Figure 8.

Neutralization of collagen XIII blocks transmigration of VLA1-positive monocytes in transwell assays. Bone marrow derived leukocytes were subjected to biopanning on plates coated with anti-VLA1 antibodies. The panned cells were mixed with either anti-collagen XIII neutralizing antibodies (Ab2) or isotype matched control antibodies (IgM) and then added to the top layer of transwells coated with a monolayer of irradiated embryonic fibroblasts, and allowed to migrate for 14 hours. Cells were collected from the top (nonmigrated) and the bottom (migrated) compartments of the chamber and analyzed by dual-fluorescence-activated cell sorting for both CD11b (abscissa) and VLA1 (ordinate). The dual immunofluorescent VLA1-positive monocyte population in Q2 shows a clear reduction in this population in the Ab2-treated cells compared with cells treated with the isotype control mAb. Ab2 reduced VLA1 migration by 40.3% ± 1.7% over three independent transwell experiments (see Table 1).