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A new mouse strain for the analysis of invariant NKT cell function

Nature Immunology volume 16, pages 799–800 (2015)Cite this article

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Invariant natural killer T cells (iNKT cells) are a T lymphocyte population with restricted T cell antigen receptor (TCR) diversity. They make up a distinct subset of T lymphocytes that is activated by glycolipid antigens presented by CD1d, a non-classical major histocompatibility complex class I–like molecule. The invariant TCR of iNKT cells consists of a specific α-chain rearrangement, of α-chain variable region 14 and α-chain joining region 18 (Vα14-Jα18), encoded by Tcra gene segments Trav11-Traj18 in mice, and Vα24-Jα18 (TRAV10-TRAJ18) in humans, and a limited β-chain repertoire1.

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Figure 1: Traj18-deficient mice lack iNKT cells but have a normal Jα-region repertoire and show significantly fewer eosinophils in an airway hyper-reactivity model.

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Acknowledgements

Supported by the US National Institutes of Health (R37 AI71922 and RO1 105215 to M.K.).

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Author notes

  1. Shilpi Chandra and Meng Zhao: These authors contributed equally to this work.

Authors and Affiliations

  1. La Jolla Institute for Allergy & Immunology, La Jolla, California, USA

    Shilpi Chandra, Meng Zhao, Alvaro de Mingo Pulido, Jeremy Day, Zheng Fu & Mitchell Kronenberg

  2. Department of Inflammation Research, Amgen, Seattle, Washington, USA

    Alison Budelsky, Lori Siegel & Dirk Smith

Authors
  1. Shilpi Chandra
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  2. Meng Zhao
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  3. Alison Budelsky
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  4. Alvaro de Mingo Pulido
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  5. Jeremy Day
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  6. Zheng Fu
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  7. Lori Siegel
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Contributions

S.C., M.Z., A.B., J.D., Z.F., L.S., D.S. and M.K. designed and analyzed the experiments, which were performed by S.C., M.Z., A.d.M.P., J.D., L.S. and D.S.; and S.C., A.B., D.S. and M.K. wrote the paper.

Corresponding author

Correspondence to Mitchell Kronenberg.

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The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 Traj18-deficient mice lack iNKT cells.

The figure represents the percentage of iNKT cells in WT and Traj18-deficient mice in different tissues. The figure represents data from individual mice analyzed in three independent experiments (mean (SD) of n = 7 for all tissues except liver where n = 5) (****, P <0.0001, unpaired t test).

Supplementary Figure 2 Traj18-deficient mice do not produce cytokines upon α-GalCer injection.

WT or Traj18-deficient mice were injected IV with α-GalCer and blood was collected either 2 h or 24 h post injection. Data (mean (SD)) are representative of two experiments (n = 2 mice each group and n = 2 wells each time point, ****, P < 0.0001; **, P < 0.05, analyzed by 2-way ANOVA).

Supplementary Figure 3 Frequency of Jα use for Trav11 and Trav12 rearrangements.

Thymocytes were sorted and analyzed as described in Figure 1. (a,b) The frequencies of Jα region usage for Trav11 (a) and Trav12 (b) in WT (blue bars) or Traj18-deficient (red bars) mice are shown. Data are representative from one of two mice analyzed.

Supplementary Figure 4 Traj18-deficient mice showed less airway resistance in OVA model of pulmonary inflammation.

Mice were sensitized with OVA for two weeks and later challenged with OVA for three consecutive days. Airway resistance IV saline or methacholine (1 and 5 mg/kg) challenge was assessed 48 h after the last OVA challenge (mean (SD) of n = 4/group) (P < 0.0001 analyzed by 2-way ANOVA).

Supplementary Figure 5 Traj18-deficient mice showed less airway resistance in CRA model of pulmonary inflammation.

Mice were challenged with CRA antigen for two weeks. Airway resistance to IV methacholine (5 mg/kg) challenge was assessed 24 h after the last CRA challenge (mean of n = 5/group) (P < 0.0001 unpaired t test).

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Chandra, S., Zhao, M., Budelsky, A. et al. A new mouse strain for the analysis of invariant NKT cell function. Nat Immunol 16, 799–800 (2015). https://doi.org/10.1038/ni.3203

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