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Normal and Malignant Muscle Cell Transplantation into Immune Compromised Adult Zebrafish
Published on: December 26, 2014
Engineering Tregs-mediated immune tolerance via foxp3a overexpression to evade allograft transplantation barriers in
Junwen Zhu1, Yongkang Hao2, Fenghua Zhang3
1State Key Laboratory of Breeding Biotechnology and Sustainable Aquaculture, Hubei Hongshan Laboratory, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei 430072, China; School of Marine Biology and Fisheries, Hainan University, Haikou, Hainan 570228, China; College of Advanced Agricultural Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.
In mammals, regulatory T cells (Tregs) are widely exploited to promote immune tolerance in organ transplantation. In zebrafish, although germline stem cell (GSC) or gonadal primordium transplantation into immunodeficient hosts can accelerate gamete production, maintaining immunocompromised lines presents substantial practical challenges. To overcome this limitation, this study generates a Tg(CMV:foxp3a) zebrafish line through systemic overexpression of Forkhead box P3a (Foxp3a), the lineage-defining transcription factor of Tregs. Transcriptomic and in situ hybridization analysis reveal downregulation of the Treg negative regulator cd127 and upregulation of multiple immunosuppressive factors in the head kidney and thymus. Single-cell RNA sequencing further demonstrates a reduction in effector T and B cell populations, accompanied by an increase in quiescent T cells exhibiting resting Treg-like features. Importantly, using Tg(CMV:foxp3a) fish as hosts for subcutaneous gonadal primordium transplantation and intraperitoneal GSC transplantation markedly accelerates germ cell maturation and enables efficient establishment of stable transgenic lines. Post-transplantation analysis indicates delayed and attenuated immune activation, enhanced graft survival, and rapid induction of immunosuppressive states. Together, foxp3a overexpression reshapes the immune landscape to confer immune tolerance, providing a practical Tregs-based alternative to immunodeficient hosts for fish genome manipulation and transplantation.
In mammals, regulatory T cells (Tregs) are widely exploited to promote immune tolerance in organ transplantation. In zebrafish, although germline stem cell (GSC) or gonadal primordium transplantation into immunodeficient hosts can accelerate gamete production, maintaining immunocompromised lines presents substantial practical challenges. To overcome this limitation, this study generates a Tg(CMV:foxp3a) zebrafish line through systemic overexpression of Forkhead box P3a (Foxp3a), the lineage-defining transcription factor of Tregs. Transcriptomic and in situ hybridization analysis reveal downregulation of the Treg negative regulator cd127 and upregulation of multiple immunosuppressive factors in the head kidney and thymus. Single-cell RNA sequencing further demonstrates a reduction in effector T and B cell populations, accompanied by an increase in quiescent T cells exhibiting resting Treg-like features. Importantly, using Tg(CMV:foxp3a) fish as hosts for subcutaneous gonadal primordium transplantation and intraperitoneal GSC transplantation markedly accelerates germ cell maturation and enables efficient establishment of stable transgenic lines. Post-transplantation analysis indicates delayed and attenuated immune activation, enhanced graft survival, and rapid induction of immunosuppressive states. Together, foxp3a overexpression reshapes the immune landscape to confer immune tolerance, providing a practical Tregs-based alternative to immunodeficient hosts for fish genome manipulation and transplantation.
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