Invariant natural killer T cells (iNKT cells) are a highly conserved lymphocyte population in mice and humans that express an invariant TCR ? chain. These lymphocytes make rapid, innate-like responses, and as a consequence, there are a number of commercial and clinical efforts to activate these cells in vivo, or expand them in vitro, in order to combat cancer, provide adjuvant or stimulating effects for vaccines, and to prevent immune-mediated diseases. It is surprising that in some cases iNKT cells stimulate Th1 or Th2 immunity and inflammation, while in other instances they are anti-inflammatory. The diversity of their effects on the immune response could be related to the selective activation of functional subsets of iNKT cells, called NKT1, NKT2 and NKT17 cells. These subsets are analogous to the well known Th1, Th2 and Th17 cells, but they differentiate in the thymus without exogenous antigenic stimulation. Our preliminary population-based and single cell RNA-Seq data show that the transcriptomes of the NKT1, NKT2 and NKT17 subsets are highly divergent, despite their similar specificity. The experiments in this application are designed to identify the differentiation steps leading to the subsets and the relationships between them, and their stability. To do this, in Aim 1 we will analyze the developmental potential of individual subsets, including iNKT cells with an intermediate or possibly transitional phenotype, using both organ cultures and by intrathymic injection. Additionally, we will use cytokine transcript fate-mapping mice to trace lineage relationships.
In aims 2 and 3, we will determine what are the cell intrinsic and extrinsic factors that drive the differentiation of the iNKT cell subsets. We will analyze nonproductive ? rearrangements by next generation sequencing to provide a clonal marker, in order to determine if a single iNKT cell clone can give rise to different functional subsets. Additionally, using different types of mice in which expression of a single TCR ? is enforced, we will determine if variations in the ? chain can drive preferential subset differentiation.
In aim 4, we will challenge mice with antigen, infection or chronic inflammation to determine how stable the thymus generated NKT17 subset is when confronted with inflammation or antigenic challenge. Additionally, we will analyze the transcriptomes of NKT1, NKT2 and NKT17 cells in spleen and liver, to assess the stability of the thymus-generated gene programs. Because iNKT cells do not recirculate, the transcriptome data also will provide information on the imprint of long-term residence in different organs on the iNKT cell subsets. Different populations of innate-like lymphocytes, which in addition to iNKT cells; include ?? T cells and innate lymphoid cells (ILCs), acquire during their differentiation gene programs that enforce polarized patterns of cytokine secretion. Undoubtedly there will be common themes between these cell types, and therefore our proposed work will have broader implications for understanding the differentiation of innate lymphocytes.

Public Health Relevance

Natural killer T cells (NKT cells) are a type of white blood cell that stimulate and carry out potent immune responses and that may be useful for immune therapy for cancer and for preventing immune mediated diseases. Categories or subsets of these cells can affect the immune response in different and sometimes opposing ways. The research in this grant is designed to understand how these different subsets originate, what factors cause them to form, and how stable they are in order so that the power of NKT cells may be harnessed more effectively for immune therapies.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
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Cellular and Molecular Immunology - B Study Section (CMIB)
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Kelly, Halonna R
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La Jolla Institute
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La Jolla
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Crosby, Catherine M; Kronenberg, Mitchell (2018) Tissue-specific functions of invariant natural killer T cells. Nat Rev Immunol 18:559-574
Chandra, Shilpi; Wingender, Gerhard; Greenbaum, Jason A et al. (2018) Development of Asthma in Inner-City Children: Possible Roles of MAIT Cells and Variation in the Home Environment. J Immunol 200:1995-2003
Hartmann, Nadine; McMurtrey, Curtis; Sorensen, Michelle L et al. (2018) Riboflavin Metabolism Variation among Clinical Isolates of Streptococcus pneumoniae Results in Differential Activation of Mucosal-associated Invariant T Cells. Am J Respir Cell Mol Biol 58:767-776
de Mingo Pulido, Álvaro; de Gregorio, Estefanía; Chandra, Shilpi et al. (2018) Differential Role of Cathepsins S and B In Hepatic APC-Mediated NKT Cell Activation and Cytokine Secretion. Front Immunol 9:391
Chandra, Shilpi; Gray, James; Kiosses, William B et al. (2018) Mrp1 is involved in lipid presentation and iNKT cell activation by Streptococcus pneumoniae. Nat Commun 9:4279
Hartmann, Nadine; Harriff, Melanie J; McMurtrey, Curtis P et al. (2018) Role of MAIT cells in pulmonary bacterial infection. Mol Immunol 101:155-159
Zhao, Meng; Svensson, Mattias N D; Venken, Koen et al. (2018) Altered thymic differentiation and modulation of arthritis by invariant NKT cells expressing mutant ZAP70. Nat Commun 9:2627
Sag, Duygu; Özkan, Müge; Kronenberg, Mitchell et al. (2017) Improved Detection of Cytokines Produced by Invariant NKT Cells. Sci Rep 7:16607
Hartmann, Nadine; Kronenberg, Mitchell (2017) MAITs onstage in mice and men with three acts for development. Immunol Cell Biol 95:3-4
Nowyhed, Heba N; Chandra, Shilpi; Kiosses, William et al. (2017) ATP Binding Cassette Transporter ABCA7 Regulates NKT Cell Development and Function by Controlling CD1d Expression and Lipid Raft Content. Sci Rep 7:40273

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