In-vitro generation and characterisation of murine CD4+CD25+ regulatory T cells with indirect allospecificity

Int Immunopharmacol. 2006 Dec 20;6(13-14):1883-8. doi: 10.1016/j.intimp.2006.07.032. Epub 2006 Sep 1.

Abstract

Naturally arising CD4(+)CD25(+) regulatory T cells play a pivotal role in the prevention of autoimmunity and in the induction of donor-specific transplantation tolerance. Harnessing regulatory cells for potential adoptive cell therapy is hampered by their lack of antigen-specificity and their limited numbers. Here we describe the generation and expansion of murine CD4(+)CD25(+) T cells with antigen-specificity for an K(d) peptide as potential reagents for adoptive cell therapy in promoting donor-specific transplantation tolerance. Using bone marrow-derived autologous dendritic cells pulsed with the K(d) peptide, we generated T cell lines from purified CD4(+)CD25(+) T cells from C56BL/6 mice. The T cell lines expressed high level of CD25 and low level of CD45RB and CD69. They maintained the expression of CD62L, GITR, CTLA-4 and more importantly FoxP3. The CD4(+)CD25(+) T cell lines were anergic after TCR stimulation and produced little cytokine such as IL-2 and IFN-gamma. Importantly, they were more potent than freshly isolated CD4(+)CD25(+) T cells in suppressing proliferation and cytokine secretion by effector CD4(+) T cells. Furthermore, the CD4(+)CD25(+) T cell lines could be expanded to large cell numbers and maintained in culture up to 1 year. The K(d)-specific CD4(+)CD25(+) T cell lines will be invaluable in devising a strategy for the induction of cardiac transplantation tolerance in wild-type B6 mice carrying a full mismatch BALB/c heart.

MeSH terms

  • Animals
  • Antibodies / immunology
  • Antibodies / pharmacology
  • Antigens, CD / metabolism
  • Antigens, Differentiation / metabolism
  • CD3 Complex / immunology
  • CD4-Positive T-Lymphocytes / cytology
  • CD4-Positive T-Lymphocytes / immunology*
  • CD4-Positive T-Lymphocytes / metabolism
  • CTLA-4 Antigen
  • Cell Culture Techniques / methods
  • Cell Line
  • Coculture Techniques
  • Cytokines / metabolism
  • Dendritic Cells / immunology
  • Flow Cytometry
  • Forkhead Transcription Factors / metabolism
  • Glucocorticoid-Induced TNFR-Related Protein
  • H-2 Antigens / immunology
  • Interleukin-2 / pharmacology
  • Interleukin-2 Receptor alpha Subunit / immunology*
  • Isoantigens / immunology*
  • L-Selectin / metabolism
  • Leukocyte Common Antigens / metabolism
  • Lymphocyte Activation / drug effects
  • Lymphocyte Activation / immunology
  • Mice
  • Mice, Inbred C57BL
  • Peptide Fragments / immunology
  • Receptors, Antigen, T-Cell, alpha-beta / immunology
  • Receptors, Nerve Growth Factor / metabolism
  • Receptors, Tumor Necrosis Factor / metabolism
  • T-Lymphocytes, Regulatory / drug effects
  • T-Lymphocytes, Regulatory / immunology*
  • T-Lymphocytes, Regulatory / metabolism

Substances

  • Antibodies
  • Antigens, CD
  • Antigens, Differentiation
  • CD3 Complex
  • CTLA-4 Antigen
  • Ctla4 protein, mouse
  • Cytokines
  • Forkhead Transcription Factors
  • Foxp3 protein, mouse
  • Glucocorticoid-Induced TNFR-Related Protein
  • H-2 Antigens
  • Interleukin-2
  • Interleukin-2 Receptor alpha Subunit
  • Isoantigens
  • Peptide Fragments
  • Receptors, Antigen, T-Cell, alpha-beta
  • Receptors, Nerve Growth Factor
  • Receptors, Tumor Necrosis Factor
  • Tnfrsf18 protein, mouse
  • L-Selectin
  • Leukocyte Common Antigens