Effects of some anti-asthma drugs on human eosinophil superoxide anions release and degranulation.

BACKGROUND: Eosinophil infiltration of bronchial tissues and subsequent release of inflammatory mediators by them are the hallmarks of bronchial asthma but it has not yet been clarified whether anti-asthma drugs affect these cells directly. In this study, we investigated the direct effects of 8 clinically used anti-asthma drugs [salbutamol, salmeterol, theophylline, denbufylline, disodium cromoglycate (DSCG), azelastine, ketotifen and dexamethasone] on superoxide anions (O2-) and eosinophil peroxidase (EPO) release from human blood eosinophils in vitro. METHODS: Highly purified eosinophils were stimulated for O2- release with platelet-activating factor (PAF) or interleukin-5 (IL-5), while for EPO release complement fragment (C5a) or N-formyl-methionyl-leucyl-phenylalanine (FMLP) was employed. Generated products were assayed by standard techniques. RESULTS: All the drugs, except ketotifen and dexamethasone, inhibited PAF-induced O2- release in a dose-dependent manner. The IC50 values were 0.7, 5.8, 330, 3,500, 4,200 and 6,250 nM for DSCG, denbufylline, salmeterol, azelastine, salbutamol and theophylline, respectively. On IL-5-induced release, the effects were similar except that salbutamol completely failed to inhibit the release induced by this stimulus. In contrast, EPO release was generally poorly inhibited, especially when the release was induced by C5a. Only theophylline and azelastine (both at 10(-4) M or more) were able to inhibit EPO release by both C5a and FMLP. Salbutamol and, to a lesser extent, salmeterol inhibited FMLP-, but not C5a-induced EPO release, while all the other drugs tested were inactive. CONCLUSIONS: The results show that some of the anti-asthma drugs, but not all, do exert direct effects on human blood eosinophils but these effects may be stimulus-dependent and by far more pronounced against O2- release than against degranulation.

Differential effects of salbutamol and salmeterol on human eosinophil responses.

In the treatment of bronchial asthma, salmeterol is believed to have a greater anti-inflammatory activity than salbutamol. To determine whether the comparative effects of these drugs on eosinophil function are the basis of their differential anti-inflammatory properties, we studied the effect of the two drugs on interleukin-5 (IL-5) and 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine (PAF)-induced O2- release and adherence to fibronectin-coated plates, as well as the C5a- and N-formylmethionyl-leucyl-phenylalanine (FMLP)-induced degranulation of purified human blood eosinophils in vitro. Salmeterol significantly inhibited IL-5-induced O2- release in a concentration-dependent manner with an IC50 of 2.2 X 10(-6) M (95% CI, 1.6-2.7 X 10(-6) M) and a maximal inhibition of about 70%. In contrast, salbutamol had no significant effect even at 10(-5) M. Both drugs significantly inhibited PAF-induced O2- generation, but salmeterol was approximately 20 times more potent than salbutamol. Salmeterol also significantly inhibited adherence induced by both IL-5 and PAF, whereas salbutamol had no significant effect on adherence induced by both agents. Both drugs failed to block C5a-induced eosinophil peroxidase release, whereas for FMLP-induced release, salbutamol, but not salmeterol, produced significant inhibition. Unlike salbutamol, all the actions of salmeterol were independent of beta-2 adrenoceptors. These results confirm that human eosinophils can be modulated directly by beta-2 adrenoceptor agonists, but that salmeterol and salbutamol have differential effects which depend on both the stimulus used and the response being measured and that the reportedly greater in vivo anti-inflammatory effect of salmeterol may reflect its superior ability to inhibit eosinophil O2- release and adherence, rather than degranulation.

The effect of salmeterol on human eosinophils is both stimulus- and response-dependent.

Salmeterol, a long-acting beta 2-adrenoceptor agonist, also possesses some anti-inflammatory properties, but whether eosinophils are the target of such action has been equivocal. To clarify the direct effect of salmeterol on eosinophil functions, we have studied the effect of the drug on the various responses of purified human eosinophils. Superoxide anions (O2-) release and adherence to fibronectin-coated plastic plates induced by platelet-activating factor (PAF), interleukin-5 (IL-5), leukotriene B4 (LTB4) and phorbol myristate acetate (PMA), as well as degranulation induced by C5a and formyl methionyl leucyl phenylalanine (FMLP), in the presence of cytochalasin B (CB) were studied. In the concentration range 10(-8)-10(-5) M, the drug inhibited PAF- and IL-5-induced O2- release, with an IC50 values of 3.2 +/- 1.2 x 10(-7) M and 2.2 +/- 0.4 x 10(-6) M, respectively, Superoxide anion release by LTB4 was only modestly inhibited while that due to PMA was completely unaffected. On the other hand, eosinophil adherence induced by all the 4 stimuli were significantly inhibited within the same concentration range. On eosinophil degranulation, the drug failed to significantly inhibit the release of eosinophil peroxidase (EPO) induced by either C5a or FMLP. In contrast, beta-hexoseaminidase (beta-HA) release by the same agents was significantly inhibited, the inhibition being more pronounced for FMLP-induced, than C5a-induced release. None of the effects of the drug was reversed by the selective beta 2-adrenoceptor antagonist ICI 118551 at a concentration of 10(-7) M. These results show that salmeterol may have some direct inhibitory effects on human eosinophil functions but that these effects are both stimulus- and response-dependent, and are unlikely to be mediated via beta 2 adrenoceptors.

Prevalence of allergic sensitization to inhalant allergens among blood donors in Kuwait--a desert country.

Kuwait is a desert country where the prevailing high temperatures, low humidity, and scant vegetation suggest a low prevalence of allergy. We evaluated the prevalence of atopic sensitization (presence of allergen-specific IgE) among young adult blood donors by screening a total of 505 subjects (male: female ratio 1.6) with mean age of 28.4 years (range 18-50 years). The Pharmacia CAP-Phadiatop test, which detects serum IgE specific to most common airborne allergens, was used. Some of the specific sensitizing allergens were also identified by the related CAP-RAST method. Sensitization was detected in 223 of the 505 subjects (44.2%) screened. Kuwaiti nationals had a significantly higher prevalence rate (50.2%) than non-Kuwaitis (34.2%) (chi 2 = 8.6, P < 0.003). The highest prevalence rate was found among male Kuwaitis (53.8%). The prevalence of current or previous allergic disease (subject-reported) was 20.6%. Bermuda grass, house-dust mite (Dermatophagoides pteronyssinus), and Chenopodium album were the most prevalent sensitizing allergens, with frequencies of 53.6%, 52.7%, and 50.9%, respectively, among the sensitized subjects (corresponding to 23.7%, 23.3%, and 22.5%, respectively) for the entire population. Sensitization increased with age, but only among the expatriates, younger Kuwaitis being as frequently sensitized as the older ones. Polysensitization was found to be common. Of the 109 CAP-RAST-positive subjects, 71 (65.1%) were sensitized to more than one allergen, and 30 of these (42.3%) were sensitized to four or more allergens. These results show that atopy is highly prevalent among young adults in Kuwait, and the higher prevalence rate among nationals than expatriates suggests the involvement of genetic or local environmental factors. The results also confirm that mite and plant pollens may be major sensitizing allergens even in a desert environment.