绵羊痒螨钙网蛋白对兔外周血单个核细胞Th1/Th2和Th17/Treg免疫平衡的影响

doi:10.11843/j.issn.0366-6964.2025.04.038

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (4): 1910-1918.doi: 10.11843/j.issn.0366-6964.2025.04.038

• 预防兽医 • 上一篇

绵羊痒螨钙网蛋白对兔外周血单个核细胞Th1/Th2和Th17/Treg免疫平衡的影响

李艳娥¹, 梁友萍¹, 樊洁¹, 吴芳燕¹, 尧香悦¹, 李毛却乎², 次仁仓决³, 郝桂英^4*, 古小彬^1*

1. 四川农业大学动物医学院, 成都 611130;
2. 青海省黄南藏族自治州泽库县畜牧兽医站, 黄南 811499;
3. 西藏日喀则市定日县加措乡农牧综合服务中心, 定日 858200;
4. 西昌学院动物科技学院, 西昌 615013

收稿日期:2024-06-14 发布日期:2025-04-28
通讯作者: 四川省科技计划重点研发项目(2019YFN0155)；四川农业大学学科双支计划(2221993268)
作者简介:李艳娥(1999-),女,硕士,主要从事动物寄生虫病学研究,E-mail:2022203037@stu.sicau.edu.cn
基金资助:
国家肉鸡产业技术体系(CARS-41)；湖北省重点研发计划项目(2021BBA252)

Effects of Recombinant Calreticulin of Psoroptes ovis on Th1/Th2 and Th17/Treg Immune Balance in Rabbit Peripheral Blood Mononuclear Cells

LI Yan'e¹, LIANG Youping¹, FAN Jie¹, WU Fangyan¹, YAO Xiangyue¹, LIMAO Quehu², CIREN Cangjue³, HAO Guiying^4*, GU Xiaobin^1*

1. College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China;
2. Zeku Animal Husbandry and Veterinary Medical Station, Huangnan Tibetan Autonomous Prefecture, Qinghai Province, Huangnan 811499, China;
3. Agriculture and Animal Husbandry Comprehensive Service Center, Gacuo Township, Dingri County, Shigaze City, Tibet, Dingri 858200, China;
4. College of Animal Science and Technology, Xichang University, Xichang 615013, China

Received:2024-06-14 Published:2025-04-28

摘要/Abstract

摘要： 旨在探讨绵羊痒螨钙网蛋白(PsoCRT)对健康新西兰兔外周血单个核细胞(PBMC) Th1/Th2和Th17/Treg免疫平衡的影响。采用RT-PCR技术从绵羊痒螨混合阶段虫体总RNA中扩增得到去除信号肽的PsoCRT基因，经原核表达、纯化后得到重组蛋白(rPsoCRT)；分别以5、10、20、40、80 μg·mL^-1 rPsoCRT刺激兔PBMC，利用荧光定量PCR检测细胞中Th1/Th2、Th17/Treg主效细胞因子(IFN-γ、IL-4、IL-17A、IL-10)和关键转录因子(T-bet、GATA-3、RORc、Foxp3)的转录水平。结果显示：PsoCRT基因全长1 254 bp，属于典型的钙网蛋白家族；原核表达获得重组蛋白rPsoCRT大小约64 ku，可促进兔PBMC中Th1细胞IFN-γ和T-bet转录水平的升高，但抑制Th2细胞IL-4转录，而对GATA-3转录无影响，导致IFN-γ/IL-4和T-bet/GATA-3比值升高，平衡向Th1发生偏移；Th17细胞IL-17A和Treg细胞Foxp3 mRNA转录水平下降，而IL-10和RORc转录水平无显著变化，导致IL-17A/IL-10和RORc/Foxp3比值降低，平衡向Treg发生偏移。综上，PsoCRT可引起兔PBMC中Th1/Th2和Th17/Treg主效细胞因子和转录因子转录水平失衡，导致宿主免疫平衡偏向Th1和Treg免疫应答。

关键词: 绵羊痒螨, 钙网蛋白, Th1/Th2, Th17/Treg, 免疫平衡

Abstract: The aim of this study was to investigate the effects of Psoroptes ovis calreticulin (PsoCRT) on Th1/Th2 and Th17/Treg immune balance in the peripheral blood mononuclear cells (PBMC) of healthy New Zealand rabbits. In this study, PsoCRT gene without signal peptide was amplified from mixed-stage P. ovis total RNA by reverse transcription PCR, and its recombinant protein (rPsoCRT) was further expressed in prokaryotic expression and purified. Moreover, healthy rabbit PBMC were isolated and cultured, then added the different concentrations of rPsoCRT (5, 10, 20, 40, and 80 μg·mL^-1). Finally, the transcription levels of the primary cytokines (IFN-γ, IL-4, IL-17A, IL-10) and transcription factors (T-bet, GATA-3, RORc, Foxp3) of Th1, Th2, Th17 and Treg cells of rabbit PBMC were measured by qRT-PCR. Our results showed that the full-length of PsoCRT was 1 254 bp, and PsoCRT belonged to the type calreticulin family. The purified rPsoCRT was of the expected molecular size of ~64 ku. Rabbit PBMC treated with rPsoCRT upregulated the IFN-γ and T-bet mRNA expressions of Th1 cells, and decreased IL-4 mRNA expression while no different of GATA-3 mRNA expression in Th2 cells, which leaded to shifting T-cell differentiation towards Th1 bias with a drop in the IFN-γ/IL-4 and T-bet/GATA-3 ratios. Additionally, the mRNA expression of IL-17A in Th17 cells and Foxp3 in Treg cells were decreased, while the expressions of IL-10 and RORc remained unchanged, which resulted in shifting T-cell differentiation towards Treg bias with a drop in the IL-17A/IL-10 and RORc/Foxp3 ratios. These results indicated that rPsoCRT could cause imbalance in the transcription levels of Th1/Th2 and Th17/Treg primary cytokines and transcription factors in rabbit PBMC, resulting in host immunity shift to Th1 and Treg responses, respectively.

Key words: Psoroptes ovis, calreticulin, Th1/Th2, Th17/Treg, immune balance

中图分类号:

S582.746

李艳娥, 梁友萍, 樊洁, 吴芳燕, 尧香悦, 李毛却乎, 次仁仓决, 郝桂英, 古小彬. 绵羊痒螨钙网蛋白对兔外周血单个核细胞Th1/Th2和Th17/Treg免疫平衡的影响[J]. 畜牧兽医学报, 2025, 56(4): 1910-1918.

LI Yan'e, LIANG Youping, FAN Jie, WU Fangyan, YAO Xiangyue, LIMAO Quehu, CIREN Cangjue, HAO Guiying, GU Xiaobin. Effects of Recombinant Calreticulin of Psoroptes ovis on Th1/Th2 and Th17/Treg Immune Balance in Rabbit Peripheral Blood Mononuclear Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(4): 1910-1918.

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绵羊痒螨钙网蛋白对兔外周血单个核细胞Th1/Th2和Th17/Treg免疫平衡的影响

Effects of Recombinant Calreticulin of Psoroptes ovis on Th1/Th2 and Th17/Treg Immune Balance in Rabbit Peripheral Blood Mononuclear Cells

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