异位胸腺类器官的发育以培育和诱导T淋巴细胞

Development of Thymic Organoids Heterotopically to Educate and Induce T Lymphocytes.

作者信息

Wang Xiuxia, Yu Shun, Qiu Yucheng, Yang Jun, Liu Fei, Zhou Xianyu

机构信息

Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, China.

Department of Burns and Plastic Surgery, Affiliated Hospital of Jiangnan University, Wuxi, China.

出版信息

Immun Inflamm Dis. 2025 Aug;13(8):e70229. doi: 10.1002/iid3.70229.

BACKGROUND

Vascularized composite allotransplantation (VCA) is a potential treatment for extensive injuries that replaces defects like-with-like, however allografts are immune-rejectable.

METHODS

This study developed in vitro thymic organoids and examined whether donor-derived HSCs could be educated in vivo into T lymphocytes via central tolerance. TECs, TMCs, and HSCs from C57BL/7 (CD45.2) or SJL/L (CD45.1) mice were labeled with cell surface markers and examined by flow cytometry. Co-culturing three cell lines in vitro created thymic aggregates. Aggregates transplanted to C57BL/7 (CD45.2) mice's inguinal regions developed thymic organoids. Immunorejection genes were identified bioinformatically. Western blot, immunofluorescence, and flow cytometry were utilized to measure rejection-related protein levels and T cell surface markers in thymic organoids to determine T cell inducement and immunomodulation.

RESULTS

In vitro, TECs, TMCs, and HSCs created thymic aggregates, which became thymic organoids after in vivo transplantation and produced CD8 and CD4 Tregs. Bioinformatics showed high correlations between transplanted rejection and IFNG, IL2RG, FCGR3A, and ICAM1 genes. Immunofluorescence and Western blot showed increased protein expression of IFNG, IL2RG, FCGR3A (immunomodulation biomarker), and decreased protein expression of CK8, CK14, and ICAM1 (TEC biomarker) in thymic organoids.

CONCLUSION

Thymic organoids heterotopically implanted in vivo can promote heterologous HSC-derived T cell development.

背景

血管化复合组织异体移植（VCA）是一种治疗大面积损伤的潜在方法，它以相似组织替换缺损组织，然而异体移植会引发免疫排斥。

方法

本研究构建了体外胸腺类器官，并检测供体来源的造血干细胞（HSCs）能否通过中枢耐受在体内被诱导分化为T淋巴细胞。对来自C57BL/7（CD45.2）或SJL/L（CD45.1）小鼠的胸腺上皮细胞（TECs）、胸腺基质细胞（TMCs）和造血干细胞进行细胞表面标记，并通过流式细胞术进行检测。将这三种细胞系在体外共培养以形成胸腺聚集体。将聚集体移植到C57BL/7（CD45.2）小鼠的腹股沟区域，使其发育成胸腺类器官。通过生物信息学方法鉴定免疫排斥相关基因。利用蛋白质免疫印迹法、免疫荧光法和流式细胞术检测胸腺类器官中与排斥相关的蛋白质水平和T细胞表面标志物，以确定T细胞的诱导和免疫调节情况。

结果

在体外，TECs、TMCs和HSCs形成了胸腺聚集体，体内移植后成为胸腺类器官，并产生了CD8和CD4调节性T细胞（Tregs）。生物信息学分析显示，移植排斥与IFNG、IL2RG、FCGR3A和ICAM1基因之间存在高度相关性。免疫荧光和蛋白质免疫印迹结果显示，胸腺类器官中IFNG、IL2RG、FCGR3A（免疫调节生物标志物）的蛋白表达增加，而细胞角蛋白8（CK8）、细胞角蛋白14（CK14）和ICAM1（TEC生物标志物）的蛋白表达降低。