vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJ20.659vavilov-2781Research ArticleМОЛЕКУЛЯРНАЯ И КЛЕТОЧНАЯ БИОЛОГИЯMOLECULAR AND CELL BIOLOGYCpG олигонуклеотиды с модифицированными фосфатными группами индуцируют созревание миелоидных дендритных клеток человека in vitroPhosphate-modified CpG oligonucleotides induce in vitro maturation of human myeloid dendritic cellshttps://orcid.org/0000-0001-6895-938XОстанинА. А.OstaninA. A.

Новосибирск

Novosibirsk

ostanin62@mail.ruhttps://orcid.org/0000-0003-3169-8643ЛеплинаО. Ю.LeplinaO. Y.

Новосибирск

Novosibirsk

https://orcid.org/0000-0001-6644-5959БураковаЕ. А.BurakovaE. A.

Новосибирск

Novosibirsk

https://orcid.org/0000-0002-7987-2017ТыриноваТ. В.TyrinovaT. V.

Новосибирск

Novosibirsk

https://orcid.org/0000-0001-5692-2483ФокинаА. А.FokinaA. A.

Новосибирск

Novosibirsk

https://orcid.org/0000-0002-7650-4331ПроскуринаА. С.ProskurinaA. S.

Новосибирск

Novosibirsk

https://orcid.org/0000-0002-2019-9382БогачевС. С.BogachevS. S.

Новосибирск

Novosibirsk

https://orcid.org/0000-0002-5215-5704СтеценкоД. А.StetsenkoD. A.

Новосибирск

Novosibirsk

https://orcid.org/0000-0003-2346-6279ЧерныхЕ. Р.ChernykhE. R.

Новосибирск

Novosibirsk

Научно-исследовательский институт фундаментальной и клинической иммунологииРоссияResearch Institute of Fundamental and Clinical ImmunologyRussian FederationНовосибирский национальный исследовательский государственный университет; Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наукРоссияNovosibirsk State University; Institute of Cytology and Genetics of Siberian Branch of the Russian Academy of SciencesRussian FederationНаучно-исследовательский институт фундаментальной и клинической иммунологии; Новосибирский национальный исследовательский государственный университетРоссияResearch Institute of Fundamental and Clinical Immunology; Novosibirsk State UniversityRussian FederationФедеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наукРоссияInstitute of Cytology and Genetics of Siberian Branch of the Russian Academy of SciencesRussian FederationНовосибирский национальный исследовательский государственный университет; Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наукРоссияNovosibirsk State University; Institute of Cytology and Genetics of Siberian Branch of the Russian Academy of SciencesRussian Federation202028102020246653660Copyright © Останин А.А., Леплина О.Ю., Буракова Е.А., Тыринова Т.В., Фокина А.А., Проскурина А.С., Богачев С.С., Стеценко Д.А., Черных Е.Р., 20202020Останин А.А., Леплина О.Ю., Буракова Е.А., Тыринова Т.В., Фокина А.А., Проскурина А.С., Богачев С.С., Стеценко Д.А., Черных Е.Р.Ostanin A.A., Leplina O.Y., Burakova E.A., Tyrinova T.V., Fokina A.A., Proskurina A.S., Bogachev S.S., Stetsenko D.A., Chernykh E.R.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/2781

Миелоидные дендритные клетки (ДК) играют важную роль в иммунном ответе, поэтому актуальной задачей является поиск соединений, способных эффективно активировать ДК. Целью настоящей работы было изучение влияния синтетических CpG олигодезоксинуклеотидов (CpG-ODN) на созревание и аллостимуляторную активность миелоидных ДК в сравнении с другими PAMP и DAMP молекулами. Для исследований были синтезированы CpG-ODN класса С (SD-101 и D-SL03), содержащие тиофосфатные межнуклеотидные группы, а также получены их оригинальные фосфат-модифицированные аналоги (SD-101М и D-SL03М) с мезилфосфорамидными межнуклеотидными группами (М = μ-модификация). Эффекты CpG-ODN и других активаторов оценивали в культурах ДК, генерированных из моноцитов крови в присутствии GM-CSF и IFN-α (IFN-ДК) или IL-4 (IL4-ДК). Оценка внутриклеточной экспрессии TLR-9 показала, что оба типа ДК (IFN-ДК и IL4-ДК) содержали в среднем 52 и 80 % TLR-9-позитивных клеток соответственно. Исследуемые CpG-ODN усиливали аллостимуляторную активность IFN-ДК, причем эффект μ-модифицированных CpG-ODN был выше, чем тиофосфатных CpG-ODN. Стимулирующий эффект CpG-ODN в дозе 1.0 мкг/мл был сопоставим (для D-SL03, D-SL03М, SD-101) или превышал (для SD-101М) действие липополисахарида (LPS) в дозе 10 мкг/мл. При этом IFN-ДК характеризовались большей чувствительностью к действию CpG-ODN, чем IL4-ДК. Усиление аллостимуляторной активности ДК в присутствии CpG-ODN было связано с индукцией конечного созревания клеток, что подтверждалось значимым снижением количества СD14+ ДК, увеличением доли зрелых CD83+ ДК и тенденцией к возрастанию CD86+ ДК. Интересно, что характерная для LPS способность усиливать экспрессию костимуляторной молекулы OX40L на ДК была выявлена только для μ-аналога SD-101М. Кроме того, CpG-ODN (SD-101 и SD-101М) оказывали стимулирующий эффект на продукцию IFN-γ, сопоставимый с действием LPS. Полученные в целом данные свидетельствуют о стимулирующем действии CpG-ODN на созревание и аллостимуляторную активность миелоидных ДК человека, которое более выражено для μ-модифицированных аналогов.

Myeloid dendritic cells (DCs) play an important role in the immune response; therefore, the search for compounds that can effectively activate DCs is a needful goal. This study was aimed to investigate the effect of synthetic CpG oligodeoxynucleotides (CpG-ODN) on the maturation and allostimulatory activity of myeloid DCs in comparison with other PAMP and DAMP molecules. For the research, we synthesized known CpG-ODN class C (SD-101 and D-SL03) containing thiophosphate internucleotide groups, and their original phosphate-modified analogues (SD-101M and D-SL03M) with mesylphosphoramide internucleotide groups (M = μ-modification). The effects of CpG-ODN and other activators were evaluated on DCs generated from blood monocytes in the presence of GM-CSF and IFN-α (IFN-DC) or IL-4 (IL4-DC). Evaluation of the intracellular TLR-9 expression showed that both types of DCs (IFN-DC and IL4-DC) contained on average 52 and 80 % of TLR-9-positive cells, respectively. The CpG-ODNs studied enhanced the allostimulatory activity of IFN-DCs, and the effect of μ-modified CpG-ODNs was higher than that of CpG-ODNs with thiophosphate groups. The stimulating effect of CpG-ODN at a dose of 1.0 μg/ml was comparable (for D-SL03, D-SL03M, SD-101) with or exceeded (for SD-101M) the effect of LPS at a dose of 10 μg/ml. At the same time, IFN-DCs were characterized by greater sensitivity to the action of CpG-ODNs than IL4-DCs. The enhancement of DC allostimulatory activity in the presence of CpG-ODNs was associated with the induction of final DC maturation, which was confirmed by a significant decrease in the number of CD14+DC, an increase in mature CD83+DC and a trend towards an increase in CD86+DC. Interestingly, the characteristic ability of LPS to enhance the expression of the co-stimulatory molecule OX40L on DCs was revealed only for the μ-analogue SD-101M. In addition, CpG-ODNs (SD-101 and SD-101M) had a stimulatory effect on IFN-γ production comparable to the action of LPS. The data obtained indicate a stimulating effect of CpG-ODN on the maturation and allostimulatory activity of human myeloid DCs, which is more pronounced for μ-modified analogs.

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