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.     

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.     

Rush allergen specific immunotherapy protocol in feline atopic dermatitis: a pilot study of four cats

Rush immunotherapy has been shown to be as safe as conventional immunotherapy in canine atopic patients. Rush immunotherapy has not been reported in the feline atopic patient. The purpose of this pilot study was to determine a safe protocol for rush immunotherapy in feline atopic patients. Four atopic cats diagnosed by history, physical examination and exclusion of appropriate differential diagnoses were included in the study. Allergens were identified via liquid phase immunoenzymatic testing (VARL: Veterinary Allergy Reference Labs, Pasadena, CA). Cats were premedicated with 1.5 mg triamcinolone orally 24 and 2 h prior to first injection and 10 mg hydroxyzine PO 24, 12 and 2 h prior to first injection. An intravenous catheter was placed prior to first injection. Allergen extracts (Greer Laboratories, Lenoir, North Carolina) were all administered subcutaneously at increasing protein nitrogen units (pnu) every 30 minutes for 5 h to maintenance dose of 15,000 pnus ml-1. Vital signs were assessed every 15 minutes. Two cats developed mild pruritus and the subsequent injection was delayed 30 minutes. No changes in either cat's vital signs were noted, nor was there any further pruritus. All four cats successfully completed rush immunotherapy. Two cats developed a dermal swelling on the dorsal neck one week later. In these four cats, this protocol appeared to be a safe regimen to reach maintenance therapy. A larger sample of feline patients is needed to determine the incidence of adverse reactions and to follow the success of ASIT based upon this method of induction.     

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.     

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.     

Plethysmographic comparison of breathing pattern in heaves (recurrent airway obstruction) versus experimental bronchoconstriction or hyperpnea in horses

BACKGROUND: Horses with recurrent airway obstruction (heaves) are described as exhibiting "increased abdominal effort," but it is unknown whether this translates to an effective contribution to ventilation. HYPOTHESIS: We hypothesized that heaves is characterized by asynchrony between rib cage and abdominal motions, and that the abdominal component is the major contributor to ventilation. ANIMALS: The rib cage versus abdominal motion in naturally occurring heaves (n = 15) was compared to controls at rest (n = 7) and during hyperpnea because of lobeline treatment, and the effects of histamine-induced bronchoconstriction in controls (n = 10). METHODS: Flow patterns, phase angle (theta) between the rib and abdominal compartments, abdominal (Vabd) contribution to tidal volume (VT), and lung mechanics were measured. RESULTS: Findings unique to the heaves group included the loss of biphasic expiratory flow, severely increased theta with the abdomen consistently lagging behind the rib cage, and a reduced contribution of the abdomen to ventilation. A subgroup of heaves (n = 5) with abdominal paradox showed a significant (P < .05) reduction in tidal volume, and increased respiratory rate. Bronchodilation reduced theta in heaves (P = .06), but theta remained significantly higher after bronchodilation than found in controls. CONCLUSIONS AND CLINICAL IMPORTANCE: We conclude that breathing pattern in horses with heaves is characterized by severe rib cage/abdominal asynchrony, with the rib cage motion in synchrony with flow, therefore dominating ventilation. In a subset of heaves, the abdominal compartment (diaphragm, abdominal muscles) was completely out of synchrony with flow ("abdominal paradox") despite the clinical appearance of "increased abdominal effort."     

The effect of user experience and inflation technique on endotracheal tube cuff pressure using a feline airway simulator

Objective

The effect of user experience and inflation technique on endotracheal tube cuff pressure using a feline airway simulator.

Pneumothorax in cats with a clinical diagnosis of feline asthma: 5 cases (1990–2000)

Objective: This paper characterizes the clinical findings in 5 cats with feline asthma complicated by concurrent pneumothorax. Design: Retrospective study. Medical records of cats with concurrent diagnoses of asthma and pneumothorax that were presented to the Veterinary Hospital of the University of Pennsylvania from 1990 to 2000 were reviewed. Results: Of 421 cases of feline asthma, 5 cats fulfilled the inclusion criteria (1.2%). All 5 had respiratory distress at presentation. One cat was panting, and the other 4 cats had respiratory rates of 28, 52, 58 and 120 breaths per minute (bpm), respectively (mean RR 65±39 bpm). Historical findings included untreated chronic cough (n=3), previously treated asthma (n=1), and no previous illness (n=1). Thoracocentesis was performed in 4/5 cats, and 3 of those cats required thoracostomy tubes. Four cats required immediate oxygen supplementation, and 1 of those cats required ventilation. All 5 cats had evidence of pneumothorax on initial radiographs. Follow‐up radiographs revealed partial or complete resolution of pneumothorax in 4 cats which were discharged alive with total hospitalization of 2–7 days, but were then lost to follow‐up. One cat was euthanized because it could not be weaned off mechanical ventilation, and necropsy confirmed end‐stage feline asthma and emphysema. Conclusion: Small airway obstruction can predispose asthma patients to increased alveolar pressure, emphysema, and spontaneous pneumothorax, which can lead to dyspnea in affected cats. The short‐term outcome in these cats was good despite the severity of dyspnea at presentation.     

Equine atopic skin disease and response to allergen-specific immunotherapy: a retrospective study at the University of California-Davis (1991-2008)

This retrospective study reports on the clinical presentation of equine atopic skin disease and evaluates response to treatment with allergen-specific immunotherapy (ASIT) based on intradermal testing and/or serum testing. Computerized medical records from January 1991 to December 2008 yielded 54 horses included in the study. Presenting clinical signs (CS) included urticaria (n=28), pruritus (n=8) or both (n=18). Forty-one of 54 horses received ASIT, and response to ASIT (n=32) was evaluated via telephone survey. Eighty-four per cent (n=27) of owners reported that ASIT reduced their horse's CS; 59% (n=19) were able to manage CS by ASIT alone. Three horses (9%) were managed with ASIT in combination with doxepin and discontinued use of corticosteroids. There was no statistical significance between type of test performed and reported success of ASIT (χ(2) analysis, P=0.53). Ninety-three per cent (n=30) of owners reported use of antipruritic medications prior to starting ASIT; 57% (n=17) of these owners reported discontinuing those medications due to success of ASIT. Adverse effects were limited to swelling at the injection site, seen in 16% (n=5). Seventy-five per cent (n=24) of owners elected to discontinue ASIT after 6 months to 8 years (mean 2.2 years): 15 due to resolution of CS, six due to persistent CS, two because the horse was sold, and one due to cost. Ten owners reported no recurrence of CS after discontinuing ASIT; five had recurrence within a median of 2 years of discontinuing ASIT (range 1-12 years). Allergen-specific immunotherapy is a safe and effective way to manage equine atopic skin disease.     

Tissue distribution of a feline AGP related protein (fAGPrP) in cats with feline infectious peritonitis (FIP)

Feline alpha(1)-acid glycoprotein (fAGP) increases during feline infectious peritonitis (FIP). We have recently identified a 29 kDa protein that we named feline AGP-related protein (fAGPrP) due to its cross-reactivity with an anti-human AGP monoclonal antibody. In this work we describe the tissue distribution of fAGPrP during FIP, and its relationship with feline coronavirus (FCoV) and myeloid cells. Tissues from five control cats and from 15 cats with FIP were examined by immunohistochemistry using monoclonal antibodies against human AGP, FCoV and myeloid antigens. Diffuse fAGPrP positivity within the lesions, likely due to vascular plasma leakage, endothelial and epithelial lining were detectable. Compared to controls, fAGPrP-expressing cells often increased in number and were diffusely distributed in lymph nodes, as usually occurs for IgM-producing plasma cells during early immune responses. These findings did not depend on the presence of FCoVs or of myeloid cells, suggesting that fAGPrP is not directly involved in the pathogenesis of FIP.     

S-adenosylmethionine (SAMe) in a feline acetaminophen model of oxidative injury

S-adenosylmethionine (SAMe) is reported to have hepatoprotective and antioxidant functions. Acetaminophen (paracetamol) was used to induce oxidative damage in cats, and to then determine the effect of SAMe treatment on erythrocyte morphology, PCV, liver histopathology, thiobarbituate reacting substances (TBARS), reduced glutathione (GSH), and oxidised glutathione (GSSG).Cats receiving acetaminophen had a significant increase in methemoglobin and Heinz body production. A significant effect for the interaction of time and treatment was found for Heinz body production and changes in PCV. No significant changes were found in blood or hepatic TBARS. Blood GSH increased significantly in all cats, while the blood GSH:GSSG ratio tended to increase the most in cats given acetaminophen only. The hepatic GSH:GSSG ratio tended to increase in cats given SAMe and decrease in cats given acetaminophen, but this effect was not significant. SAMe protected erythrocytes from oxidative damage by limiting Heinz body formation and erythrocyte destruction and maybe useful in treating acetaminophen toxicity.     

Management of chronic gangrenous mastitis in a 3-year-old cow using partial (quarter) mastectomy

Bovine gangrenous mastitis is an acute or peracute condition involving one or more quarters of the cow’s udder. It occurs infrequently, but when it occurs, mortality of the affected cows is high. A partial mastectomy of one quarter using a cranial epidural analgesia with 2% lignocaine is described to manage a gangrenous mastitis affecting only one quarter caused by Proteus mirabilis (a gram-negative bacteria) which was not amenable to medical treatment. Partial mastectomy can be a safe and effective procedure for ruminants with udder disease in genetically or otherwise valuable cattle.     

Seroprevalence of feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) in shelter cats on the island of Newfoundland,Canada

Feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) are retroviruses found within domestic and wild cat populations. These viruses cause severe illnesses that eventually lead to death. Housing cats communally for long periods of time makes shelters at high risk for virus transmission among cats. We tested 548 cats from 5 different sites across the island of Newfoundland for FIV and FeLV. The overall seroprevalence was 2.2% and 6.2% for FIV and FeLV, respectively. Two sites had significantly higher seroprevalence of FeLV infection than the other 3 sites. Analysis of sequences from the FeLV env gene (envelope gene) from 6 positive cats showed that 4 fell within the FeLV subtype-A, while 2 sequences were most closely related to FeLV subtype-B and endogenous feline leukemia virus (en FeLV). Varying seroprevalence and the variation in sequences at different sites demonstrate that some shelters are at greater risk of FeLV infections and recombination can occur at sites of high seroprevalence.     

Characterization of Env antigenicity of feline foamy virus (FeFV) using FeFV-infected cat sera and a monoclonal antibody

To characterize neutralizing antigenicity in relation to env genotypes of feline foamy virus (FeFV), serological analyses were performed using FeFV-infected cat sera and several field isolates including two env genotypes (F17- and FUV-types). Since three cats from which FeFV were isolated were found to have undetectable titers of virus neutralization (VN) antibodies, even to the homologous virus, VN antibodies were further examined with complement supplementation as an enhancement factor. With the presence of complement, the VN titers of FeFV-infected cat sera increased drastically. Although most of serum samples neutralized strains of either env genotype, sera sampled from two cats neutralized all the strains examined at similar titers, suggesting that superinfection with both env genotypes of FeFV might have occurred in the two cats. Further, we produced a monoclonal antibody (mAb) specifically neutralizing FeFV strains of FUV-type. The mAb was shown to have higher affinity to an epitope on Env of FUV-type than that of F17-type by immunoprecipitation assay. This study supplies basic information important for studies on FeFV vector development as well as on the relationship between the virus and the host immune response.     

Semen collection in the ostrich (Struthio camelus) using a dummy or a teaser female

1. We investigated the feasibility of training male ostriches to ejaculate into an artificial cloaca (AC) using a teaser female or a dummy. 2. Male ostriches develop desirable behaviour patterns that allow them to be trained to the teaser and the dummy female. Training success can be high as both methods rely on natural stimulation and voluntary ejaculation into the AC provided suitable individuals are identified. 3. The key factors to successful training appear to be: the teaser method - temperament of male ostriches, the crouching behaviour of the female ostriches for the semen collector and acceptance of the collector by the male; dummy method - courtship of humans, learning ability of ostriches to habituate and then to mount a dummy. 4. Both methods are reliable, yield ejaculates of high quality and give reproducible results. The ejaculate volume was 1.09 +/- 0.13 ml, the concentration of spermatozoa 4.21 +/- 0.27 x 10(9)/ml, the total number of spermatozoa 4.67 +/- 0.62 x 10(9) and motility 4.3 +/- 0.1. 5. Commercial ostrich production that relies on natural mating can take advantage of those methods to develop artificial insemination technologies.     

Evaluation of 9-(2-phosphonylmethoxyethyl) adenine therapy for feline immunodeficiency virus using a quantitative polymerase chain reaction.

To determine the efficacy of 9-(2-phosphonylmethoxyethyl)adenine (PMEA) as a prophylactic chemotherapeutic agent for the treatment of lentivirus infections, three groups of specific pathogen free cats were treated with 0, 3, or 6 mg kg-1 twice daily doses of PMEA beginning 24 h prior to virus challenge with feline immunodeficiency virus Petaluma strain. Treatment was continued for 7 weeks post challenge. During this time cats were monitored for drug toxicity, virus specific antibody response, circulating viral antigen and infectious recoverable virus. To determine the long-term influence of PMEA therapy the cats were monitored for 1 year following the cessation of treatment. The low levels of infectious virus present in blood prompted the development of quantitative polymerase chain reaction assay to enumerate viral DNA burdens in the peripheral blood mononuclear cells of the infected cats and thereby assess drug efficacy. The results indicate that, although prophylactic PMEA did not prevent infection, it did substantially limit feline immunodeficiency virus replication. Furthermore, viral DNA levels remained low in the cats receiving drug a full year (the duration of the study) after cessation of treatment.     

Molecular cloning of feline lung resistance-related protein (LRP) cDNA and its expression in a feline lymphoma cell line and adriamycin-resistant subline

Molecular cloning of feline lung resistance-related protein (LRP) was performed to evaluate the relationship between its expression level and drug resistance against chemotherapeutics. The nucleotide sequence of the coding region of feline LRP cDNA was found to be 2670-bp long and to show 84.2-92.6% homology to its human, mouse, and rat counterparts. The expression level of feline LRP mRNA was relatively high in lung, jejunum, and colon. An adriamycin (ADM)-resistant feline lymphoma subline, FT-1/ADM, showed a high level of MDR1 mRNA expression compared with parental FT-1 cells. However, no relationship was observed between the drug-resistant phenotype and the LRP mRNA expression level. Although no direct contribution of LRP to the development of the drug-resistant phenotype was observed, further investigation is advisable.