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* Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka, Japan;
PRESTO, Japan Science and Technology Agency, Kawaguchi, Japan;
Tumor Microenvironment Program, Cancer Research Center, Burnham Institute for Medical Research, La Jolla, CA; and
Division of Mammalian Development, National Institute of Genetics, Mishima, Japan
High endothelial venules (HEVs) are specialized blood vessels of secondary lymphoid organs composed of endothelial cells with a characteristic cuboidal morphology. Lymphocytes selectively adhere to and migrate across HEVs to initiate immune responses. In this study, we established a novel transgenic mouse line expressing Cre recombinase under the transcriptional control of the gene encoding HEV-expressed sulfotransferase, N-acetylglucosamine-6-O-sulfotransferase 2 (GlcNAc6ST-2), using bacterial artificial chromosome recombineering. Crossing these transgenic mice with the ROSA26 reporter strain, which expresses lacZ following Cre-mediated recombination, and staining the resulting progeny with 5-bromo-4-chloro-5-indolyl-β-D-galactoside indicated that Cre recombinase was specifically expressed in mAb MECA79-reactive HEVs in secondary lymphoid organs but not in any other blood vessels of the transgenic mice. The expression of Cre recombinase correlated with a developmental switch, from immature, mAb MECA367-reactive HEVs to mature, mAb MECA79-reactive HEVs in neonatal lymph nodes. In addition to the HEVs, Cre recombinase was also strongly expressed in the colonic villi, which recapitulated the intrinsic expression of GlcNAc6ST-2 as confirmed in GlcNAc6ST-2GFP/GFP knock-in mice and by RT-PCR. Furthermore, treatment with an antimicrobial agent revealed that the colonic expression of Cre recombinase in the transgenic mice was regulated by commensal bacteria in the colon. In addition, Cre recombinase was expressed in a small subset of cells in the brain, testis, stomach, small intestine, and lung. In view of the restricted expression of Cre recombinase, this transgenic mouse line should be useful for elucidating tissue-specific gene functions using the Cre/loxP system.
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1 This work was supported in part by Grants-in-Aid for Scientific Research, Category (B) and Grants-in-Aid for Scientific Research on Priority Areas, Dynamics of Extracellular Environments from the Ministry of Education, Culture, Sports, Science and Technology, Japan (Grants 18390029 and 18060034, respectively, to H.K.), the 25th Anniversary Research Grant from the Research Foundation for Pharmaceutical Sciences, Japan (to H.K.), National Institute of Genetics Cooperative Research Program 2006-B9 (to H.K.), and National Institutes of Health Grant P01 CA71932 (to M.F.).
2 Address correspondence and reprint requests to Dr. Hiroto Kawashima, Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Shizuoka 422-8526, Japan. E-mail address: kawashih{at}u-shizuoka-ken.ac.jp
3 Abbreviations used in this paper: HEV, high endothelial venule; PNAd, peripheral node addressin; PLN, peripheral lymph node; MLN, mesenteric lymph node; GlcNAc6ST, N-acetylglucosamine-6-O-sulfotransferase; MAdCAM-1, mucosal addressin cell adhesion molecule 1; PP, Peyer’s patch; BAC, bacterial artificial chromosome; X-gal, 5-bromo-4-chloro-3-indolyl-β-D-galactoside; EGFP, enhanced GFP; LB, Luria-Bertani; pAb, polyclonal Ab; Plzf, promyelocytic leukemia zinc finger protein; DDX4, DEAD-box protein 4; Muc2, mucin 2; Sox9, SRY box containing gene 9; AMPC, amoxicillin; NALT, nasal-associated lymphoid tissue.
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