Bronchoconstriction and Airway Microvascular Leakage in Guinea Pigs Sensitized with Trimellitic Anhydride 1 , 2

JAMES P. HAYES, JAN O. LOTVALL, JIM BARANIUK, ROB DANIEL, PETER J. BARNES, ANTHONY J. NEWMAN TAYLOR, and K. FAN CHUNG

Introduction

Trimellitic anhydride (TMA) is a reactive chemical widely used in industry as a curing agent for epoxy resins. Exposure to TMA in the workplace may result in the development of occupational asthma (1). Asthma caused by the acid anhydrides has been associated with the development of a specific IgE or Ig04 antibody response (2). Nonspecific bronchial hyperreactivity is a characteristic feature of occupational asthma caused by low molecular weight chemicals including the acid anhydrides and specific inhalation tests in sensitizedindividuals can provoke changes in airway caliber and in nonspecific airway responsiveness (3). In a preliminary study, we have shown that intradermal injection of TMA induces a specific immune response to TMA conjugated to guinea pig serum albumin (OPSA) and that an intradermal injection of TMA is an effective method of sensitization in the guinea pig for the induction of bronchoconstrictor responses to intravenous TMA-OPSA (4). Plasma exudation into the airway lumen may cause airway edema, a feature of asthma, and this may playa substantial role in increased airway responsiveness found in asthmatics (5). Many of the inflammatory mediators implicated in asthma have been shown to induce airway microvascular leakage (6, 7), which is believed to occur at the postcapillary venules (8). Airway microvascular leakage has been shown to occur following allergen challenge in sensitized guinea pigs. In this study, we assessed the effect of TMA-OPSA instilled into the trachea on lung resistance in guinea pigs previously sensitized to the free hapten and the degree and site of airway microvascular leakage associated with this response. Methods Sensitization Female Dunkin-Hartley guinea pigs, weigh1306

SUMMARY We have developed a guinea pig model of immediate airway responses following Intradermal sensitization with free trimellltic anhydride (TMA). Guinea pigs were given an Intradermal injection with either 0.1 ml of 0.3% TMA In corn 011 (n = 8) or 0.1 ml of corn 011 alone (n = 6). A guinea pig serum albumin conjugate of trimellltic anhydride (TMA-OPSA) was prepared with a substitution ratio of 21:1. All sensitized guinea pigs had raised specific serum IgG1antibodies (ELISA), and IgE antibodies were detected in six of the eight sensitized guinea pigs by passive cutaneous anaphylaxis. On Days 21 to 28, guinea pigs were anesthetized, tracheostomlzed, and ventilated. Evans blue dye (20 mglml), an albumin marker,was Injected Intravenously to quantify airway microvascular leakage (MVL). TMA-GPSA(50 111; 1%) in saline was instilled Into the trachea. Lung resistance (RL) was measured for 6 min. The guinea pigs were killed, and the lungs were removed. Peak RL (cm H20/mi x S-l) was significantly Increased in sensitized guinea pigs from 0.26 ± 0.01, mean ± SEM to 21.3 ± 6.9 (p < 0.05), compared with nonsensltlzed guinea pigs. There was a significant increase in Evans blue at all levels of the tracheobronchial tree In sensitized guinea pigs compared with the controls (p < 0.005). The site of MVL was localized to the postcaplllary venules as assessed by extravasation of Intravascular Monastral blue dye. We conclude that Intradermal sensitization of guinea pigs to TMA Induces a polyclonallmmune response, associated with bronchoconstrlctlon and airway microvascular leakage, when challenged specifically with TMA-GPSA. This guinea pig model may be useful In examining the pathogenesis of TMA-Induced occupational asthma. AM REV RESPIR DIS 1992; 146:1306-1310

ing 200 to 250 g (n = 8), were injected intradermally with 0.1ml of 0.3070 TMA in corn oil. Controls (n = 6) were injected with 0.1 ml of corn oil alone.

Preparation of Trimellitic Anhydride Conjugates TMA was conjugated to guinea pig serum albumin (GPSA) as follows. Four aliquots of 250 mg GPSA (Sigma, Poole, UK) weremixed with 50 ml of 9070 NaHC0 3 on ice and 250 mg of ground TMA (Aldrich, Gillingham, UK) dust was added and allowed to stand for 10 min. Each aliquot was stirred separately for 1 h. The aliquots were pooled and spun for 10 min at 600 g (MSE Centaur 2). The supernatant was separated and dialysed with 0.02 M NH 4HC03 for 48 h with six changes. This was further dialyzed with distilled water for 24 h and lyophilized. The degree of substitution was assessed by UV spectrophotometry. Briefly, 1.0 mg TMA-GPSA was added to 1 ml of 0.1 M borate buffer at pH 9.3. A 0.03 M solution of 2,4,6,trinitrobenzenesulfonic acid (TNBS) (Aldrich, Gillingham, UK) in borate buffer was prepared. Then 25 III of TNBS solution was added to 1 ml ofthe TMA-GPSA solution and incubated for 20 min at room temperature. The sample was read on the spectrophotometer at an

optical density of 420 nm. The substitution ratio for TMA-GPSA was 21:1.

ELISA for Assessment of IgOl Antibodies Plates werecoated with 5 ug/ml TMA-GPSA in coating buffer and incubated overnight at 4° C. On the next day, the plates were washed four times in PBS/tween. Serum was diluted to 1:50solution, and plates were coated with PBS/tween. Serum was added in doubling dilutions across the plate and incubated for 3 hat 27° C. The plates were again washed four times with PBS/tween and 1:2,500 of rabbit anti-guinea pig IgGl (Aldrich, Gillingham, UK) was added and incubated for 2 h at 27° C. Again the plates were washed with PBS/

(Received in original form July 22, 1991 and in revised form May 5, 1992) 1 From the Departments of Thoracic Medicine and Occupational and Environmental Medicine, National Heart and Lung Institute, London, United Kingdom. 2 Correspondence and requests for reprints should be addressed to Dr. K. F. Chung, Department of Thoracic Medicine, National Heart and Lung Institute, Dovehouse Street, London SW3 6LY, United Kingdom.

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tween. Anti-rabbit IgG-peroxidase (Aldrich, Gillingham, UK) was added at 1:5,000dilution and incubated at 27 0 C for 2 h. Finally the plates were washed in PBS/tween and chromogen was added. The reaction was stopped after 6 min with 1 M H 2S04 and read at an optical density of 492 nm. The titers of IgGI were calculated from the last dilution that was double the control sera.

Passive Cutaneous Anaphylaxis Passive cutaneous anaphylaxis (PCA) was performed to detect IgE antibodies as described by Ovary and colleagues(9). Sera from the guinea pigs were injected into the flanks of naive guinea pigs in increasing dilutions. Six days later, TMA-GPSA (5 mg), together with Evans blue dye (10mg), was injected intravenously. Samples of sera preheated to 56 0 C for 1 h were also injected. The animals were killed and the cutaneous responses read on the outer surface of the skin 30 min after the intravenous injection. A blue lesion of greater than 5 mm in diameter was considered positive. Measurement of Lung Resistance On Days 21 to 28, guinea pigs were anesthetized with pentabarbitone 60 mg/ml intraperitoneally. A tracheal cannula (10 mm length and 2.7 mm internal diameter) was inserted into the lumen of the trachea through a tracheostomy. A polyethylene catheter was inserted into the left carotid artery to monitor blood pressure and heart rate with a pressure transducer. Transpulmonary pressure was measured with a pressure transducer (Model Fe040; ± 1,000mm H 20, Furness Controls Ltd, Bexhill, Sussex, UK) with one side attached to a catheter inserted into the right pleural cavity and the other side attached to a catheter connected to a side port of the intratracheal cannula. The ventilatory circuit had a total volume of 201 ml. Airflow was measured with a pneumotachograph (Model FIL; Mercury Electronics Ltd., Glasgow,UK) connected to a transducer (Model FCO 40; ± 20 mm H 20, Furness Controls Ltd.). The signals from the transducers were digitalized with a 12-bit analog-digital board (NB-MIO16; National Instruments, Austin, TX) connected to a Macintosh II computer (Apple Computer Inc., Cupertino, CA) and analyzed with a software (Labview, National Instruments, Austin, TX), which was programmed to instantaneously calculate lung resistance (RL) by the method of von Neergaard and Wirz (10). 'Iranspulmonary pressure and mean blood pressure werealso monitored throughout the experiment. Measurement of Airway Microvascular Leakage Evans blue dye (20 mg/ml) was injected intravenouslyover 1 min. After another minute, 50 ul of 1070 TMA-GPSA in saline was instilled into the trachea though a polyethylene cannula (internal diameter 0.9 mm) that was inserted into the trachea having passed

through the length of the tracheal cannula. Lung resistance was monitored for 5 min, the guinea pigs werehyperinflated manually with twice the tidal volume, and 60 s later, the guinea pigs were disconnected from the ventilator. The chest was opened and a cannula was inserted into the aorta through the left ventricle. This cannula wasperfused with normal saline at a pressure of 100to 120 mg Hg to eliminate dye from the bronchial circulation. After 90 s of perfusion, a second cannula was inserted into the pulmonary artery through the right ventricle and perfused with 20 ml of saline over 60 s to eliminate dye from the pulmonary circulation. The trachea and lungs were dissected out en bloc, the parenchyma carefully scraped off, and extraneous tissue removed. The trachea, major bronchi, and intrapulmonary airways were then separated from each other, and the intrapulmonary airways were divided into two portions, arbitrarily named proximal and distal. All tissues were weighed after having been freezedried for 36 h. Evans blue dye was extracted by placing each sample of tissue in 2 ml of formamide in a 40 0 C water bath overnight. Absorption at 620 nm wasmeasured in a spectrophotometer (Model 8480; Philips, Cambridge, UK). Extravasated Evans blue dyewas quantified by interpolation on a standard curve of dye concentrations in the range of 0.5 to 10 ug/ml and expressed as ng dye/mg dry tissue. Evans blue dye measurements have been shown to correlate highly with the extravasation of radio labeled albumin into guinea pig airways. In order to demonstrate the site of airway microvascular leakage, a guinea pig sensitized to TMA and a control guinea pig injected with corn oil alone were given an intravenous injection of Monastral blue (l mllkg of3% solution; Sigma, Poole, UK), and after 1 min, 50 ul of TMA-GPSA was instilled into the trachea. After 6 min of exposure, the animals were killed by exsanguination and perfused as described previously with saline. The trachea and lungs were removed. The left lung was fixed in 4% paraformaldehyde in PBS and paraffin sections prepared. The right lung parenchyma was scraped from the right bronchi. The anterior trachea and right main stem and segmental bronchi weredivided with scissors and spread apart. This tracheobronchial preparation was placed between two microscope slides and fixed in 4% paraformaldehyde in PBS at 4 0 C. The preparation was cleared in ethanol and zylene before mounting in DBX mountant (BDH, Poole, UK).

Statistical Analysis Data are reported as mean ± SEM. MannWhitney U test was used to assess significance between the two groups. The Spearman-Rank correlation coefficient was used to determine any relationship in the immune response, the degree of bronchoconstriction, and airway microvascular leakage. The degree of bronchoconstriction was assessed either as the peak RLor as the area under the time course

curve of RL over 6 min. Data were analyzed on a Macintosh computer (Apple Computer Inc., Cupertino, CA) using a standard statistical package. Results

All guinea pigs injected intradermally with TMA developed specific IgGl antibodies to TMA-GPSA (figure 1, bottom). Six of the eight guinea pigs had evidence of IgE antibodies to TMA-GPSA as determined by passive cutaneous anaphylaxis (figure 1, top). Preheating the sera to 56° C inhibited this response. There was no evidence of specific IgGl or IgE antibodies in the control guinea pigs. All sensitized guinea pigs demonstrat-

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Fig. 1. (bottom) Individual IgG1 antibodies to TMAGPSA measured by ELISA (see text) in guinea pigs injected intradermally with 0.3% TMA compared with control guinea pigs injected with corn oil alone. (top) Individual IgE antibody titers to TMA-GPSA assessed by passive cutaneous anaphylaxis (peA) in guinea pigs injected intradermally with 0.3% TMA compared with control guinea pigs injected with corn oil alone.

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ed a significant increase in maximal following instillation of TMA-GPSA, although there was considerable variability in the maximal response seen (p < 0.05; figure 2, bottom). Similarly, there was a significant increase in the overall increase in lung resistance (area under the curve) in sensitized guinea pigs (86.9 ± 22.4 units of area; mean ± SEM) compared with control guinea pigs (8.1 ± 0.5; p < 0.02). This was characterized by a rapid increase in lung resistance starting 1 min after instillation of TMA-GPSA in six of the eight sensitized guinea pigs (figure 2, top). In two of the eight guinea pigs, the response did not commence until 3 min after instilling TMA-GPSA and continued to increase up to 6 min after the challenge. There was a transient increase in RL found in the control group after a similar challenge that was followed by a return to baseline. This effect was small when compared with that seen in the sensitized group. The maximal and total changes in RL in the control group after instillation of TMA-GPSA were compared with changes in RLseen in the sensitized group. We have found a simi-

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Fig. 3. Effect of tracheal instillation of 1% TMA-GPSAon airway microvascular leakage as measuredbyextravasation of Evans blue dye in sensitized (hatched bars) and control (closed bars) guinea pigs at various levels of the respiratory tract. Data are expressed as mean ± SEM. * indicates p < 0.005 compared with the control group. Tr = trachea; MB = main bronchi; PIP = proximal intrapulmonary airways; DIP = distal intrapulmonary airways.

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Fig. 2. (bottom) Individual peak lung resistance after tracheal instillation of 1% TMA-GPSA (50Ill) in sensitizedand control guinea pigs. * indicates p < 0.05. (top) Time course of changes in lung resistance induced by tracheal instillation of 1% TMA-GPSA in sensitized (open circles) and control (closed circles) guinea pigs. Data are expressed as mean ± SEM.

lar increase in RL in sensitized animals after tracheal instillation of GPSA alone. There was no change in the mean blood pressure in the sensitized or control guinea pigs after instillation of the TMA-GPSA. All sensitized guinea pigs demonstrated a significant increase in airway microvascular leakage as measured by extraction of the Evans blue dye at all four levels of the bronchial tree examined when compared with the nonsensitized guinea pigs (p < 0.005) (figure 3). There was a dense pattern of Monastral blue labeling in postcapillary venulesin the trachea, carina, main stem bronchi, and subsegmental bronchi. The labeled vesselsended abruptly in the subsegmentallocation. There were no labeled vessels in the smaller airways or the parenchyma (figure 4). By contrast, there was no Monastral blue dye visible in the airways of nonsensitized guinea pigs challenged with TMA-GPSA. There was no significant correlation between peak RL response and the degree of Evans blue dye extravasation airway in sensitized guinea pigs. There was also no significant correlation among peak RL,the degree of Evans blue dye extravasation, and the level of IgGl or IgE titers. Discussion

We have developed in the guinea pig a model of sensitization to trimellitic anhydride following intradermal injection.

AIRWAY RESPONSES TO TMA IN SENSITIZED GUINEA PIGS

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Fig. 4. Photomicrographs of guinea pig tracheobronchial wholemount preparations after tracheal instillation of TMA-GPSA (magnification x 100). (A): In a sensitized guinea pig, the tracheal posterior membranous region between the two cartilage rings (car) is shown. A dense pattern of Monastral blue B in the postcapillary venules can be seen. (B): The trachea of a nonsensitized animal is shown. There is no staining with Monastral Blue B. (C): Monastral blue B is shown in the venules of the bronchi (diameter 0.5 mm) in a sensitized guinea pig. (0): The bronchi (diameter 0.5 mm) in a nonsensitized guinea pig with no staining with Monastral blue.

In this model, an acute, immediate bronchoconstrictor response associated with airway microvascular leakage was demonstrated following direct instillation of a protein conjugate of trimellitic anhydride (TMA-GPSA). No response was observed in nonsensitized animals challenged in a similar manner. We have previously shown that these antibodies are specific for TMA-GPSA, the protein conjugate of TMA. All guinea pigs injected with intradermal TMA developed IgGl antibodies to TMA-GPSA. We have demonstrated previously that this antibody response is specific for TMA-GPSA (4). Serum IgE was detected only in only six of the eight guinea pigs sensitized to TMA. The measurement of serum IgE by passive cutaneous anaphylaxis is at best a semiquantitative measurement, and low titers of IgE may not have been detected by this method because much of the IgE may have been bound to cells. There was no correlation between the magnitude of the antibody responses and the degree of bronchocon stricti on or airway microvascular leak in sensitized animals. There are few animal models of airway responses to low molecular weight

chemicals that are known to cause occupational asthma. Botham and coworkers (11), using a similar method of sensitization but a higher dose of TMA, induced an increase in respiratory rate (used as an index of respiratory function) in sensitized guinea pigs exposed to free TMA dust. However, because they did not measure changes in airway caliber and because TMA dust may have different effects on the airways when inhaled (12), the exact nature of this response is unknown. A lower sensitizing dose of TMA appears to be more effective at inducing detectable IgE immune responses than higher doses and causes significant increases in bronchopulmonary responses following an intravenous injection of TMA-GPSA (13). Biagnini and colleagues (14)sensitized cynomolgus monkeys to ammonium hexachloroplatinate inhaled concurrently with ozone, and subsequently inhalation of hexachloroplatinate provoked immediate airway responses. Bernstein and coworkers (15) provoked changes in airway caliber in guinea pigs passively sensitized by antisera to hexamethylene diisocyanate (HDI) conjugated to human serum albumin and challenged intrave-

nously with HDI conjugated to transferrin. Chan and colleagues (16) also provoked an IgE response to plicatic acid in rabbits injected with plicatic acid conjugated to ovalbumin and induced airway responses by intravenous challenge with plicatic acid conjugated to human serum albumin. In our model, we have sensitized guinea pigs to unconjugated TMA and induced an immune response similar to that seen in humans. In addition, we have demonstrated acute bronchoconstriction with concomitant airway microvascular leakage after challenge to a protein conjugate of TMA. TMA-GPSA was introduced directly into the airways because of difficulties in effectively nebulizing the viscous TMA-GPSA suspension. We chose to conjugate TMA to GPSA because the introduction of TMA dust into a tracheal cannula in an anesthetized animal would be technically very difficult and because the reactivity of TMA might result in direct airway damage associated with nonspecific changes in airway caliber. TMAGPSA itself did not cause any significant changes in lung resistance or airway microvascular leakage in nonsensitized guinea pigs.

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Airway microvascular leakage is likely to be an important feature of asthma causes extravasation of plasma proteins such as albumin into the airway wall and lumen. Also in asthmatic subjects, increased concentration of albumin can be recovered from the bronchoalveolar fluid (17). This observation has also been reported in sensitized workers challenged with Western Red Cedar (18). Microvascular leakage has also been reported in ovalbumin-sensitized guinea pigs exposed to ovalbumin intravenously or by aerosol (19, 20).In TMA-sensitizedguinea pigs, we found that increased airway microvascular leakage, as observed by the Evans blue technique, occurred throughout the respiratory tract in a uniform fashion, similar to the distribution seen in ovalbumin-challenged guinea pigs. In addition, we showed that the site of airway microvascular leakage could be visualized beneath the basement membrane of the vascular endothelium (21, 22). The mechanism by which increased airway microvascular leakage occurs in our model has not been studied. It seems likely that vasoactive mediators are released following challenge with TMAGPSA, causing both bronchoconstriction and airway microvascular leakage. In ovalbumin-sensitized guinea pigs, we have shown that a 5-lipoxygenase inhibitor (A-63162) attenuates both aspects of the response provoked by inhalation of ovalbumin (23). It has been suggested that increased airway microvascular leakage by causing airway edema may lead to narrowing of the intraluminal airway caliber. The contribution of such a process to changes in pulmonary resistance is unclear, and we found no clear relationship between changes in pulmonary resistance. It is possible that those changes may be independent of each other, despite being the result of the effect of similar inflammatory mediators. In summary, we have developed a model of immediate airway responses in

HAYES, LOTVALL, BARANIUK, ET AL.

guinea pigs sensitized to free trimellitic anhydride who develop both IgGl and IgE antibody responses. The provoked airway response is associated with significant microvascular leakage. This suggests that airway microvascular leakage may contribute to asthma caused by trimellitic anhydride. This model may be of value in the further investigation of the underlying mechanisms of occupational asthma caused by the acid anhydrides and possibly other low molecular weight chemicals. References 1. Fawcett IW, Newman TaylorAJ, Pepys J. Asthma due to inhaled chemical agents - epoxy resin systems containing phthalic anhydride and triethylene tetramine. Clin Allergy 1977; 7:1-14. 2. VenablesKM. Lowmolecular chemicals, hypersensitivity, and direct toxicity: the acid anhydrides. Br J Ind Med 1989; 46:222-32. 3. Newman TaylorAJ, Venables KM, Durham SR, Graneek BJ, Topping MD. Acid anhydrides and asthma. Int Arch Allergy Appl Immunol 1987; 82:435-9. 4. Hayes JP, Daniel R, Tee RD, Barnes PJ, Newman Taylor AJ, Chung KF. Immunological and bronchial anaphylactoid responses to intradermal trimellitic anhydride in guinea pigs. Eur Respir J 1990; 3(Suppl 1O:111s). 5. Persson CGA. Roleof plasma exudation in asthmatic airways. Lancet 1986; 2:1126-9. 6. Evans TW, RogersDF, Aursudkij B, Chung KF, Barnes PJ. Regional and time-dependent effects of inflammatory mediators on airway microvascular permeability in the guinea pig. Clin Sci 1989; 76:61-6. 7. RogersDF, BelvisiMG, Aursudkij B, Evans TW, Barnes PJ. Effects and interactions of sensory neuropeptides on airway microvascular leakage in guinea pigs. Br J Pharmacol 1988; 95:1109-16. 8. Majno G, Shea SM, Levanthal M. Endothelial contraction induced by histamine-type mediators. J Cell BioI 1969; 42:647-52. 9. Ovary Z, Kaplan B, Kojima S. Characteristics of guinea pig 19B.Int Arch Allergy Appl Immunol 1976; 51:416-8. 10. von Neergaard K, Wirz K. Die messung den stromungswiderstande in den atemwegen des menschen; inbesondere bei asthma und emphysema. Z Klin Med 1927; 105:51-82. 11. BothamPA,RattrayNJ, Woodcock DR, Walsh ST, Hext PM. The induction of respiratory allergy in guinea pigs following intradermal injection of trimellitic anhydride: a comparison with the re-

sponse to 2,4,-dinitrochlorobenzene. Toxicol Lett 1989; 47:25-39. 12. Zeiss CR, Patterson R, Pruzansky J J, Miller MM, Rosenberg M, Levitz D. Trimellitic-anhydride induced airway syndromes: clinical and immunologic syndromes. J Allergy Clin Immunol 1977; 60:96-103. 13. Hayes JP, Daniel R, Tee RD, Barnes PJ, Newman-Taylor AJ. Airway responses followingintravenous challenge with a protein conjugate of trimellitic anhydride in guinea pigs sensitized to the free hapten; comparison of 3 methods. Thorax 1991; 46:28Op. 14. Biagnini RE, Moorman WJ, LewisTR, Bernstein IL. Ozone enhancement of platinum asthma in a primate model. Am Rev Respir Dis 1986; 134:719-25. 15. Bernstein IL, Splansky GL, Chen SE, Vinegar A. The guinea pig model of diisocyanate sensitization II physiologic studies. J Allergy Clin Immunol 1982; 70:393-8. 16. Chan H, Kam ST, Van Oostdam J, Moreno R, Pare PD, Chan-Yeung M. A rabbit model of hypersensitivity to plicatic acid, the agent responsible for red cedar asthma. J Allergy Clin Immunol 1987; 79:762-7. 17. Lam S, Leriche J, Kijek K, Phillips RT.Effect of bronchial lavage volume on cellular and protein recovery. Chest 1985; 88:856-9. 18. Lam S, LeRiche J, Phillips D, Chan-YeungM. Cellular and protein changes in bronchial lavage fluid after late asthmatic reaction in patients with red cedar asthma. J Allergy Clin Immunol 1987; 80:44-50. 19. Evans TW, Rogers DF, Aursudkij B, Chung KF, Barnes PJ. Inflammatory mediators involved in antigen-induced airway microvascular leakage in guinea pigs. Am Rev Respir Dis 1988;138:395-9. 20. Hui KP, Lotvall JO, Rogers DF, Barnes PJ, Chung KF.Airway aerosol challengein activelysensitized guinea pigs: relationship between airway microvascular leakage and airflow obstruction. Allergy 1992; (In press). 21. Kowalaski ML, Kaliner MA. Neurogenic inflammation, vascular permeability and mast cells. J Immunol 1988; 140:3905-11. 22. McDonald DM. Neurogenic inflammation in the respiratory tract: actions of sensory nerve mediators on blood vessels and epithelium of the airwaymucosa. Am RevRespirDis 1987; 136:S65-S72. 23. Hui KP, Lotvall JO, Chung KF, Barnes PJ. Attenuation of inhaled allergeninduced airflow obstruction and airway microvascular leakage in actively sensitized guinea pigs by 5-lipoxygenase inhibition (A-63162). Am Rev Respir Dis 1991; 143:1015-9.

Bronchoconstriction and airway microvascular leakage in guinea pigs sensitized with trimellitic anhydride.

We have developed a guinea pig model of immediate airway responses following intradermal sensitization with free trimellitic anhydride (TMA). Guinea p...
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