A.B. Kay

Neutrophil Chemotactic Activity in Antigen-Induced Late Asthmatic Reactions

Nine patients with bronchial asthma who had early and late falls in forced expiratory volume in one second (FEV1)(10 minutes and six hours, respectively) after inhalational challenge with specific antigens were studied for the presence of circulating neutrophil chemotactic activity (NCA). NCA was detected during both the early and the late asthmatic responses; the time course of appearance of NCA in the circulation paralleled that of the falls in FEV1. In contrast, five patients with asthma who had early reactions had only a single early peak of NCA, with no further rise for up to 24 hours. The NCA detected during early and late reactions eluted with macromolecules of an estimated molecular weight greater than 500,000 daltons when applied separately to columns of Sephadex G-200. Since high-molecular-weight NCA is believed to be associated with mast cells, these observations support the view that mediators of hypersensitivity are released in both the late and the early asthmatic responses.

Disodium cromoglycate inhibits activation of human inflammatory cells in vitro

Recent clinical studies indicate that disodium cromoglycate (DSCG) may have a direct effect on inflammatory cells because the drug reversed various changes in leukocyte function, such as increased membrane-receptor expression and enhanced cytotoxic capacity observed in peripheral white blood cells from subjects with asthma undergoing allergen-inhalation challenge. In the present study, we have demonstrated that DSCG, at low concentrations (a concentration of drug required to produce 50% inhibition, approximately 10(-8) mol/L) and in a time-dependent fashion, directly inhibited the activation in vitro of human neutrophils, eosinophils, and monocytes. Peripheral blood leukocytes were incubated with the synthetic chemoattractant, formyl-methionyl-leucyl-phenylalanine (at an optimal concentration of 10(-8) mol/L), and activation was assessed by measuring increases in the percentages of complement and IgG (Fc) rosettes as well as the enhanced capacity of these cells to kill target organisms (schistosomula of Schistosoma mansoni). DSCG at a concentration of 10(-7) mol/L totally inhibited both the formyl-methionyl-leucyl-phenylalanine-induced enhancement of complement and IgG rosettes, as well as increased schistosomular killing. These observations indicate that DSCG directly inhibits the secretory properties of inflammatory cells and that in turn might have important implications in modulating mechanisms contributing to the inflammatory component of asthma and allergic disease. It may also help to explain why compounds with considerably greater mast cell stabilizing properties than DSCG have been so disappointing when they are evaluated clinically.

Exercise-Induced Late Asthmatic Reactions with Neutrophil Chemotactic Activity

Two adults and 13 children with exercise-induced asthma had both immediate and late reductions in forced expiratory volume in one second (FEV1) after treadmill exercise. The late reactions developed 4 to 10 hours after exercise and in each instance were associated with wheezing or chest tightness (or both). Increases in neutrophil chemotactic activity, measured in the 2 adults and in 11 of the children, accompanied the reductions in FEV1 in all these subjects. In contrast, four other adults with only an immediate fall in FEV1 after exercise had only an initial elevation in neutrophil chemotactic activity, with no subsequent increase for the remaining 24-hour period. The agent responsible for the neutrophil chemotactic activity released during exercise-induced late reactions appeared to be identical to that released during immediate reactions. These observations suggest that some patients with exercise-induced asthma have late reactions that, as in the case of antigen-induced bronchoconstriction, are accompanied by the release of neutrophil chemotactic activity.

Leukocyte Activation in Allergen-Induced Late-Phase Asthmatic Reactions

Some patients with allergen-induced asthma have both an early and late reaction to allergen (dual asthmatic reactions). To investigate the role of leukocyte activation in dual asthmatic reactions, we measured neutrophil chemotactic activity, percentages of neutrophil and monocyte complement rosettes, and one-second forced expiratory volume (FEV1) in 11 patients with allergen-induced dual asthmatic reactions after a challenge with allergen. To control for the effects of bronchoconstriction, the same studies were done after a challenge with methacholine. In all subjects there was a biphasic increase in neutrophil chemotactic activity and the percentages of neutrophil and monocyte complement rosettes, accompanied by a reduction in the FEV1. After methacholine, there were no significant changes in neutrophil chemotactic activity or percentages of complement rosettes, despite bronchoconstriction. Six patients with single-phase allergen-induced asthma had similar responses, but they were monophasic. We conclude that allergen-induced early and late asthmatic reactions are accompanied by activation of leukocytes and that these alterations probably reflect the release of mediators from mast cells rather than an effect of bronchoconstriction.

Activation of neutrophils and monocytes after allergen- and histamine-induced bronchoconstriction

To determine whether neutrophils and monocytes are activated after allergen-induced asthma, changes in the expression of complement (C3b) receptors were measured by use of the rosette technique. There was a time-dependent increase in the percentages of neutrophil and monocyte rosettes in 13 asthmatic patients for up to 60 min after allergen-inhalation challenge. This was preceded by elevations in the concentrations of serum neutrophil chemotactic activity and reductions in the FEV1. These changes were not observed in seven asthmatic patients who had histamine-induced bronchoconstriction. The present findings support the view that inflammatory cells are activated after allergen-induced asthma and that this may be the result of the release of mast cell-associated mediators rather than the consequence of bronchoconstriction per se.

Mediators in exercise-induced asthma

Circulating concentrations of the mast cell-associated mediators, histamine and neutrophil chemotactic factor (NCF) of high molecular weight, were measured in atopic and nonatopic asthmatics after treadmill exercise. Elevations in the concentrations of both mediators accompanied the development of exercise-induced asthma (EIA). Normal individuals did not release mediators or develop bronchoconstriction after an identical exercise. The elaboration of mediators was not due to the onset of airflow obstruction, the postexercise basophilia, or the exercise task per se. A treadmill exercise undertaken while inhaling fully conditioned air inhibited EIA and NCF release; in contrast the same exercise undertaken while breathing cold, dry air elicited EIA and the production of mediators. This suggests that the stimulus for EIA and mediator release may be identical. Late-phase asthmatic reactions occur 3 to 9 hr after exercise in some asthmatics and are accompanied by the appearance of circulating NCF, as previously reported in allergen-induced late responses. In addition to the contribution of mediators to the spasmogenic reaction in EIA, mediators may contribute to bronchial inflammation by activating circulating leukocytes. There was a kinetic increase in the expression of neutrophil C3b receptors in EIA (+) asthmatics for up to 60 min after treadmill exercise. The enhancement of C3b receptors, as evidence of neutrophil activation, was preceded by release of NCF and reductions in peak expiratory flow rates. The prior administration of cromolyn inhibited EIA, NCF release, and enhancement of C3b receptors. These changes were not observed in EIA (-) asthmatics after an identical exercise task. These findings support the view that mediators are released in EIA and may play an important role in its pathogenesis.

COMPLEMENT RECEPTOR ENHANCEMENT AS EVIDENCE OF NEUTROPHIL ACTIVATION AFTER EXERCISE-INDUCED ASTHMA

To determine whether neutrophils are activated after exercise-induced asthma, increases in neutrophil complement receptor numbers (complement receptor enhancement) were measured by the rosette technique. In twelve asthmatic patients there was a time-dependent rise in complement receptor numbers for up to 60 min after treadmill exercise. This enhancement of complement receptors was preceded by a rise in plasma neutrophil chemotactic activity and a reduction in the peak expiratory flow rate. These changes could be inhibited by prior administration of disodium cromoglycate. The changes were not observed in seven asthmatic patients in whom asthma was not induced by an identical exercise task. Complement receptor enhancement was also observed in vitro when partially purified neutrophil chemotactic activity from a patient with exercise-induced asthma was incubated with normal neutrophils. These findings suggest that inflammatory cells are activated after exercise-induced asthma, possibly as a result of the release of neutrophil chemotactic activity and other mast-cell-associated mediators.

Neutrophil chemotactic activity in milk-induced asthma☆

Four subjects with clinical histories of milk-induced asthma were studied (three allergic to cows milk; one to soya milk). In each instance, skin prick tests, RAST (IgE and IgG4), the basophil histamine release, and serum precipitins, using appropriate milk extracts, were negative. After the ingestion of milk all the subjects developed a reproducible and dose-dependent increase in airflow limitation. Three patients (two allergic to cows milk; one to soya milk) gave a characteristic immediate-type reaction, which was maximal at 30 min after challenge. The fourth individual developed an isolated late-phase response, with maximal airways obstruction 3 hr after ingesting milk. In the three subjects who gave an early reaction, wheezing was accompanied by an elevation in circulating neutrophil chemotactic activity (NCA). This was not observed in the individual with the isolated late reaction. By Sephacryl S-400 gel-filtration chromatography it was shown that NCA of the early reactions eluted with proteins having an estimated molecular weight of 600,000 daltons. The immediate asthmatic response in peak expiratory flow rate and the elevation in NCA were inhibited by the prior oral administration of either disodium cromoglycate (DSCG) or oral beclomethasone dipropionate (BDP). In contrast, DSCG had no effect on airways obstruction in the subject with the isolated late asthmatic response, although inhibition was achieved by BDP.