Effects of age and allergen-induced airway inflammation in cats: radiographic and cytologic correlation

Thoracic radiography is an important diagnostic tool for feline respiratory medicine. The aim of this study was (1) to assess age-related changes of thoracic radiographic views in healthy young cats and (2) to test if experimentally-induced bronchial inflammation by inhaling Ascaris suum (AS) allergens leads to radiographic changes after single or repeated exposures. Healthy cats (n=15-30) aged between 6 and 30 months were evaluated. Eight healthy cats and eight AS-sensitised cats, respectively, inhaled sterile saline or allergen. Radiographs were taken 24h before, and 6, 24 and 48 h after the challenge. Bronchoalveolar lavage (BAL) was performed after the last radiographic examination. AS-sensitised cats underwent three further allergen challenges at 3-month intervals. The radiographic evaluation was based on a scoring system considering bronchial, interstitial and alveolar patterns. A significant age-related increase in interstitial and total radiographic score was detected in healthy cats older than 18 months and in healthy cats older than 24 months. Whilst saline inhalation did not affect radiographic scores, a single AS challenge induced significant changes of all scores within 6-24h. A significant positive correlation between radiographic scores and BAL neutrophils and eosinophils was found. Repeated AS challenges did not induce irreversible changes in radiographic scores.     

Bronchodilators in bronchoscopy-induced airflow limitation in allergen-sensitized cats

This study investigated the effect of bronchoscopy and bronchoalveolar lavage (BAL) on respiratory function, determined by barometric whole-body plethysmography (BWBP), of healthy and allergen-sensitized cats. Furthermore, the efficacy of inhaled bronchodilators in preventing changes in respiratory function was determined. For test 1, 18 healthy experimental cats were investigated on day 1 by BWBP. On day 2, the cats underwent BWBP after sedation (medetomidine), after anesthesia induction (propofol), and after bronchoscopy and BAL. Enhanced pause (Penh) was significantly increased after bronchoscopy and BAL (1.64 +/- 0.17 versus 1.23 +/- 0.07, P < .05). For test 2, 6 cats were sensitized to ovalbumin (OVA), 6 cats were sensitized to Ascaris suum (AS), and 6 cats served as controls. On day 0, OVA- and AS-sensitized cats underwent an inhaled allergen challenge, whereas controls were exposed to saline. On days 1 and 2, the same protocol as described for test 1 was repeated. Post-BAL Penh of the AS-sensitized cats was significantly higher than at test 1 (2.28 +/- 0.22 versus 1.69 +/- 0.33, P < .05) and was correlated with BAL fluid neutrophil count (r = 0.55, P < .05). During tests 3, 4, and 5, the same protocol as used for test 2 was applied to each cat group, with the animals being randomly treated before sedation with inhaled salbutamol (200 microg), ipratropium bromide (40 microg), or a combination of both (200 + 40 microg). Post-BAL Penh of the AS-sensitized group was significantly decreased after the salbutamol + ipratropium bromide treatment (1.56 +/- 0.18 versus 2.28 +/- 0.22, P < .05). This study suggests that bronchoscopy and BAL induce airflow limitation in cats, which is more severe in the presence of lower airway inflammation. Inhaled salbutamol + ipratropium bromide reduce BAL-induced bronchoconstriction in AS-challenged cats and might be recommended as preventive treatment of asthmatic cats undergoing bronchoscopy.     

Thoracic radiography,bronchoalveolar lavage cytopathology,and pulmonary parenchymal histopathology: a comparison of diagnostic results in 11 cats

The purpose of this study was to compare the diagnostic results and value of thoracic radiography, bronchoalveolar lavage (BAL) fluid cytopathology, and lung histopathology in 11 cats with spontaneous respiratory disease in which radiography and cytopathology were inadequate in establishing a definitive diagnosis. In these cats, radiographic patterns were characterized as bronchial (n=6), interstitial (n=3), and alveolar (n=2); other features included hyperinflation (n=3), bronchiectasis (n=2), pleural fissure lines (n=2), pulmonary nodules (n=2), atelectasis (n=1), and a tracheal mass (n=1). Bronchoalveolar lavage fluid was unremarkable in two cats. Abnormal BAL fluid showed inflammation (n=5), hemorrhage (n=2), epithelial hyperplasia (n=1), or was suspicious for neoplasia (n=1). Histopathological evaluation revealed inflammation (n=8), neoplasia (n=2), and vascular congestion (n=1). The predominant radiographic location of disease correlated with the same histopathological location in seven cats, and the cytopathological class of BAL fluid was consistent with the histopathological class of disease in seven cats. There was poor correlation between the types of cells found in the BAL fluid and the pathologist's prediction of the types of cells likely to be found in the BAL fluid based on the amount and type of airway cellularity seen on histopathological examination. The results of this study suggest that in some cats, BAL fluid cytopathology does not always correlate with the type of pulmonary disease identified on histopathology. In respiratory diseases where radiography and cytopathology fail to provide a definitive diagnosis, histopathological examination of the lung may be necessary.     

Inhaled fluticasone reduces bronchial responsiveness and airway inflammation in cats with mild chronic bronchitis

This study investigated the effect of inhaled fluticasone on lower airway inflammation and bronchial responsiveness (BR) to inhaled carbachol in cats with very mild, chronic bronchitis (n = 5) that were compared with healthy cats serving as controls (n = 6). Chest radiographs, BR tests performed non-invasively by barometric whole body plethysmography (BWBP) and bronchoalveolar lavage (BAL) were performed before and after treatment. BR was quantified by calculating the concentration of carbachol inducing bronchoconstriction (C-Penh300%), defined as a 300% increase of baseline Penh, an index of bronchoconstriction obtained by BWBP. BAL fluid was analyzed cytologically and the oxidant marker 8-iso-PGF2alpha was determined. At test 1, healthy cats and cats with bronchitis were untreated, whereas for test 2 inhalant fluticasone (250 microg once daily) was administrated for 2 consecutive weeks to cats with bronchitis. Control cats remained untreated. Inhaled fluticasone induced a significant increase in C-Penh300% and a significant decrease of BAL fluid total cells, macrophages, neutrophils and 8-iso-PGF2alpha in cats with bronchitis, whilst untreated control cats did not show significant changes over time. This study shows that a 2-week fluticasone treatment significantly reduced lower airway inflammation in very mild bronchitis. BR could be successfully monitored in cats using BWPB and decreased significantly in response to inhaled fluticasone. 8-Iso-PGF2alpha in BAL fluid was responsive to treatment and appeared as a sensitive biomarker of lower airway inflammation in cats.     

Bronchoscopic findings in 48 cats with spontaneous lower respiratory tract disease (2002-2009)

Background: Diagnosis of lower respiratory disease requires collection of airway samples to confirm the etiology of disease. Bronchoscopic evaluation is commonly performed in dogs but less information is available in cats. Hypothesis: The presence and number of bronchoscopic abnormalities visualized during bronchoscopic evaluation of cats with lower respiratory disease will correlate with the type of disease and total and differential cell counts in bronchoalveolar lavage (BAL) fluid. Animals: Forty‐eight cats prospectively evaluated by a single bronchoscopist. Methods: Bronchoscopy was performed during clinical evaluation of cats presenting with cough, respiratory distress, or both. Cats were evaluated for airway hyperemia, stenosis, or collapse, mucus accumulation, bronchiectasis, and epithelial irregularities. Cats were placed into groups of bronchitis/“asthma,” pneumonia, or neoplasia based on BAL findings, histopathology, and response to appropriate medical therapy. Summation of bronchial abnormalities and total and differential cell counts were compared among groups. Results: Endobronchial abnormalities were common in cats with feline bronchitis/asthma, pneumonia, and neoplasia and no differentiating features were found. Excessive mucus accumulation was common (83%), followed by stenosis of bronchial openings and nodular epithelial irregularities (56%), airway hyperemia (54%), airway collapse (48%), and bronchiectasis (27%). Total bronchoscopic score and total cell count did not differ among groups, although differential cell counts were significantly different. A weak correlation (R²= 0.16, P= .006) between age and total bronchoscopic score was noted. Conclusions and Clinical Relevance: Bronchoscopic abnormalities are common in cats with lower respiratory tract disease, and visualization of the airways provides additional nonspecific clinical information in cats.     

Increased adenosine concentration in bronchoalveolar lavage fluid of horses with lower airway inflammation

Several reports have suggested a role for adenosine in the pathogenesis of chronic airway conditions and this has led to new therapeutic strategies to limit airway inflammation. In this study, detectable levels of adenosine in bronchoalveolar lavage (BAL) samples from 11 horses with non-infectious lower-airway inflammation and 14 healthy controls are reported, with significantly higher values in horses with airway inflammation. Although these increased levels did not correlate with changes in neutrophil percentage in BAL, a positive association between adenosine levels and signs of lower airway inflammation (clinical score) was observed. These novel findings support the hypothesis that adenosine may contribute to bronchoconstriction and also act as a pro-inflammatory mediator in the bronchoalveolar milieu of horses with airway inflammation. Further investigation of this axis could lead to new approaches for the treatment of highly prevalent lower airway inflammatory conditions in the horse.     

Effects of cyproheptadine and cetirizine on eosinophilic airway inflammation in cats with experimentally induced asthma

OBJECTIVE: To determine whether oral administration of cyproheptadine or cetirizine blocks the action of serotonin and histamine, respectively, and results in diminished eosinophilic airway inflammation in cats with experimentally induced asthma. ANIMALS: 9 cats in which asthma was experimentally induced through exposure to Bermuda grass allergen (BGA) during a 3-month period. PROCEDURES: Cats were randomized to receive monotherapy with each of 3 treatments for 1 week: placebo (flour in a gelatin capsule, PO, q 12 h), cyproheptadine (8 mg, PO, q 12 h), or cetirizine (5 mg, PO, q 12 h). A 1-week washout period was allowed to elapse between treatments. Prior to and following each 1-week treatment period, blood and bronchoalveolar lavage fluid (BALF) samples were collected. The percentage of eosinophils in BALF was evaluated to determine treatment efficacy. Serum and BALF BGA-specific immunoglobulin contents and plasma and BALF histamine concentrations were determined via ELISAs. Plasma and BALF serotonin concentrations were measured by use of a fluorometric method. RESULTS: The mean +/- SD percentage of eosinophils in BALF did not differ significantly among treatment groups (placebo, 40 +/- 22%; cyproheptadine, 27 +/- 16%; and cetirizine, 31 +/- 20%). Among the treatment groups, BGA-specific immunoglobulin content and histamine and serotonin concentrations were not significantly different. CONCLUSIONS AND CLINICAL RELEVANCE: In cats with experimentally induced asthma, cyproheptadine and cetirizine were not effective in decreasing airway eosinophilic inflammation or in altering several other measured immunologic variables. Neither cyproheptadine nor cetirizine can be advocated as monotherapy for cats with allergen-induced asthma.     

Non-invasive assessment of airway responsiveness in healthy and allergen-sensitised cats by use of barometric whole body plethysmography

This study aimed at determining whether airway responsiveness (AR) tests performed by use of barometric whole body plethysmography (BWBP) were repeatable in cats and to what extent AR was affected by the nebulization protocol used, the age of the animals, the inflammatory status of the airways and prior bronchodilator treatment. Repeatability of AR was tested on two occasions in 30 healthy cats. The concentration of carbachol inducing a 300% increase of the enhanced pause (Penh)--an estimator of airflow limitation--was calculated (C-Penh300) and did not differ significantly between the two tests (0.035+/-0.017% compared to 0.034+/-0.016%) and was significantly and positively correlated. The comparison between rapidly and slowly increasing carbachol concentrations was performed in ten healthy cats and showed a significantly lower C-Penh300 (%) when slowly increasing concentrations were used (0.037+/-0.013% compared to 0.039+/-0.015%, P<0.05). A significant age-related increase of C-Penh300 was evidenced by performing AR tests in 15 healthy cats at 12, 18, 24 and 30 months (12 months: 0.026+/-0.008%, 18 months: 0.031+/-0.009%, 24 months: 0.038+/-0.01%, 30 months: 0.043+/-0.014%, P<0.05). C-Penh300 significantly decreased in 12 Ascaris suum-sensitised cats after allergen exposure (0.026+/-0.016% compared to 0.033+/-0.016%, P<0.05) and was negatively correlated with the granulocyte percentage of bronchoalveolar lavage fluid (r=-0.36, P<0.01). Compared with a placebo inhalation, pre-treatment with inhaled salbutamol significantly increased C-Penh300 in four healthy cats (0.093+/-0.021% compared to 0.036+/-0.004%, P<0.05). This study provides evidence that AR determination by use of BWBP is promising as non-invasive indicator of lower airway inflammation or for monitoring response to bronchodilator treatment.     

Effect of chronic airway inflammation and exercise on pulmonary and systemic antioxidant status of healthy and heaves-affected horses

In heaves-affected horses the relation between oxidant status, airway inflammation (AI) and pulmonary function (PF) is unknown. The oxidant status of blood and pulmonary epithelial lining fluid (PELF) of healthy (H, n = 6) and heaves-affected horses in clinical remission (REM, n = 6) and in crisis (CR, n = 7) was assessed at rest, during and after standardised exercise test by measurement of reduced and oxidised glutathione, glutathione redox ratio [GRR%]; uric acid and 8-epi-PGF2alpha. Oxidant status was related to PF parameters (mechanics of breathing and arterial blood gas tension) and Al parameters (bronchoalveolar lavage [BAL] neutrophil % and AI score). Haemolysate glutathione was significantly different between groups and was correlated with PF and AI parameters; GRR in PELF was increased during CR and was correlated with PF and AI parameters. Exercise induced an increase of plasma uric acid that was significantly higher both in REM and CR. PELF 8-epi-PGF2alpha was significantly increased in CR and correlated with PF and AI parameters. These results suggest that oxidative stress occurring in heaves is correlated with PF and AI and may be locally assessed by PELF glutathione status, uric acid and 8-epi-PGF2alpha. Systemic repercussions are reflected by assay of GSH in resting horses and by uric acid in exercising horses.     

Flexible bronchoscopy and bronchoalveolar lavage in 68 cats (2001-2006)

BACKGROUND: Bronchoscopy is an important tool for identifying an underlying etiology for respiratory disease in cats. However, the procedure is challenging, because feline airways are small and prone to bronchoconstriction. HYPOTHESIS: Bronchoscopy and bronchoalveolar lavage (BAL) are appropriate and safe diagnostic procedures in the cat. ANIMALS: Sixty-eight cats. METHODS: Flexible bronchoscopy was performed in all cats with the cats under propofol infusion with jet ventilation. The procedures were reviewed for BAL volumes instilled and recovered and for the number and type of complications with the use of 3 flexible endoscopes < 5.0-mm outer diameter. The BAL procedure was compared among scopes by using a one-way analysis of variance. Complication rates were compared by using chi-square analysis. Significance was set at P < .05. RESULTS: Clinical diagnoses included inflammatory airway disease in 46 of 68 cats, pneumonia in 10 of 68, neoplastic disease in 8 of 68, and other conditions in 4 of 68 cats. Mean lavage volumes instilled for the 3 scopes were 2.62-5.05 mL/kg (range, 0.77-9.38 mL/kg). Mean percent fluid recovered for the 3 scopes was 51-73%, (range, 0-140%). BAL cell counts were adequate for cytologic assessment (> 300 cells/microL) in 61 of 64 cats (97%), and in 107 of 120 samples (89%) collected. Complications occurred in 38% of procedures; however, these were mild in 24% of cats; 6% of cats died or were euthanized after the procedure. Complications were not associated with fluid volume instilled or recovered, and could not be related to the underlying disease process. CONCLUSIONS AND CLINICAL IMPORTANCE: Flexible bronchoscopy with BAL was well tolerated in most cats examined.     

Subclinical airway inflammation despite high-dose oral corticosteroid therapy in cats with lower airway disease

Management of feline chronic lower airway disease focuses on controlling clinical signs and decreasing airway inflammation. This retrospective study evaluated the correlation between the resolution of clinical signs in cats with lower airway disease receiving oral glucocorticoids with the resolution of inflammation based on bronchoalveolar lavage fluid (BALF) cytology. Ten cats diagnosed with lower airway disease based on characteristic clinical signs and inflammatory BALF cytology received oral glucocorticoids for at least 3 weeks. They were required to have resolution of clinical signs and BALF collected while asymptomatic and still receiving glucocorticoids. Cats received prednisolone or prednisone (average dose of 1.8±0.2mg/kg daily) for 35.7±5.5 days. Three cats had resolution of clinical signs and lacked inflammatory BALF cytology; seven had persistent inflammatory BALF cytology despite resolution of clinical signs. Given that subclinical inflammation during high-dose glucocorticoid treatment was common, current recommendations to taper therapy based on resolution of clinical signs should be re-evaluated.     

Effect of nutritional antioxidant supplementation on systemic and pulmonary antioxidant status,airway inflammation and lung function in heaves-affected horses

An oxidant/antioxidant imbalance in favour of oxidants has been identified as playing a decisive role in the pathogenesis of chronic inflammatory airway diseases. Nutritional antioxidant supplementation might reduce oxidative damage by enhancement of the antioxidant defence, thereby modulating inflammatory processes. In a placebo-controlled, blind study, it was tested whether a dietary antioxidant supplement administered for 4 weeks would improve lung function and reduce airway inflammation in heaves-affected horses. Eight horses in clinical remission of heaves were investigated at rest and after a standardised exercise test before and after treatment with an antioxidant supplement (consisting of a mixture of natural antioxidants including vitamins E and C and selenium from a variety of sources) or placebo (oatfeed pellets without additive). Pulmonary function and exercise tolerance were monitored; systemic and pulmonary lining fluid uric acid, glutathione and 8-epi-PGF(2alpha) were analysed, and bronchoalveolar lavage (BAL) cytology and inflammatory scoring of the airways were performed. The antioxidant treatment significantly improved exercise tolerance and significantly reduced endoscopic inflammatory score. Plasma uric acid concentrations were significantly reduced, suggesting downregulation of the xanthine-dehydrogenase and xanthine-oxydase pathway. Haemolysate glutathione showed a nonsignificant trend to increase, while plasma 8-epi-PGF(2alpha) remained unchanged. Pulmonary markers and BAL cytology were not significantly affected by antioxidant supplementation. The present study suggests that the antioxidant supplement tested modulated oxidant/antioxidant balance and airway inflammation of heaves-affected horses.     

Comparison of tracheal aspirates and bronchoalveolar lavage in racehorses. 2. Evaluation of the diagnostic significance of neutrophil percentage

OBJECTIVE: To determine whether diagnosis of airway inflammation, using cut-off percentages for neutrophils, differs when based on samples from tracheal aspirate (TA) and bronchoalveolar lavage (BAL) collected concomitantly from the same racehorse. DESIGN: Retrospective case series of 48 young Thoroughbred and Standardbred racehorses in race training, but showing poor performance. PROCEDURE TA and BAL samples were collected from all horses 1 to 2 h after high-speed treadmill exercise. Aliquots of the retrieved fluid were cytocentrifuged and smears stained with Diff-Quik. The mean percentage of neutrophils was calculated. Diagnostic cut-off points were set at 20% for TA samples and 5% for BAL samples. Agreement in the interpretations between the two techniques was analysed. RESULTS: In 19 of 51 paired samples (37%) there were differences in diagnostic interpretation between TA and BAL samples. Of these, airway inflammation was indicated only by the TA sample in 13 and only by the BAL in 6. CONCLUSION: TA and BAL samples give important information about different regions of the airway, but neither should be used alone for the diagnosis of inflammation of the entire lung. The limitations of these procedures mean that both samples should be collected when it is desired to cytologically evaluate the entire lower airway.     

Effects of drug treatment on inflammation and hyperreactivity of airways and on immune variables in cats with experimentally induced asthma

OBJECTIVE: To compare the effects of an orally administered corticosteroid (prednisone), an inhaled corticosteroid (flunisolide), a leukotriene-receptor antagonist (zafirlukast), an antiserotonergic drug (cyproheptadine), and a control substance on the asthmatic phenotype in cats with experimentally induced asthma. ANIMALS: 6 cats with asthma experimentally induced by the use of Bermuda grass allergen (BGA). PROCEDURES: A randomized, crossover design was used to assess changes in the percentage of eosinophils in bronchoalveolar lavage fluid (BALF); airway hyperresponsiveness; blood lymphocyte phenotype determined by use of flow cytometry; and serum and BALF content of BGA-specific IgE, IgG, and IgA determined by use of ELISAs. RESULTS: Mean +/- SE eosinophil percentages in BALF when cats were administered prednisone (5.0 +/- 2.3%) and flunisolide (2.5 +/- 1.7%) were significantly lower than for the control treatment (33.7 +/- 11.1%). We did not detect significant differences in airway hyperresponsiveness or lymphocyte surface markers among treatments. Content of BGA-specific IgE in serum was significantly lower when cats were treated with prednisone (25.5 +/- 5.4%), compared with values for the control treatment (63.6 +/- 12.9%); no other significant differences were observed in content of BGA-specific immunoglobulins among treatments. CONCLUSIONS AND CLINICAL RELEVANCE: Orally administered and inhaled corticosteroids decreased eosinophilic inflammation in airways of cats with experimentally induced asthma. Only oral administration of prednisone decreased the content of BGA-specific IgE in serum; no other significant local or systemic immunologic effects were detected among treatments. Inhaled corticosteroids can be considered as an alternate method for decreasing airway inflammation in cats with asthma.     

Pulmonary Eosinophilia Associated with Increased Airway Responsiveness in Young Racing Horses

Horses are known to acquire small airway disease (SAD), an allergen-induced naturally occurring syndrome of reversible obstructive lung disease accompanied by airway hyperresponsiveness and increased inflammatory cell numbers on bronchoalveolar lavage (BAL). This disorder has received scant attention in young racehorses. The purpose of the present report was to examine the effect of BAL eosinophilia in young racehorses on clinical examination, BAL, hematology, airway responsiveness, and on pulmonary function at rest and after a standardized exercise challenge. Five (3 males, 2 females; age 2.6 ± 0.9 years) with a history of respiratory compromise and BAL eosinophil differential count <5% and 6 controls (4 males, 2 females; age 3.5 ± 1.0 years) training and performing to expectation with normal BAL cell differential (eosinophils < 1%) were studied. Respiratory system clinical examination was performed and expressed as a clinical score. Arterial blood gas measurements, CBC, and pulmonary function testing were performed at rest. Pulmonary mechanics measurements were repeated 1 hour and 20 hours after a standardized treadmill exercise challenge. Incremental histamine inhalation challenge was performed and the concentration of histamine effecting a 35% decrease in dynamic compliance (PC35C_Dyn) was determined. Significant differences were noted between and controls with regard to clinical score ( P = .01), blood eosinophils ( P = .04), BAL cell count ( P = .04), BAL macrophage differential ( P = .04), PC35C_Dyn ( P = .008), and tidal volume and respiratory rate at 20 hours following exercise challenge ( P = .05). We conclude that pulmonary eosinophilia and airway hyperresponsiveness are manifest in some young horses without overt airway obstruction at rest. We speculate that these may be early events in the natural progression of heaves.     

Rush immunotherapy in an experimental model of feline allergic asthma

Specific allergen immunotherapy represents the only curative treatment of allergy. No studies have evaluated its efficacy in feline allergic asthma. We hypothesized that an abbreviated course of immunotherapy (rush immunotherapy, RIT) would blunt eosinophilic airways inflammation in experimental feline asthma induced with Bermuda grass allergen (BGA). The 6-month study included asthmatic-RIT treated cats; asthmatic-no RIT treated cats; and non-asthmatic cats. RIT involved increasing parenteral doses (20-200 microg) of BGA over 2 days. Numbers of eosinophils in bronchoalveolar lavage fluid (BALF), serum and BALF immunoglobulins, lymphocyte blastogenesis assays, and cytokines in blood and BALF were evaluated. BALF eosinophils decreased (P=0.048) only in asthmatic-RIT treated cats (baseline 1.1 x 10(6); Month 6, 2.4 x 10(5)). Serum BGA-specific IgG was higher (P<0.001) at all time points after baseline within the asthmatic-RIT group, and was higher (P<0.001) than asthmatic-no RIT cats at Months 1 and 3. No differences (P=0.133) in BGA-specific IgE levels over time were noted among asthmatic-RIT cats, but this group had lower IgE levels (P<0.001) levels than asthmatic no-RIT cats at Months 3 and 6. Differences in BGA-specific IgA levels over time and between the two groups did not reach the traditional level of significance. The mean BGA stimulation index in the asthmatic-RIT cats was biologically insignificant at 6 months, reflecting BGA-specific lymphocyte hypoproliferation. Preliminary results of cytokine profiles were not significantly different; however, BAL cytokine profiles favoring a Th2 response prior to RIT shifted to increased IFN-g and IL-10 thereafter. RIT dampens eosinophilic airways inflammation in cats with experimental asthma. The mechanism of RIT may involve changes in allergen-specific immunoglobulins, induction of hyporesponsive lymphocytes, or alteration of cytokine profiles.     

Beneficial cross-protection of allergen-specific immunotherapy on airway eosinophilia using unrelated or a partial repertoire of allergen(s) implicated in experimental feline asthma

The study hypothesis was that in experimentally asthmatic cats rush immunotherapy (RIT) using allergens not completely matched with sensitizing allergen(s) would at least partially attenuate the asthmatic phenotype and modulate the aberrant immune response. In phase I, cats sensitized to Bermuda grass allergen (BGA), house dust mite allergen (HDMA) or placebo received BGA RIT. In phase II, cats dually sensitized to BGA and HDMA received RIT using BGA, HDMA or placebo. Efficacy of RIT was assessed using percentage bronchoalveolar lavage fluid (BALF) eosinophils. Additionally, a variety of immunologic assays were performed. Eosinophilic airway inflammation significantly decreased over time in asthmatic cats given RIT using sensitizing allergen or unrelated allergen (P<0.001). In dually sensitized cats, single allergen RIT but not placebo reduced airway eosinophilia (P=0.038). Differences in allergen-specific lymphocyte proliferation, in the number of IL-10 producing cells and in the percentage T regulatory cells were detected between asthmatic cats getting RIT and controls. Cross-protection manifested by reduced airway eosinophilia was noted in cats treated with RIT allergens which did not completely match allergen used in asthma induction. However, the mechanism of immunologic tolerance may differ when improperly matched allergens to the sensitizing allergens are used in RIT.     

Age-related changes in protein concentrations in serum and respiratory tract lavage fluid obtained from cats.

Total protein concentrations was determined in serum, bronchoalveolar lavage (BAL) fluid, and nasal flush fluid obtained from specific-pathogen-free cats from birth to maturity and from adult conventionally raised cats. Protein components were analyzed by immunoelectrophoresis and isoelectric focusing. Albumin, and alpha-, beta-, and gamma-globulins were among the proteins identified in BAL fluid, and their isoelectric point ranged from 3.1 to 5.1. gamma-Globulin was not detected in serum or BAL fluid of newborn kittens before they had ingested colostrum. By day 3 after ingestion of colostrum, IgG was detected in high concentration in serum and was the predominant immunoglobulin in serum and BAL fluid of older cats. Nasal flush fluid from cats greater than 6 months old contained albumin, and alpha-, beta-, and gamma-globulins, with IgA being the predominant immunoglobulin. Total protein concentration in nasal flush fluid increased progressively with increasing age, and albumin was the predominant protein. Protein concentration was significantly (P less than 0.01) higher in BAL fluid from conventionally raised adult cats than in that from specific-pathogen-free cats.     

Cytologic, microbiologic, and biochemical analysis of bronchoalveolar lavage fluid obtained from 24 healthy cats

Twenty-four healthy cats underwent bronchoscopy and bronchoalveolar lavage to determine the normal cytologic environment of the lower respiratory tract of cats. Initial screening to ensure the health of the study population included complete histories, physical examinations, thoracic radiography, CBC, serologic tests for feline leukemia virus, feline immunodeficiency virus, and occult heartworm, and sugar and Baermann fecal flotation. In 18 cats, protected catheter brush samples of airway secretions from the lavaged lung segment were taken for culture of aerobic and anaerobic bacteria and mycoplasma. Bronchial lavage fluid (5 sequential 10-ml aliquots of normal saline solution) was pooled and filtered with cotton gauze. The unspun sample was used for determination of a total nucleated cell count. Lavage fluid was cytocentrifuged and 500 cells/slide were scored for determination of the cellular differential. Activity of lactate dehydrogenase and concentrations of total protein and IgG within the supernatant were measured, and assays were performed to detect the presence of IgA and IgM. Complete histologic evaluation of the lavaged lung of each of 6 random-source cats was performed after differential cell counting revealed 18% eosinophils within bronchoalveolar lavage fluid recovered from this group. Alveolar macrophages were the predominant cells encountered; however, a quarter of all cells recovered were eosinophils. A significant relationship was not found between the abundance of eosinophils in the lavage fluid, and either isolation of aerobic bacteria, high total nucleated cell counts, total protein concentrations, or activity of lactate dehydrogenase. Histologic evaluation of the lungs of 5 of 6 random-source cats revealed normal lungs in 2 cats, and minimal abnormal change in 3 others. Evaluation of the lungs from 1 random source cat revealed acute, mild eosinophilic bronchiolitis. We conclude that large numbers of eosinophils may be retrieved from the bronchoalveolar lavage fluid of healthy cats.