In vitro selection of RNA aptamers that block CCL1 chemokine function.
CCL1, the CCR8 ligand, is a CC chemokine secreted by activated monocytes and lymphocytes and is a potent chemoattractant for these cell types. The in vivo role of the CCL1/CCR8 axis in Th2-mediated inflammation is far from clear. Ligand neutralisation studies reported discrepancies in the effect of CCL1/CCR8 and CCR8 knockout studies showed very different insights into the functional role of the CCR8. To further study the biological function of CCL1, we focused on the generation and characterisation of RNA aptamers. We report here the in vitro isolation of the first nuclease resistant and selective RNA aptamer (T48) with high-binding affinity for human and mouse CCL1. The T48 aptamer but not a random control aptamer antagonises CCL1 function in a dose-dependent fashion in both heparin binding and chemotaxis assays. To our knowledge, the T48 aptamer constitutes one of the most potent CCL1 antagonists reported to date and is an excellent tool to dissect CCL1-specific function in vivo. The T48 aptamer may also have potential as new generation of therapeutic tools.
Marro
Martin L
ML
Gene Expression and Protein Biochemistry, GlaxoSmithKline R&D, Gunnels Wood Road, Stevenage, Herts SG1 2NY, UK.
Daniels
Dion A
DA
Andrew
David P
DP
Chapman
Trevor D
TD
Gearing
Katy L
KL
D000818:Animals; D001483:Base Sequence; D001671:Biochemistry / Q000379:methods*; D054402:Chemokine CCL1; D018925:Chemokines / Q000737:chemistry / Q000378:metabolism; D019742:Chemokines, CC / Q000737:chemistry*; D002630:Chemotactic Factors / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D006801:Humans; D066298:In Vitro Techniques; D007249:Inflammation; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry*
C065971:CCL1 protein, human; D054402:Chemokine CCL1; D018925:Chemokines; D019742:Chemokines, CC; D002630:Chemotactic Factors; D008024:Ligands; D012313:RNA