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J. Biol. Chem., Vol. 281, Issue 15, 10222-10229, April 14, 2006

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Ephrin-B1 Is Critical in T-cell Development^*

Guang Yu¹, Jianning Mao¹, Yulian Wu, Hongyu Luo², and Jiangping Wu, A National Scholar of the FRSQ^¶3

From the Laboratory of Immunology and the ^¶Nephrology Service of Notre Dame Hospital, Centre Hospitalier de l'Université de Montréal (CHUM), Montréal, Quebec H2L 4M1, Canada and the Second Affiliated Hospital of Zhejiang University Medical College, Hangzhou 310009, China

Eph kinases are the largest family of receptor tyrosine kinases, and their ligands, ephrins (EFNs), are also cell surface molecules. In this study, we investigated the role of EFNB1 and the Ephs it interacts with (collectively called EFNB1 receptors) in mouse T-cell development. In the thymus, CD8 single positive (SP) and CD4CD8 double positive (DP) cells expressed high levels of EFNB1 and EFNB1 receptors, whereas CD4 SP cells had moderate expression of both. Soluble EFNB1-Fc in fetal thymus organ culture caused significant subpopulation ratio skew, with increased CD4 SP and CD8 SP and decreased DP percentage, while the cellularity of the thymus remained constant. Moreover, in EFNB1-treated fetal thymus organ culture, CD117⁺, CD25⁺, DP, CD4 SP, and CD8 SP cells all had significantly enhanced proliferation history, according to bromodeoxyuridine uptake. In vitro culture of isolated thymocytes revealed that EFNB1-Fc on solid-phase protected thymocytes from anti-CD3-induced apoptosis, with concomitant augmentation of several antiapoptotic factors, particularly in CD4 SP and CD8 SP cells; on the other hand, soluble EFNB1-Fc promoted anti-CD3-induced apoptosis, as was the case in vivo. This study reveals that EFNB1 and EFNB1 receptors are critical in thymocyte development.

Received for publication, September 20, 2005 , and in revised form, February 7, 2006.

^* This work was supported by Canadian Institutes of Health Research (CIHR) Grants MOP57697 and MOP69089, the Kidney Foundation of Canada, the Heart and Stroke Foundation of Quebec, Juvenile Diabetes Research Foundation USA Grant 1-2005-197, and the J.-Louis Levesque Foundation (to J. W.). This work was also supported by group grants from the CIHR for New Emerging Teams in Transplantation and from Fonds de la Recherche en Santé du Québec (FRSQ) for Transfusional and Hemovigilance Medical Research. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ These two authors contributed equally to this work.

²To whom correspondence may be addressed: Laboratory of Immunology, Research Center, Notre-Dame Hospital, CHUM, Pavilion DeSève, Rm. Y-5616, 1560 Sherbrooke St. E., Montreal, Quebec H2L 4M1, Canada. Tel.: 514-890-8000 (ext. 27421); Fax: 514-412-7596; E-mail: hongyu.luo{at}umontreal.ca. ³To whom correspondence may be addressed: Laboratory of Immunology, Research Center, Notre-Dame Hospital, CHUM, Pavilion DeSève, Rm. Y-5616, 1560 Sherbrooke St. E., Montreal, Quebec H2L 4M1, Canada. Tel.: 514-890-8000 (ext. 25164); Fax: 514-412-7596; E-mail: jianping.wu{at}umontreal.ca.

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