mimmunМедицинская иммунологияMedical Immunology (Russia)1563-06252313-741XSPb RAACI10.15789/1563-0625-2015-5-455-460mimmun-933Research ArticleКРАТКИЕ СООБЩЕНИЯSHORT COMMUNICATIONSМЕХАНИЗМЫ АКТИВАЦИИ КИШЕЧНОАССОЦИИРОВАННОЙ ЛИМФОИДНОЙ ТКАНИ В УСЛОВИЯХ ХРОНИЧЕСКОГО СОЦИАЛЬНОГО СТРЕССАACTIVATION MECHANISMS OF GUT-ASSOCIATED LYMPHOID TISSUE UNDER CHRONIC SOCIAL STRESS CONDITIONSКамышныйА. М.KamyshnyiA. M.

Доктор медицинских наук, доцент, заведующий кафедрой микробиологии, вирусологии и иммунологии.

Адрес для переписки: Камышный Александр Михайлович Запорожский государственный медицинский университет 69035, Украина, г. Запорожье, пр. Маяковского, 26. Тел.: +38 (061) 234-26-31. Е-mail: alexkamyshny@yandex.ru

PhD, MD (Medicine), Associate Professor, Chief, Department of Microbiology, Virology and Immunology.

Address for correspondence: Kamyshnyi Alexandr M. Zaporozhye State Medical University, 69035,Ukraine, Zaporozhye, Mayakovsky ave, 26. Phone: +38 (061) 234- 26-31. Е-mail: alexkamyshny@yandex.ru

alexkamyshny@yandex.ruТополИ. А.TopolI. A.

Ассистент кафедры микробиологии, вирусологии и иммунологии

Assistant Professor, Department of Microbiology, Virology, and Immunology

alexkamyshny@yandex.ruЗапорожский государственный медицинский университетУкраинаZaporozhye State Medical UniversityUkraine201507102015175455460Copyright © Камышный А.М., Топол И.А., 20152015Камышный А.М., Топол И.А.Kamyshnyi A.M., Topol I.A.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.mimmun.ru/mimmun/article/view/933

Стресс-индуцированная иммунная дисрегуляция является фактором риска развития аутоиммунных и воспалительных заболеваний, однако механизмы такого влияния на сегодняшний момент в полной мере не известны.

Мы изучили уровень экспрессии мРНК NR3C1и Adrβ2-рецепторов, провоспалительных цитокинов IL-1β, IL-17α, Nlrp3-субъединицы инфламмасомы, а также выявили особенности распределения RORγt +, FoxP3+, LMP2+, XBP1+ лимфоцитов в КАЛТ крыс в условиях ХСС.

Эксперимент проводили на самках крыс линии Вистар в возрасте 5-6 месяцев. Для определения уровня экспрессии мРНК генов-мишеней проводили ОТ-ПЦР в реальном времени на амплификаторе CFX96™ Real-Time PCR Detection Systems («Bio-Rad Laboratories, Inc.», США). Относительный уровень экспрессии вышеуказанных генов оценивали по методу ΔΔCt, нормализуя по референс-гену GAPDH. Иммунопозитивные лимфоциты были идентифицированы с помощью метода непрямой иммунофлюоресценции с использованием моноклональных антител.

Развитие ХСС приводит к снижению уровня экспрессии мРНК Nr3c1 иAdrβ2-рецепторов в клетках КАЛТ и сопровождается однонаправленной динамикой по увеличению транскрипционной активности генов провоспалительных цитокинов IL-1β, IL-17α и Nlrp3-инфламмасомы. Данные изменения сопровождаются снижением соотношения FoxP3+/RORγt + клеток, свидетельствуя о доминировании Th17-дифференцировки на фоне супрессорной недостаточности. Развитие ХСС сопровождалось однонаправленной тенденцией по увеличению общего количества LMP2+ лимфоцитов и снижению плотности популяции ХВР1+ клеток в лимфоидных структурах подвздошной кишки крыс.

События, происходящие в КАЛТ в условиях ХСС, противоречат классической парадигме стресса и провоцируют не иммуносупрессию, а выраженную активацию иммунной системы и воспалительный процесс. 

Stress-induced immune disregulation is a risk factor of autoimmune and inflammatory diseases, but, so far, the mechanisms for this effect are not fully known. Expression levels of specific mRNAs were assessed in gut-associated lymphoid tissue (GALT) from Wistar rats subjected to chronic social stress (CSS). Gene expression was evaluated for NR3C1, Adrβ2, as well as IL-1β, IL-17α pro-inflammatory cytokines, and Nlrp, an inflammasome gene. Under the CSS conditions, we have shown altered distribution of RORγt +, FoxP3+, LMP2+, XBP1+ lymphocytes in GALT.

The experiments were carried out with female Wistar rats aged 5–6 months. Specific mRNA expression for the target genes was determined by means of real-time PCR performed in a CFX96™ thermocycler («BioRadLaboratories, Inc»,USA). Relative levels of a target gene expression were quantified by the ΔΔCt method, being compared with rat GAPDH reference gene expression. Statistical analysis was performed with available «BioRad СFX Manager 3.1» software. Specific monoclonal rat antibodes were used for detection of immunopositive lymphocytes by means of indirect immunofluorescence technique.

CSS development leads to decreased levels of mRNA expression for Nr3c1 and Adrβ2-genes in the GALT cells, being accompanied with unidirectional changes, i.e., increased transcription of pro-inflammatory cytokine mRNAs (IL-1β, IL-17α) and Nlrp3-inflammasome genes. These changes are accompanied by decreased FoxP3+/RORγt + cell ratio and predominant Th17 differentiation accompanied by suppressor failure. In addition, CSS development was characterized by unidirectional tendency for increasing total number of LMP2+ lymphocytes and reduced ХВР1+ cell population density in lymphoid structures of rat ileum.

The events observed in GALT cell populations under CSS conditions are opposing classical paradigm of the stress response. The CSS-associated effects do not promote immunosuppression, however, are able to cause activation of immune system and inflammatory process. 

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