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Inhibition of neutrophil respiratory burst and degranulation responses by CVT-E002, the main active ingredient in COLD-FX


Human peripheral blood neutrophils contribute to the first line of defence in the immune system and are critical for maintaining health and immunity against opportunistic infections. Neutrophils and their granule-derived mediators are frequently found elevated in patient samples in viral infections, asthma exacerbations, and other respiratory ailments. COLD-FX has been shown to reduce the symptoms and severity of respiratory tract viral infections. Our hypothesis is that COLD-FX modulates neutrophil activity. To determine the effects of COLD-FX on neutrophils, peripheral blood neutrophils (>97% purity) were isolated from healthy human volunteers.


Neutrophils were preincubated with varying doses of CVT-E002 (0.01-1 mg/ml), the active ingredient of COLD-FX, for 30, 60, and 120 min. Extracellular ROS production was measured by cytochrome c reduction from neutrophils stimulated with 50 ng/ml phorbol myristate acetate for up to 60 min. Degranulation was measured by the presence of extracellular myeloperoxidase, a marker of the azurophilic granules, in neutrophils stimulated with cytochalasin B and f-Met-Leu-Phe for 15 min.


CVT-E002 (1 mg/ml) had no significant effect on viability at up to 120 min of incubation. At 60 min of incubation with CVT-E002, neutrophils showed a 30% reduction in ROS generation (p < 0.001) which was maintained for up to 120 min. Preliminary experiments also showed that incubation of neutrophils with CVT-E002 for 30 min inhibited myeloperoxidase release.


These novel findings demonstrate that COLD-FX significantly reduces activation of neutrophils. The implications of this study are that COLD-FX may reduce oxidative stress and tissue-damage triggered by neutrophilic inflammation and activation.

Funding source: Afexa Life Sciences, Inc.

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Pillai, R., Lacy, P. Inhibition of neutrophil respiratory burst and degranulation responses by CVT-E002, the main active ingredient in COLD-FX. All Asth Clin Immun 7 (Suppl 2), A31 (2011).

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