Preparation of functionally preserved CD4+ CD25high regulatory T cells from leukapheresis products from ulcerative colitis patients, applicable to regulatory T-cell transfer therapy

Y. Sumida, Kazuhiko Nakamura, K. Kanayama, H. Akiho, T. Teshima, R. Takayanagi

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Background: Ulcerative colitis (UC) is an intractable disease; therefore new therapies need to be developed. CD4+ CD25high regulatory T cells (Treg) significantly ameliorate colitis in animal models. In active UC patients, although Treg are functionally preserved, their proportion in peripheral blood decreases. Thus Treg transfer therapy is expected to be efficacious for UC. During leukapheresis for UC, Treg are depleted, as well as colitogenic effector leukocytes. We therefore designed a leukapheresis/Treg transfer therapy in which Treg are isolated from leukapheresis products and transfused to patients, and studied large-scale germ-free methods of Treg preparation. Methods: Using the CliniMACS cell selection system, we conducted Treg isolation experiments from leukapheresis products in which B and CD8+ T cells were depleted, followed by positive selection of CD25+ cells. In some experiments, isolated Treg or non-Treg were expanded with interleukin-2 (IL-2) ± transforming growth factor (TGF)-β 1. Expression of a Treg-specific marker, FOXP3, and gut-homing receptors, and suppressor activity of isolated or cultured cells, were analyzed. Results: CD4+ CD25high T cells were collected and efficiently enriched with a good recovery rate. Isolated cells preferentially expressed FOXP3 and significantly suppressed T-cell proliferation in vitro. In addition, isolated Treg could be efficiently expanded, and Treg could be induced from non-Treg with TGF-β 1 in vitro. TGF-β 1 significantly up-regulated αEβ7 and αEβ7 integrins. Discussion: We have established a method of Treg isolation from leukapheresis products that can be used clinically; therefore, Treg transfer therapy is feasible in combination with leukapheresis for UC. Expansion or induction of Treg in vitro may be another approach to Treg-based immunotherapy.

Original languageEnglish
Pages (from-to)698-710
Number of pages13
JournalCytotherapy
Volume10
Issue number7
DOIs
Publication statusPublished - Jan 1 2008

Fingerprint

Leukapheresis
Regulatory T-Lymphocytes
Cell- and Tissue-Based Therapy
Ulcerative Colitis
Transforming Growth Factors
T-Lymphocytes
Colitis
Therapeutics
Integrins
Immunotherapy
Interleukin-2
Cultured Cells
Leukocytes
Animal Models
Cell Proliferation
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology
  • Oncology
  • Genetics(clinical)
  • Cell Biology
  • Transplantation
  • Cancer Research

Cite this

Preparation of functionally preserved CD4+ CD25high regulatory T cells from leukapheresis products from ulcerative colitis patients, applicable to regulatory T-cell transfer therapy. / Sumida, Y.; Nakamura, Kazuhiko; Kanayama, K.; Akiho, H.; Teshima, T.; Takayanagi, R.

In: Cytotherapy, Vol. 10, No. 7, 01.01.2008, p. 698-710.

Research output: Contribution to journalArticle

Sumida, Y. ; Nakamura, Kazuhiko ; Kanayama, K. ; Akiho, H. ; Teshima, T. ; Takayanagi, R. / Preparation of functionally preserved CD4+ CD25high regulatory T cells from leukapheresis products from ulcerative colitis patients, applicable to regulatory T-cell transfer therapy. In: Cytotherapy. 2008 ; Vol. 10, No. 7. pp. 698-710.
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AU - Nakamura, Kazuhiko

AU - Kanayama, K.

AU - Akiho, H.

AU - Teshima, T.

AU - Takayanagi, R.

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N2 - Background: Ulcerative colitis (UC) is an intractable disease; therefore new therapies need to be developed. CD4+ CD25high regulatory T cells (Treg) significantly ameliorate colitis in animal models. In active UC patients, although Treg are functionally preserved, their proportion in peripheral blood decreases. Thus Treg transfer therapy is expected to be efficacious for UC. During leukapheresis for UC, Treg are depleted, as well as colitogenic effector leukocytes. We therefore designed a leukapheresis/Treg transfer therapy in which Treg are isolated from leukapheresis products and transfused to patients, and studied large-scale germ-free methods of Treg preparation. Methods: Using the CliniMACS cell selection system, we conducted Treg isolation experiments from leukapheresis products in which B and CD8+ T cells were depleted, followed by positive selection of CD25+ cells. In some experiments, isolated Treg or non-Treg were expanded with interleukin-2 (IL-2) ± transforming growth factor (TGF)-β 1. Expression of a Treg-specific marker, FOXP3, and gut-homing receptors, and suppressor activity of isolated or cultured cells, were analyzed. Results: CD4+ CD25high T cells were collected and efficiently enriched with a good recovery rate. Isolated cells preferentially expressed FOXP3 and significantly suppressed T-cell proliferation in vitro. In addition, isolated Treg could be efficiently expanded, and Treg could be induced from non-Treg with TGF-β 1 in vitro. TGF-β 1 significantly up-regulated αEβ7 and αEβ7 integrins. Discussion: We have established a method of Treg isolation from leukapheresis products that can be used clinically; therefore, Treg transfer therapy is feasible in combination with leukapheresis for UC. Expansion or induction of Treg in vitro may be another approach to Treg-based immunotherapy.

AB - Background: Ulcerative colitis (UC) is an intractable disease; therefore new therapies need to be developed. CD4+ CD25high regulatory T cells (Treg) significantly ameliorate colitis in animal models. In active UC patients, although Treg are functionally preserved, their proportion in peripheral blood decreases. Thus Treg transfer therapy is expected to be efficacious for UC. During leukapheresis for UC, Treg are depleted, as well as colitogenic effector leukocytes. We therefore designed a leukapheresis/Treg transfer therapy in which Treg are isolated from leukapheresis products and transfused to patients, and studied large-scale germ-free methods of Treg preparation. Methods: Using the CliniMACS cell selection system, we conducted Treg isolation experiments from leukapheresis products in which B and CD8+ T cells were depleted, followed by positive selection of CD25+ cells. In some experiments, isolated Treg or non-Treg were expanded with interleukin-2 (IL-2) ± transforming growth factor (TGF)-β 1. Expression of a Treg-specific marker, FOXP3, and gut-homing receptors, and suppressor activity of isolated or cultured cells, were analyzed. Results: CD4+ CD25high T cells were collected and efficiently enriched with a good recovery rate. Isolated cells preferentially expressed FOXP3 and significantly suppressed T-cell proliferation in vitro. In addition, isolated Treg could be efficiently expanded, and Treg could be induced from non-Treg with TGF-β 1 in vitro. TGF-β 1 significantly up-regulated αEβ7 and αEβ7 integrins. Discussion: We have established a method of Treg isolation from leukapheresis products that can be used clinically; therefore, Treg transfer therapy is feasible in combination with leukapheresis for UC. Expansion or induction of Treg in vitro may be another approach to Treg-based immunotherapy.

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