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.     

Biological availability of inhaled fluticasone propionate in horses

Healthy horses received aerosolised, intranasal or oral doses of 3 mg of fluticasone propionate evenly divided over morning and evening treatments for seven days. The bioavailability of the drug was determined in terms of the suppression of the endogenous cortisol concentrations in the horses during the period of treatment. The horses which received the aerosolised drug had significantly lower concentrations of endogenous cortisol on days 5 and 8 than the horses which received aerosolised placebo. The horses which received nasal and oral doses of fluticasone propionate showed no significant changes in their endogenous cortisol concentrations.     

Allergen-specific IgG and IgA in serum and bronchoalveolar lavage fluid in a model of experimental feline asthma

Allergic asthma, a Th2 cell driven response to inhaled allergens, has classically been thought of as predominantly mediated by IgE antibodies. To investigate the role of other immunoglobulin classes (e.g., IgG and IgA) in the immunopathogenesis of allergic asthma, levels of these allergen-specific immunoglobulins were measured in serum and mucosal fluids. Bermuda grass allergen (BGA)-specific IgG and IgA ELISAs in serum and bronchoalveolar lavage fluid (BALF) were developed and optimized in an experimental model of BGA-induced feline asthma. Levels of BGA-specific IgG and IgA significantly increased over time in serum and BALF after allergen sensitization. Additionally, these elevated levels of BGA-specific IgG and IgA were seen in conjunction with the development of an asthmatic phenotype indicated by positive intradermal skin tests, enhanced airways hyperreactivity, and increased eosinophil percentages in the BALF.     

Adjuvanted rush immunotherapy using CpG oligodeoxynucleotides in experimental feline allergic asthma

Allergic asthma is driven by relative overexpression of Th2 cell-derived cytokines in response to aeroallergens. In independent studies, both allergen-specific rush immunotherapy (RIT) and CpG oligodeoxynucleotides (ODN) showed promise in blunting eosinophilic inflammation in a model of feline allergic asthma. We hypothesized that RIT using allergen and CpG ODN would work synergistically to dampen the asthmatic phenotype in experimentally asthmatic cats. Twelve cats with asthma induced using Bermuda grass allergen (BGA) were studied. Of these, six were administered adjuvanted BGA RIT using CpG ODN #2142; six were administered placebo (saline) RIT and later crossed over to adjuvanted RIT. Over 2 days, subcutaneous CpG ODN (0.5ng/kg) with BGA (increasing doses every 2h from 20 to 200microg) was administered. Adverse events were recorded and compared with historical controls. Percentage of eosinophils in bronchoalveolar lavage fluid (BALF), % peripheral CD4+CD25+ T regulatory cells (Tregs), lymphocyte proliferation in response to ConA, and cytokine concentrations in BALF were measured over 2 months. Group mean BALF % eosinophils for the adjuvanted RIT cats were significantly lower at week 1 and month 1 (p=0.03 for both), and marginally significantly lower at month 2 (p=0.09) compared with placebo RIT cats. By the end of the study, 8/12 treated cats had BALF % eosinophils within the reference range for healthy cats. Adjuvanted RIT, but not placebo RIT, cats had significant decreases in the ConA stimulation index over time (p=0.05). BALF IL-4 concentrations were significantly higher at week 1 in adjuvanted RIT cats compared with baseline and month 2, and also with placebo RIT cats at week 1. No significant differences were detected between treatments or over time for IL-10 or IFN-gamma concentrations in BALF or for %Tregs cells in peripheral blood. Adjuvanted RIT using CpG ODN in experimental feline asthma dampens eosinophilic airway inflammation. Adverse effects associated with adjuvanted RIT were less severe compared with a historical, non-adjuvanted RIT protocol. The exact mechanism(s) by which adjuvanted RIT alters the aberrant allergic immune response were not elucidated in this study.     

Evaluation of Key's hypothesis in the feline tibia: an experimental model for augmented bone healing studies

Key's hypothesis states that a segmental long bone defect 1.5 times the diaphyseal diameter exceeds the regenerative capacity of bone in skeletally mature dogs and results in nonunion. This hypothesis was evaluated in 5 adult cats with rigidly fixated segmental tibial ostectomies ranging from 1.25 to 1.52 times the diaphyseal diameter. Clinical, radiographic, and histologic data were obtained over a 12-week period. Healing was classified as mature bony union, clinical union, delayed union, or nonunion. Absence of a consolidating callus and instability after removal of fixation devices was found for all cats at 12 weeks. Scant formation of new bone within the gap was histologically evident for only 1 cat. In the remaining 4 cats, fibrous tissue and striated muscle predominated within the gap, and independent healing of the proximal and distal cut ends of the bone were observed. The occurrence of 4 nonunions and 1 equivocally delayed union indicated that Key's hypothesis overestimates the regenerative capacity of bone in the cat. Failure of this experimental model to produce clinical union within 12 weeks demonstrated that this model is a valid method of investigating augmented bone healing techniques that promote union within this period in the cat.     

Development of an in vitro model of feline cartilage degradation

Osteoarthritis is the most common arthropathy of mammalian species including cats. Cartilage degradation is central to the disorder and here we present, for the first time, an in vitro model of feline cartilage degradation which will be useful for further studies in this target species. Feline articular cartilage explant cultures were maintained for 28 days and in the presence of oncostatin M with and without interleukin (IL)-17, tumour necrosis factor (TNF), IL-1α, or IL-1β. Media samples and digested cartilage explants were analysed for glycosaminoglycan (GAG) and collagen content. The combination of IL-1β and OSM, both at 20 ng/ml, was able to promote GAG release to the greatest extent at 14 days. At 28 days, all groups showed relatively high release of GAG. At 14 days, only IL-1β and OSM in combination were associated with a statistically significant increase in collagen release over and above control tissue. IL-1β dose-response studies showed that an IL-1β dose of 10 ng/ml promotes a statistically significant increase in GAG breakdown when used with OSM, and higher doses of IL-1β did not result in significantly greater response. The model demonstrated both GAG and collagen degradation and will be of use for further understanding of feline cartilage metabolism and for screening of potential structure-modifying agents to be used in cats.     

Rotating dome trochleoplasty: an experimental technique for correction of patellar luxation using a feline model.

The purpose of this study was to compare a trochlear block recession to a rotating dome trochleoplasty, a novel technique for the correction of patellar luxation in small animals. Twenty-eight limbs were used from 14 feline cadavers. With the stifles in flexion and extension, computed tomography was utilized to compare width and depth of the trochlea, medial trochlear ridge height, trochlear articular surface area preserved, patellar contact articular surface area, patellar area covered by the trochlear ridges and patellar tilt angle. The results of this study demonstrated that a rotating dome trochleoplasty is superior to a trochlear block recession with regard to medial trochlear height, trochlear width, trochlear depth and trochlear surface area preservation. The results of this study support further biomechanical evaluation of this technique which eventually may lead to clinical trials.     

Influence of dosage and chemical restraints on feline excretory urography

Three series of trials involving 10 domestic short-haired cats were carried out to determine the influence of dosage of contrast media or type of chemical restraint on feline excretory urography. The 1st series (group A) involved 5 cats sedated with 2.0 mg/kg intramuscular (i.m) injection of 2% xylazine and receiving 800 mg/kg of 76 % meglumine diatrizoate (urografin). The 2nd series (group B) involved another 5 cats sedated with 2.0 mg/kg (i.m) injection of 2% xylazine and receiving 1200 mg/kg of 76% urografin. The 3rd series (group C) involved the repeat urography of the group B cats but sedated with 15 mg/kg (i.m) injection of 5% ketamine hydrochloride. Ventrodorsal radiographs were obtained immediately, 5, 15 and 40 minutes after the injection of 76% urografin. Scores were assigned to nephrographic opacification as described in the literature. The heart rates, respiratory rates and rectal temperatures of the cats were also determined before sedation, after sedation, immediately after the injection of 76% urografin and at 15-minute intervals over a period of 60 minutes. In this study, there were significant differences (P < 0.05) in the nephrographic opacification scores between the group A and group B cats at times 0 and 40 minutes post-administration of urografin. Group A cats had good initial nephrographic opacification which faded later while the nephrographic opacification of group B cats progressively increased. Similarly, nephrographic opacification was significantly (P < 0.05) higher in the xylazine-sedated cats (groups A and B) than the ketamine-sedated cats (group C). However, there were no significant differences (P > 0.05) in heart rates, respiratory rates and rectal temperatures between the 3 groups of cats. It was therefore concluded that increasing the dosage of urografin above 800 mg/kg in cats does not provide additional beneficial effects on the nephrograms produced. Xylazine sedation was observed to produce better nephrographic opacification, however, with delayed nephrographic fading compared to ketamine sedation.     

Cytokine induction in pulmonary airways of horses with heaves and effect of therapy with inhaled fluticasone propionate

Work in humans and laboratory animals has identified a central role for cytokines and chemokines in development and persistence of lower airway inflammation. The objectives of this study were to determine interleukin (IL)-1 beta, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha induction in bronchoalveolar lavage (BAL) of control horses and horses with heaves both during remission and exacerbation of the disease, and to determine the effect of therapy with inhaled fluticasone propionate on the cytokine profile of horses with heaves. IL-1 beta and TNF-alpha mRNA expression was significantly higher in horses with heaves after exposure to moldy hay compared to either values obtained during clinical remission or to healthy controls. IL-8 mRNA expression and protein concentrations were significantly higher in horses with heaves than in controls. Both IL-4 and IFN-gamma mRNA expression was increased at various times in heaves-susceptible horses compared to controls. IL-2, IL-5 and IL-10 mRNA expression was not detected in BAL cells of either group. Therapy with inhaled fluticasone propionate after induction of a severe heaves exacerbation resulted in complete resolution of clinical signs, normalization of pulmonary function tests, and significant decrease in BAL neutrophilia. This was associated with a significant decrease in IL-4 mRNA expression and increase in IFN-gamma/IL-4 ratio in horses with heaves. These results demonstrate the clinical efficacy of inhaled fluticasone propionate for the treatment of heaves and suggest a role for cytokines in the development of lower airway inflammation in heaves-susceptible horses.     

Effect of short-term oral and inhaled corticosteroids on airway inflammation and responsiveness in a feline acute asthma model

The objective of this study was to investigate whether high-dose inhaled fluticasone propionate (FP), alone or in combination with salmeterol (SAL), is as effective as oral prednisolone in reducing airway inflammation and obstruction in cats with experimentally-induced acute asthma. Six cats sensitised to Ascaris suum (AS) were enrolled in a prospective controlled therapeutic trial and underwent four aerosol challenges, at 1-month intervals with AS allergen. The allergen - stimulated animals received four consecutive days treatment with either oral prednisolone at 1mg/kg twice daily, 500 μg of FP inhaled twice daily, or a combination of FP/SAL at 500 μg/50 μg inhaled twice daily, respectively, according to a randomised cross-over design. Treatment-related changes in lung function, airway responsiveness (AR) and bronchoalveolar lavage fluid (BALF) cytology were assessed. Barometric whole-body plethysmography (BWBP) was used for the assessment of respiratory variables and AR. No significant differences in respiratory rate or Penh (an estimate of airflow limitation measured by BWBP) were detected among treatment groups. Allergen-induced airway hyper-responsiveness was significantly inhibited by all three steroid treatments (P<0.05). The mean BALF eosinophil percentage (±SEM) was lower after oral and inhaled corticosteroid treatment and these changes were significant for groups receiving prednisolone and the FP/SAL combination. Findings suggest high-dose FP, particularly in combination with SAL, is effective in ameliorating airway inflammation and hyper-responsiveness in this model of acute feline asthma, and highlight the potential use of these drugs in cats experiencing acute exacerbations of the naturally occurring disease.     

Production of polyclonal antisera against feline immunoglobulin E and its use in an ELISA in cats with experimentally induced asthma

Serum samples from six cats with experimentally induced asthma were used to purify feline IgE using gel filtration and affinity chromatography. The resultant IgE, evaluated for purity by immunoelectrophoresis (IEP) and reactivity by Prausnitz-Kustner (PK) testing, was used to develop polyclonal rabbit anti-feline IgE antisera. Using reverse cutaneous anaphylaxis (RCA), the antisera were determined to be specific for feline IgE. The polyclonal rabbit anti-feline IgE antiserum was then validated in an allergen-specific ELISA. Serum samples from an additional five asthmatic cats sensitized with Bermuda grass allergen (BGA) were evaluated prior to sensitization, after parenteral sensitization, and after aerosol sensitization and challenge. A significant increase in serum BGA-specific IgE was noted over time.     

Stunting syndrome in turkeys. Development of an experimental model

Experiments were conducted to establish a stunting syndrome (SS) model to facilitate research on nutritional aspects of enteric disorders of poults. One-day-old turkeys were dosed per os with tryptose phosphate broth (TPB) (controls) or inoculum (inoculated). The inoculum was prepared by homogenizing intestines from 11-day-old commercial poults diagnosed to have SS in TPB (1:0.5 [wt:wt]). Subsequently, intestines from 8-day-old inoculated poults from the previous experiment were used. Inoculation reduced growth (P less than 0.001) and feed consumption (P less than 0.001) at 8 and 14 days of age. In Expts. 1, 2, and 3, gain of inoculated poults was 60.9%, 58.8%, and 52.6% that of controls up to 8 days of age and 77.9%, 76.6%, and 80.9% that of controls from 8 to 15 days of age, respectively. Feed conversion was impaired (P less than 0.001) up to 8 days of age. The activity of maltase and sucrase in the jejunum and of pancreatic enzymes was determined every 2 days up to 13 days of age. Inoculation decreased (P less than 0.001) maltase and sucrase starting at 3 days of age (i.e., maltase activity was 17.45 and 1.70 mumols maltose hydrolyzed/hr.mg protein in control and inoculated poults, respectively). Inoculation had no effect on pancreatic lipase, amylase, or trypsin.     

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.     

Sheep as an experimental model ofTaenia saginata cysticercosis

Summary Taenia saginata cysticercosis was experimentally produced in 4 Soay sheep, 1 four-horned sheep and 1 Texel sheep by oral administration of eggs of the parasite. Some of these sheep were given dexamethasone therapy. The evolution ofT. saginata cysticercosis in this host is characterised by a rapid immune reaction so that by 6 weeks after infection almost all the cysts were dead. There were indications that the dexamethasone therapy may have assisted establishment of the infection and to some extent, postponed the process of degeneration.

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Ovejas Como Modelo Experimental De Cisticercosis PorTaenia Saginata

Resumen Se produjo cisticercosis experimental mediante la administración oral de huevos deTaenia saginata a 4 ovejas Soay, 1 oveja cuatro cuernos y 1 oveja Texel. Algunas ovejas recibieron terapia con dexametasona. La evolución de la cisticercosis en estos huéspe des se caracteriza por una reacción inmunitaria rápida, con muerte de los quistes a la sexta semana después de la infección. Hubo indicaciones de que la dexametasona ayudó en el establecimiento de la infección, posponiendo el proceso degenerativo de los quistes.

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Le Mouton Comme Modele Experimental De Cysticerques DeTaenia Saginata

Résumé Des cysticerques deTaenia saginata ont été expérimentalement obtenus chez 4 moutons Soay, un mouton à quatres cornes et un mouton Texel à la suite de l'administration d'oeufs de ce parasite. Certains de ces moutons ont fait l'objet d'un traitement à la dexamethasone. L'évolution de cysticerques deT. saginata chez l'hôte est caractèrisée par une réaction immunitaire rapide qui fait que 6 semaines après l'infection la presque totalité des cysticerques sont morts. Il existe des indications montrant que le traitement à la dexamethasone peut avoir contribué à l'établissement de l'infestation et à un retard dans le processus de dégénerescence.

     

Clinical and pathological findings in feline immunodeficiency virus experimental infection.

A study is described of the clinical and pathological findings in 20 specific pathogen free cats infected when 1 year old with feline immunodeficiency virus and monitored over 12 months. Cats were divided into two groups (A and B). The clinical and clinicopathological features were studied in Group A. In Group B, at 1, 2, 4, 9 and 12 months post infection two cats were necropsied. Clinically all cats developed generalised lymphadenopathy, six cats were neutropenic and five cats lymphopenic. Three cats became febrile with conjunctivitis and anterior uveitis and one of these cats ultimately developed jaundice. Postmortem examinations confirmed a generalised lymphadenopathy involving peripheral and visceral lymph nodes with concurrent stimulation of splenic white matter and mucosal lymphoid tissue of the digestive tract and conjunctiva. Within the lymph nodes there was a reactive follicular hyperplasia accompanied by a paracortical hyperplasia with an increased paracortical vascularity. Unusual features were the presence of lymphoid follicles in the bone marrow, thymus and parathyroid tissue. In addition, aggregates of lymphoid cells were found within salivary glands, kidneys, sclera and choroid of the eye. One cat developed a lymphosarcoma affecting the liver and kidneys at 36 weeks post infection. The cat with jaundice had a cholangitis with marked biliary epithelial hyperplasia.