IMMUNE NETWORK

  • 제8권4호
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  • Pages.137-142
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  • 2008
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  • 1598-2629(pISSN)
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  • 2092-6685(eISSN)
대한면역학회 (The Korean Association of Immunobiologists)
권호 표지

생쥐 수지상세포에서 발현하는 CD11c 프로모터의 규명

Characterization of the CD11c Promoter Which Is Expressed in the Mouse Dendritic Cells

  • 김봉긔 (서울대학교 의과대학 미생물학교실, 바이오이종장기 개발 사업단, 종양면역의과학센터, 장기이식연구소) ;
  • 김정식 (서울대학교 의과대학 미생물학교실, 바이오이종장기 개발 사업단, 종양면역의과학센터, 장기이식연구소) ;
  • 박정규 (서울대학교 의과대학 미생물학교실, 바이오이종장기 개발 사업단, 종양면역의과학센터, 장기이식연구소)
  • Kim, Bon-Gi (Department of Microbiology and Immunology, Cancer Research Institute, Tumor Immunity Medical Research Center, Transplantation Research Institute, Xenotransplantation Research Center, Seoul National University College of Medicine) ;
  • Kim, Jung-Sik (Department of Microbiology and Immunology, Cancer Research Institute, Tumor Immunity Medical Research Center, Transplantation Research Institute, Xenotransplantation Research Center, Seoul National University College of Medicine) ;
  • Park, Chung-Gyu (Department of Microbiology and Immunology, Cancer Research Institute, Tumor Immunity Medical Research Center, Transplantation Research Institute, Xenotransplantation Research Center, Seoul National University College of Medicine)
  • 발행 : 2008.12.30
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초록

Background: CD11c, also known as integrin alpha x, is one of the optimum markers of dendritic cells. However, the regulation of the CD11c expression in mouse has not been identified yet. In this study, in order to analyze the regulation of CD11c expression, the promoter of CD11c was cloned and characterized. Methods: To identify the promoter portion, various sizes of what are considered to be CD11c promoter fragments was amplified by polymerase chain reaction (PCR), using mouse genomic DNA as a template. After sequence was obtained, these fragments were transfected into various cell lines including mouse dendritic cell lines such as JAWSII and DC2.4 and L929 as control cell line.. The promoter activity of three promoter fragments was measured and compared by luciferase activity in the transfected cells. Results: Three clones with size of 1kb, 3kb and 6kb were obtained from mouse genomic DNA. Flow cytometry analysis of JAWSII cells revealed that 52% of the cells expressed CD11c, which was confirmed by RT-PCR analysis. On the contrary, L929 and DC 2.4 cells did not express CD11c. The CD11c+ JAWSII cells were enriched from 52% to 90% with cell sorter. The comparative luciferase activity analyisis demonstrated that the region responsible for tissue specific expression was contained within -3 kb and the clone with size of 3 kb particularly showed higher luciferase activity than 6 kb and 1 kb clones. Conclusion: The CD11c promoter region containing the region responsible for tissue specificity was successfully cloned and -3 kb region showed the highest activity.

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