Characterization of NKT-cell Hybridomas Expressing Invariant T-cell Antigen Receptors

Dashtsoodol Nyambayar; Kazuya Iwabuchi; Emma Hedlund; Satoko Murakawa; Keiko Shirai; Chikako Iwabuchi; Yujiroh Kon; Yusei Miyazaki; Yoshiki Yanagawa; Kazunori Onoé

doi:10.3960/jslrt.47.1

Original Articles

Characterization of NKT-cell Hybridomas Expressing Invariant T-cell Antigen Receptors

Dashtsoodol Nyambayar, Kazuya Iwabuchi, Emma Hedlund, Satoko Murakawa, Keiko Shirai, Chikako Iwabuchi, Yujiroh Kon, Yusei Miyazaki, Yoshiki Yanagawa, Kazunori Onoé

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Dashtsoodol Nyambayar
Division of Immunobiology, Research Section of Pathophysiology, Institute for Genetic Medicine, Hokkaido University
RIKEN Research Center for Allergy and Immunology
Kazuya Iwabuchi
Division of Immunobiology, Research Section of Pathophysiology, Institute for Genetic Medicine, Hokkaido University
Emma Hedlund
Division of Immunobiology, Research Section of Pathophysiology, Institute for Genetic Medicine, Hokkaido University
Lund University School of Medicine
Satoko Murakawa
Division of Immunobiology, Research Section of Pathophysiology, Institute for Genetic Medicine, Hokkaido University
Keiko Shirai
Division of Immunobiology, Research Section of Pathophysiology, Institute for Genetic Medicine, Hokkaido University
Chikako Iwabuchi
Division of Immunobiology, Research Section of Pathophysiology, Institute for Genetic Medicine, Hokkaido University
Yujiroh Kon
Division of Immunobiology, Research Section of Pathophysiology, Institute for Genetic Medicine, Hokkaido University
Yusei Miyazaki
Division of Immunobiology, Research Section of Pathophysiology, Institute for Genetic Medicine, Hokkaido University
Yoshiki Yanagawa
Division of Immunobiology, Research Section of Pathophysiology, Institute for Genetic Medicine, Hokkaido University
Kazunori Onoé
Division of Immunobiology, Research Section of Pathophysiology, Institute for Genetic Medicine, Hokkaido University

Keywords: NKT-cell, Hybridoma, Invariant TCR, IL-4, IFN-γ

JOURNAL FREE ACCESS

2007 Volume 47 Issue 1 Pages 1-8

DOI https://doi.org/10.3960/jslrt.47.1

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Abstract

Two natural killer T (NKT)-cell hybridomas were established by fusing sorted NKT cells with BW1100 thymoma cells. The first hybridoma line, 1B6, was CD4⁺8^-, whereas the second one, 2E10, was CD4^low8^-. Initial characterizations revealed that both cell lines expressed an invariant T cell antigen receptor, which could be readily detected with α-galactosylceramide-loaded CD1d : Ig fusion protein (α-GalCer/CD1d). Sequence analyses of the α and β chains of the T cell receptor V genes revealed that 1B6 and 2E10 cells expressed Vα14Jα18/Vβ8.2Dβ2Jβ2.7 and Vα14Jα18/Vβ8.1Dβ1Jβ1.1, respectively. When these hybridoma cells were stimulated with immobilized anti-CD3 monoclonal antibodies, α-GalCer/CD1d, or α-GalCer in the presence of antigen-presenting cells, they produced IL-4 and IFN-γ. The expression levels of CD69, CD154, and CD178 were concomitantly up-regulated on both hybridomas upon stimulation. Because it is difficult to isolate a sufficient number of NKT cells, these hybridomas should provide useful platforms to study a variety of functions of NKT cells. [J Clin Exp Hematopathol 47(1) : 1-8, 2007]

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