Background Adiponectin isoforms (HMW/MMW) appears to be more involved in the pathogenesis of rheumatoid arthritis (RA) than LMW isoform.
Objectives The purpose of this study was to generate monoclonal antibodies (MAbs) specific to different adiponectin isoforms to develop a therapeutic agent for RA patients.
Methods Hybridoma cells producing monoclonal antibody were generated by fusing FO myeloma cells (ATCC) with splenic B cells from female Balb/c mice that were immunized with recombinant human adiponectin monomer. Hybridoma cells that produced MAbs recognizing human adiponectin isoforms were screened by ELISA and Western blots.
Results The MAb from hybridoma clone KH8-26 recognized mainly the middle molecular weight (MMW, hexamer) and weakly low molecular weight (LMW, trimer) isoforms of adiponectin in human serum while the KH8-3 MAb detected only MMW (hexamer) adiponectin. Also, the KH4-7 clone recognized the high molecular weight (HMW, multimer) and MMW adiponectin. These MAbs also recognized adiponectin in adipose tissue. In addition, they recognized the MMW adiponectin mostly in mouse, rat, horse and human serum. The MAb from clone KH4-7 effectively inhibited the increased IL-6 and IL-8 expression in adiponectin-stimulated human osteoblasts and human umbilical vein endothelial cells (HUVEC). This result suggests that MAb treatment may decrease the inflammation induced by adiponectin.
Conclusions In conclusion, MAbs against recombinant human adiponectin monomer can potentially be developed as therapeutic antibodies to target a detrimental isoform of adiponectin while maintaining the beneficial isoforms.
Arthritis Research & Therapy 2009, 11:R161
Ann Rheum Dis 2012;71:1724–1732
Acknowledgements The present study was supported by the Basic Science Research Program through the National Research Foundation of Korea and funded by the Ministry of Education, Science and Technology (Korea; grant nos. 2011-0009061 and 2010-0024089).
Disclosure of Interest None declared