Characterization of NKT-cell hybridomas expressing invariant T-cell antigen receptors.
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(low)8(-). Initial characterizations revealed that both cell lines expressed an invariant T cell antigen receptor, which could be readily detected with alpha-galactosylceramide-loaded CD1d : Ig fusion protein (alpha-GalCer/CD1d). Sequence analyses of the alpha and beta chains of the T cell receptor V genes revealed that 1B6 and 2E10 cells expressed V alpha 14J alpha 18/V beta 8.2D beta 2J beta 2.7 and V alpha 14J alpha 18/V beta 8.1D beta 1J beta 1.1, respectively. When these hybridoma cells were stimulated with immobilized anti-CD3 monoclonal antibodies, alpha-GalCer/CD1d, or alpha-GalCer in the presence of antigen-presenting cells, they produced IL-4 and IFN-gamma. 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.
Nyambayar
Dashtsoodol
D
Division of Immunobiology, Research Section of Pathophysiology, Institute for Genetic Medicine, Hokkaido University, Sapporo, Japan.
Iwabuchi
Kazuya
K
Hedlund
Emma
E
Murakawa
Satoko
S
Shirai
Keiko
K
Iwabuchi
Chikako
C
Kon
Yujiroh
Y
Miyazaki
Yusei
Y
Yanagawa
Yoshiki
Y
Onoé
Kazunori
K
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism; D023201:CD40 Ligand / Q000378:metabolism; D045744:Cell Line, Tumor; D053222:Fas Ligand Protein / Q000378:metabolism; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism*; D007371:Interferon-gamma / Q000378:metabolism*; D015847:Interleukin-4 / Q000378:metabolism*; D007694:Killer Cells, Natural / Q000145:classification / Q000276:immunology / Q000378:metabolism*; D037181:Lectins, C-Type; D016131:Lymphocyte Subsets / Q000145:classification / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D013945:Thymoma / Q000378:metabolism; D013953:Thymus Neoplasms / Q000378:metabolism; D015854:Up-Regulation; D000099192:CD69 Antigens
D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D023201:CD40 Ligand; D053222:Fas Ligand Protein; D007371:Interferon-gamma; D015847:Interleukin-4; D037181:Lectins, C-Type; D011948:Receptors, Antigen, T-Cell; D000099192:CD69 Antigens