不同发育阶段BTBR T+ Itpr3tf孤独症小鼠T细胞亚群异常研究
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昆明医科大学第二附属医院

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国家自然科学基金资助项目(82460281、82060306),云南省李斌专家工作站(202305AF150068),云南省科技厅-昆明医科大学联合专项(202401AY070001-086)


Abnormal T cell subsets of BTBR T+Itpr3tf autistic mice at different developmental stages
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The Second Affiliated Hospital of Kunming Medical University

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The National Natural Science Foundation of China (82460281、82060306), the Yunnan Province Li Bin Expert Workstation (202305AF150068),the Yunnan Provincial Department of Science and Technology Kunming Medical University Joint Special Project(202401AY070001-086)

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    摘要:

    目的:为了研究4周、8周和12周龄BTBR T+ Itpr3tf孤独症小鼠脾脏T细胞亚群表达情况,明确用于研究免疫异常与孤独症关系的最适BTBR孤独症小鼠年龄。方法:随机选取5-6只4周、8周和12周龄雄性BTBR小鼠与对应年龄的同性别C57小鼠进行三箱社交实验、理毛实验和埋珠实验;取8周龄和12周龄小鼠脾脏制成单细胞悬液,通过流式细胞技术检测8种T细胞亚群(TH1、TH2、TH17、TC1、TC2、TC17、TFH和Treg)细胞。结果:4周、8周和12周龄BTBR小鼠相较于同龄C57小鼠,社交时间减少(p<0.0001,p<0.0001,p<0.0001),理毛时间增加(p=0.004,p=0.0001,p<0.0001),8周龄和12周龄BTBR小鼠埋珠数量增多(p=0.001,p<0.0001);与同龄C57小鼠相比,8周龄BTBR小鼠TH1(p=0.0001)、TH2(p=0.006)、Tc1(p=0.026)、Tc2(p=0.0001)、TFH(p=0.001)细胞表达明显增加,Treg细胞(p=0.0001)明显降低;12周龄 BTBR小鼠TH1(p=0.002)、TH2(p=0.009)、TH17(p=0.016)、Tc1(p=0.001)、Tc2(p=0.0001)、Tc17(p=0.005)、TFH(p=0.0001)细胞表达增加,Treg细胞(p=0.01)表达降低。不同年龄阶段,8周龄BTBR小鼠较12周龄小鼠,p<0.05,TH1/Treg,TC1/Treg比例明显升高,而Tc17/Treg比例降低。结论:不同发育阶段BTBR小鼠均表现出不同程度的Teff/Treg比例异常增高;结合行为学数据,建议使用8周龄BTBR小鼠用于开展孤独症与免疫异常的研究。

    Abstract:

    Objective: To investigate the expression of T cell subsets in the spleen of BTBR T+Itpr3tf autistic mouse at 4, 8, and 12 weeks of age, and to determine the optimal age for studying the relationship between immune abnormalities and autism in BTBR autistic mice. Methods: It randomly selected 5-6 male BTBR mouse at 4 weeks, 8 weeks, and 12 weeks of age and C57 mouse of the same gender at corresponding ages for the three-box social interaction test, the self-grooming test, and the marble-burying test; Single cell suspensions were prepared from the spleens of mouse at 8 and 12 weeks of age, and flow cytometry was used to detect 8 subsets of T cells (TH1, TH2, TH17, TC1, TC2, TC17, TFH, and Treg). Results: Compared with C57 mouse of the same age,p<0.05, BTBR mouse at 4 weeks, 8 weeks, and 12 weeks of age showed a decrease in social time(p<0.0001,p<0.0001,p<0.0001), an increase in grooming time(p=0.004,p=0.0001,p<0.0001), and an increase in the number of marbles buried(p=0.001,p<0.0001) in BTBR mouse at 8 weeks and 12 weeks of age; As well , the expression of TH1(p=0.0001), TH2p=0.006), TC1(p=0.026), TC2(p=0.0001), and TFH(p=0.001)cells in 8-week-old BTBR mouse were significantly increased, while the expression of Treg(p=0.0001) cells were significantly decreased; The expression of TH1(p=0.002), TH2(p=0.009), TH17(p=0.016), TC1(p=0.001), TC2(p=0.0001), TC17(p=0.005), and TFH (p=0.0001)increased in 12-week-old BTBR mouse, while the expression of Treg (p=0.01)cells decreased. At different age stages, p<0.05,the ratio of TH1/Treg and TC1/Treg in 8-week-old BTBR mouse were significantly higher than those in 12 week old mouse, while the TC17/Treg ratio decreased. Conclusions: BTBR mouse at different developmental stages exhibit varying degrees of abnormal increase in Teff/Treg ratio; Based on results of behavioral test, it is recommended to use 8-week-old BTBR mice for research on autism and immune abnormalities.

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  • 收稿日期:2024-08-16
  • 最后修改日期:2024-10-15
  • 录用日期:2025-04-25
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