siRNA knockdown of ADAM-10, but not ADAM-17, significantly reduces fractalkine shedding following pro-inflammatory cytokine treatment in a human adult brain endothelial cell line.
Fractalkine shedding is believed to occur constitutively and following induction via the activity of two membrane-bound enzymes, ADAM-10 and ADAM-17. However, our previous work suggested that ADAM-17 is not involved in the proteolytic release of fractalkine under TNF treatment of a human adult brain endothelial cell line, hCMEC/D3. The pro-inflammatory cytokine, TNF, has previously been shown to be expressed in the perivascular cuffs in multiple sclerosis. Here we sought to identify, using siRNAs to silence the expression of ADAM-10 and ADAM-17, whether ADAM-10 is responsible for TNF-induced shedding of fractalkine from the cell membrane in hCMEC/D3. Our findings suggest that ADAM-10, and not ADAM-17, is the major protease involved in fractalkine release under pro-inflammatory conditions in this human adult brain endothelial cell model.
Biomedical Research Centre, Sheffield Hallam University, Howard St., Sheffield S1 1WB, UK.
Louise A
LA
Hurst
Rowena A D
RA
Bunning
Basil
B
Sharrack
M Nicola
MN
Woodroofe
D051722:ADAM Proteins / Q000235:genetics*; D000072197:ADAM10 Protein; D000072198:ADAM17 Protein; D000328:Adult; D053829:Amyloid Precursor Protein Secretases / Q000235:genetics*; D001921:Brain / Q000098:blood supply*; D002460:Cell Line; D054428:Chemokine CX3CL1 / Q000378:metabolism*; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D054428:Chemokine CX3CL1; D008565:Membrane Proteins; D034741:RNA, Small Interfering; D014409:Tumor Necrosis Factor-alpha; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human; D000072198:ADAM17 Protein; C000606462:ADAM17 protein, human