Comparison of the CD11b activation epitope on peripheral and transmigrated neutrophils was analysed by Wilcoxon matched pairs test. Correlations between IL-8, both endogenous and recombinant, and the expression of CD11b were analysed by Spearman’s rank order analysis. Significant correlations with a regression coefficient (R) ≥0.7

were further analysed. A P < 0.05 was considered significant. The median number of transmigrated cells per skin chamber was 2.14 (1.59–3.96) 106 cells with 82.1 (77.6–84.8) % granulocytes, 13.2 (10.3–16.4) % monocytes and 2.82 (2.37–4.37) % lymphocytes. In the peripheral circulation, the number of leucocytes was 4.85 (3.6–6.1)*109 leucocytes/l with 54.1 (50.5–58.9) % granulocytes, 8.51 (7.61–10.5) % monocytes and 33.3 (30.8–37.9) % lymphocytes. From the original skin blister, analysed for CD11b activation after 14 h of incubation, Opaganib price the median number of LY2109761 solubility dmso extravasated leucocytes was 0.11 (0.04–0.14) million cells per skin blister. The expression of CD11b activation epitope on extravasated neutrophils from the original 14-h blister was 73.2 (18.9–83.4) % and corresponding expression on circulating neutrophils was 1.96 (1.29–2.14) %, P = 0.04. The concentration of IL-8 in serum was 8.5 (1.9–11) pg/ml and in the 14-h blister fluid 338 (194–10,627) pg/ml, P = 0.04. The concentration of soluble mediators in serum and

in the skin chamber fluid was assessed by Milliplex multi-analysis and ELISA and

is presented in Fig. 1. Significantly higher concentrations of soluble markers were detected in the skin chamber fluid compared to that in serum for all markers Liothyronine Sodium except eotaxin (P < 0.01 for IL-4 and IL-10 and P < 0.001 for the remaining markers). TCC was analysed in chamber fluid, and median concentration was 28 (17–40) AU/ml. Figure 2 demonstrates the correlation between the number of in vivo extravasated neutrophils and the concentration of IL-8 in the chamber fluid at P < 0.05 and R = 0.79. In addition, the number of in vivo extravasated neutrophils also correlated with IL-1β, R = 0.83; IL-6, R = 0.73; IL-7 R = 0.71; and TNF-α, R = 0.71, all at P < 0.05. When the total number of extravasated leucocytes were analysed, the corresponding numbers were IL-8, R = 0.83; IL-1β, R = 0.81; IL-6, R = 0.72; IL-7 R = 0.71 and TNF-α, R = 0.70, also at P < 0.05. Following in vitro transmigration, the mean percentage of neutrophils that migrated towards chamber fluid was 34.2 ± 5.4% and towards cell culturing medium was 1.16 ± 0.55%. The percentage of transmigrated neutrophils correlated with the concentration of IL-8 (R = 0.79), IL-1β (R = 0.77) and TNFα (R = 0.79) at P < 0.05. The expression of CD11b activation epitope was measured following in vitro incubation with serum and skin chamber fluid. Figure 3 displays the expression of CD11b activation epitope following incubation with 50% serum, 50% skin camber fluid or IL-8 at 100 ng/ml.