IMMUNE NETWORK

The Korean Association of Immunobiologists (대한면역학회)
권호 표지

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

생쥐 수지상세포에서 발현하는 CD11c 프로모터의 규명

  • 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)
  • 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)
  • Published : 2008.12.30
Download PDF
⟨ Previous Next ⟩

Abstract

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.

Keywords

References

  1. Lanier LL, Arnaout MA, Schwarting R, Warner NL, Ross GD: p150/95, third member of the LFA-1/CR3 polypeptide family identified by anti-Leu M5 monoclonal antibody. Eur J Immunol 15;713-718, 1985 https://doi.org/10.1002/eji.1830150714
  2. Springer TA, Miller LJ, Anderson DC: p150,95, the third member of the Mac-1, LFA-1 human leukocyte adhesion glycoprotein family. J Immunol 136;240-245, 1986
  3. Sanchez-Madrid F, Nagy JA, Robbins E, Simon P, Springer TA: A human leukocyte differentiation antigen family with distinct alpha-subunits and a common beta-subunit: the lymphocyte function-associated antigen (LFA-1), the C3bi complement receptor (OKM1/Mac-1), and the p150,95 molecule. J Exp Med 158;1785-1803, 1983 https://doi.org/10.1084/jem.158.6.1785
  4. Corbi AL, Miller LJ, O'Connor K, Larson RS, Springer TA: cDNA cloning and complete primary structure of the alpha subunit of a leukocyte adhesion glycoprotein, p150,95. EMBO J 6;4023-4028, 1987
  5. Malhotra V, Hogg N, Sim RB: Ligand binding by the p150,95 antigen of U937 monocytic cells: properties in common with complement receptor type 3 (CR3). Eur J Immunol 16;1117-1123, 1986 https://doi.org/10.1002/eji.1830160915
  6. Keizer GD, Te Velde AA, Schwarting R, Figdor CG, De Vries JE: Role of p150,95 in adhesion, migration, chemotaxis and phagocytosis of human monocytes. Eur J Immunol 17;1317-1322, 1987 https://doi.org/10.1002/eji.1830170915
  7. Hogg N, Takacs L, Palmer DG, Selvendran Y, Allen C: The p150,95 molecule is a marker of human mononuclear phagocytes: comparison with expression of class II molecules. Eur J Immunol 16;240-248, 1986 https://doi.org/10.1002/eji.1830160306
  8. Winkel K, Sotzik F, Vremec D, Cameron PU, Shortman K: CD4 and CD8 expression by human and mouse thymic dendritic cells. Immunol Lett 40;93-99, 1994 https://doi.org/10.1016/0165-2478(94)90178-3
  9. Metlay JP, Witmer-Pack MD, Agger R, Crowley MT, Lawless D, Steinman RM: The distinct leukocyte integrins of mouse spleen dendritic cells as identified with new hamster monoclonal antibodies. J Exp Med 171;1753-1771, 1990 https://doi.org/10.1084/jem.171.5.1753
  10. Agger R, Crowley MT, Witmer-Pack MD: The surface of dendritic cells in the mouse as studied with monoclonal antibodies. Int Rev Immunol 6;89-101, 1990 https://doi.org/10.3109/08830189009056621
  11. Shelley CS, Teodoridis JM, Park H, Farokhzad OC, Bottinger EP, Arnaout MA: During differentiation of the monocytic cell line U937, Pur alpha mediates induction of the CD11c beta 2 integrin gene promoter. J Immunol 168;3887-3893, 2002 https://doi.org/10.4049/jimmunol.168.8.3887
  12. Lopez-Cabrera M, Nueda A, Vara A, Garcia-Aguilar J, Tugores A, Corbi AL: Characterization of the p150,95 leukocyte integrin alpha subunit (CD11c) gene promoter. Identification of cis-acting elements. J Biol Chem 268;1187-1193, 1993
  13. Lopez-Rodriguez C, Botella L, Corbi AL: CCAAT-enhancer-binding proteins (C/EBP) regulate the tissue specific activity of the CD11c integrin gene promoter through functional interactions with Sp1 proteins. J Biol Chem 272;29120-29126, 1997 https://doi.org/10.1074/jbc.272.46.29120
  14. Lopez-Rodriguez C, Corbi AL: PU.1 negatively regulates the CD11c integrin gene promoter through recognition of the major transcriptional start site. Eur J Immunol 27;1843-1847, 1997 https://doi.org/10.1002/eji.1830270804
  15. Bell D, Young JW, Banchereau J: Dendritic cells. Adv Immunol 72;255-324, 1999 https://doi.org/10.1016/S0065-2776(08)60023-1
  16. Jung S, Unutmaz D, Wong P, Sano G, De los Santos K, Sparwasser T, Wu S, Vuthoori S, Ko K, Zavala F, Pamer EG, Littman DR, Lang RA: In vivo depletion of CD11c(+) dendritic cells abrogates priming of CD8(+) T cells by exogenous cell-associated antigens. Immunity 17;211-220, 2002 https://doi.org/10.1016/S1074-7613(02)00365-5
  17. Brocker T, Riedinger M, Karjalainen K: Targeted expression of major histocompatibility complex (MHC) class II molecules demonstrates that dendritic cells can induce negative but not positive selection of thymocytes in vivo. J Exp Med 185;541-550, 1997 https://doi.org/10.1084/jem.185.3.541
  18. Arthur JF, Butterfield LH, Roth MD, Bui LA, Kiertscher SM, Lau R, Dubinett S, Glaspy J, McBride WH, Economou JS: A comparison of gene transfer methods in human dendritic cells. Cancer Gene Ther 4;17-25, 1997
  19. Diebold SS, Kursa M, Wagner E, Cotten M, Zenke M: Mannose polyethylenimine conjugates for targeted DNA delivery into dendritic cells. J Biol Chem 274;19087-19094, 1999 https://doi.org/10.1074/jbc.274.27.19087
  20. Diebold SS, Lehrmann H, Kursa M, Wagner E, Cotten M, Zenke M: Efficient gene delivery into human dendritic cells by adenovirus polyethylenimine and mannose polyethylenimine transfection. Hum Gene Ther 10;775-786, 1999 https://doi.org/10.1089/10430349950018535
  21. Strobel I, Berchtold S, Gotze A, Schulze U, Schuler G, Steinkasserer A: Human dendritic cells transfected with either RNA or DNA encoding influenza matrix protein M1 differ in their ability to stimulate cytotoxic T lymphocytes. Gene Ther 7;2028-2035, 2000 https://doi.org/10.1038/sj.gt.3301326
  22. Irvine AS, Trinder PK, Laughton DL, Ketteringham H, McDermott RH, Reid SC, Haines AM, Amir A, Husain R, Doshi R, Young LS, Mountain A: Efficient nonviral transfection of dendritic cells and their use for in vivo immunization. Nat Biotechnol 18;1273-1278, 2000 https://doi.org/10.1038/82383
  23. Lohmann S, Galle K, Knop J, Enk AH: CD83+ human dendritic cells transfected with tumor peptide cDNA by electroporation induce specific T-cell responses: a potential tool for gene immunotherapy. Cancer Gene Ther 7;605-614, 2000 https://doi.org/10.1038/sj.cgt.7700187
  24. Van Tendeloo VF, Snoeck HW, Lardon F, Vanham GL, Nijs G, Lenjou M, Hendriks L, Van Broeckhoven C, Moulijn A, Rodrigus I, Verdonk P, Van Bockstaele DR, Berneman ZN: Nonviral transfection of distinct types of human dendritic cells: high-efficiency gene transfer by electroporation into hematopoietic progenitor- but not monocyte-derived dendritic cells. Gene Ther 5;700-707, 1998 https://doi.org/10.1038/sj.gt.3300626
  25. Awasthi S, Cox RA: Transfection of murine dendritic cell line (JAWS II) by a nonviral transfection reagent. Biotechniques 35;600-602, 604, 2003
  26. Shen Z, Reznikoff G, Dranoff G, Rock KL: Cloned dendritic cells can present exogenous antigens on both MHC class I and class II molecules. J Immunol 158;2723-2730, 1997