Production of monoclonal antibodies that recognize the extracellular domain of mouse langerin/CD207.
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.
Cheong
Cheolho
C
Laboratory of Cellular Physiology and Immunology and Chris Browne Center for Immunology and Immune Diseases, The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA.
Idoyaga
Juliana
J
Do
Yoonkyung
Y
Pack
Maggi
M
Park
Sung Ho
SH
Lee
Haekyung
H
Kang
Young-Sun
YS
Choi
Jae-Hoon
JH
Kim
Jae Y
JY
Bonito
Anthony
A
Inaba
Kayo
K
Yamazaki
Sayuri
S
Steinman
Ralph M
RM
Park
Chae Gyu
CG
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000378:metabolism; D000954:Antigens, Surface / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D001666:Binding Sites, Antibody; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D007075:Immunoglobulin M / Q000096:biosynthesis / Q000378:metabolism; D037181:Lectins, C-Type / Q000172:deficiency / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D037241:Mannose-Binding Lectins / Q000172:deficiency / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D009924:Organ Culture Techniques; D011485:Protein Binding / Q000276:immunology; D017434:Protein Structure, Tertiary; D051381:Rats; D011920:Rats, Inbred WF
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; C498497:Cd207 protein, mouse; D007075:Immunoglobulin M; D037181:Lectins, C-Type; D037241:Mannose-Binding Lectins