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
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Animals
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Antibodies / immunology
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Antibodies / pharmacology
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Antigens, CD / metabolism
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Antigens, Differentiation / metabolism
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CD3 Complex / immunology
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CD4-Positive T-Lymphocytes / cytology
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CD4-Positive T-Lymphocytes / immunology*
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CD4-Positive T-Lymphocytes / metabolism
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CTLA-4 Antigen
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Cell Culture Techniques / methods
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Cell Line
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Coculture Techniques
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Cytokines / metabolism
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Dendritic Cells / immunology
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Flow Cytometry
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Forkhead Transcription Factors / metabolism
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Glucocorticoid-Induced TNFR-Related Protein
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H-2 Antigens / immunology
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Interleukin-2 / pharmacology
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Interleukin-2 Receptor alpha Subunit / immunology*
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Isoantigens / immunology*
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L-Selectin / metabolism
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Leukocyte Common Antigens / metabolism
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Lymphocyte Activation / drug effects
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Lymphocyte Activation / immunology
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Mice
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Mice, Inbred C57BL
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Peptide Fragments / immunology
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Receptors, Antigen, T-Cell, alpha-beta / immunology
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Receptors, Nerve Growth Factor / metabolism
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Receptors, Tumor Necrosis Factor / metabolism
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T-Lymphocytes, Regulatory / drug effects
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T-Lymphocytes, Regulatory / immunology*
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T-Lymphocytes, Regulatory / metabolism
Substances
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Antibodies
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Antigens, CD
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Antigens, Differentiation
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CD3 Complex
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CTLA-4 Antigen
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Ctla4 protein, mouse
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Cytokines
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Forkhead Transcription Factors
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Foxp3 protein, mouse
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Glucocorticoid-Induced TNFR-Related Protein
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H-2 Antigens
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Interleukin-2
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Interleukin-2 Receptor alpha Subunit
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Isoantigens
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Peptide Fragments
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Receptors, Antigen, T-Cell, alpha-beta
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Receptors, Nerve Growth Factor
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Receptors, Tumor Necrosis Factor
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Tnfrsf18 protein, mouse
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L-Selectin
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Leukocyte Common Antigens