经 DNA 构建体转染的树突状细胞编码 CCR9、IL-10 和 II 型胶原在关节炎模型中显示诱导免疫耐受。

Dendritic cells transfected with DNA constructs encoding CCR9, IL-10, and type II collagen demonstrate induction of immunological tolerance in an arthritis model.

机构信息

Laboratory of Molecular Immunology, Federal State Budgetary Scientific Institution Research Institute of Fundamental and Clinical Immunology, Novosibirsk, Russia.

Department of Microscopic Research, State Research Centre for Virology and Biotechnology «Vektor», Koltsovo, Russia.

出版信息

Front Immunol. 2024 Aug 5;15:1447897. doi: 10.3389/fimmu.2024.1447897. eCollection 2024.

DOI:10.3389/fimmu.2024.1447897

PMID:39161770

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11330828/

Abstract

INTRODUCTION

Restoring immune tolerance is a promising area of therapy for autoimmune diseases. One method that helps restore immunological tolerance is the approach using tolerogenic dendritic cells (tolDCs). In our study, we analyzed the effectiveness of using dendritic cells transfected with DNA constructs encoding IL-10, type II collagen, and CCR9 to induce immune tolerance in an experimental model of arthritis.

METHODS

Dendritic cell cultures were obtained from bone marrow cells of Balb/c mice. Dendritic cells (DCs) cultures were transfected with pmaxCCR9, pmaxIL-10, and pmaxCollagen type II by electroporation. The phenotype and functions of DCs were studied using enzyme-linked immunosorbent assay (ELISA) and flow cytometry. Migration of electroporated DCs was assessed . Induction of antigen-collagen induced arthritis (ACIA) was carried out according to the protocol in Balb/c mice. DCs were then administered to ACIA mice. The development of arthritis was monitored by measuring paw swelling with a caliper at different time points. The immunological changes were assessed by analyzing the content of antibodies to type II collagen using enzyme immunoassay. Additionally, a histological examination of the joint tissue was conducted, followed by data analysis.

THE RESULTS ARE AS FOLLOWS

DCs were obtained, characterized by reduced expression of CD80, CD86, and H-2Db (MHC class I), increased expression of CCR9, as well as producing IL-10 and having migratory activity to thymus cells. Transfected DCs induced T-regulatory cells (T-reg) and increased the intracellular content of IL-10 and TGF-β in CD4T cells in their co-culture, and also suppressed their proliferative activity in response to antigen. The administration of tolDCs transfected with DNA constructs encoding type II collagen, IL-10, and CCR9 to mice with ACIA demonstrated a reduction in paw swelling, a reduction in the level of antibodies to type II collagen, and a regression of histological changes.

CONCLUSION

The study presents an approach by which DCs transfected with DNA constructs encoding epitopes of type II collagen, IL-10 and CCR9 promote the development of antigen-specific tolerance, control inflammation and reduce the severity of experimental arthritis through the studied mechanisms: induction of T-reg, IL-10, TGF-β.

摘要

介绍

恢复免疫耐受是治疗自身免疫性疾病的一个有前途的治疗领域。一种有助于恢复免疫耐受的方法是使用致耐受性树突状细胞（tolDC）的方法。在我们的研究中，我们分析了使用转染 DNA 构建体编码 IL-10、II 型胶原和 CCR9 的树突状细胞诱导关节炎实验模型中免疫耐受的效果。

方法

从 Balb/c 小鼠的骨髓细胞中获得树突状细胞培养物。通过电穿孔将 pmaxCCR9、pmaxIL-10 和 pmaxCollagen type II 转染到树突状细胞（DC）培养物中。使用酶联免疫吸附试验（ELISA）和流式细胞术研究 DC 的表型和功能。评估电穿孔的 DC 的迁移。根据 Balb/c 小鼠的方案进行抗原诱导胶原诱导关节炎（ACIA）。然后将 DC 给予 ACIA 小鼠。在不同时间点使用卡尺测量爪肿胀来监测关节炎的发展。通过酶免疫测定分析 II 型胶原抗体的含量来评估免疫变化。此外，还进行了关节组织的组织学检查，然后进行数据分析。

结果如下

获得了树突状细胞，其特征在于 CD80、CD86 和 H-2Db（MHC 类 I）的表达减少，CCR9 的表达增加，产生 IL-10 并具有向胸腺细胞迁移的活性。转染的 DC 诱导 T 调节细胞（T-reg），并增加共培养物中 CD4T 细胞内的 IL-10 和 TGF-β含量，并且还抑制它们对抗原的增殖活性。将转染编码 II 型胶原、IL-10 和 CCR9 的 DNA 构建体的 tolDC 给予 ACIA 小鼠可减少爪肿胀，降低 II 型胶原抗体水平，并使组织学变化消退。