Fig. 6.
Peripheral blood leukocyte (PBL) migration was examined by using a dual-chamber motility system, in which the upper chamber was separated from the lower chamber by a polycarbonate track-etch membrane with 3-μm pores. Cell migration was allowed to proceed from the upper to the lower chamber for 60 min at 37°C in a CO2 incubator. The lower chamber contained materials to be tested for their effect on PBL motility. PBL motility was measured with an ATP luminescence-based motility-invasion assay. PBL migration (60 min at 37°C) was significantly increased by IL-8 (0.5 × 10–9 M), as expected, and by the supernatant (Sup) of the HCl (pH 5, 3 h)-filled mucosal sac (*P < 0.05 ANOVA). When using antagonists (Antag) or antibodies, cells were pretreated with the antagonist or antibody for 30 min before being placed in the upper chamber. The lower chamber contained the same concentration of antagonists or antibodies as the upper chamber. The increased PBL migration was significantly reduced by IL-8 immunoneutralization by an IL-8 antibody (1:200), by a PAF receptor antagonist (CV3988, 10−5 M), and by an NK-1 receptor (NK1R) antagonist (10−5 M) (*P < 0.05 ANOVA). The increased PBL migration was not affected by a CGRP antagonist (CGRP8–37, 10−6 M). Data represent means + SE of 3 experiments.