[In vitro study on the amelioration of T cell-mediated osteoclastogenesis by anti-RANKL-polyclonal antibody].
OBJECTIVE
To characterize the effect of receptor activator of NF-kB ligand (RANKL) polyclonal antibody on T cell-mediated osteoclasogenesis.


METHODS
Rabbit anti-rat RANKL polyclonal IgG antibody was performed, performing recombinated mouse RANKL-induced osteoclastogenesis in different antibody doses (1microg/mL,5microg/mL and 10microg/mL), the number of TRAP positive multinuclear cells (cells/well) was calculated under microscopy. The cervical lymph nodes in rats were enriched T-cells for in vitro experiments. The T cell-mediated osteoclastogenesis was tested by co-culture assay of RAW 264.7 cells with formalin-fixed T cell or T cell supernatant, the number of TRAP positive multinuclear cells (cells/well) was calculated under microscopy. In some experiments, the concentration of sRANKL (soluble RANKL) was measured in supernatant by ELISA (pg/mL). SPSS 11.5 software package was used for statistical analysis.


RESULTS
Rabbit anti-rat RANKL antibody inhibited osteoclastogenesis by recombined mouse RANKL-induced, and dose-dependent; Anti-RANKL antibody inhibited T cell and culture supernatant induced osteoclastogensis, significantly different from that in the group without antibody(P<0.05), and dose-dependent; Anti-RANKL specific antibody also markedly reduced sRANKL concentration in supernatant, significantly different from that in the control group without antibody(P<0.05).


CONCLUSIONS
Anti-RANKL specific antibody blocks direct effect of RANKL and interferes with immune T cell-mediated bone resorption by reduction of sRANKL level.
Lin
Xiao-ping
XP
Department of Stomatology, Shengjing Hospital of China Medical University, Shenyang 110004, Liaoning Province, China. Xiaoping_ba@yahoo.com
Han
Xiao-zhe
XZ
Wei
Wei
W
Martin
A Taubman
AT
D000818:Animals; D000888:Antibodies, Anti-Idiotypic; D002352:Carrier Proteins; D018920:Coculture Techniques; D066298:In Vitro Techniques; D008562:Membrane Glycoproteins; D051379:Mice; D010010:Osteoclasts; D053245:RANK Ligand; D011817:Rabbits; D051381:Rats; D053246:Receptor Activator of Nuclear Factor-kappa B; D013601:T-Lymphocytes
D000888:Antibodies, Anti-Idiotypic; D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D053245:RANK Ligand; D053246:Receptor Activator of Nuclear Factor-kappa B; C048844:anti-IgG