Orthopedic & Muscular System: Current Research
Open Access

Can a functional assay on cytokine kinetics be used for the identification of a disease-related role for Single Nucleotide Polymorphisms (SNPs) in Ankolysing spondylitis?

3^rd International Conference and Exhibition on Orthopedics & Rheumatology

July 28-30, 2014 DoubleTree by Hilton Hotel San Francisco Airport, USA

Thomas Gelsing Carlsen, Pernille Kjærsgaard, Trine Jørgensen, Gunna Christiansen and Svend Birkelund

Accepted Abstracts: Orthop Muscul Syst

Abstract :

Interleukin 1α(IL-1α) is a proinflammatory cytokine that belongs to the IL-1 family. It is produced mainly by macrophages at sites of infection and regarded as an essential regulator of acute inflammation.IL-1α is synthesized as a 33 kDa precursor peptide that is cleaved by a calpain-like protease to a nuclear -associated 16 kDa propiece and a secreted 17 kDa mature IL-1α peptide. However, the full understanding of its dual function is missing. Recently, SNPs in the gene for IL-1α was also associated with the risk of developing ankylosing spondylitis (AS), a subgroup of the spondyloarthropathies. These findings lead us to produce antibodiestowards the N- and C-terminal region of IL-1α to investigate IL-1α kinetics in human macrophages. This would eventually be used to asses any correlation between a defect in the production of the cytokine and a disease related SNP found in the IL-1α gene in patients suffering from AS. In the present study, human macrophages (Mf) from blood monocytes, stimulated the cells with lipopolysaccharide (LPS) and analysed the production and localization of IL-1α by use of monoclonal antibodies (MAbs) was generated against recombinant precursor IL-1α. It was obtained a MAb specific for the N-terminal propiece and for the C-terminal mature form of IL-1α, respectively. Assays, including DNA sequencing, immunofluorescence microscopy, qPCR and FACS are now available for the analysis of IL-1α kinetics in blood samples from AS patients.