Regulatory T cells expanded by rapamycin in vitro suppress colitis in an experimental mouse model

J Gastroenterol. 2012 Apr;47(4):366-76. doi: 10.1007/s00535-011-0502-y. Epub 2011 Dec 22.

Abstract

Background: To provide rapid immunosuppression without side effects, we analyzed whether rapamycin alone, and regulatory T cells (Tregs) expanded ex vivo by rapamycin, suppressed colitis in a mouse model.

Methods: Severe combined immunodeficiency (SCID) mice reconstituted with naive CD4(+) T cells were treated with or without intraperitoneal rapamycin. Body weight was evaluated. CD4(+) T cells were cultured in the presence of rapamycin for three 7-day rounds of stimulation. The ratio of Tregs to CD4(+) T cells was analyzed by flow cytometry. Naive CD4(+) T cells were transferred into SCID mice with CD4(+) T cells expanded in the presence or absence of rapamycin. Clinical symptoms of colitis, histological changes, and cytokine expression were investigated.

Results: Systemic rapamycin partially prevented the development of colonic inflammation in a transfer model of colitis, but decreased body weight in control mice. With rapamycin, stimulated CD4(+) T cells expanded eightfold in 3 weeks in vitro, and the proportion of Tregs increased to about 40%. Without rapamycin, CD4(+) T cells expanded 20-fold in 3 weeks, but the proportion of Tregs remained at about 15%. CD4(+) T cells expanded with rapamycin prevented the development of colitis in a naïve CD4(+) T-cell transfer model, in association with the downregulation of Th1 and Th17 responses.

Conclusions: We demonstrated, for the first time, that CD4(+) T cells expanded with rapamycin in vitro suppressed colitis. Therefore, rapamycin-expanded Treg transfer therapy is expected to be efficacious for inflammatory bowel disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CD4 Lymphocyte Count
  • Cell Culture Techniques
  • Colitis / drug therapy*
  • Colitis / immunology
  • Colon / immunology*
  • Colon / metabolism
  • Colon / pathology
  • Cytokines / metabolism*
  • Immunosuppressive Agents / pharmacology
  • Immunosuppressive Agents / therapeutic use*
  • Mice
  • Mice, Inbred BALB C
  • Mice, SCID
  • Models, Animal
  • RNA, Messenger / metabolism
  • Real-Time Polymerase Chain Reaction
  • Sirolimus / pharmacology
  • Sirolimus / therapeutic use*
  • T-Lymphocytes, Regulatory / drug effects*
  • T-Lymphocytes, Regulatory / metabolism

Substances

  • Cytokines
  • Immunosuppressive Agents
  • RNA, Messenger
  • Sirolimus