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Purification and Expansion of Mouse Invariant Natural Killer T Cells for in vitro and in vivo Studies
08:37

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Published on: February 15, 2021

A novel mouse model for invariant NKT cell study.

Hiroshi Wakao1, Hiroshi Kawamoto, Sakura Sakata

  • 1Environmental Biology, School of Medicine, Hokkaido University, Sapporo, Japan. hwakao@med.hokudai.ac.jp

Journal of Immunology (Baltimore, Md. : 1950)
|September 6, 2007
PubMed
Summary

A novel mouse model with rearranged Valpha14-Jalpha18 T-cell receptor (TCR) enhances invariant NKT (iNKT) cell populations. These mice exhibit distinct iNKT cell characteristics, aiding research into iNKT cell development and function.

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Area of Science:

  • Immunology
  • Genetics
  • Cell Biology

Background:

  • Invariant natural killer T (iNKT) cells are crucial immune regulators.
  • Understanding iNKT cell development and function is vital for immunology.
  • Existing models may not fully recapitulate endogenous iNKT cell biology.

Purpose of the Study:

  • To generate and characterize a novel mouse model with an in-frame rearranged Valpha14-Jalpha18 T-cell receptor (TCR) allele.
  • To investigate the impact of this specific TCR configuration on iNKT cell development and characteristics.
  • To explore the functional responses of iNKT cells in this new model.

Main Methods:

  • Generation of a novel mouse model by crossing NKT cell clones with wild-type mice to achieve an in-frame Valpha14-Jalpha18 TCR.
  • Analysis of iNKT cell populations in various tissues (thymus, liver, spleen, bone marrow).
  • Stimulation with alpha-galactosylceramide (alphaGalCer) to assess cytokine production (Th1, Th2) and serum IgE levels.
  • Flow cytometry to analyze TCR Vbeta repertoire, Rag2 expression, and cell cycle progression.
  • Crossing with CD1d knockout mice to identify novel iNKT cell populations.

Main Results:

  • The Valpha14-Jalpha18 mouse model showed increased iNKT cell numbers across multiple organs.
  • Stimulation revealed balanced Th1/Th2 cytokine production without affecting IgE levels, distinguishing it from transgenic models.
  • iNKT cells displayed a biased TCRVbeta8 usage and reduced Rag2 expression in immature stages.
  • A novel iNKT cell precursor population was identified in Valpha14-Jalpha18 x CD1d knockout mice.

Conclusions:

  • The novel Valpha14-Jalpha18 mouse model accurately reflects endogenous iNKT cell development and function.
  • This model provides a valuable tool for studying iNKT cell biology, including development, repertoire selection, and functional responses.
  • The findings offer new insights into the heterogeneity and regulation of iNKT cell populations.