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Defining a novel subset of CD1d-dependent type II natural killer T cells using natural killer cell-associated markers

Journal article
Authors Avadhesh Kumar Singh
Sara Rhost
Linda Löfbom
Susanna Cardell
Published in Scandinavian Journal of Immunology
Volume 90
Issue 3
ISSN 0300-9475
Publication year 2019
Published at Institute of Biomedicine, Department of Microbiology and Immunology
Language en
Keywords cytokines, experimental animals, natural killer T cells, transcription factors, nkt cells, receptor, plzf, distinct, mice, identification, repertoire, prevention, population, expression, Immunology
Subject categories Immunology in the medical area


Natural killer T (NKT) cells are alpha beta T cell receptor (TCR) expressing innate-like T cells that display natural killer (NK) cell markers. Based on TCR characteristics, they are divided into two groups restricted to the MHC class I-like molecule CD1d. Type I NKT cells, most extensively studied, are identified by a semi-invariant V alpha 14-J alpha 18 (mouse, V alpha 24-J alpha 18 in humans) TCR reactive to the prototypic ligand alpha-galactosylceramide presented on CD1d. In contrast, type II NKT cells display diverse TCR reacting to different CD1d-presented ligands. There are no reagents that identify all type II NKT cells, limiting their exploration. Here, we searched for novel type II NKT cells by comparing J alpha 18(-/-)MHCII(-/-) mice that harbour type II but not type I NKT cells, and CD1d(-/-)MHCII(-/-) mice, lacking all NKT cells. We identified significantly larger populations of CD4(+) and CD4(-)CD8(-) (double negative, DN) TCR beta(+) cells expressing NKG2D or NKG2A/C/E in J alpha 18(-/-)MHCII(-/-) mice compared with CD1d(-/-)MHCII(-/-) mice, suggesting that 30%-50% of these cells were type II NKT cells. They expressed CD122, NK1.1, CXCR3 and intermediate/low levels of CD45RB. Further, the CD4(+) subset was CD69(+), while the DN cells were CD49b(+) and CD62L(+). Both subsets expressed the NKT cell-associated promyelocytic leukaemia zinc finger (PLZF) transcription factor and Tbet, while fewer cells expressed ROR gamma t. NKG2D(+) CD4(+) and DN populations were producers of IFN-gamma, but rarely IL-4 and IL-17. Taken together, we identify a novel subset of primary CD4(+) and DN type II NKT cells that expresses NKG2 receptors have typical NKT cell phenotypes and a TH1-like cytokine production.

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