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1.
Poult Sci ; 101(2): 101596, 2022 Feb.
Article in English | MEDLINE | ID: mdl-34929441

ABSTRACT

Parasitism is a divesting problem that is frequently overlooked and may result in severe prominent clinical manifestation. This study aimed to investigate the seasonal and sexual prevalence of the gastrointestinal nematode Ascaridia columbae (A. columbae) infection among domestic pigeons in Giza governorate, Egypt, during the period from 2020 to 2021. One hundred and sixty suspected pigeons were clinically investigated. Blood & tissue samples were collected from infected birds to estimate serum zinc concentration, malondialdehyde (MDA), and nitric oxide levels. As well as tumor necrosis factor-alpha (TNF-α), interleukin 1ß (IL1ß) activity, and histopathological examination were estimated; also, worms were collected for morphological identification using electron microscope (SEM) and molecularly identified using polymerase chain reaction (PCR), further sequenced, and submitted in GenBank with accession number MZ343369. The average ascarid (length × breadth) were 72.4 ± 3.3 µm (70.5 - 79.9 µm) × 39.9 ± 2.5 µm (37.6 - 42.3 µm). The distinguishing morphological characteristics that have been noticed in ascarid worms were creamy white, cylindrical worm with triradiate lips with wide cephalic alae extending on both the lateral sides and filariform esophagus. In males, spicules were almost equal with the presence of precloacal chitinous-rimmed sucker. The prevalence of A. columbae infection was (63.1%) with a higher incidence in females (79.2%) than males (46.1%). The highest seasonal prevalence was observed in winter (92.5%), followed by summer and spring (87.5% and 55%), respectively while, the lowest prevalence was observed in autumn (17.5%). The intensity of worms in the infected intestine varied from 5 to 120 adult worms. The histopathological examination revealed the presence of chronic diffuse moderate catarrhal enteritis with roundworms in the lumen. Infected birds showed a significant increase in nitric oxide and MDA levels while serum zinc levels were lowered in infected pigeons. Infected pigeons revealed a marked increase in IL1-ß and TNFα than apparently healthy ones.


Subject(s)
Ascaridia/anatomy & histology , Ascaridiasis/veterinary , Bird Diseases , Columbidae , Animals , Bird Diseases/immunology , Bird Diseases/parasitology , Columbidae/immunology , Columbidae/parasitology , Egypt , Female , Gastrointestinal Tract , Male , Seasons
2.
Int Arch Allergy Immunol ; 182(11): 1017-1025, 2021.
Article in English | MEDLINE | ID: mdl-34139709

ABSTRACT

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.


Subject(s)
Bird Fancier's Lung , Disease Models, Animal , Administration, Inhalation , Aerosols , Allergens/administration & dosage , Animals , Bird Fancier's Lung/immunology , Bird Fancier's Lung/pathology , Columbidae/immunology , Dander/immunology , Feathers/immunology , Feces , Female , Freeze Drying , Granuloma/immunology , Granuloma/pathology , Lung/immunology , Lung/pathology , Lymphocytes/immunology , Macrophages/immunology , Male , Powders , Pulmonary Fibrosis/immunology , Pulmonary Fibrosis/pathology , Rats, Sprague-Dawley
3.
Immunobiology ; 226(3): 152086, 2021 05.
Article in English | MEDLINE | ID: mdl-33784545

ABSTRACT

BACKGROUND: Certain urban areas could contain many pigeon's allergens, which may play an imperative role in the exacerbation of asthma in pigeon allergen sensitive asthma patients. The circulating form of MUC1 in human serum has been considered as a biomarker for some allergic diseases. The study aimed to investigate the role of MUC1 in pigeon allergens positive asthma patients. METHODS: We were enrolled 200 asthma patients including 81 males and 119 females. After positive pigeon exposure history, 108 patients underwent SPT testing against pigeon allergens (dropping and feather). A total of 17 patients, who had exposure history with SPT positive were undergone detail clinical examination. Serum MUC1expression analysis was done by western blotting method. RESULTS: Out of 200 asthmatic patients, 108 (54%) patients had a history of exposure to pigeons. Skin prick test against pigeon (feather & dropping) allergens was positive in 17 (15.7%) patients among exposure asthmatics. The mean age of the study population was 28.8 ± 10.4 years with 9 males and 8 females. Baseline airway obstruction was seen in 58.8% cases. Out of 17 pigeons expose and sensitive asthmatic the MUC1 expression was up-regulated in 15 (88.2%) and down-regulated in 2 (11.8%). The mean value MUC1 fold change of 15 patients with up-regulation was 4.63 ± 3.00 fold. CONCLUSION: MUC1 expression was up-regulated in 88.2% of patients, who were exposed and sensitive to pigeon allergen (dropping and feather). MUC1 may consider as a biomarker in pigeon sensitive asthma patients.


Subject(s)
Allergens/immunology , Asthma/diagnosis , Asthma/etiology , Columbidae/immunology , Gene Expression , Mucin-1/genetics , Adult , Animals , Biomarkers , Disease Susceptibility , Female , Humans , Male , Middle Aged , Mucin-1/metabolism , Skin Tests , Young Adult
4.
J Appl Microbiol ; 130(5): 1695-1704, 2021 May.
Article in English | MEDLINE | ID: mdl-33048404

ABSTRACT

AIMS: In this study, we aimed to isolate and evaluate the efficacy of Bacillus velezensis as a probiotic and to assess its activity towards pigeons infected with pigeon circovirus (PiCV). METHODS AND RESULTS: Bacillus velezensis, isolated from pigeon faeces, was orally administered to pigeons for 60 days. After pigeons were challenged with PiCV, the PiCV viral load and expression of indicator genes for innate immunity were detected in spleen tissue and faeces of pigeons. Bacillus velezensis significantly reduced the PiCV viral load in the faeces and spleen of pigeons 5 days post-challenge (dpc). The mRNA expression levels of treated pigeons showed that interferon-gamma (IFN-γ), myxovirus resistance 1 (Mx1), and signal transducers and activators of transcription 1 (STAT1) genes were upregulated, whereas no expression of interleukin-4 (IL-4) was detected. Moreover, toll-like receptor 2 (TLR2) and 4 (TLR4) were significantly upregulated in probiotic-treated pigeons (P < 0·05). CONCLUSIONS: This is the first report showing that probiotic supplementation can effectively enhance the T-helper type 1 immune response and decrease the PiCV viral loads in pigeons. SIGNIFICANCE AND IMPACT OF THE STUDY: This study proposes that the administration of a probiotic strain, B. velezensis, to pigeons can protect against PiCV infection.


Subject(s)
Bacillus , Circoviridae Infections/immunology , Circovirus/immunology , Columbidae/immunology , Immunity, Innate/genetics , Probiotics/pharmacology , Animals , Antiviral Agents/pharmacology , Bird Diseases/immunology , Bird Diseases/virology , Circoviridae Infections/veterinary , Circovirus/drug effects , Columbidae/genetics , Columbidae/virology , Cytokines/genetics , Cytokines/metabolism , DNA, Viral , Dietary Supplements/microbiology , Feces/microbiology , Gene Expression Regulation , Interferon-gamma , Spleen , Viral Load
5.
Allergol Int ; 70(2): 208-214, 2021 Apr.
Article in English | MEDLINE | ID: mdl-33041192

ABSTRACT

BACKGROUND: Bird antigens are some of the most relevant antigens in hypersensitivity pneumonitis (HP). Possible sources of bird antigens are bird breeding, feather products and fertilizer with fowl droppings. For the screening and diagnosis of HP, the measurement of bird-specific antibodies should be standardized. The aim of this study was to clarify the utility of serum IgG (sIgG) and IgA (sIgA) antibodies to bird antigens in screening and diagnosing acute/chronic bird-related HP with ImmunoCAP® in multi-centre clinical research. METHODS: We executed a clinical performance test by conducting a multi-institutional study to measure the levels of sIgG/sIgA against pigeon, parrot and budgerigar antigens by the ImmunoCAP® system in 29 acute and 46 chronic bird-related HP patients. RESULTS: The levels of sIgG/sIgA against the bird antigens of the three species were significantly higher in subjects with acute bird-related HP and chronic bird-related HP with acute episodes (recurrent type) than in the control subjects. For sIgG, the optimal cutoff values by receiver operating characteristic (ROC) analysis were 24.6 mgA/L for pigeon, 14.0 mgA/L for parrot, and 8.7 mgA/L for budgerigar. By measuring multiple bird antigens and combining sIgG values of two species, the sensitivity and specificity for acute and recurrent-type chronic bird-related HP patients were 85-91% and 73-80%, respectively. For recurrent and insidious types of chronic bird-related HP, the sensitivity and specificity were 48-61% and 73-80%, respectively. CONCLUSIONS: Measurement of the levels of sIgG/sIgA against pigeon, budgerigar and parrot antigens by ImmunoCAP® was useful for screening and diagnosis in bird-related HP.


Subject(s)
Allergens/immunology , Bird Fancier's Lung/diagnosis , Columbidae/immunology , Immunoglobulin A/blood , Immunoglobulin G/blood , Parrots/immunology , Acute Disease , Aged , Animals , Bird Fancier's Lung/blood , Bird Fancier's Lung/immunology , Chronic Disease , Female , Humans , Immunoassay , Male , Middle Aged
6.
BMC Pulm Med ; 20(1): 288, 2020 Nov 07.
Article in English | MEDLINE | ID: mdl-33160347

ABSTRACT

BACKGROUND: The number of reports on sarcoidosis complicated by hypersensitivity pneumonitis (HP) is limited, and most describe cases complicated by chronic bird-related HP. Here, we present for the first time a case with Propionibacterium acnes-associated sarcoidosis complicated by acute bird-related HP. CASE PRESENTATION: A 62-year-old man with a past medical history of sarcoidosis was admitted to our department, and chest computed tomography showed diffuse ground-glass opacities, which appeared as he rapidly increased the number of pigeons he kept for a competition. Random transbronchial lung biopsy revealed well-formed non-caseating epithelioid granulomas, which contained positively stained substances on immunohistochemistry using the PAB antibody, a specific monoclonal antibody against P. acnes lipoteichoic acid. Poorly formed non-caseating granulomas without positively stained substances were also detected. CONCLUSION: We describe the successful identification of this exceptionally rare case of sarcoidosis complicated by acute bird-related HP in which two morphologically and immunohistologically different types of granulomas were present in the same lung.


Subject(s)
Bird Fancier's Lung/etiology , Columbidae/immunology , Granuloma/microbiology , Propionibacterium acnes/isolation & purification , Sarcoidosis/microbiology , Acute Disease , Animals , Antibodies, Bacterial/blood , Biopsy , Bird Fancier's Lung/pathology , Granuloma/pathology , Humans , Immunohistochemistry , Lung/microbiology , Lung/pathology , Male , Middle Aged , Sarcoidosis/pathology , Tomography, X-Ray Computed
7.
Clin Exp Allergy ; 50(12): 1381-1390, 2020 12.
Article in English | MEDLINE | ID: mdl-33010071

ABSTRACT

BACKGROUND: Chronic hypersensitivity pneumonitis (CHP) remains a diagnostic challenge. The process of collecting and extracting serum and droppings from causative animals for the inhalation challenge test is complicated and the risk of inducing disease progression exists. OBJECTIVE: To investigate the utility and safety of an inhalation challenge test using pigeon eggs. METHODS: Pigeon eggs were pasteurized and mixed with a saline solution to produce an inhalation fluid. An inhalation challenge test was conducted on 19 patients with bird-related CHP and 17 patients with interstitial lung disease other than bird-related CHP. To identify antigens in pigeon eggs, the antigen-antibody responses of the pigeon eggs and serum from patients were evaluated using Western blotting. RESULTS: The mean changes in C-reactive protein, alveolar-arterial oxygen difference, erythrocyte sedimentation rate, and lactate dehydrogenase significantly increased by 0.32 mg/dL (P = .014), 7.8 Torr (P = .002), 1.4 mm/h (P = .012), and 5.4 U/mL (P = .0019), respectively, in bird-related CHP group compared to the control 24 hours after the inhalation challenge test. Furthermore, within 24 hours of the inhalation test, the mean forced vital capacity decreased by 2.3% in the bird-related CHP group compared with a decline of 0.05% in the control group (P = .035). Serum collected from seven bird-related CHP patients who underwent the inhalation challenge test and reacted to antigens with molecular weights of 37-75 KDa, and these molecular weights were consistent with egg albumin and globulin. CONCLUSION: Since a mild response was observed after the inhalation challenge test using pigeon eggs, this test was an obvious candidate for diagnosing bird-related CHP.


Subject(s)
Allergens/administration & dosage , Bird Fancier's Lung/diagnosis , Bronchial Provocation Tests , Columbidae/immunology , Egg Proteins/administration & dosage , Immunologic Tests , Lung/immunology , Administration, Inhalation , Aged , Allergens/immunology , Animals , Bird Fancier's Lung/blood , Bird Fancier's Lung/immunology , Bird Fancier's Lung/physiopathology , Case-Control Studies , Egg Proteins/immunology , Female , Humans , Lung/physiopathology , Male , Middle Aged , Predictive Value of Tests , Time Factors , Vital Capacity
8.
Clin Exp Allergy ; 50(12): 1391-1399, 2020 12.
Article in English | MEDLINE | ID: mdl-32966647

ABSTRACT

BACKGROUND: Identifying early stages of hypersensitivity pneumonitis (HP) is hampered by variable presentation, heterogeneous or undetected causal antigens and lack of gold-standard biomarkers. Krebs von den Lungen (KL)-6 is pathophysiological biomarker of alveolar epithelial damage. Pigeon fanciers, susceptible to HP, provide a model to investigate early HP. OBJECTIVE: To test the hypothesis that plasma concentrations of KL-6 are increased in early-stage acute HP. METHODS: Clinical history, spirometry and blood samples were obtained from pigeon fanciers, 20 with intermittent acute symptoms indicative of developing HP, 27 with no symptoms and 10 healthy subjects with no avian exposure. Plasma KL-6 (units/mL) and pigeon antigen-specific IgG antibody were quantified by enzyme immunoassay. Blood lymphocytes were quantified by flow cytometry and antigen specificity by in vitro cytokine production. RESULTS: KL-6 was higher in fanciers than controls, median (IQR) 452 (244, 632) vs 274 (151, 377), P = .01. Although fanciers with symptoms had similar antigen exposure and lung function, they had higher KL-6 than those without, 632 (468, 1314) vs 320 (200, 480), P < .001. KL-6 correlated with IgG antibody titre in those with symptoms, r = .591, P = .006. High KL-6, irrespective of symptom category, was associated with higher antibody (P = .006) and lymphocyte proliferation (P = .041), and lower CD4+ T lymphocyte proportion (P = .032). CONCLUSION AND CLINICAL RELEVANCE: Raised KL-6 is associated with acute symptoms of early-stage HP, and its correlation with antibody may support therapeutic strategies when HP is suspected. KL-6 may act as a mechanistic biomarker of early pathogenesis by linking lung pathophysiological changes with an endotype of immune hypersensitivity.


Subject(s)
Bird Fancier's Lung/diagnosis , Columbidae/immunology , Mucin-1/blood , Adult , Animals , Biomarkers/blood , Bird Fancier's Lung/blood , Bird Fancier's Lung/immunology , Bird Fancier's Lung/physiopathology , CD4-Positive T-Lymphocytes/immunology , Case-Control Studies , Cell Proliferation , Cells, Cultured , Cross-Sectional Studies , Early Diagnosis , Humans , Immunoglobulin G/blood , Lung/immunology , Lung/physiopathology , Lymphocyte Activation , Middle Aged , Predictive Value of Tests , Up-Regulation
9.
Innate Immun ; 26(6): 490-504, 2020 08.
Article in English | MEDLINE | ID: mdl-32393097

ABSTRACT

TNF receptor-associated factor 6 (TRAF6) is a signal transducer, which plays a pivotal role in triggering a variety of signalling cascades. Here, we cloned and identified the TRAF6 gene from the King pigeon. The open reading frame sequence of pigeon TRAF6 (piTRAF6) is 1638 bp long and encodes a 545 aa protein, including a low-complexity domain, RING finger, Zinc finger, coiled coil domain, and meprin and TRAF homology domain. The aa sequence of piTRAF6 shared a strong identity with that of other birds. PiTRAF6 transcripts were broadly expressed in all the tested tissues; piTRAF6 levels were the highest and lowest in the heart and stomach, respectively. Overexpression of piTRAF6 activated NF-κB in a dose-dependent manner and induced IFN-ß expression. Upon piTRAF6 knockdown by small interfering RNAs, NF-κB activation was markedly inhibited in HEK293T cells. The expression of piTRAF6, as well as pro-inflammatory cytokines and antiviral molecules, were obviously increased after TLR ligand stimulation and Newcastle disease virus or Salmonella Pullorum inoculation. These results suggest that piTRAF6 may play a key immunoregulatory role in the innate immune response against viral and bacterial infections.


Subject(s)
Avian Proteins/genetics , Columbidae/genetics , Newcastle Disease/immunology , Newcastle disease virus/physiology , Salmonella Infections/immunology , Salmonella/physiology , TNF Receptor-Associated Factor 6/genetics , Animals , Avian Proteins/metabolism , Cloning, Molecular , Columbidae/immunology , HEK293 Cells , Humans , Immunity, Innate , Inflammation/genetics , NF-kappa B/genetics , NF-kappa B/metabolism , RNA, Small Interfering/genetics , Sequence Alignment , Signal Transduction , TNF Receptor-Associated Factor 6/metabolism , Zinc Fingers/genetics
10.
Vet Microbiol ; 242: 108591, 2020 Mar.
Article in English | MEDLINE | ID: mdl-32122595

ABSTRACT

Pigeon circovirus (PiCV) is the most diagnosed virus in pigeons (Columba livia) and have been studied and reported globally. PiCV infections can lead to immunosuppression and pigeons infected with PiCV can result to lymphocyte apoptosis and atrophy of immune organs. Young pigeon disease syndrome (YPDS) is a complex disease and believed that PiCV could be one of the agents leading to this syndrome. An effective treatment regimen is needed to control the spread of PiCV in pigeons. In this study pigeon interferon alpha (PiIFN-α) was cloned and expressed and its antiviral effects were tested against fowl adenovirus type 4 (FAdV-4) in vitro and PiCV in vivo. No detectable levels of FAdV-4 viral genome in LMH cells stimulated with 300 µg/mL PiIFN-α were found. Additionally, PiIFN-α was stable at different temperature and pH for 4 h, and no reduction in antiviral activity was observed in untreated and treated cells. In pigeons naturally and experimentally infected by PiCV, no detectable levels of PiCV virus titers were found after treatment with PiIFN-α. Cytokine and ISG expression levels in liver and spleen samples were detected and IFN-γ and Mx1 genes were dominantly up-regulated following PiIFN-α treatment (p < 0.05). This study demonstrated that PiCV can be inhibited by administration of PiIFN-α and PiFN-α can be used as a therapeutic approach to prevent the spread of PiCV in pigeons.


Subject(s)
Bird Diseases/virology , Circoviridae Infections/veterinary , Circovirus/physiology , Cytokines/immunology , Interferon-alpha/pharmacology , Virus Replication/immunology , Animals , Bird Diseases/immunology , Cell Line , Circoviridae Infections/immunology , Circovirus/genetics , Circovirus/immunology , Columbidae/immunology , Columbidae/virology , Escherichia coli/genetics , Female , Genome, Viral , Hydrogen-Ion Concentration , Liver/immunology , Liver/virology , Male , Protein Stability , Spleen/immunology , Spleen/virology , Temperature , Viral Load/immunology , Virus Replication/drug effects
11.
Microb Pathog ; 139: 103905, 2020 Feb.
Article in English | MEDLINE | ID: mdl-31790792

ABSTRACT

Pigeon circovirus (PiCV) is able to infect racing and meat pigeons of all ages and is a key factor that triggers young pigeon disease syndrome (YPDS). PiCV vaccine research has been impeded because PiCV cannot be grown or propagated in cell cultures. Virus-like particles (VLPs), which can be generated by a wide range of expression systems, have been shown to have outstanding immunogenicity and constitute promising vaccines against a wide range of pathogens. Cap protein, which contains neutralizing antibody epitopes, is the only capsid protein of PiCV. In this study, the baculovirus expression system was utilized to express the PiCV Cap protein, which was self-assembled into VLPs with a spherical morphology and diameters of 15-18 nm. Specific antibodies against the Cap protein were induced after BALB/c mice immunized intramuscularly (i.m.) with VLPs combined with adjuvant. Based on these findings, PiCV VLPs may be a promising candidate vaccine against PiCV.


Subject(s)
Bird Diseases/virology , Circoviridae Infections/veterinary , Circovirus/physiology , Columbidae/virology , Animals , Antibodies, Viral/immunology , Baculoviridae/genetics , Baculoviridae/metabolism , Bird Diseases/immunology , Bird Diseases/prevention & control , Capsid Proteins/administration & dosage , Capsid Proteins/genetics , Capsid Proteins/immunology , Circoviridae Infections/immunology , Circoviridae Infections/prevention & control , Circoviridae Infections/virology , Circovirus/genetics , Circovirus/immunology , Columbidae/immunology , Female , Gene Expression , Immunization , Mice , Mice, Inbred BALB C , Viral Vaccines/administration & dosage , Viral Vaccines/genetics , Viral Vaccines/immunology
12.
Comp Med ; 69(3): 179-183, 2019 05 01.
Article in English | MEDLINE | ID: mdl-30885287

ABSTRACT

Pigeons (Columba livia) are used in biomedical research for studies of vision, cognition, neuronal pathways, and spatial orientation. Because there are few commercial laboratory sources, research pigeons are typically acquired from local fancier breeders or bred onsite. For acquired pigeons, the health and vaccine status is often unknown. A juvenile pigeon, born onsite and living in an enclosed outdoor loft, presented with small, bleeding, wart-like lesions on the medial aspects of digits 1 and 4. Topical treatment was initiated. Within a week, 4 fledglings were reported for small, dark papular lesions on the face, head, neck, and beak, and shortly thereafter, 2 additional juvenile pigeons developed similar lesions. The fledglings were euthanized, and histologic examination revealed numerous intralesional eosinophilic cytoplasmic viral inclusions (Bollinger bodies) confirming a diagnosis of poxvirus infection, likely pigeon pox. Although usually self-limiting, pigeon pox can cause moderate to severe lesions in fledgling and juvenile birds. Vaccination with a modified live poxvirus labeled for chickens was used to create herd immunity to pigeon poxvirus. Since vaccination of our entire flock and implementation of more stringent health protocols, all lesions have resolved, and no new lesions have been noted.


Subject(s)
Avipoxvirus , Bird Diseases/virology , Columbidae/virology , Poxviridae Infections/veterinary , Animals , Animals, Laboratory/virology , Avipoxvirus/immunology , Avipoxvirus/pathogenicity , Bird Diseases/pathology , Bird Diseases/prevention & control , Chickens , Columbidae/immunology , Disease Outbreaks/prevention & control , Disease Outbreaks/veterinary , Poxviridae Infections/pathology , Poxviridae Infections/prevention & control , Skin/pathology , Vaccination/veterinary , Viral Vaccines/administration & dosage
14.
Dev Comp Immunol ; 90: 29-40, 2019 01.
Article in English | MEDLINE | ID: mdl-30170033

ABSTRACT

The regulation of interferon-α signaling pathways is essential to protect the host from infection with a broad range of viruses. However, information regarding antiviral response and the specific molecular mechanism of Columba livia interferon-α (CoIFN-α) has not been reported to date. In this study, we cloned a 723bp complete ORF of CoIFN-α gene. The specific antiviral activity of CoIFN-α in VSV (TCID50 = 10-5.87/100 µL)-infected CEFs reached 5.5 × 105 U/mg. Moreover, our result indicated that the anti-VSV efficient of CoIFN-α might depend on the expression of NF-κB. CoIFN-α also showed high sensitivity to trypsin and relatively stable after acid, alkali or heat treatment. Moreover, CoIFN-α activated STAT/Jak signaling and autophagy to inhibit VSV-induced apoptosis. Although the expression of p53 was further increased, apoptosis was not involved in CoIFN-α against VSV. Notably, although STAT signaling was efficiently activated, knockdown p53 did inhibit the antiviral activity of the CoIFN-α via decreasing the expression of Mx1 but not weakened Jak phosphorylation. Moreover, VSV aggravated the apoptosis and the expression of cleaved Mdm2 in knockdown p53 under preincubated CoIFN-α. Taken together, p53 might as a highly interconnected regulator in IFN-α antiviral response and cleaved Mdm2 might as a dominant-negative regulator by competing with full length Mdm2 for p53 binding in virus infection. Overall, our research not only enriches CoIFN-α antiviral features but also helps explain that p53 enhance the CoIFN-α antiviral response against pigeon viral diseases.


Subject(s)
Avian Proteins/genetics , Bird Diseases/immunology , Columbidae/immunology , Interferon-alpha/genetics , Rhabdoviridae Infections/immunology , Vesicular stomatitis Indiana virus/physiology , Animals , Apoptosis , Autophagy , Avian Proteins/metabolism , Cells, Cultured , Cloning, Molecular , Columbidae/genetics , Gene Knockdown Techniques , Immunity, Innate/genetics , Interferon-alpha/metabolism , NF-kappa B/metabolism , Proto-Oncogene Proteins c-mdm2/genetics , Proto-Oncogene Proteins c-mdm2/metabolism , STAT Transcription Factors/metabolism , Signal Transduction , Tumor Suppressor Protein p53/genetics
15.
Viruses ; 10(11)2018 10 31.
Article in English | MEDLINE | ID: mdl-30384424

ABSTRACT

Pigeon circovirus (PiCV) is the most frequently diagnosed virus in pigeons and is thought to be one of the causative factors of a complex disease called the young pigeon disease syndrome (YPDS). The development of a vaccine against this virus could be a strategy for YPDS control. Since laboratory culture of PiCV is impossible, its recombinant capsid protein (rCP) can be considered as a potential antigen candidate in sub-unit vaccines. The aim of this basic research was to evaluate the immune response of pigeons to PiCV rCP. Sixty six-week-old carrier pigeons were divided into two groups (experimental immunized with PiCV rCP mixed with an adjuvant, and control immunized with an adjuvant only), and immunized twice in a 21-day interval. On the day of immunization and on two, 23, 39, and 46 days post first immunization (dpv), samples of blood, spleen, and bursa of Fabricius were collected from six birds from each group to examine anti-PiCV rCP IgY, anti-PiCV rCP IgY-secreting B cells (SBC), IFN-γ gene expression, and percentage of T CD3⁺, CD4⁺, CD8⁺, and B IgM⁺ lymphocytes. The results indicated a correct immune response to PiCV rCP both in humoral and cell-mediated immunity, which was manifested by seroconversion since 23 dpv, by a significantly higher anti-PiCV rCP IgY-SBC number on two and 23 dpv, and significantly higher IFN-γ gene expression since two dpv. There were no significant differences or trends noted between particular T and B lymphocyte subpopulations. To conclude, PiCV rCP may be deemed immunogenic and could be considered as an antigen candidate in sub-unit vaccines against PiCV infections in pigeons.


Subject(s)
Bird Diseases/immunology , Bird Diseases/virology , Capsid Proteins/immunology , Circoviridae Infections/veterinary , Circovirus/immunology , Columbidae/immunology , Columbidae/virology , Host-Pathogen Interactions/immunology , Animals , Enzyme-Linked Immunosorbent Assay , Enzyme-Linked Immunospot Assay/methods , Flow Cytometry , Immunity , Immunoglobulin M/immunology , Immunoglobulins/immunology , Interferon-gamma/genetics , Recombinant Proteins , T-Lymphocyte Subsets/immunology , T-Lymphocyte Subsets/metabolism
16.
Dev Comp Immunol ; 89: 66-72, 2018 12.
Article in English | MEDLINE | ID: mdl-30096338

ABSTRACT

Previous studies on immunoglobulin light chain (IgL) genes in avian species are limited to Galloanseres, and few studies have investigated IgL genes in Neoaves, which includes most living birds. Based on published genome data, we demonstrate that the pigeon (Columba livia) IgL locus spans approximately 24 kb of DNA and contains twenty Vλ segments located upstream of a single pair of Jλ-Cλ. Among the identified Vλ gene segments, four segments are structurally intact and all four segments are able to recombine with Jλ. Moreover, the four functional Vλ segments are preferentially utilized in VλJλ recombination. Phylogenetic analysis suggests that the presence of the four functional Vλ segments in pigeon was likely generated by gene duplication that occurred after the divergence of pigeon and other birds. Our study provides insight into IgL gene evolution and evolutionary diversity of Ig genes in birds.


Subject(s)
Columbidae/genetics , Columbidae/immunology , Genes, Immunoglobulin Light Chain , Animals , Avian Proteins/genetics , Avian Proteins/immunology , Base Sequence , Evolution, Molecular , Gene Duplication , Gene Expression , Genetic Variation , Genome , Immunoglobulin Light Chains/genetics , Immunoglobulin Light Chains/immunology , Phylogeny , Sequence Homology, Nucleic Acid , V(D)J Recombination
17.
Poult Sci ; 97(11): 4031-4039, 2018 Nov 01.
Article in English | MEDLINE | ID: mdl-29945253

ABSTRACT

Toll-like receptors (TLRs) are pattern recognition receptors that are vital for the recognition of pathogen-associated molecular patterns. TLR5 is responsible for the recognition of bacterial flagellin to induce the NF-κB activation and innate immune responses. In this study, we cloned and identified the TLR5 gene from the King pigeon (Columba livia) designated as PiTLR5. Full-length PiTLR5 cDNA (2583 bp) encoded an 860-amino acid protein containing a signal peptide sequence, 10 leucine-rich repeat domains, a leucine-rich repeat C-terminal domain, a transmembrane domain, and an intracellular Toll-interleukin-1 receptor domain. Pigeon TLR5 mRNA expression was quantified by performing quantitative real-time PCR (qRT-PCR), which showed that PiTLR5 was broadly expressed in all examined tissues, with the highest expression in the liver, peripheral blood mononuclear cells, and spleen. PiTLR5-mediated innate immune responses were measured by determining its effects on NF-κB activation and cytokine expression. The results showed that HEK293T cells transfected with PiTLR5 robustly activated the NF-κB response to flagellin, but not other TLR stimuli, and induced significant upregulation of IL-1ß, IL-8, TNF-α, and IFN-γ, indicating that PiTLR5 is a functional TLR5 homolog. Additionally, following flagellin stimulation of pigeon splenic lymphocytes, the levels of TLR5, NF-κB, IL-6, IL-8, CCL5, and IFN-γ mRNA, assessed using qRT-PCR, were significantly upregulated. Besides, TLR5 knockdown resulted in the significantly downregulated expression of NF-κB and related cytokines/chemokines. Triggering pigeon TLR5 contributes to significant upregulation of inflammatory cytokines and chemokines, suggesting that pigeon TLR5 plays an important role in the innate immune responses.


Subject(s)
Avian Proteins/genetics , Columbidae/genetics , Flagellin/pharmacology , Gene Expression Profiling/veterinary , Gene Expression Regulation/immunology , Immunity, Innate/genetics , Toll-Like Receptor 5/genetics , Amino Acid Sequence , Animals , Avian Proteins/metabolism , Columbidae/immunology , Phylogeny , Sequence Alignment/veterinary , Toll-Like Receptor 5/metabolism
19.
BMC Vet Res ; 14(1): 148, 2018 May 02.
Article in English | MEDLINE | ID: mdl-29716604

ABSTRACT

BACKGROUND: The aim of the study was to evaluate the impact of herbal extracts on selected immunity mechanisms in clinically healthy pigeons and pigeons inoculated with the pigeon paramyxovirus type 1 (PPMV-1). For the first 7 days post-inoculation (dpi), an aqueous solution of Aloe vera or licorice extract was administered daily at 300 or 500 mg/kg body weight (BW). The birds were euthanized at 4, 7 and 14 dpi, and spleen samples were collected during necropsy. Mononuclear cells were isolated from spleen samples and divided into two parts: one part was used to determine the percentage of IgM+ B cells in a flow cytometric analysis, and the other was used to evaluate the expression of genes encoding IFN-γ and surface receptors on CD3+, CD4+ and CD8+ T cells. RESULTS: The expression of the IFN-γ gene increased in all birds inoculated with PPMV-1 and receiving both herbal extracts. The expression of the CD3 gene was lowest at 14 dpi in healthy birds and at 7 dpi in inoculated pigeons. The expression of the CD4 gene was higher in uninoculated pigeons receiving both herbal extracts than in the control group throughout nearly the entire experiment with a peak at 7 dpi. A reverse trend was observed in pigeons inoculated with PPMV-1 and receiving both herbal extracts. In uninoculated birds, increased expression of the CD8 gene was noted in the pigeons receiving a lower dose of the Aloe vera extract and both doses of licorice extracts. No significant differences in the expression of this gene were found between inoculated pigeons receiving both herbal extracts. The percentage of IgM+ B cells did not differ between any of the evaluated groups. CONCLUSIONS: This results indicate that Aloe vera and licorice extracts have immunomodulatory properties and can be used successfully to prevent viral diseases, enhance immunity and as supplementary treatment for viral diseases in pigeons.


Subject(s)
Aloe/chemistry , Bird Diseases/virology , Glycyrrhiza/chemistry , Paramyxoviridae Infections/veterinary , Paramyxoviridae/drug effects , Plant Extracts/therapeutic use , Animals , Bird Diseases/drug therapy , Bird Diseases/immunology , Columbidae/immunology , Columbidae/virology , Flow Cytometry/veterinary , Immunity, Cellular/drug effects , Immunity, Humoral/drug effects , Interferon-gamma/metabolism , Paramyxoviridae Infections/drug therapy , Paramyxoviridae Infections/immunology , Spleen/cytology , Spleen/drug effects , T-Lymphocyte Subsets/drug effects , T-Lymphocyte Subsets/immunology
20.
J Immunol ; 200(11): 3720-3728, 2018 06 01.
Article in English | MEDLINE | ID: mdl-29686053

ABSTRACT

Compared with mammals, the bird Ig genetic system relies on gene conversion to create an Ab repertoire, with inversion of the IgA-encoding gene and very few cases of Ig subclass diversification. Although gene conversion has been studied intensively, class-switch recombination, a mechanism by which the IgH C region is exchanged, has rarely been investigated in birds. In this study, based on the published genome of pigeon (Columba livia) and high-throughput transcriptome sequencing of immune-related tissues, we identified a transcriptionally forward α gene and found that the pigeon IgH gene locus is arranged as µ-α-υ1-υ2. In this article, we show that both DNA deletion and inversion may result from IgA and IgY class switching, and similar junction patterns were observed for both types of class-switch recombination. We also identified two subclasses of υ genes in pigeon, which share low sequence identity. Phylogenetic analysis suggests that divergence of the two pigeon υ genes occurred during the early stage of bird evolution. The data obtained in this study provide new insight into class-switch recombination and Ig gene evolution in birds.


Subject(s)
Columbidae/genetics , Genes, Immunoglobulin/genetics , Transcription, Genetic/genetics , Animals , Columbidae/immunology , DNA/genetics , Evolution, Molecular , Gene Deletion , Genes, Immunoglobulin/immunology , Genome/genetics , Genome/immunology , Immunoglobulin Class Switching/genetics , Immunoglobulin Class Switching/immunology , Phylogeny , Transcription, Genetic/immunology
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