Blocking antibodies against tTG and β1
- or β3-integrins differentially affect migration of TPA-induced THP-1 cells on the 110-kd and the 42-kd fragments of Fn. The 5 × 104 THP-vector cells in serum-free AIM-V medium containing 0.5% BSA were stimulated with 150 ng/mL TPA and placed for 4 hours at 37°C into upper chambers of Transwells (Costar) where undersurface of the inserts was precoated with Fn (░) or its 110-kd (■) or 42-kd (▪) fragments. Then 125 ng/mL MCP-1 was added to lower chambers. Before adding cells to the Transwells, cell samples were preincubated for 1 hour with control nonimmune IgG, function-blocking polyclonal anti-tTG antibody, anti-tTG mAb 4G3 or mAb CUB7402, antibody against FXIIIA, or blocking mAbs against β1-, β2-, or β3-integrins. The antibodies were present in the medium during the assay. The numbers of transmigrated cells on each substrate in the presence of control nonimmune IgG (23.6 × 103 on Fn, 7.9 × 103 on the 110-kd fragment, and 13.7 × 103 on the 42-kd fragment) were taken as 100%. Bars depict the means of 3 separate experiments performed in duplicate.
