Cytokines and inflammationCytokines and inflammationЦитокины и воспаление1684-7849Eco-Vector64633110.17816/CI646331Original Study ArticlesОригинальные исследованияResearch ArticleThe effect of streptococcal arginine deiminase on Th17 lymphocyte differentiation in vitroВлияние стрептококковой аргининдеиминазы на дифференцировку Th17-лимфоцитов in vitrohttps://orcid.org/0000-0002-9687-74346488-4036StarikovaEleonora A.СтариковаЭлеонора А.
Russian Federation

Cand. Sci. (Biology)

канд. биол. наук

starickova@yandex.ruhttps://orcid.org/0000-0003-4381-69931418-6373MammedovaJennet T.МаммедоваДженнет Т.
Russian Federation
jennet_m@mail.ruhttps://orcid.org/0000-0001-9033-05377427-7395SokolovAlexey V.СоколовАлексей В.
Russian Federation

Dr. Sci. (Biology)

д-р биол. наук

biochemsokolov@gmail.comhttps://orcid.org/0000-0002-8493-52116025-1790RubinsteinArtem A.РубинштейнАртём А.
Russian Federation
arrubin6@mail.ruhttps://orcid.org/0000-0002-5716-08363449-0970EgidarovaElena Yu.ЕгидароваЕлена Ю.
Russian Federation
lena.egidarova.97@mail.ruhttps://orcid.org/0009-0007-0829-3978YakupovArslan M.ЯкуповАрслан М.
Russian Federation
arsl@n-yakupov.ruhttps://orcid.org/0009-0000-4345-4967AbdulkadyrovaNazkhanum Sh.АбдулкадыроваНазханум Ш.
Russian Federation
naza.v2003@gmail.comhttps://orcid.org/0000-0001-7204-78504903-7636KudryavtsevIgor V.КудрявцевИгорь В.
Russian Federation

Cand. Sci. (Biology), Assistant Professo

канд. биол. наук, доцент

igorek1981@yandex.ruInstitute of Experimental MedicineИнститут экспериментальной медициныI.P. Pavlov First Saint Petersburg State Medical UniversityПервый Санкт-Петербургский государственный медицинский университет им. академика И.П. ПавловаA. Almazov National Medical Research CenterНациональный медицинский исследовательский центр им. В.А. АлмазоваSaint-Petersburg State Institute of Technology (Technical University)Санкт-Петербургский государственный технологический институт (технический университет)Smorodintsev Research Institute of InfluenzaНаучно-исследовательский институт гриппа им. А.А. Смородинцева20022025150620242121111201701202512022025Copyright ©; 2024, Starikova E.A., Mammedova J.T., Sokolov A.V., Rubinstein A.A., Egidarova E.Y., Yakupov A.M., Abdulkadyrova N.S., Kudryavtsev I.V.Copyright ©; 2024, Старикова Э.А., Маммедова Д.Т., Соколов А.В., Рубинштейн А.А., Егидарова Е.Ю., Якупов А.М., Абдулкадырова Н.Ш., Кудрявцев И.В.2024Starikova E.A., Mammedova J.T., Sokolov A.V., Rubinstein A.A., Egidarova E.Y., Yakupov A.M., Abdulkadyrova N.S., Kudryavtsev I.V.Старикова Э.А., Маммедова Д.Т., Соколов А.В., Рубинштейн А.А., Егидарова Е.Ю., Якупов А.М., Абдулкадырова Н.Ш., Кудрявцев И.В.https://cijournal.ru/1684-7849/article/view/646331

BACKGROUND: The mTOR/S6K metabolic intracellular signaling cascade plays a key role in regulating the activation and differentiation of Th17/Treg lymphocyte populations. Pathogens can disrupt T-lymphocyte differentiation programs and weaken immune responses through arginine depletion mechanisms.

AIM: To evaluate the effect of bacterial arginine-hydrolyzing enzyme arginine deiminase on the balance of Th17/Treg helper T-cell polarization in vitro.

MATERIALS AND METHODS: Experiments were conducted using mononuclear leukocytes from healthy donors. Lymphocyte activation and differentiation were induced with activating antibodies and a cytokine cocktail. Recombinant Streptococcus pyogenes arginine deiminase was used. The enzyme’s arginine-hydrolyzing activity was confirmed by a modified Sakaguchi method. Mitochondrial oxidative phosphorylation activity was analyzed via MTT assay. The proportion of Th17, Treg, and IL-17A+/Foxp3+ transitional lymphocyte populations was assessed using monoclonal antibodies and flow cytometry.

RESULTS: The enzyme caused a significant sevenfold decrease in arginine concentration in mononuclear leukocyte cultures. The arginine deficiency induced by arginine deiminase activity could not be compensated even by supplementing supraphysiological concentrations of the amino acid. Differentiation was accompanied by a significant reduction in oxidative phosphorylation (p <0.001). Arginine deiminase inhibited mitochondrial respiration in both intact (p <0.001) and differentiated cells (p <0.05). The enzyme’s activity suppressed Th17 cell differentiation (p <0.05) but did not affect IL-17A+/Foxp3+ transitional cells, Treg lymphocytes, or the Th17/Treg ratio.

CONCLUSIONS: Arginine deiminase may disrupt adaptive immune development and diminish the effectiveness of protective immune responses.

Обоснование. mTOR/S6K-метаболический внутриклеточный сигнальный каскад играет ключевую роль в регуляции активации и дифференцировки Th17-/Тreg-популяций лимфоцитов. Патогенные микробы, используя механизм истощения аргинина, могут нарушать программу дифференцировки Т-лимфоцитов и снижать эффективность иммунного ответа.

Цель исследования — оценка влияния бактериального аргинин-гидролизующего фермента аргининдеиминазы на баланс поляризации Th17-/Тreg-популяций Т-хелперов in vitro.

Материалы и методы. Эксперименты проводили с использованием мононуклеарных лейкоцитов здоровых доноров. Активацию и дифференцировку лимфоцитов осуществляли с помощью активирующих антител и коктейля цитокинов. В работе использовали рекомбинантную аргининдеиминазу Streptococcus pyogenes. Аргинин-гидролизующую активность фермента подтверждали с помощью модифицированного метода Сaкагучи. В ММТ-тесте анализировали активность процессов окислительного фосфорилирования. Долю клеток Тh17, Treg, а также клеток переходной популяции IL-17A+/Foxp3+ лимфоцитов оценивали с использованием моноклональных антител и проточной цитометрии.

Результаты. Было установлено, что фермент вызывал достоверное семикратное снижение концентрации аргинина в культуре мононуклеарных лейкоцитов. Дефицит аргинина, вызванный активностью аргининдеиминазы, не удалось компенсировать даже за счёт добавки супрафизиологической концентрации аминокислоты. В ходе дифференцировки происходило достоверное снижение процессов окислительного фосфорилирования (p <0,001). Аргининдеиминаза подавляла митохондриальное дыхание как в интактных (p <0,001), так и в дифференцированных клетках (p <0,05). Активность фермента ингибировала дифференцировку Th17-клеток (p <0,05), но не влияла на долю переходных IL-17A+/Foxp3+ клеток, Treg-лимфоцитов и соотношения Th17/Treg.

Заключение. Аргининдеиминаза может нарушать программу развития адаптивного иммунитета и снижать эффективность защитных реакций.

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