Establishment of Rat Models of Different Pathological Types of Hypersensitivity Pneumonitis Using Pigeon Droppings.

BACKGROUND: The pathogenesis and pulmonary histopathological characteristics of hypersensitivity pneumonitis (HP) are not yet fully understood. Therefore, we established animal models of HP of different stages, aiming to provide support for research on this disease. METHODS: We established rat models of pigeon breeder's lung of different pathological types by creating freeze-dried allergen powder from fresh pigeon feathers, dander, and other droppings. Freeze-dried allergen powder suspensions of pigeon droppings were used to establish 2 rat models of HP, one by aerosol inhalation and one by airway instillation, and the rats were sacrificed after different lengths of time to observe the pathological changes in their lung tissues. RESULTS: By the 40th week after allergen inhalation, granulomas were the main changes in the model, without fibrotic changes. When using airway instillation to establish the model, at the 20th week, group 1 (low dose + twice/week) and group 2 (medium dose + twice/week) showed granuloma changes, but no fibrosis; group 3 (high dose + once/week) and group 4 (high dose + twice/week) both showed obvious pulmonary fibrotic changes, but the death rate of rats in group 4 was greater. CONCLUSIONS: Both aerosol inhalation and airway instillation of freeze-dried pigeon allergen powder can successfully establish an HP model. The airway instillation method can cause pulmonary fibrotic changes in a short time, and the pulmonary pathological changes of animal models manifest with an obvious time-dose effect.

Frequency of goose and duck down causation of hypersensitivity pneumonitis within an 80-patient cohort.

BACKGROUND: A variety of antigens have been identified as causative of hypersensitivity pneumonitis (HP), which is characterized by inflammation to the lung parenchyma that is induced by exposure. Goose and duck down (GDD) bedding is often overlooked by physicians as a potential cause, yet the use of GDD has markedly increased in recent years, paralleling an increased frequency of reports of GDD-induced HP. OBJECTIVE: To determine the frequency of GDD as the causative antigen in patients with HP who use bedding that contains GDD. METHODS: Patients referred with a working diagnosis of HP underwent a detailed environmental history. Those who were using GDD were asked to remove it as an avoidance procedure. Signs, symptoms, spirometry, and inflammatory markers were followed up at weekly intervals for up to 1 month to determine the effect of remediation. RESULTS: Eighty patients with HP were seen during an 8-year period. Thirty-two patients (40%) were using GDD bedding. Of these 32 patients, 12 (37.5% of those exposed and 15% of the total HP population experienced remission (or nonprogression) of disease by simply avoiding GDD bedding. Eleven (92%) of these 12 patients were female. In patients with GDD-induced HP, lung biopsy patterns were varied. CONCLUSION: Approximately one-third of patients with HP, who slept with GDD, had persistent improvement or remission with simple avoidance. The higher incidence of GDD-induced HP in females may be hormonal and/or sociocultural related. Lung biopsy findings were across the spectrum of histopathologic patterns. Avoidance-challenge techniques were effective in confirming diagnoses and causation and mitigating the need for additional remediation.

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.

T lymphocytes dominate local leukocyte infiltration in response to intradermal injection of functionalized graphene-based nanomaterial.

Graphene-based nanomaterials (GBN) have many potential biomedical applications. However, information regarding their biological properties and interactions with cells and/or soluble factors within a complex tissue is limited. The objective of this study was to use the growing feather (GF) of chickens as a minimally invasive cutaneous test-site to assess and monitor leukocyte recruitment in response to intradermal GBN injection. Specifically, the dermis of 20 GFs per chicken was injected with 10 µl of phosphate-buffered saline (PBS)-vehicle or 10 µl of 300 µg ml-1 oxygen-functionalized (f) GBN (6 chickens/treatment). GFs were collected before- (0) and at 0.25, 1, 2, 3, 4, 5, and 7 days post-injection and used for leukocyte-population analysis of immunofluorescently stained pulp cell suspensions or histological examination. Based on flow-cytometric cell population analysis, lymphocytes and macrophages were the major leukocyte-populations infiltrating GFs in response to f-GBN presence. Compared with PBS-controls, levels of T cells (γδ-, αß-, CD4- and CD8-T cells) were greatly elevated in f-GBN-injected GFs within 6 h and remained elevated throughout the 7-day examination period. f-GBN's effects on local tissue leukocyte recruitment were not reflected in the blood, except for a higher percentage of lymphocytes on 7 days. These observations together with a visual examination of f-GBN-injected GF tissue-sections suggest a delayed-type hypersensitivity-like, inflammatory cell-mediated response to the non-biodegradable f-GBN. The GF 'in vivo test-tube'system together with blood sampling provided unique insight into the time-course, qualitative, and quantitative aspects of immune system activities initiated by the presence of f-GBN in a complex tissue of a living animal. Copyright © 2017 John Wiley & Sons, Ltd.

Quantification of rHVT-F genome load in feather follicles by specific real-time qPCR as an indicator of NDV-specific humoral immunity induced by day-old vaccination in SPF chickens.

The purpose of this study was to look for a reliable molecular method for confirmation of uptake of recombinant turkey herpesvirus vaccine against Newcastle disease (rHVT-F) and for use as a valuable prediction tool of Newcastle disease virus (NDV)-specific immune response in chickens deprived of maternally derived antibody (MDA). A quantitative real-time polymerase chain reaction (real-time qPCR) specific to rHVT-F was developed. The method was applied to various tissue samples taken from specific pathogen free (SPF) chickens experimentally inoculated at day-old with one dose of rHVT-F vaccine over a 6-week period. Among the tested tissues, the rHVT-F vaccine was detected predominantly in the bursa of Fabricius (BF) and the lung for the first week, followed by a progressive decline from 9 days onwards. Then, an increase of genome load was observed in the feather follicles (FF) with a peak at 2 weeks, rising to a level almost 10(3)-fold greater than in the other tissues. Importantly, the rHVT-F genome load in FF appeared to be strongly correlated to the humoral immunity specific to NDV as evaluated by haemagglutination inhibition (HI) test and NDV-specific IgG, IgM and IgA ELISAs. This is the first report of quantification of rHVT-F vaccine in FF and its correlation with the induction of ND-specific immune response in chickens with no MDA. Our data indicate that the application of this real-time qPCR assay on FF samples taken from chickens in the field may be used to confirm rHVT-F vaccine administration and uptake with the important added benefit of offering a non-disruptive sampling procedure.

Sensitization to aeroallergens in atopic dermatitis patients: association with concomitant allergic diseases.

BACKGROUND: A few reports demonstrate the relationship between IgE sensitization to aeroallergens in atopic dermatitis (AD) patients and other allergic diseases and parameters. OBJECTIVES: The objective of this study was to evaluate, if there is a significant relationship between the sensitization to common aeroallergens in AD patients and the occurrence of asthma bronchiale, rhinitis and other atopic parameters. METHODS: Sensitization to dust, mites, animal dander and bird feather was examined (skin prick test, specific IgE) and the relationship with the occurrence of asthma bronchiale, rhinitis, duration of AD, family history and onset of AD was evaluated. RESULTS: Two hundred and eighty-eight patients were examined - 90 men and 198 women. According to our results, IgE sensitization to animal dander, dust and mites may increase the risk of developing asthma or rhinitis. Persistent lesions of AD occur more often in patients with sensitization to animal dander, mites and dust. Patients with the sensitization to bird feather have the onset of AD more often above 5 years of age and in these patients, there is no relationship with the positive data about atopy in the family history. CONCLUSION: There is a greater likelihood of developing other allergic diseases in atopic dermatitis patients who suffer from sensitisation to animal dander, mites, and dust. Thus, prompt management of atopic dermatitis and allergy to inhallant allergens that develop in early infancy may be a successful method for preventing of atopic march.

[Improved IgG Antibody Diagnostics of Feather Duvet Lung by an Antibody Screening Test]. / Optimierte IgG-Antikörperdiagnostik der Bettfedern-Alveolitis mittels EAA-Suchtest.

The underdiagnosed feather duvet lung, an extrinsic allergic alveolitis (hypersensitivity pneumonitis) caused by duck and goose feathers, can be more frequently diagnosed, if duck and goose feather antibodies are included in the panel of the routinely applied IgG antibody screening test. This does not necessarily require extending the screening test to include duck and goose feather antigens. By analysing 100 sera with duck and goose antibodies we found that the commonly used pigeon and budgerigar antibodies can also screen for feather duvet antibodies. All examined sera lacking pigeon and budgerigar antibodies also lacked clear-cut duck and goose feather antibodies. The examined sera with strong pigeon or budgerigar antibodies always also contained feather duvet antibodies. However, sera with medium or low concentrated pigeon or budgerigar antibodies are not always associated with feather duvet antibodies. In the light of these observations, we find that 71% of the duck and goose antibody analyses would be dispensable without essential loss of quality, if the results of screening for pigeon and budgerigar antibodies were incorporated into the procedure of a step-by- step diagnostics.

Spontaneous immunological activities in the target tissue of vitiligo-prone Smyth and vitiligo-susceptible Brown lines of chicken.

Introduction: Vitiligo is an acquired de-pigmentation disorder characterized by the post-natal loss of epidermal melanocytes (pigment-producing cells) resulting in the appearance of white patches in the skin. The Smyth chicken is the only model for vitiligo that shares all the characteristics of the human condition including: spontaneous post-natal loss of epidermal melanocytes, interactions between genetic, environmental and immunological factors, and associations with other autoimmune diseases. In addition, an avian model for vitiligo has the added benefit of an easily accessible target tissue (a growing feather) that allows for the repeated sampling of an individual and thus the continuous monitoring of local immune responses over time. Methods: Using a combination of flow cytometry and gene expression analyses, we sought to gain a comprehensive understanding of the initiating events leading to expression of vitiligo in growing feathers by monitoring the infiltration of leukocytes and concurrent immunological activities in the target tissue beginning prior to visual onset and continuing throughout disease development. Results: Here, we document a sequence of immunologically significant events, including characteristic rises in infiltrating B and αß T cells as well as evidence of active leukocyte recruitment and cell-mediated immune activities (CCL19, IFNG, GZMA) leading up to visual vitiligo onset. Examination of growing feathers from vitiligo-susceptible Brown line chickens revealed anti-inflammatory immune activities which may be responsible for preventing vitiligo (IL10, CTLA4, FOXP3). Furthermore, we detected positive correlations between infiltrating T cells and changes in their T cell receptor diversity supporting a T cell-specific immune response. Conclusion: Collectively, these results further support the notion of cell-mediated immune destruction of epidermal melanocytes in the pulp of growing feathers and open new avenues of study in the vitiligo-prone Smyth and vitiligo-susceptible Brown line chickens.

The proteomics of feather development in pied flycatchers (Ficedula hypoleuca) with different plumage coloration.

The genetic theory of morphological evolution postulates that form evolves largely by changing the expression proteins that are functionally conserved. It follows that understanding the function of proteins during different phases of development as well as the mechanisms by which the functions are modified is a prerequisite for understanding evolutionary change. Male pied flycatchers exhibit marked phenotypic variation in their breeding plumage. This variation has repeatedly been shown to have adaptive significance, but the molecular basis of this variation is not known. Here, we characterize the proteome of developing pied flycatcher feathers from differently pigmented males and also introduce a new method for examining the effect sizes of expression differences in protein interaction networks. Approximately 300 proteins were identified in the developing feathers of males. Gene products associated with cellular transport, cell metabolism and protein synthesis formed a large part of the developing feather proteome. Sixty-five proteins associated with the development of the epidermis and/or pigmentation were detected in the data. The examination of expression level differences of protein-protein interaction networks revealed an immunological signalling-related network to exhibit significantly higher expression in black compared to brown males. Additionally, indications of differences in energy balance and oxidative stress related characteristics were detected. Together, these results provide new insight into the molecular mechanisms and evolutionary significance of plumage colour variation.

[A case of acute hypersensitivity pneumonia in a worker at a feather duvet factory].

A 44-year-old woman was admitted to our hospital because of a 15-month history of exertional dyspnea, nonproductive cough and fever. Thoracic high-resolution computed tomography (HRCT) showed centrilobular ground-glass opacities distributed in bilateral lung fields. She had worked at a down quilt factory and had been exposed to a large amount of feathers for 5 years. A peripheral lymphocyte proliferation test by positive was positive for pigeon serum. We diagnosed bird-related hypersensitivity pneumonia. After quitting her job, improvement of her clinical symptoms and chest imaging findings were observed and she has been free of relapse.

Antigen-specific IgG antibodies in feather duvet lung.

BACKGROUND: Feather duvet lung (FDL) is a rare subgroup of bird fancier's lung (BFL). We were interested in determining antigen-specific IgG antibodies in patients with FDL and comparing them with those with BFL. MATERIAL AND METHODS: Specific IgG antibodies against goose and duck feathers, analysed with an automated fluorimetric enzyme immunoassay, were measured in healthy subjects (group A, n = 30), in patients with FDL (group B, n = 10) and with BFL (group C, n = 35); typical specific IgG antibodies of BFL in groups B and C. RESULTS: An optimal threshold value for antibodies against goose or duck feathers to differentiate patients with either BFL or FDL from healthy subjects was determined at 10.85 mg L(-1) for goose feathers and at 8.81 mg L(-1) for duck feathers, respectively. Specific IgG antibodies against goose feathers were significantly higher in group B compared with group C. A ratio of specific IgG antibodies against goose feathers and budgerigar antigens with a threshold value of 0.91 could discriminate between patients with FDL and BFL with a specificity of 97% and a sensitivity of 90%. CONCLUSIONS: We were able to demonstrate the significant difference in IgG antibodies in patients with FDL and BFL and their contribution to discriminate between these similar kinds of extrinsic allergic alveolitis.

Presenting features of feather duvet lung.

BACKGROUND: Feather duvet lung (FDL) is a rare subgroup of bird fancier's lung. It is caused by inhalation of organic dust due to goose or duck feathers in duvets or pillows. METHODS: A retrospective review of the medical records of 13 patients with FDL was performed to assess the specific history and review clinical characteristics of patients with this disease. RESULTS: All patients were female with a mean age of 53 years (26-71). They were recently exposed to feather duvets (6), pillows (1) or both (6). Specific histories were duvets or pillows filled with raw goose feathers from their own farms (4), intensive contact with goose feathers in youth (3), and bird exposure prior to symptom onset (5). In all patients specific IgG antibodies to goose and/or duck feathers were detected. Pulmonary function tests revealed a moderate to severe reduced diffusion capacity and a mild restrictive pattern. High-resolution computed tomography was performed in 11 patients and demonstrated predominantly ground-glass opacities (10) and fibrosis (6). In bronchoalveolar lavage fluid, lymphocytic alveolitis was demonstrated in all patients. Lung biopsies were obtained in 9 patients and demonstrated lymphocytic alveolitis (8), granulomas (3), bronchiolitis obliterans organizing pneumonia pattern (2), and usual interstitial pneumonia pattern (1). CONCLUSIONS: The clinical findings of FDL are typical of extrinsic allergic alveolitis. Primary sensitization could be due to former exposure to bird antigens at home or goose/duck feather exposure in youth. In view of the increasing popularity of feather duvets, FDL should be considered in the differential diagnosis of patients with extrinsic allergic alveolitis.

Local and systemic inflammatory responses to lipopolysaccharide in broilers: new insights using a two-window approach.

The inflammatory response involves a complex interplay of local tissue activities designed to recruit leukocytes and proteins from the blood to the infected tissue. For egg-type chickens, we established the growing feather (GF) as an accessible tissue test site to monitor tissue responses to injected test-material. For commercial broilers, whose health depends to a large extent on innate immune system functions, the GF test system offers an important novel window to directly assess their natural defenses. This study was conducted to adapt the GF test system for use in broilers, and use it to simultaneously examine local (GF) and systemic (blood) inflammatory responses initiated by GF pulp injection of lipopolysaccharide (LPS). Specifically, GF of 12 male and 12 female, 5-week-old broilers were injected with LPS (16 GF/chicken; 1 µg LPS/GF). Blood and GF were collected at 0 (before), 6, and 24 h after GF injection. GF pulp was used to determine leukocyte-infiltration and gene-expression profiles, reactive-oxygen-species generation, and superoxide dismutase (SOD) activity. Blood was used to determine blood cell profiles and SOD activity. A time effect (P ≤ 0.05) was observed for most aspects examined. In GF, LPS injection resulted in heterophil and monocyte infiltration reaching maximal levels at 6 and 24 h, respectively. Reactive-oxygen-species generation, SOD activity, and mRNA levels of IL-1ß, IL-8, IL-6, IL-10, and cathelicidin B1 were elevated, whereas those of TNF-α, LITAF, SOD1, and SOD2 decreased after LPS injection. In blood, levels of heterophils and monocytes were elevated at 6 h, lymphocytes and RBC decreased at 6 h, and thrombocytes and SOD activity increased at 24 h. Assessment of LPS-induced activities at the site of inflammation (GF) provided novel and more relevant insights into temporal, qualitative, and quantitative aspects of inflammatory responses than blood. Knowledge generated from this dual-window approach may find direct application in identification of individuals with robust, balanced innate defenses and provide a platform for studying the effects of exogenous treatments (e.g., nutrients, probiotics, immunomodulators, etc.) on inflammatory responses taking place in a complex tissue.

Expression of cell-adhesion molecules in embryonic induction. II. Morphogenesis of adult feathers.

The developmental appearance of cell-adhesion molecules (CAMs) was mapped during the morphogenesis of the adult chicken feather. Neural CAM (N-CAM), liver CAM (L-CAM), and neuron-glia CAM (Ng-CAM), as well as substrate molecules (laminin and fibronectin), were compared in newborn chicken skin by immunohistochemical means. N-CAM was found to be enriched in the dermal papilla, which was closely apposed to L-CAM-positive papillar ectoderm. The two CAMs were then co-expressed in cells of the collar epithelium. Subsequently generated barb epithelia expressed only L-CAM, but N-CAM reappeared periodically on cells between developing barbs and barbules. N-CAM first appeared on a single L-CAM-positive basilar cell located in each valley flanked by two adjacent barb ridges. Subsequently, the expression of N-CAM extended one cell after another to include the whole basilar layer. N-CAM also appeared in the L-CAM-positive axial-plate epithelia, beginning in a single cell located at the ridge base. The two collectives of N-CAM-positive epithelia constituting the marginal and axial plates then disintegrated, leaving interdigitating spaces between keratinized structures that had previously expressed L-CAM. The morphological transformation from an epithelial cylinder to a three-level branched feather pattern is thus achieved by coupling alternating CAM expression in linked cell collectives with specific differentiation events, such as keratinization. During all of these morphogenetic processes, laminin and fibronectin formed a continuous basement membrane separating pulp from feather epithelia, and were excluded from the sites involved in periodic appearances of N-CAM. The same staining pattern described for developing chickens persisted in the feather follicles of adult chicken tissue that have gone through several cycles of molting. Cyclic expression of the two different CAMs underlies each of the different morphological events that are generated epigenetically during feather morphogenesis.

Feather duvet lung.

A 43-year-old non-smoker was referred with a 3-month history of malaise, fatigue and breathlessness. Blood avian precipitins were strongly positive. Lung function testing confirmed a restrictive pattern with impaired gas transfer. A 'ground glass' mosaic pattern was seen on CT imaging, suggestive of hypersensitivity pneumonitis. Although he had no pet birds, on closer questioning he had recently acquired a duvet and pillows containing feathers. His symptoms, chest radiograph and lung function tests improved after removal of all feather bedding, and he was also started on oral corticosteroid therapy. Our case reinforces the importance of taking a meticulous exposure history and asking about domestic bedding in patients with unexplained breathlessness. Prompt recognition and cessation of antigen exposure may prevent the development of irreversible lung fibrosis.

Cellular and cytokine responses in feathers of chickens vaccinated against Marek's disease.

In Marek's disease virus infection, feather follicle epithelium (FFE) constitutes the site of formation of infectious virus particles and virus shedding. The objective of this study was to characterize cellular and cytokine responses as indicators of cell-mediated immune response in FFE and associated feather pulp following immunization against Marek's disease. Analysis of feather tips collected between 4 and 28 days post-immunization (d.p.i.) from chickens vaccinated post-hatch with either CVI988/Rispens or herpesvirus of turkeys revealed that replication of these vaccine viruses started at 7d.p.i., peaked by 21d.p.i., and subsequently, showed a declining trend. This pattern of viral replication, which led to viral genome accumulation in feather tips, was associated with infiltration of T cell subsets particularly CD8+ T cells into the feather pulp area and the expression of cytokine genes such as interferon-gamma, which is an indication of elicitation of cell-mediated immune responses at the site of virus shedding.

Preservation potential of keratin in deep time.

Multiple fossil discoveries and taphonomic experiments have established the durability of keratin. The utility and specificity of antibodies to identify keratin peptides has also been established, both in extant feathers under varying treatment conditions, and in feathers from extinct organisms. Here, we show localization of feather-keratin antibodies to control and heat-treated feathers, testifying to the repeatability of initial data supporting the preservation potential of keratin. We then show new data at higher resolution that demonstrates the specific response of these antibodies to the feather matrix, we support the presence of protein in heat-treated feathers using ToF-SIMS, and we apply these methods to a fossil feather preserved in the unusual environment of sinter hot springs. We stress the importance of employing realistic conditions such as sediment burial when designing experiments intended as proxies for taphonomic processes occurring in the fossil record. Our data support the hypothesis that keratin, particularly the ß-keratin that comprises feathers, has potential to preserve in fossil remains.

Are cutaneous hypersensitivity tests to inhalant allergens a severity marker for vernal keratoconjunctivitis?

PURPOSE: The purpose of this study was to analyze the cutaneous sensitivity to a variety of allergens in patients with vernal keratoconjunctivitis (VKC) and to demonstrate the relation between skin response and clinical aspects of the disease. METHODS: Twenty patients with vernal keratoconjunctivitis were randomly chosen from the External Disease and Cornea Sector. They were clinically evaluated, and a score ranging from 0 to 20 was applied based on signs and symptoms on ophthalmic examination. All subjects underwent a skin prick test against standardized allergens, such as house dust mites D. pteronyssinus, D. farinae, and Blomia tropicalis, as well as allergens from cat, dog, fungi and feather. RESULTS: Seventy-five per cent of patients were positive for at least one of the allergens tested. House dust mites were responsible for the majority of the cases (75%). There was a poor correlation between the clinical score and sensitivity to allergens (r= - 0.119 for fungi; r= - 0.174 for dog; r= - 0.243 for house dust mites; r= - 0.090 for feather). A significant correlation was found only for cat allergen extract (r = - 0.510; p=0.024). CONCLUSIONS: Our study demonstrated poor correlation between cutaneous hypersensitivity tests and clinical findings in patients with vernal keratoconjuntivitis. We concluded that skin response to inhalant allergens is not a useful test to identify clinical severity and chronicity of inflammatory process in this disease.

Pathogenic and serologic studies of Japanese quail infected with JM strain of Marek's disease herpesvirus.

Japanese quail were inoculated with Marek's disease herpesvirus at 1 day of age, and a second group of the same age was infected by contact with inoculated quail. A third group raised in our laboratory as a source of hatching eggs was accidentally infected with Marek's disease (MD). All quail were observed and examined for periods up to 360 days; the usual clinical signs of MD were not observed. However, the birds were susceptible to MD infection because of the evidence that: 1) The MD-specific agar gel-preceipitation (AGP) antigen was found in the feather tips or feather follicular epithelium of some of them; 2) the MD-specific AGP antibody was found in 6 of 57 exposed to MD; and 3) the virus was recovered from 4 MD-exposed birds by subpassage of their kidney cultures into quail embryo fibroblasts. The specificity of the AGP antigen or antibody in MD-infected quail was examined by the line of identity and cross-absorption tests. The antigen specificity was tested against similar to 700 sera from chickens and quail. Although most of these sera reacted with quail-positive antigen (QPA) prepared from MD-infected quail, some sera from infected or uninfected normal chickens reacted nonspecifically with QPA and with an antigen prepared from normal quail. By gel filtration on Sephadex G-150, the specific antigenic component could be separated from the nonspecific component(s) in QPA.