Abstract
CD1d-restricted Valpha14-Jalpha281 invariant alphabetaTCR(+) (NKT) cells are well defined in the C57BL/6 mouse strain, but they remain poorly characterized in non-NK1.1-expressing strains. Surrogate markers for NKT cells such as alphabetaTCR(+)CD4(-)CD8(-) and DX5(+)CD3(+) have been used in many studies, although their effectiveness in defining this lineage remains to be verified. Here, we compare NKT cells among C57BL/6, NK1.1-congenic BALB/c, and NK1.1-congenic nonobese diabetic mice. NKT cells were identified and compared using a range of approaches: NK1.1 expression, surrogate phenotypes used in previous studies, labeling with CD1d/alpha-galactosylceramide tetramers, and cytokine production. Our results demonstrate that NKT cells and their CD4/CD8-defined subsets are present in all three strains, and confirm that nonobese diabetic mice have a numerical and functional deficiency in these cells. We also highlight the hazards of using surrogate phenotypes, none of which accurately identify NKT cells, and one in particular (DX5(+)CD3(+)) actually excludes these cells. Finally, our results support the concept that NK1.1 expression may not be an ideal marker for CD1d-restricted NKT cells, many of which are NK1.1-negative, especially within the CD4(+) subset and particularly in NK1.1-congenic BALB/c mice.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antigens / biosynthesis
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Antigens, CD / biosynthesis
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Antigens, CD1 / biosynthesis
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Antigens, CD1 / genetics*
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Antigens, CD1 / metabolism
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Antigens, CD1d
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Antigens, Ly*
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Antigens, Surface
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Binding Sites / immunology
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CD24 Antigen
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CD3 Complex / biosynthesis
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CD8 Antigens / biosynthesis
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Carrier Proteins / biosynthesis
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Cytokines / biosynthesis
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Galactosylceramides / metabolism
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Immunophenotyping
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Killer Cells, Natural / cytology
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Killer Cells, Natural / immunology*
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Killer Cells, Natural / metabolism
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L-Selectin / biosynthesis
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Lectins, C-Type
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Lymphocyte Count
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Membrane Glycoproteins*
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Membrane Proteins / biosynthesis
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Mice
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Mice, Congenic
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Mice, Inbred BALB C
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Mice, Inbred C57BL
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Mice, Inbred NOD
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NK Cell Lectin-Like Receptor Subfamily B
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Organ Specificity / genetics
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Organ Specificity / immunology
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Protein Biosynthesis
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Proteins*
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Receptors, Antigen, T-Cell, alpha-beta / biosynthesis
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Receptors, Interleukin-2 / biosynthesis
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Receptors, NK Cell Lectin-Like
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Species Specificity*
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T-Lymphocyte Subsets / cytology
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T-Lymphocyte Subsets / immunology*
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T-Lymphocyte Subsets / metabolism
Substances
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Antigens
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Antigens, CD
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Antigens, CD1
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Antigens, CD1d
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Antigens, Ly
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Antigens, Surface
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CD24 Antigen
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CD3 Complex
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CD8 Antigens
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Carrier Proteins
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Cd24a protein, mouse
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Cytokines
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Galactosylceramides
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Klrb1c protein, mouse
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Lectins, C-Type
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Membrane Glycoproteins
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Membrane Proteins
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NK Cell Lectin-Like Receptor Subfamily B
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Proteins
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Receptors, Antigen, T-Cell, alpha-beta
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Receptors, Interleukin-2
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Receptors, NK Cell Lectin-Like
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L-Selectin