Characterization and application of two RANK-specific antibodies with different biological activities.
Antibodies play an important role in therapy and investigative biomedical research. The TNF-family member Receptor Activator of NF-κB (RANK) is known for its role in bone homeostasis and is increasingly recognized as a central player in immune regulation and epithelial cell activation. However, the study of RANK biology has been hampered by missing or insufficient characterization of high affinity tools that recognize RANK. Here, we present a careful description and comparison of two antibodies, RANK-02 obtained by phage display (Newa, 2014 [1]) and R12-31 generated by immunization (Kamijo, 2006 [2]). We found that both antibodies recognized mouse RANK with high affinity, while RANK-02 and R12-31 recognized human RANK with high and lower affinities, respectively. Using a cell apoptosis assay based on stimulation of a RANK:Fas fusion protein, and a cellular NF-κB signaling assay, we showed that R12-31 was agonist for both species. R12-31 interfered little or not at all with the binding of RANKL to RANK, in contrast to RANK-02 that efficiently prevented this interaction. Depending on the assay and species, RANK-02 was either a weak agonist or a partial antagonist of RANK. Both antibodies recognized human Langerhans cells, previously shown to express RANK, while dermal dendritic cells were poorly labeled. In vivo R12-31 agonist activity was demonstrated by its ability to induce the formation of intestinal villous microfold cells in mice. This characterization of two monoclonal antibodies should now allow better evaluation of their application as therapeutic reagents and investigative tools.
['Chypre|Mélanie|M|', 'Seaman|Jonathan|J|', 'Cordeiro|Olga G|OG|', 'Willen|Laure|L|', 'Knoop|Kathryn A|KA|', 'Buchanan|Andrew|A|', 'Sainson|Richard C A|RC|', 'Williams|Ifor R|IR|', 'Yagita|Hideo|H|', 'Schneider|Pascal|P|', 'Mueller|Christopher G|CG|']
D000818:Animals D000911:Antibodies, Monoclonal D000915:Antibody Affinity D002454:Cell Differentiation D061505:Cell Surface Display Techniques D004847:Epithelial Cells D000939:Epitopes D057809:HEK293 Cells D006801:Humans D007117:Immunization, Secondary D056747:Immunomodulation D007422:Intestines D019169:Jurkat Cells D007801:Langerhans Cells D051379:Mice D008810:Mice, Inbred C57BL D016328:NF-kappa B D053246:Receptor Activator of Nuclear Factor-kappa B D015398:Signal Transduction
Langerhans cell Monoclonal antibody Epithelial microfold cell RANK (TNFRSF11a)