNAПроскуринаА. С.ProskurinaA. S.likhacheva@bionet.nsc.ruОрищенкоК. Е.OrishchenkoK. E.likhacheva@bionet.nsc.ruПоттерЕ. А.PotterE. A.likhacheva@bionet.nsc.ruДолговаЕ. В.DolgovaE. V.likhacheva@bionet.nsc.ruСпасельниковаА. В.SpaselnikovaA. V.likhacheva@bionet.nsc.ruРиттерГ. С.RitterG. S.likhacheva@bionet.nsc.ruВараксинН. А.VaraksinN. A.likhacheva@bionet.nsc.ruРябичеваТ. Г.RyabichevaT. G.likhacheva@bionet.nsc.ruЛеплинаО. Ю.LeplinaO. Y.likhacheva@bionet.nsc.ruОстанинА. А.OstaninA. A.likhacheva@bionet.nsc.ruЧерныхЕ. Р.ChernykhE. R.likhacheva@bionet.nsc.ruБогачевС. С.BogachevS. S.likhacheva@bionet.nsc.ruФедеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук, НовосибирскРоссияInstitute of Cytology and Genetics SB RAS, NovosibirskRussian FederationФедеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук, Новосибирск; Новосибирский национальный исследовательский государственный университет, НовосибирскРоссияInstitute of Cytology and Genetics SB RAS, Novosibirsk; Novosibirsk State University, NovosibirskRussian FederationАкционерное общество «Вектор-Бест», р. п. Кольцово, Новосибирская областьРоссияState Research Center of Virology and Biotechnology “Vector”, Koltsovo, Novosibirsk regionRussian FederationНаучно-исследовательский институт фундаментальной и клинической иммунологии, НовосибирскРоссияInstitute of Fundamental and Clinical Immunology, NovosibirskRussian Federation201729112017216717727Copyright © Проскурина А.С., Орищенко К.Е., Поттер Е.А., Долгова Е.В., Спасельникова А.В., Риттер Г.С., Вараксин Н.А., Рябичева Т.Г., Леплина О.Ю., Останин А.А., Черных Е.Р., Богачев С.С., 20172017Проскурина А.С., Орищенко К.Е., Поттер Е.А., Долгова Е.В., Спасельникова А.В., Риттер Г.С., Вараксин Н.А., Рябичева Т.Г., Леплина О.Ю., Останин А.А., Черных Е.Р., Богачев С.С.Proskurina A.S., Orishchenko K.E., Potter E.A., Dolgova E.V., Spaselnikova A.V., Ritter G.S., Varaksin N.A., Ryabicheva T.G., Leplina O.Y., Ostanin A.A., Chernykh E.R., Bogachev S.S.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/1193

Одно из важнейших свойств экстраклеточной двуцепочечной ДНК, имеющее отношение к терапии различных заболеваний, – это ее способность через дендритные клетки (ДК) активировать эффекторные клетки иммунной системы (противоопухолевый и вакцинальный иммунитет). Стимулирующий эффект ДНК на ДК опосредуется через TLR9 сигнальный путь и/или через систему цитозольных сенсоров и проявляется усилением экспрессии МНС антигенов II класса и костимулирующих молекул, а также синтеза иммунорегуляторных цитокинов. В настоящей работе была исследована экспрессия генов цитокинов, антигенов дифференцировки и факторов транскрипции в ДК, активированных двуцепочечной ДНК человека: (1) без использования каких-либо дополнительных факторов, (2) при использовании липофильного агента и (3) при блокировании TLR9 хлорохином. Оценка эффекта ДНК проводилась после 6- и 24-часовой экспозиции. Показано, что препарат двуцепочечной ДНК при трансфекции липофектамином активирует ДК на таком же уровне, как и Poly(dA : dT), синтетический аналог двуцепочечной ДНК. Установлено, что совместная обработка ДНК и хлорохином усиливает экспрессию генов ИФН-α, ИФН-β, ИФН-γ, ИЛ­8, МСР1, VEGF, CD25 и CD83 к 24 ч инкубации. Впервые показано, что геномная «self» двуцепочечная ДНК как монопрепарат активирует синтез мРНК ИФН-α, ИФН-β, ИФН-γ, ИЛ­8, ИЛ­10 и VEGF в ДК к 6 часам индукции.

 

 

 

 

One of the most important properties of extracellular double-stranded DNA related to the treatment of various diseases is its ability to activate effector cells of the immune system (anti-tumor and vaccinal immunity) through dendritic cells (DCs). The stimulatory effect of DNA on DCs is mediated by the TLR9 signaling pathway and/or through a system of cytosolic sensors and is manifested by increased expression of MHC class II antigens and costimulatory molecules and by increased synthesis of immunoregulatory cytokines. In this work, the expression of cytokines, differentiation antigens and transcription factor genes has been investigated in DCs activated by double-stranded human DNA (i) without any additional factors, (ii) using a lipophilic agent, and (iii) by blocking TLR9 with chloroquine. Evaluation of the DNA effect was carried out after the 6- and 24-hour exposure. It was found that the preparation of double-stranded DNA transfected by Lipofectamine 2000 boosts DCs at the same level as Poly(dA : dT), a synthetic equivalent of double-stranded DNA. It was discovered that combined application of DNA and chloroquine enhances expression of the IFN-α, IFN-β, IFN-γ, IL­8, МСР1, VEGF, CD25, and CD83 genes by hour 24 of incubation. It was for the first time shown that genomic “self” double-stranded DNA as a mono agent activates mRNA synthesis of cytokines IFN-α, IFN-β, IFN-γ, IL­8, IL­10, and VEGF in DCs at 6 hours of induction.

 

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The authors declare that there are no conflicts of interest present.