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Expansion of CD4+CD25+ regulatory T cells from cord blood CD4+ cells using the common γ-chain cytokines (IL-2 and IL-15) and rapamycin

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Abstract

Rapamycin has important roles in the modulation of regulatory T cells. We tried to expand CD4+CD25+ regulatory T cells (Treg cells) from umbilical cord blood (CB) CD4-positive cells using interleukin (IL)-15 or IL-2 with transforming growth factor (TGF)-β and rapamycin. We were able to obtain more than 500-fold expansion of CD4+CD25+ cells from CB CD4+ cells using IL-15 and TGF-β with rapamycin. These expanded CD4+CD25+ cells expressed forkhead box P3 (FoxP3) mRNA at a level about 100-fold higher and could suppress allogeneic mixed lymphocyte culture (MLC) by more than 50%. Early after rapamycin stimulation, CB CD4+ cells showed increased expression of FoxP3 and a serine/threonine kinase Pim2 and sustained expression of negative phosphoinositide 3-kinase regulator phosphatase and tensin homolog deleted on chromosome 10 (PTEN). On the other hand, CD4+CD25+ cells expanded with rapamycin for 8 days showed much higher levels of FoxP3 mRNA expression and decreased expression of PTEN. A comparison of IL-15 stimulation and IL-2 stimulation showed slightly higher efficiency of IL-15 for expansion of CD4+CD25+ cells, and for FoxP3 expression, IL-15 also showed significantly higher efficacy for inhibition of MLC. The combination of the common γ-chain cytokine IL-15, TGF-β, and rapamycin may be a useful means for expanding Treg cells. Pim2 expression early after stimulation with rapamycin may be important for conferring rapamycin resistance for growth of Treg cells. IL-15 is not less useful than IL-2 for expansion of Treg cells.

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Acknowledgments

We thank Ms. M. Yamane, Ms. M. Mayanagi, and Ms. Y. Ishimaru for their technical assistance. This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture, Japan (Tokyo, Japan).

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Authors and Affiliations

  1. Department of Hematology and Oncology, Hokkaido University Graduate School of Medicine, N15 W7, Kita-Ku, Sapporo, 060-8638, Japan

    Shinsuke Asanuma, Junji Tanaka, Junichi Sugita, Mizuha Kosugi, Souichi Shiratori, Kentarou Wakasa, Yusuke Shono, Akio Shigematsu & Masahiro Imamura

  2. Third Department of Internal Medicine, Hokkaido University Graduate School of Medicine, N15 W7, Kita-Ku, Sapporo, 060-8638, Japan

    Takeshi Kondo, Takahiko Kobayashi & Masahiro Asaka

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  1. Shinsuke Asanuma
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  2. Junji Tanaka
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Correspondence to Junji Tanaka.

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Asanuma, S., Tanaka, J., Sugita, J. et al. Expansion of CD4+CD25+ regulatory T cells from cord blood CD4+ cells using the common γ-chain cytokines (IL-2 and IL-15) and rapamycin. Ann Hematol 90, 617–624 (2011). https://doi.org/10.1007/s00277-010-1121-z

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