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The Journal of Immunology, 2006, 176: 3855.
Copyright © 2006 by The American Association of Immunologists

LETTERS TO THE EDITOR

Comment on "CD226 Is Specifically Expressed on the Surface of Th1 Cells and Regulates Their Expansion and Effector Functions"

Kazuko Shibuya, Kai Shibata, Satoko Tahara-Hanaoka and Akira Shibuya

Department of Immunology, Institute of Basic Medical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Japan

The Th1/Th2 cell plays a distinct role in immune responses. However, cell surface markers specific for each cell type have not yet been identified. In a recent article, Dardalhon et al. (1) reported that CD226 (DNAM-1) is a cell surface molecule expressed selectively on differentiated Th1, but not Th2 or Th0, cells from BALB/c mice, using the 10E5 mAb generated in their laboratory. They also described that CD226 was expressed on a lower percentage (~40%) of fresh CD4+ T cells (1). In our laboratory, we generated three different rat mAbs, designated TX-42, TX-43, and TX-44, against mouse CD226 (2, 3). Using these three mAbs, we observed that >80% of fresh CD4+ T cells in spleen of BALB/c as well as C57BL/6 mice expressed CD226 (Fig. 1A). We also found that CD226 was expressed on most naive CD4+ T cells, as characterized by the high expression of CD62L (data not shown). Moreover, CD226 was expressed on the majority of both polarized Th1 and Th2 cells generated by in vitro culture of naive CD4+ T cells with IL-12 and IL-4, respectively, in addition to stimulation with anti-CD3 mAb. We finally found that not only polarized IFN-{gamma}-producing but also IL-4-producing CD4+ T cells expressed CD226 in spleen from both BALB/c and C57BL/6 mice (Fig. 1B). Thus, our studies indicate that CD226 is indeed expressed on Th2 cells and thereby not a specific marker for Th1 cells.


Figure 1
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  		FIGURE 1. Expression of CD226 on CD4+ helper T cells. A, Spleen cells from BALB/c or C57BL/6 mice were stained with biotin-conjugated anti-CD226 mAb (TX42 or TX43) (solid lines) or biotin-conjugated isotype-matched control Ig (rat IgG2a) (dotted lines). Cells were then stained with FITC-conjugated anti-CD4 mAb and PE-conjugated streptavidin and analyzed by flow cytometry (A). Similar results were obtained by using TX44 mAb. B, Spleen cells from BALB/c or C57BL/6 mice were stimulated with PMA and ionomycin for 4 h, and intracellular cytokines and CD226 expressions, by using TX42 mAb, on spleen cells from BALB/c or C57BL/6 mice were analyzed by three-color flow cytometry as described previously (4 ).

 

References

  1. Dardalhon, V., A. S. Schubart, J. Reddy, J. H. Meyers, L. Monney, C. A. Sabatos, R. Ahuja, K. Nguyen, G. J. Freeman, E. A. Greenfield, et al 2005. CD226 is specifically expressed on the surface of Th1 cells and regulates their expansion and effector functions. J. Immunol. 175: 1558-1565. [Abstract/Free Full Text]
  2. Tahara-Hanaoka, S., A. Miyamoto, A. Hara, S. Honda, K. Shibuya, A. Shibuya. 2005. Identification and characterization of murine DNAM-1 (CD226) and its poliovirus receptor family ligands. Biochem. Biophys. Res. Commun. 329: 996-1000. [Medline]
  3. Tahara-Hanaoka, S., K. Shibuya, H. Kai, A. Miyamoto, Y. Morikawa, N. Ohkochi, S. I. Honda, A. Shibuya. 2006. Tumor rejection by the poliovirus receptor family ligands of the DNAM-1 (CD226) receptor. Blood 107: 1491-1496. [Abstract/Free Full Text]
  4. Shibuya, K., J. Shirakawa, T. Kameyama, S. Honda, S. Tahara-Hanaoka, A. Miyamoto, M. Onodera, T. Sumida, H. Nakauchi, H. Miyoshi, A. Shibuya. 2003. CD226 (DNAM-1) is involved in lymphocyte function-associated antigen 1 costimulatory signal for naive T cell differentiation and proliferation. J. Exp. Med. 198: 1829-1839. [Abstract/Free Full Text]



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V. Dardalhon, A. S. Schubart, and V. K. Kuchroo
Response to Comment on "CD226 Is Specifically Expressed on the Surface of Th1 Cells and Regulates Their Expansion and Effector Functions"
J. Immunol., April 1, 2006; 176(7): 3856 - 3856.
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