Influence of antigen challenge on platelet responsiveness in horses with chronic obstructive pulmonary disease.

A role for platelets in allergic airways disease has been postulated and changes in the responsiveness of circulating platelets have been demonstrated following antigen challenge of asthmatic human subjects. In this study agonist-induced aggregation of equine platelets in vitro has been compared before and after exposure of horses to a controlled hay and stray challenge. Prior to challenge the response of platelets, from horses with chronic obstructive pulmonary disease (COPD) and normal animals, to adenosine diphosphate (ADP) and platelet activating factor (PAF) did not differ. Five hours after initiation of the challenge, there was a modest but significant decrease in the response of platelets from the COPD horses to PAF, but not to ADP. Platelets from normal horses were not less sensitive to either agonist at this time. Twenty four hours after challenge the responses of platelets from COPD horses to both agonists were the same as pre-challenge values. These results demonstrate that antigen challenge alters the responsiveness of platelets from allergic horses to PAF and this change is suggestive of PAF release accompanying allergen exposure in the horse.

Effects of platelet activating factor on the distribution of radiolabelled leucocytes and platelets in normal horses and asymptomatic horses with chronic obstructive pulmonary disease.

Antigen challenge is known to cause the recruitment of neutrophils to the lungs of horses with chronic obstructive pulmonary disease (COPD). To evaluate a possible role of platelet activating factor (PAF) in this process, the effects of PAF on the distribution of radiolabelled neutrophils were compared in normal horses and asymptomatic horses with COPD. Changes in lung function, heart rate and the distribution of platelets and eosinophils were also measured. PAF (5 ng kg-1 intravenously) caused immediate but transient increases in the number of radiolabelled neutrophils in the lungs and a concomitant decrease in the peripheral neutrophil count. The total numbers of circulating leucocytes and neutrophils were also significantly decreased by PAF. Rapid and reversible increases in heart rate, respiratory rate and pleural pressure were also observed. In separate experiments, the numbers of radiolabelled eosinophils and platelets in the lungs increased transiently after the administration of PAF. The responses to PAF were qualitatively and quantitatively similar in normal horses and asymptomatic COPD horses. The PAF receptor antagonist WEB2086 (3 mg kg-1 intravenously) inhibited the effects of PAF. These results suggest that PAF, if released in the lungs of horses with COPD during an antigen challenge, might contribute to the recruitment of leucocytes and the respiratory changes.

A study of the effect of a platelet activating factor (PAF) receptor antagonist on antigen challenge of horses with chronic obstructive pulmonary disease.

Antigen challenge involving exposure to straw and mouldy hay for 7 h produced lung function changes and neutrophil recruitment to the lungs in horses with chronic obstructive pulmonary disease (COPD). During the challenge, an increase in radiolabelled neutrophils in the lungs occurred, together with increased respiratory rate and pleural pressure. The role of platelet activating factor (PAF) in antigen-induced neutrophil accumulation, and increased pleural pressure and respiratory rate was investigated by administering the PAF receptor antagonist WEB 2086 to asymptomatic COPD horses prior to antigen challenge. WEB 2086 (3 mg/kg i.v.) did not affect antigen-induced changes in either neutrophil accumulation or respiratory function. These results suggest that PAF may not be an important mediator of the response to antigen in equine COPD.

Studies on the mechanisms involved in the inflammatory response in a reversed passive Arthus reaction in guinea-pig skin: contribution of neutrophils and endogenous mediators.

1. Mediators of inflammation can increase vascular permeability in at least two different ways: by acting directly on endothelial cells or, indirectly, through an incompletely understood mechanism, dependent on circulating neutrophils. Neutrophil-dependent oedema formation has been described in the skin of rabbits, rats, hamsters, mice and man. In contrast, we presented evidence in a previous study that local oedema formation induced by i.d. injection of chemoattractants in guinea-pig skin was neutrophil-independent. In the present study, we sought evidence of neutrophil-dependent oedema formation in immune-complex-mediated vasculitis, the reversed passive Arthus (RPA) reaction, in guinea-pig skin. We also investigated whether haemorrhage in the RPA reaction was neutrophil-dependent (as it is in other species) and the role of endogenous mediators of inflammation (prostaglandins, nitric oxide, histamine, PAF and leukotrienes) in contributing to the local inflammatory response. 2. In the RPA reaction, most oedema formation occurred over the first 60 min whereas 111In-neutrophil accumulation was still increasing from 60 to 240 min. The different kinetics of these two events suggested that they may be dissociated. 3. Oedema formation was partially inhibited by a long-acting PAF antagonist (UK-74,505) and an H1 histamine receptor antagonist (mepyramine) but not by a 5-lipoxygenase inhibitor (ZM 230487). A nitric oxide synthesis inhibitor (NG-nitro-L-arginine methyl ester, L-NAME) suppressed oedema formation by 68% whereas a cyclo-oxygenase inhibitor suppressed oedema by 27%. 4. 111In-neutrophil accumulation in the RPA reaction was partially suppressed by UK-74,505. In contrast, ZM 230487 was without effect at doses which abrogated arachidonic acid-induced 111In-neutrophil accumulation. 5. The anti-CD18 monoclonal antibody, (mAb) 6.5E F(ab')2, effectively inhibited 111In-neutrophil accumulation induced by PAF, zymosan-activated plasma (ZAP) and in the RPA reaction. However, oedema formation measured in the same sites was not altered. In contrast, oedema formation in the RPA reaction was partially suppressed by 6.5E whole mAb which was 2.5 times more potent than 6.5EF(ab')2 at inhibiting guinea-pig neutrophil adhesion to protein-coated plastic. Haemorrhage induced by PAF and in the RPA reaction was significantly inhibited by 6.5E F(ab')2 pretreatment.6. We conclude that in the RPA reaction in guinea-pig skin, oedema formation is partially neutrophil dependent as assessed by using an anti-CD18 mAb, whereas ZAP-induced oedema formation is neutrophil-independent. Haemorrhage was also dependent on neutrophil accumulation. In addition, our studies support a role for PAF in mediating both oedema formation and "'In-neutrophil accumulation in the RPA reaction. Endogenous release of histamine also appears to be important in mediating oedema formation suggesting that mast cells play a critical role in increases of vascular permeability in inflammatory reactions in guinea-pig skin. Moreover, our results confirm previous findings which suggest a dominant role for nitric oxide in maintaining cutaneous blood flow in the guinea-pig.

Duration of antigen-induced hyperresponsiveness in horses with allergic respiratory disease and possible links with early airway obstruction.

Antigen-induced airway hyperresponsiveness in allergic horses has previously been demonstrated when clinical signs of acute airway obstruction were apparent, as a consequence of exposure of animals to hay and straw for variable periods of time, and repeat measurements of hyperresponsiveness have been made no earlier than 1 week after challenge. In the present study airway responsiveness to methacholine has been measured in normal horses and allergic horses in clinical remission before and 24, 48 and 72 h after a hay and straw challenge of fixed, short, duration (7 h). Correlations between early increases in interpleural pressure and hyperresponsiveness have also been investigated. As in other studies, the mean airway responsiveness of groups of normal and allergic horses in clinical remission was not significantly different. The responsiveness to methacholine of allergic, but not normal, horses was increased after antigen challenge and was significantly greater than that of normal horses at 48 and 72 h after challenge (log PD8 cm: -0.77 +/- 0.28 vs. 0.27 +/- 0.14 at 48 h and -0.6 +/- 0.25 vs. 044 +/- 01 at 72 h; P < 0.05). There was also a significant correlation between interpleural pressure at the end of the 7-h challenge in allergic horses and the increase in responsiveness to methacholine at 24, 48 and 72 h. These results demonstrate that antigen induces an increase in airway responsiveness in allergic horses that persists for up to 3 days and which may be linked to the initial increase in interpleural pressure.

Early neutrophil but not eosinophil or platelet recruitment to the lungs of allergic horses following antigen exposure.

Previous studies have shown that bronchoalveolar lavage fluid from horses with allergic respiratory disease and showing clinical symptoms contains increased numbers of neutrophils. In some cases, the eosinophil count is also increased. In this study the time course of changes in lung function and the accumulation of radiolabelled leucocytes and platelets in the lungs of allergic and normal horses has been examined during a 7 hr allergen exposure. Antigen challenge had no effect on pleural pressure or the distribution of radiolabelled neutrophils, eosinophils or platelets in normal horses. In contrast, in 6/8 allergic horses, there was an increase in pleural pressure and neutrophil accumulation in the lungs, both of which were evident after 4-5 hr. However, during the 7 hr challenge period radiolabelled eosinophils were detected in the lungs of only 1/6 horses exhibiting an increase in pleural pressure and in 1/7 horses that failed to show a change in airway function despite a clinical history of allergic respiratory disease. Antigen challenge did not alter the distribution of radiolabelled platelets in the five allergic horses tested. These results demonstrate that increased pleural pressure is not accompanied by eosinophil or platelet accumulation in the lungs of horses with allergic respiratory disease following exposure to antigen. However, changes in airway function can be associated with neutrophil accumulation but can also take place in the absence of this cell recruitment. This raises the possibility that the presence of neutrophils in the lung is not a prerequisite for changes in lung function.