Establishment of a transgenic mouse model with musculin knockout and Foxp3-GFP knockin and role of musculin in severe trauma-induced Treg-Th17 imbalance

Jing YU; Wanqi TANG; Xue YANG; Wei MA; Jun YAN; Huaping LIANG

doi:10.16016/j.1000-5404.201802112

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Establishment of a transgenic mouse model with musculin knockout and Foxp3-GFP knockin and role of musculin in severe trauma-induced Treg-Th17 imbalance

更新时间：2026-01-09

- Establishment of a transgenic mouse model with musculin knockout and Foxp3-GFP knockin and role of musculin in severe trauma-induced Treg-Th17 imbalance
- Journal of Third Military Medical University Vol. 40, Issue 16, Pages: 1435-1443(2018)
- 作者机构：
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81471863);the Special Project of Basic Science and Frontier Technology Research of Chongqing(CSTC2016jcyjA0048)
- DOI：10.16016/j.1000-5404.201802112
  CLC：
- Received：21 February 2018，
  
  Published：30 August 2018
- 稿件说明：
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Jing YU, Wanqi TANG, Xue YANG, et al. Establishment of a transgenic mouse model with musculin knockout and Foxp3-GFP knockin and role of musculin in severe trauma-induced Treg-Th17 imbalance[J]. Journal of Third Military Medical University, 2018, 40(16): 1435-1443.
DOI：

Jing YU, Wanqi TANG, Xue YANG, et al. Establishment of a transgenic mouse model with musculin knockout and Foxp3-GFP knockin and role of musculin in severe trauma-induced Treg-Th17 imbalance[J]. Journal of Third Military Medical University, 2018, 40(16): 1435-1443. DOI： 10.16016/j.1000-5404.201802112.

摘要

Abstract

Objective

To establish a transgenic mouse model with musculin knockout (MSCKO) and Foxp3 and green fluorescent protein knockin (Foxp3-GFPKI) and investigate the role of musculin in regulatory T cell (Treg)-Th17 imbalance induced by severe trauma.

Methods

MSCKO homozygote (

-/-

) mice and Foxp3-GFPKI homozygote (

+/+

) mice were crossed to obtain the heterozygote (MSCKO

+/-

-Foxp3-GFPKI

+/-

) mice.The offspring mice were bred continuously until the homozygote mice (MSCKO-Foxp3-GFPKI) were acquired.The MSCKO-Foxp3-GFPKI mice were crossed for at least 5 generations

and the mice in each generation were assessed for genotypes

phenotypes and genetic stability.Severe trauma (fracture with blood loss) models were then established in the MSCKO-Foxp3-GFPKI and Foxp3-GFPKI mice

and 3 h after the modeling

peripheral lymph nodes (LN)

mesenteric lymph nodes (MLN)

Peyer' s patch (PP)

spleen

bone marrow (BM) and thymus samples were collected.The distribution of Treg in different immune organs and tissues was detected by flow cytometry

and the expression levels of musculin

Foxp3 and retinoic acid-related orphan receptor γt (ROR-γt) were measured using quantitative RT-PCR to explore the role of musculin in Treg-Th17 imbalance mediated by severe trauma.

Results

All the homozygote (MSCKO-Foxp3-GFPKI) mice in different generations expressed the molecular markers with normal activities

weight and breeding capacity.Fluorescence-activated cell sorting revealed GFP signals in the immune organs and tissues of the homozygote mice.MSCKO-Foxp3-GFPKI mice with severe trauma showed obvious changes in Treg percentages in the LN (5.81%)

MLN (4.81%)

PP (1.41%) and SP (3.1%) and exhibited significantly increased mRNA levels of musculin and Foxp3 and decreased ROR-γt mRNA expression in the MLN and PP (

0.05 or 0.01)

indicating a left shift of Treg-Th17 balance after t

rauma.

Conclusion

We successfully obtain MSCKO-Foxp3-GFPKI mice with stable phenotypes.Musculin affects the distribution of Treg and causes shift of Treg/Th17 balance after severe trauma

suggesting its role as a potential regulatory target in the maintenance of the internal environment and immune homeostasis after severe trauma.

关键词

Keywords

references

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