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Production of receptor activator of NF-κB ligand (RANKL) and tumour necrosis factor-α (TNF-α) and expression of macrophage colony-stimulating factor (M-CSF) and RANKL
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posted on 2011-12-31, 04:37 authored by Bert De Klerck, Isabelle Carpentier, Rik J Lories, Yvette Habraken, Jacques Piette, Geert Carmeliet, Rudi Beyaert, Alfons Billiau, Patrick MatthysCopyright information:
Taken from "Enhanced osteoclast development in collagen-induced arthritis in interferon-γ receptor knock-out mice as related to increased splenic CD11bmyelopoiesis"
Arthritis Research & Therapy 2004;6(3):R220-R231.
Published online 12 Mar 2004
PMCID:PMC416444.
Copyright © 2004 De Klerck et al., licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
RANKL and TNF-α concentrations measured in supernatant of anti-CD3 antibody-stimulated splenocytes. Splenocytes of naive and immunised mice (day 21 after immunisation) were cultured in the presence of M-CSF and stimulated with 1 μg/ml anti-CD3 antibody. RANKL (a) and TNF-α (b) detected in supernatants by enzyme-linked immunosorbent assay 6 days after stimulation. *< 0.05 compared with wild-type mice (Mann–Whitney -test). Reverse transcriptase PCR performed on RNA of isolated inflamed synovia of two interferon-γ receptor knock-out (IFN-γR KO) mice (KO1, KO2) and two wild-type mice (WT1, WT2) showing transcription of RANKL and M-CSF within the inflamed synovium. The housekeeping gene β-actin was used to normalise the levels of cDNA. Analysis of calcitonin receptor expression level by real-time quantitative PCR on synovium and spleen from three wild-type and three IFN-γR KO mice. Values are the numbers of calcitonin receptor mRNA copies per 1000 copies of hypoxanthine transferase. CIA, collagen-induced arthritis.