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Large-scale in vitro expansion of human regulatory T cells with potent xenoantigen-specific suppression

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Abstract

Xenotransplantation is a potential solution to the organ donor shortage. Immunosuppression is required for successful application of xenotransplantation but may lead to infection and cancer. Thus, strategies for immune tolerance induction need to be developed. Polyclonal regulatory T cells (Treg) play a central role in the induction and maintenance of immune tolerance and have been shown to protect against islet xenograft rejection in vivo. However, global immune suppression may be mediated by polyclonal Treg immunotherapy and a simple method for in vitro expansion of xenoantigen-specific Treg for efficient Treg application becomes necessary. Human Treg isolated from peripheral blood mononuclear cells (PBMCs) were initially cultured with anti-CD3/CD28 beads, rapamycin and IL-2 for 7 days as polyclonal expansion. Expanded Treg were then cocultured with irradiated porcine PBMC as xenoantigen stimulation for three subsequent cycles with 7 days for each cycle in the presence of IL-2 and anti-CD3/CD28 beads. Treg phenotype and suppressive capacity were assessed after each cycle of xenoantigen stimulation. Treg expanded with one cycle of xenoantigen stimulation retained Treg suppressive phenotype but acquired no xenoantigen specificity along with poor expansion efficiency, whereas expansion with two-cycle xenoantigen stimulation resulted in not only more than 800-fold Treg expansion but highly suppressive xenoantigen-specific Treg with effector Treg phenotype. However further increase of stimulation cycles resulted in reduced Treg suppressive potency. This study provides a simple approach to obtain high numbers of xenoantigen-specific Treg for immune tolerance induction in xenotransplantation.

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Acknowledgments

This study was supported by grants from the National Health and Medical Research Council of Australia (NH&MRCA), the Juvenile Diabetes Research Foundation International (JDRFI), the Key Program of National Nature Science Foundation of China (No. 30930088) and National Natural Science Foundation of China (No. 30872382).

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The authors declare no conflicts of interest.

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Authors and Affiliations

  1. Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, West China Hospital, Sichuan University, Chengdu, 610000, Sichuan, China

    Xi Jin & Yanrong Lu

  2. Regenerative Medicine Research Center, West China Hospital, Sichuan University, Chengdu, 610000, Sichuan, China

    Xi Jin & Yanrong Lu

  3. Center for Transplant and Renal Research, Westmead Hospital, Westmead, NSW, 2145, Australia

    Xi Jin, Ye Zhao & Shounan Yi

  4. Transplantation Research Center, The 2nd Xiangya Hospital, Central South University, Changsha, 410000, Hunan, China

    Shounan Yi

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  1. Xi Jin
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Correspondence to Shounan Yi.

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Jin, X., Lu, Y., Zhao, Y. et al. Large-scale in vitro expansion of human regulatory T cells with potent xenoantigen-specific suppression. Cytotechnology 68, 935–945 (2016). https://doi.org/10.1007/s10616-015-9845-1

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