Abstract
Thymic selection is controlled by the interaction between TCR and MHC/peptide. Strength and quality of the signal determine whether thymocytes are selected or deleted. The factors that contribute to this signal remain poorly defined. Here we show that fetal thymic organ cultures (FTOCs) derived from OT-I transgenic mice (the OT-I TCR is restricted by K(b)-SIINFEKL) on a K(b)D(b-/-) background support positive selection, but only when provided with soluble H-2K(b)-SIINFEKL complexes. Selection of CD8 T cells is independent of the valency of the ligand or its capability to coengage CD8 molecules. Both CD8alphaalpha and CD8alphabeta T cells are selected by H-2K(b)-SIINFEKL, but only CD8alphabeta cells are capable of releasing IFN-gamma in response to the same ligand. The alpha(4)beta(7) integrin is up-regulated on postselection thymocytes from FTOCs. After adoptive transfer, FTOC-derived OT-I CD8 T cells divide in response to the agonist peptide SIINFEKL. These results establish that CD8 T cells responsive to their nominal peptide-Ag can be generated in FTOC supplemented with soluble MHC class I molecules equipped with the same peptide.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Adoptive Transfer
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Animals
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CD8-Positive T-Lymphocytes / immunology*
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CD8-Positive T-Lymphocytes / metabolism
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CD8-Positive T-Lymphocytes / transplantation*
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Cell Division / genetics
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Cell Division / immunology
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Cell Membrane / immunology
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Cell Membrane / metabolism
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Cell Movement / genetics
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Cell Movement / immunology
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Cells, Cultured
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Egg Proteins / immunology
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Fetus
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H-2 Antigens / administration & dosage
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H-2 Antigens / genetics
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H-2 Antigens / immunology*
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Immunophenotyping
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Integrins / biosynthesis
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Interferon-gamma / biosynthesis
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Interphase / genetics
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Interphase / immunology
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Intestinal Mucosa / cytology
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Intestinal Mucosa / immunology
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Lymph Nodes / cytology
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Lymph Nodes / immunology
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Lymphocyte Activation / genetics
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Lymphocyte Activation / immunology*
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Mice
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Mice, Inbred C57BL
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Mice, Knockout
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Mice, Transgenic
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Organ Culture Techniques
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Ovalbumin / immunology
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Peptide Fragments / administration & dosage
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Peptide Fragments / genetics
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Peptide Fragments / immunology*
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Peyer's Patches / cytology
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Peyer's Patches / immunology
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Thymus Gland / cytology
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Thymus Gland / immunology*
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Thymus Gland / transplantation
Substances
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Egg Proteins
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H-2 Antigens
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H-2Kb protein, mouse
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Integrins
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OVA-8
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Peptide Fragments
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integrin alpha4beta7
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Interferon-gamma
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Ovalbumin