Advancing diagnostic precision: Unveiling sensitization relationships between cat, dog, and horse allergen molecules.

BACKGROUND: Recent advancements in molecular diagnostics have unveiled a multitude of allergen molecules (AMs) associated with animal sensitizations, revealing significant cross- and co-sensitization patterns among these seemingly distinct allergens. METHOD: We investigated the sensitization profiles of 120 children, sensitized to at least one of the 14 AMs from cat, dog, or horse using the Alex test, employing correlations and hierarchical clusters to explore relationship between sensitizations. RESULTS: Sensitizations to Fel d 1, Can f 4/5, and Equ c 4 differ from other cat, dog, and horse AM sensitizations, suggesting they may represent genuine sensitizations for their respective animals. High correlations were observed among various AMs, including lipocalins (Can f 1/2/6, Fel d 4/7, and Equ c 1), serum albumins (Fel d 2, Can f 3, and Equ c 3), and uteroglobins (Fel d 1 and Can f_Fd1). Hierarchical clustering of sensitizations identified two similarity clusters and one dissimilarity cluster, providing an estimation of the likelihood of cross-reactivity. Additionally, our method facilitated speculation regarding cross-, co-, or genuine sensitization. Moreover, we noted a potential increase in the number and level of sensitized animal AMs concurrent with increased sensitization to other aeroallergens with advancing age. No significant difference was detected for the presence or absence of various types of allergic comorbidities. CONCLUSION: Correlations and hierarchical clustering can unveil the extent and magnitude of cross-, co-, and genuine sensitization relationships among animal AMs. These insights can be leveraged to enhance artificial intelligence algorithms, improving diagnostic accuracy through the integration of other measures of sensitization.

Standardized allergen extracts for allergen immunotherapy.

Nineteen U.S. allergen extracts were standardized by the U.S. Food and Drug Administration (FDA) between 1987 and 1998, including of two house-dust mites, short ragweed, cat hair and cat pelt, seven temperate and one southern grass, and six Hymenoptera venom preparations. Relevant literature was reviewed. For each allergen, a "representative" extract was established; the potency of each representative extract was determined by measurement of the total protein content (Hymenoptera venom), radial diffusion measurement of the dominant allergen (short ragweed and cat), or, if there was no dominant allergen, then by quantitative skin testing by using the ID50EAL (intradermal dilution for 50 mm sum of erythema determines the bioequivalent allergy units) method. In vitro tests were developed to allow the manufacturer to demonstrate that each lot of its extract was statistically identical, within defined limits, to the FDA reference extract. These tests included radial immunodiffusion, competitive enzyme-linked immunosorbent assay, and isoelectric focusing. The standardized extracts offer the advantage of consistent potency from lot to lot for each manufacturer and also from manufacturer to manufacturer, and assure the presence of recognized significant allergens within the extract. Therefore, standardized extracts offer improved safety and efficacy over their nonstandardized predecessors.

Kisspeptin/Kiss1r system and angiogenic and immunological mediators at the maternal-fetal interface of domestic cats.

The Kisspeptin/Kiss1r system is a key regulator of reproduction by stimulating gonadotrophin-releasing hormone and luteinizing hormone release, and in vitro studies have shown that Kisspeptin can modulate angiogenesis and immune function, factors that are also essential for reproduction However, there are no studies on the expression of Kisspeptin/Kiss1r at the maternal-fetal interface in domestic cats and its relationship with angiogenic and immunological mediators. Thus, our objective was to evaluate the spatiotemporal expression profile of Kisspeptin/Kiss1r and angiogenic and immunological mediators in the uterus and placenta of domestic cats during pregnancy. Uterus and placenta samples were collected from cats in mid pregnancy (N = 6) and late pregnancy (N = 6), in addition to uterus from non-pregnant cats in diestrus (N = 7), to evaluate protein and gene expression of kisspeptin (Kiss1), kisspeptin receptor (Kiss1r), vascular endothelial growth factor (VEGF), tyrosine kinase receptor (Flk-1), placental growth factor (PLGF), interferon gamma (INFγ), migration inhibiting factor (MIF), tumor necrosis factor (TNFα), interleukins (IL6 and IL10) by immunohistochemistry and quantitative polymerase chain reaction. Pregnancy increased the uterine expression of Kiss1 and Kiss1r, especially at the late pregnancy, in addition to upregulating INFy, MIF, Vegf, Il10, and Tnf and downregulating Plgf. Higher placental expression of Kiss1r and Plgf mRNA occurred at the late pregnancy, while the expression of Kiss1, VEGF, Flk-1, INFy, TNFα, Il6, and IL10 was higher in the mid of pregnancy. A positive correlation between Kiss1 and Tnf was observed in the placenta, while Kiss1r had a negative correlation with Infγ, Il6, and Il10. The findings reveal that Kisspeptin/Kiss1r and angiogenic and immunological mediators at the maternal-fetal interface of pregnant cat have a gene correlation and are modulated by the gestational age. These data suggest possible functional links of Kisspeptin in placental angiogenesis and immunology.

Nasal allergen challenge and environmental exposure chamber challenge: A randomized trial comparing clinical and biological responses to cat allergen.

BACKGROUND: The direct-instillation nasal allergen challenge (NAC) and the environmental exposure chamber (EEC) are 2 methods of conducting controlled allergen provocations. The clinical and biological comparability of these methods has not been thoroughly investigated. OBJECTIVE: We sought to compare clinical and immunologic responses to cat allergen in NAC versus EEC. METHODS: Twenty-four participants were randomized to receive either NAC followed by a 2-day challenge in an EEC or a 2-day challenge in an EEC followed by NAC. Challenges were separated by 28-day washout periods. We measured total nasal symptom scores, peak nasal inspiratory flow, nasal (0-8 hours) and serum cytokines, serum antibodies, peripheral blood antigen-specific T lymphocytes, and gene expression in nasal scrapings. The primary outcome was the total nasal symptom score area under the curve for the first 3 hours after allergen exposure in NAC or after initiation of exposure in EEC. RESULTS: Both challenges increased IL-5 and IL-13 in nasal fluids and serum and resulted in altered nasal cell expression of gene modules related to mucosal biology and transcriptional regulation. Changes in gene modules, more so than cytokine measurements, showed significant associations with total nasal symptom score and peak nasal inspiratory flow. Overall, EEC exposure generated larger responses and more early terminations compared with NAC. Although the 2 challenges did not correlate in symptom magnitude or temporality, striking correlations were observed in cytokine levels. CONCLUSIONS: Although clinical outcomes of NAC and EEC were temporally different and nonequivalent in magnitude, immunologic responses were similar. Selection of a particular allergen challenge method should depend on considerations of study objectives and cost.

Topology and expressed repertoire of the Felis catus T cell receptor loci.

BACKGROUND: The domestic cat (Felis catus) is an important companion animal and is used as a large animal model for human disease. However, the comprehensive study of adaptive immunity in this species is hampered by the lack of data on lymphocyte antigen receptor genes and usage. The objectives of this study were to annotate the feline T cell receptor (TR) loci and to characterize the expressed repertoire in lymphoid organs of normal cats using high-throughput sequencing. RESULTS: The Felis catus TRG locus contains 30 genes: 12 TRGV, 12 TRGJ and 6 TRGC, the TRB locus contains 48 genes: 33 TRBV, 2 TRBD, 11 TRBJ, 2 TRBC, the TRD locus contains 19 genes: 11 TRDV, 2 TRDD, 5 TRDJ, 1 TRDC, and the TRA locus contains 127 genes: 62 TRAV, 64 TRAJ, 1 TRAC. Functional feline V genes form monophyletic clades with their orthologs, and clustering of multimember subgroups frequently occurs in V genes located at the 5' end of TR loci. Recombination signal (RS) sequences of the heptamer and nonamer of functional V and J genes are highly conserved. Analysis of the TRG expressed repertoire showed preferential intra-cassette over inter-cassette rearrangements and dominant usage of the TRGV2-1 and TRGJ1-2 genes. The usage of TRBV genes showed minor bias but TRBJ genes of the second J-C-cluster were more commonly rearranged than TRBJ genes of the first cluster. The TRA/TRD V genes almost exclusively rearranged to J genes within their locus. The TRAV/TRAJ gene usage was relatively balanced while the TRD repertoire was dominated by TRDJ3. CONCLUSIONS: This is the first description of all TR loci in the cat. The genomic organization of feline TR loci was similar to that of previously described jawed vertebrates (gnathostomata) and is compatible with the birth-and-death model of evolution. The large-scale characterization of feline TR genes provides comprehensive baseline data on immune repertoires in healthy cats and will facilitate the development of improved reagents for the diagnosis of lymphoproliferative diseases in cats. In addition, these data might benefit studies using cats as a large animal model for human disease.

Cat Allergy as a Source Intensification of Atopic Dermatitis in Adult Patients.

Atopic dermatitis (AD) is characterized by exacerbations and remissions of eczematous skin, underlain by impaired skin barrier and aberrant Th2-type and Th-22 cytokine production. A number of allergens, in particular contact with fur animals, may aggravate the disease. This study seeks to define the influence of having a regular contact with a pet cat at home on the severity of symptoms and signs of AD. We addressed the issue using the SCORing Atopic Dermatitis (SCORAD) and visual analog (VAS) scores to assess the intensity of pruritus and by measuring the blood content of specific IgE and IL-4, IL-13, and IL-22 cytokines. The study group consisted of 47 adult patients suffering from AD since childhood, 18 of whom declared having regular contact with a cat and the remaining 29 who denied it. There also was a control group consisted of 16 healthy volunteers with no AD signs. The SCORAD and VAS scores were significantly higher in patients in contact with a cat than in those without it (median SCORAD 61.0 vs. 50.4 and VAS 9.0 vs. 4.0 points, respectively). The sIgE of a majority of patients (94.4%) in contact with a cat was in Class V-VI, compared with just a few patients (3.4%) with no such contact, having sIgE in the same classes (p < 0.001). Significant correlations were revealed between SCORAD and VAS scores and the class level of serum sIgE value. In addition, IL-22 was a single elevated cytokine, only in the patients in contact with a cat, and it correlated with pruritus severity. The results of the study underline the need to beware of the cat fur allergen, and they stress forethought and caution in acquiring and keeping a pet cat by patients suffering from AD.

Asthma prevalence, but not allergic rhinitis nor atopic dermatitis, is associated to exposure to dogs in adolescents.

BACKGROUND: Exposure to pets can be a predisposing factor in the development of certain diseases, including allergic diseases. OBJECTIVE: We analyzed the role that exposure to indoor dogs and cats plays in the prevalence of allergic diseases. METHODS: We examined the cross-sectional data of 1056 women and 936 men aged 15 to 18 years; these individuals were selected through stratified and cluster random sampling. We asked all participants about their exposure to indoor dogs and cats during the year that preceded our study. The prevalence of allergic diseases was determined through core questions taken from The International Study of Asthma and Allergies in Childhood questionnaire. RESULTS: The prevalence was 12.7% (95% CI: 11.3%-14.2%) for asthma, 9.0% (95% CI: 7.8%-10.4%) for allergic rhinitis, and 5.2% (95% CI: 4.3%-6.2%) for atopic dermatitis. The multivariate analyses showed that exposure to indoor dogs, but not indoor cats, was associated with asthma prevalence (aOR 1.37; 95% CI: 1.03-1.83), as was male sex (aOR=1.42; 95% CI: 1.08-1.86), a personal history of allergic rhinitis (aOR=3.24; 95% CI: 2.25-4.66), and a maternal history of asthma (aOR=3.06; 95% CI: 1.89-4.98). The population attributable risk for exposure to indoor dogs was 18%. Notably, neither allergic rhinitis nor atopic dermatitis was found to be associated with dog or cat exposure (p> 0.05). CONCLUSION: Exposure to dogs in late adolescence is a factor associated with asthma, although its contribution to the development of asthma should be investigated in new studies.

The Major Cat Allergen Fel d 1 Binds Steroid and Fatty Acid Semiochemicals: A Combined In Silico and In Vitro Study.

The major cat allergen Fel d 1 is a tetrameric glycoprotein of the secretoglobin superfamily. Structural aspects and allergenic properties of this protein have been investigated, but its physiological function remains unclear. Fel d 1 is assumed to bind lipids and steroids like the mouse androgen-binding protein, which is involved in chemical communication, either as a semiochemical carrier or a semiochemical itself. This study focused on the binding activity of a recombinant model of Fel d 1 (rFel d 1) towards semiochemical analogs, i.e., fatty acids and steroids, using both in silico calculations and fluorescence measurements. In silico analyses were first adopted to model the interactions of potential ligands, which were then tested in binding assays using the fluorescent reporter N-phenyl-1-naphthylamine. Good ligands were fatty acids, such as the lauric, oleic, linoleic, and myristic fatty acids, as well as steroids like androstenone, pregnenolone, and progesterone, that were predicted by in silico molecular models to bind into the central and surface cavities of rFel d 1, respectively. The lowest dissociation constants were shown by lauric acid (2.6 µM) and androstenone (2.4 µM). The specific affinity of rFel d 1 to semiochemicals supports a function of the protein in cat's chemical communication, and highlights a putative role of secretoglobins in protein semiochemistry.

Systemic reactions to subcutaneous immunotherapy: Effects of dosing and aeroallergen content.

BACKGROUND: Systemic reactions are a known risk of subcutaneous immunotherapy (SCIT) for aeroallergens. OBJECTIVE: To identify the dose of SCIT that results in the most systemic reactions to SCIT (SCITSRs) and other risk factors for SCITSRs. METHODS: We performed a retrospective review of all SCIT encounters from 2013 to 2017 at a multisite allergy/immunology practice. SCITSRs were identified from the electronic health record through immunotherapy encounters in which epinephrine was administered. Collected data included patient demographics, the dose of immunotherapy at the time of the SCITSR, the presence or absence of asthma, and aeroallergen content. The control group was generated randomly from the same cohort during the same period. RESULTS: There were 86,949 SCIT visits, with 81 SCITSRs (0.9 per 1000 injections). A total of 77.8% of reactions occurred at a dose of 1:1 0.1 mL and above. The presence of cat (81.5% vs 63.0%, P = .01), dog (67.9% vs 37.0%, P < .001), and grass extracts (85.2% vs 67.5%, P = .01) were associated with SCITSRs. Asthma was not significantly associated with SCITSRs. The presence of dust mites, trees, weeds, and molds was not associated with SCITSRs. There were no months or seasons where SCITSRs were more likely to occur. Individuals who experienced SCITSRs had a mean (SD) higher number of included aeroallergenic groups compared with controls (5.86 [1.88] vs 5.00 [1.92], P < .001). CONCLUSION: Risk factors for SCITSRs in a multisite allergy/immunology practice included administration of the highest immunotherapy doses; inclusion of cat, dog, and grass extracts; and the number of aeroallergenic groups included in the extract. This information helps further characterize risk for patients receiving SCIT.

Endotoxin, cat, and house dust mite allergens in electrostatic cloths and bedroom dust.

Environmental exposure to endotoxin, Fel d I (cat) allergen and Der p I (house dust mite) allergen have been associated with asthma symptoms and have been measured in the environment using various sampling methods, including the electrostatic dust collector. The objectives of this study were to investigate whether levels of endotoxin and allergens were detectable in electrostatic dust collectors and to examine the correlation of allergen and endotoxin levels between electrostatic dust collectors and vacuum sampling methods (floor dust and mattress dust). Electrostatic cloths, bedroom floor dust and mattress dust samples from a subset of 60 homes were randomly selected from the Health of Occupants of Mouldy Homes study for allergen and endotoxin analysis. Fel d I and Der p I allergens were analyzed by double monoclonal antibody ELISA and endotoxin by the kinetic Limulus amoebocyte lysate assay. An enhanced ELISA method was used to analyze Der p I in the electrostatic cloths. Endotoxin was detected in all samples, however Fel d I and Der p I were not detected in all electrostatic dust collector samples (detection in 53% and 15% of cloths respectively). No correlations were found between cloth and dust samples for endotoxin or Der p I, but moderate-to-strong correlations were found between all three sampling methods for Fel d I (rs = 0.612-0.715, p < 0.001). Poor correlation was found between floor dust and mattress dust samples for Der p I (rs = 0.256, p = 0.048). Electrostatic dust collectors may provide a way to measure airborne dust and allergen. Given the moderate-to-low correlations with vacuum dust sampling, this may present a unique measurement system which, when collected alongside traditional vacuum dust sampling, could provide additional exposure measures. Further studies are required to correlate endotoxin and allergen levels measured by electrostatic dust collector with air sampling and to explore the relationships between these bioaerosols, environmental factors and asthma.

Evaluation of the manufacturer-dependent differences in specific immunoglobulin E results for indoor allergens.

BACKGROUND: Even though allergies are an important health issue, the detected amount of allergen-specific IgE (sIgE) has differed widely between manufacturers in the past, and even as recently as this year. These discrepancies hinder diagnostics and can even impact allergen immunotherapy. OBJECTIVE: The aim of this study was to evaluate the development and status quo of the quality of in vitro diagnostic testing for house dust mites (HDM) and cat epithelium, 2 important indoor allergen sources. METHODS: We analyzed data on the allergen sources European HDM, American HDM, and cat epithelium, which were collected by the Society for Promoting Quality Assurance in Medical Laboratories (INSTAND e.V.) during 7 years as part of External Quality Assessment schemes (EQAs). A pseudoanonymized comparison was made of the semiquantitative data and allergen-class results of the 4 main suppliers of in vitro diagnostic sIgE tests. Coefficients of variation (CV) were determined in order to evaluate interlaboratory comparability. RESULTS: In vitro allergy diagnostic testing for the major allergen sources HDM and cat epithelium still reveals manufacturer-dependent differences. Despite this, a cautious trend was found towards an alignment of the results and interlaboratory comparability, with the exception of 1 supplier. CONCLUSION: Even though these results are promising, future EQAs have to be closely monitored to ensure this positive trend is not just a snapshot.

Prevalence of antibodies against Toxoplasma gondii in pets and their owners in Shandong province, Eastern China.

BACKGROUND: Pet ownership in China has been steadily increasing over recent years. However, the risk of pet-associated zoonotic infection with the protozoan parasite Toxoplasma gondii remains poorly defined. METHODS: In a cross-sectional survey, we have determined the seroprevalence of T. gondii infection in pet dogs and cats, and pet owners. Serum samples were collected from 360 pets and 460 corresponding pet owners between March 2016 to June 2017, from Shandong province, eastern China. Sera from the animals were tested for anti-T. gondii antibodies using an indirect haemagglutination assay (IHA) and from the pet owners using an enzyme-linked immunosorbent assay (ELISA). RESULTS: Antibodies against T. gondii were detected in 67 of 360 (18.61%) pets. Seroprevalence of T. gondii in pet cats and dogs was 21.67% and 15.56%, respectively. IgG and IgM antibodies were detected in 79 (17.17%) and 4 (0.87%) of pet owners, respectively; with a total of 83 of 460 (18.04%) pet owners testing seropositive for T. gondii. Our seroprevalence data also suggest that cat owners in general and female pet owners in particular could face a higher risk of acquiring T. gondii infection. CONCLUSIONS: Significant levels of anti-T. gondii antibodies were detected in the pets and their owners in Shandong province, eastern China, indicating a potential zoonotic risk. Prophylactic measures should be implemented to reduce the risk of pet owner's exposure to T. gondii infection.

Local Allergic Rhinitis in Adult Patients with Chronic Nasal Symptoms.

BACKGROUND: Local allergic rhinitis (LAR) remains an underdiagnosed condition characterized by the local production of IgE antibodies during the natural exposure to aeroallergens. The prevalence of LAR in adult patients with a previous diagnosis of non-AR was assessed. MATERIAL AND METHODS: Eighty-four patients with perennial nasal allergy symptoms but a negative skin prick test and specific IgE antibodies against common inhalant allergens were included in the study. Nasal provocation tests were performed with the inhalant allergens Dermatophagoides pteronyssinus, Alternaria, and cat allergen, followed by the detection of nasal-specific IgE antibodies in the lavage during the challenge. RESULTS: LAR was confirmed in 21 (25%) study patients. In the remaining 63 (75%) patients, non-AR was diagnosed. In addition, LAR was found following exposure to D. pteronyssinus in 19 (22.6%) patients, Alternaria in 3 (3.6%) patients, and the cat allergen in 1 (1.2%) patient. In 2 patients, concomitant allergies to D. pteronyssinus and Alternaria were observed. CONCLUSION: LAR can be a form of chronic perennial rhinitis that has previously been considered to be non-AR.

A descriptive pilot study of cytokine production following stimulation of ex-vivo whole blood with commercial therapeutic feline hydrolyzed diets in individual healthy immunotolerant cats.

BACKGROUND: Hydrolyzed diets are used in companion animals for the diagnosis and treatment of adverse food reaction. Similarly, hydrolyzed formulas are used in human infants with severe inflammatory bowel disease or milk allergy, and these must meet the standard of hypoallergenicity through rigorous testing. Unfortunately, no standards are currently applied to hydrolyzed veterinary therapeutic diets, and data for the immunogenicity of feline diets is also not available. Therefore, the main aim of this pilot study was to determine if ex-vivo whole blood stimulation assays could be used to characterize the cytokine response to hydrolyzed commercial diets in a small number of individual healthy immunotolerant cats. This approach has also been used to investigate cytokine production in response to cow milk protein in humans and currently similar studies do not exist in companion animals. Nine healthy cats previously eating the same basal diet were divided into groups and fed one of three hydrolyzed diets exclusively for 6 weeks. Heparinized whole blood was collected from each cat before and after the feeding trial. Ex-vivo whole blood stimulation assays were performed using crude extracts of the basal diet as a positive control, as this diet contained the same proteins present in the hydrolyzed diet but were intact, saline as a negative control, and each cat's respective hydrolyzed diet. Supernatants were collected and analyzed for tumor necrosis factor-alpha, interleukin-10 (IL-10), and interleukin-4 using enzyme-linked immunosorbant assay. RESULTS: Seven cats produced detectable amounts of the anti-inflammatory cytokine IL-10 upon stimulation with the basal diet. Two cats produced detectable amounts of IL-10 upon stimulation with a hydrolyzed soy-based diet and one cat produced a detectable amount of IL-10 upon stimulation with a hydrolyzed chicken-based diet (>125 pg/mL). CONCLUSIONS: Results from this pilot study suggest that in some healthy immunotolerant cats, some hydrolyzed diets may elicit a similar cytokine response compared to their basal diet, which contained the same proteins intact. Therefore, animals may be able to recognize and react to some hydrolyzed forms of tolerated proteins, and may also suggest IL-10 as a target for investigation as a potential marker for dietary tolerance in cats, however further studies would be necessary to corroborate this. Further studies are also needed to determine if this would also be the same in immunologically naïve, sensitized and clinically hypersensitized cats.

Extensive protein hydrolyzation is indispensable to prevent IgE-mediated poultry allergen recognition in dogs and cats.

BACKGROUND: The central premise for the commercialization of diets with hydrolyzed ingredients is that the small-sized digested peptides would be unable to crosslink allergen-specific IgE at the surface of tissue mast cells and induce their degranulation. Evidence for the validity of this concept to diagnose food allergies in dogs and cats is limited, however. Our objectives were to study the recognition of standard and variably hydrolyzed poultry extracts by sera from dogs and cats with elevated chicken-specific serum IgE. RESULTS: Forty sera from dogs and 40 from cats with undetectable, low, medium or high serum levels of chicken-specific IgE were tested by ELISA on plates coated with the positive controls chicken, duck and turkey meat extracts and the negative controls beef meat (dogs) or wheat (cats). Plates were also coated with a non-hydrolyzed chicken meal, and mildly- or extensively-hydrolyzed poultry feather extracts. The frequencies of dogs with positive IgE against the various extracts were: chicken meat: 100%, duck and turkey meats: 97%, beef meat: 3%, non-hydrolyzed chicken meal: 73%, mildly-hydrolyzed poultry feathers: 37% and extensively-hydrolyzed poultry feathers: 0%. For cats, these respective percentages were (with wheat replacing beef as a negative control): 100, 84, 97, 7, 7, 0 and 0%. To detect any allergenic cross-reactivity between poultry meat-based and feather hydrolysate-derived extracts, an IgE ELISA inhibition was also done. Ten canine sera with the highest level of anti-poultry IgE in the previous experiment were incubated overnight with a previously optimized 50 µg amount of each of the extracts used above. We performed ELISA on plates coated with chicken, duck or turkey meats with or without inhibitors. The median inhibition percentages after incubation with the non-hydrolyzed chicken meal were ~22%, with the mildly-hydrolyzed poultry feathers: 14-22%, and those with the extensively-hydrolyzed poultry feathers: 5 to 10%; the last inhibition level was similar to that of the beef meat negative control. CONCLUSIONS: Altogether, these results suggest that an extensive-but not partial-hydrolyzation of the poultry feather extract is necessary to prevent the recognition of allergenic epitopes by poultry-specific IgE.

Myeloid differentiation protein 2 facilitates pollen- and cat dander-induced innate and allergic airway inflammation.

BACKGROUND: The National Health and Nutrition Examination Survey identified several pollens and cat dander as among the most common allergens that induce allergic sensitization and allergic diseases. We recently reported that ragweed pollen extract (RWPE) requires Toll-like receptor 4 (TLR4) to stimulate CXCL-mediated innate neutrophilic inflammation, which in turn facilitates allergic sensitization and airway inflammation. Myeloid differentiation protein 2 (MD2) is a TLR4 coreceptor, but its role in pollen- and cat dander-induced innate and allergic inflammation has not been critically evaluated. OBJECTIVE: We sought to elucidate the role of MD2 in inducing pollen- and cat dander-induced innate and allergic airway inflammation. METHODS: TCM(Null) (TLR4(Null), CD14(Null), MD2(Null)), TLR4(Hi), and TCM(Hi) cells and human bronchial epithelial cells with small interfering RNA-induced downregulation of MD2 were stimulated with RWPE, other pollen allergic extracts, or cat dander extract (CDE), and activation of nuclear factor κB (NF-κB), secretion of the NF-κB-dependent CXCL8, or both were quantified. Wild-type mice or mice with small interfering RNA knockdown of lung MD2 were challenged intranasally with RWPE or CDE, and innate and allergic inflammation was quantified. RESULTS: RWPE stimulated MD2-dependent NF-κB activation and CXCL secretion. Likewise, Bermuda, rye, timothy, pigweed, Russian thistle, cottonwood, walnut, and CDE stimulated MD2-dependent CXCL secretion. RWPE and CDE challenge induced MD2-dependent and CD14-independent innate neutrophil recruitment. RWPE induced MD2-dependent allergic sensitization and airway inflammation. CONCLUSIONS: MD2 plays an important role in induction of allergic sensitization to cat dander and common pollens relevant to human allergic diseases.

Determination of threshold concentrations of plant pollens in intradermal testing using fluorescein in clinically healthy nonallergic cats.

BACKGROUND: Currently the same allergen concentrations for canine intradermal testing (IDT) are recommended for feline IDT. Feline skin reactions are subtle and more difficult to read than canine reactions. This difference may be due to suboptimal allergen concentrations used for IDT in cats. HYPOTHESIS/OBJECTIVES: To determine the irritant threshold concentration (ITC) of 16 pollen allergens using serial dilutions of allergen and intravenous fluorescein. The hypothesis tested was that feline IDT currently is performed at suboptimal allergen concentrations for pollens. ANIMALS: Twenty privately owned healthy clinically nonallergic cats. METHODS: IDT was performed in duplicate using 16 pollen allergens (weeds, grasses and trees) at a dilution of 8000 PNU/mL. Two blinded investigators graded the test reactions independently using subjective and objective criteria. Intravenous fluorescein was then administered and the test reactions were re-evaluated. IDT was repeated for any allergen that was positive, using serial dilutions of allergen at a concentration of 6000 and 4000 PNU/mL. RESULTS: The ITC for 2 of 16 of the allergens was determined. The ITC of Cynodon dactylon (Bermuda grass) and Schinus spp. (Peppercorn) was determined to be between 6000 and 8000 PNU/mL. The ITC of all other allergens tested in this study was >8000 PNU/mL. CONCLUSIONS AND CLINICAL IMPORTANCE: This study confirms that suboptimal allergen concentrations currently are used for feline IDT as the ITC is >8000 PNU/mL for 14 of 16 of the grass, weed and tree pollens evaluated. The ITC of Cynodon dactylon and Schinus spp. was determined to be between 6000 and 8000 PNU/mL.

Chronic cat allergen exposure induces a TH2 cell-dependent IgG4 response related to low sensitization.

BACKGROUND: In human subjects, allergen tolerance has been observed after high-dose allergen exposure or after completed allergen immunotherapy, which is related to the accumulation of anti-inflammatory IgG4. However, the specific T-cell response that leads to IgG4 induction during chronic allergen exposure remains poorly understood. OBJECTIVE: We sought to evaluate the relationship between cat allergen-specific T-cell frequency, cat allergen-specific IgE and IgG4 titers, and clinical status in adults with cat allergy with and without cat ownership and the cellular mechanism by which IgG4 is produced. METHODS: Fel d 1-, Fel d 4-, Fel d 7-, and Fel d 8-specific T-cell responses were characterized by CD154 expression after antigen stimulation. RESULTS: In allergic subjects without cat ownership, the frequency of cat allergen (Fel d 1 and Fel d 4)-specific TH2 (sTH2) cells correlates with higher IgE levels and is linked to asthma. Paradoxically, we observed that subjects with cat allergy and chronic cat exposure maintain a high frequency of sTH2 cells, which correlates with higher IgG4 levels and low sensitization. B cells from allergic, but not nonallergic subjects, are able to produce IgG4 after cognate interactions with sTH2 clones and Fel d 1 peptide or the Fel d 1 recombinant protein. CONCLUSION: These experiments suggest that (1) allergen-experienced B cells with the capacity to produce IgG4 are present in allergic subjects and (2) cat allergen exposure induces an IgG4 response in a TH2 cell-dependent manner. Thus IgG4 accumulation could be mediated by chronic activation of the TH2 response, which in turn drives desensitization.

Allergic sensitization is age-dependently associated with rhinitis, but less so with asthma.

BACKGROUND: Epidemiologic data describing the association between allergic sensitization and asthma and allergic rhinitis in adults are scarce. OBJECTIVE: To determine the prevalence and impact of specific sensitization to airborne allergens on asthma and allergic rhinitis among adults in relation to age. METHODS: A random population sample (age 21-86 years) was examined with structured interview and analysis of specific IgE to 9 common airborne allergens. Of those invited, 692 (68%) subjects participated in blood sampling. IgE level of 0.35 U/mL or more to the specific allergen was defined as a positive test result. RESULTS: Allergic sensitization decreased with increasing age, both in the population sample and among subjects with asthma and allergic rhinitis. In a multivariate model, sensitization to animal was significantly positively associated with asthma (odds ratio [OR], 4.80; 95% CI, 2.68-8.60), whereas sensitization to both animal (OR, 3.90; 95% CI, 2.31-6.58) and pollen (OR, 4.25; 95% CI, 2.55-7.06) was significantly associated with allergic rhinitis. The association between allergic sensitization and rhinitis was consistently strongest among the youngest age group, whereas this pattern was not found for asthma. The prevalence of allergic sensitization among patients with asthma decreased by increasing age of asthma onset, 86% with asthma onset at age 6 y or less, 56% at age 7 to 19 years, and 26% with asthma onset at age 20 years or more. CONCLUSIONS: Sensitization to animal was associated with asthma across all age groups; allergic rhinitis was associated with sensitization to both pollen and animal and consistently stronger among younger than among older adults. Early onset of asthma was associated with allergic sensitization among adults with asthma.