Production of monoclonal antibodies that recognize the extracellular domain of mouse Langerin/CD207
Autor: | Sayuri Yamazaki, Anthony J. Bonito, Jae Y. Kim, Haekyung Lee, Sung Ho Park, Yoonkyung Do, Young-Sun Kang, Kayo Inaba, Ralph M. Steinman, Jae-Hoon Choi, Maggi Pack, Chae Gyu Park, Juliana Idoyaga, Cheolho Cheong |
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Rok vydání: | 2007 |
Předmět: |
Langerhans cell
Langerin medicine.drug_class Birbeck granules Immunoelectron microscopy Immunology Population Rats Inbred WF CHO Cells Monoclonal antibody Article Cell Line Mice Cricetulus Organ Culture Techniques Antigen Cricetinae medicine Animals Humans Immunology and Allergy Lectins C-Type education Mice Inbred BALB C education.field_of_study integumentary system biology Antibodies Monoclonal Dendritic cell Virology Molecular biology Protein Structure Tertiary Rats Mannose-Binding Lectins medicine.anatomical_structure Immunoglobulin M Antigens Surface biology.protein Binding Sites Antibody Protein Binding |
Zdroj: | Journal of Immunological Methods. 324:48-62 |
ISSN: | 0022-1759 |
Popis: | Langerin CD207 is a type II transmembrane protein. It is responsible for the formation of Birbeck granules, which are intracellular organelles within Langerhans cells, the dendritic cells of stratified squamous epithelia like the epidermis. Because current anti-CD207 antibodies have limitations, we prepared new monoclonals by immunizing rats with the extracellular region of mouse Langerin followed by a boost with enriched Langerhans cells (LCs). We secured a large panel of mAbs, most of which reacted with the carboxy terminal carbohydrate recognition domain. These mAbs could be used to immunoblot and immunoprecipitate mouse Langerin and to stain the cell surface and intracellular pools of CD207 by FACS analysis. Labeling of Birbeck granules was also achieved by immunoelectron microscopy. Anti-CD207 identified LCs in the epidermis and skin draining lymph nodes of BALB/c and C57BL/6 mice, but BALB/c mice had an additional Langerin(+) population in spleen, thymus and mesenteric lymph node. This additional subset had higher levels of CD8 and CD205 than epidermal LCs, and also had a less mature phenotype, i.e., lower MHC II, CD40 and CD86. Subcutaneous injection of IgG but not IgM forms of these new anti-CD207 mAbs led to rapid and selective labeling of the Langerin(+) cells in skin draining lymph nodes as well as spleen. The new IgG anti-CD207 mAbs should be useful for further research on LCs and dendritic cells including an evaluation of the consequences of antigen delivery within anti-CD207 mAbs in vivo. |
Databáze: | OpenAIRE |
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