Effect of enoxaparin fractions on <i>ex-vivo</i> cytokine release.

<p><b>(A)</b> PHA-induced release of cytokines (logarithmic scale) from PBMCs of healthy (n = 5) and allergic asthmatic (n = 10) subjects. Each sample was analysed in triplicate and the data is presented as the average of five and ten individual samples for healthy and allergic asthmatic subjects, respectively. Data is presented as mean ± SD. <b>(B)</b> Ion-chromatographic (IC) separation of enoxaparin, fractionated using a CarboPac PA100 semi-preparative column; a 32–74% NaCl gradient over 70 minutes; 2 mL/minute flow rate and a detection wavelength of 232 nm. The numbers indicate the area of all the fractions collected. Data represents a typical experiment out of five independent experiments. <b>(C)</b> Anticoagulant activity of IC-separated enoxaparin fractions. Data is presented as mean ± SD (n = 3). Anticoagulant activity was observed from fraction 6 onwards.<b>(D)</b> Inhibition of TNF-α release by different fractions of enoxaparin. TNF-α was released by PBMCs of allergic asthmatic subjects (n = 10) after <i>ex-vivo</i> stimulation with PHA. The relative percentile amount of each fraction (fraction 1 to 14) present in 500 μg/mL of intact enoxaparin was: 9.18%, 4.24%, 6.12%, 5.15%, 2.99%, 4.78%, 20.32%, 11.4%, 10.53%, 7.35%, 5.98%, 4.6%, 2.95% and 5.06% respectively. Data is presented as mean ± SD. ***<i>p</i><0.001 versus the PHA-stimulated control.