ABSTRACT
Cytotoxic T lymphocytes (CTLs) mediate host defense against viral and intracellular bacterial infections and tumors. However, the magnitude of CTL response and their function needed to confer heterosubtypic immunity against influenza virus infection are unknown. We addressed the role of CD8+ T cells in the absence of any cross-reactive antibody responses to influenza viral proteins using an adenoviral vector expressing a 9mer amino acid sequence recognized by CD8+ T cells. Our results indicate that both CD8+ T cell frequency and function are crucial for heterosubtypic immunity. Low morbidity, lower viral lung titers, low to minimal lung pathology, and better survival upon heterosubtypic virus challenge correlated with the increased frequency of NP-specific CTLs. NP-CD8+ T cells induced by differential infection doses displayed distinct RNA transcriptome profiles and functional properties. CD8+ T cells induced by a high dose of influenza virus secreted significantly higher levels of IFN-γ and exhibited higher levels of cytotoxic function. The mice that received NP-CD8+ T cells from the high-dose virus recipients through adoptive transfer had lower viral titers following viral challenge than those induced by the low dose of virus, suggesting differential cellular programming by antigen dose. Enhanced NP-CD8+ T-cell functions induced by a higher dose of influenza virus strongly correlated with the increased expression of cellular and metabolic genes, indicating a shift to a more glycolytic metabolic phenotype. These findings have implications for developing effective T cell vaccines against infectious diseases and cancer. IMPORTANCE: Cytotoxic T lymphocytes (CTLs) are an important component of the adaptive immune system that clears virus-infected cells or tumor cells. Hence, developing next-generation vaccines that induce or recall CTL responses against cancer and infectious diseases is crucial. However, it is not clear if the frequency, function, or both are essential in conferring protection, as in the case of influenza. In this study, we demonstrate that both CTL frequency and function are crucial for providing heterosubtypic immunity to influenza by utilizing an Ad-viral vector expressing a CD8 epitope only to rule out the role of antibodies, single-cell RNA-seq analysis, as well as adoptive transfer experiments. Our findings have implications for developing T cell vaccines against infectious diseases and cancer.
Subject(s)
CD8-Positive T-Lymphocytes , Orthomyxoviridae Infections , T-Lymphocytes, Cytotoxic , Animals , Orthomyxoviridae Infections/immunology , Orthomyxoviridae Infections/virology , Mice , CD8-Positive T-Lymphocytes/immunology , T-Lymphocytes, Cytotoxic/immunology , Mice, Inbred C57BL , Female , Adoptive Transfer , Interferon-gamma/immunology , Interferon-gamma/metabolism , Nucleocapsid Proteins/immunology , Lung/immunology , Lung/virology , RNA-Binding Proteins/immunology , RNA-Binding Proteins/genetics , Nucleoproteins/immunology , Nucleoproteins/genetics , Viral Core Proteins/immunology , Viral Core Proteins/geneticsABSTRACT
An adult male captive diamondback water snake (Nerodia rhombifer) was found dead after a 1-d history of lethargy and cutaneous ulcers. The snake had eaten 2 sunfish (Mola spp.) 5 d before death. Gross examination revealed white-to-tan nodules in the lung and liver and segmental intestinal impactions with digested fish. Histopathology confirmed disseminated granulomas with numerous intrahistiocytic acid-fast bacteria in the skin, skeletal muscle, lung, liver, and intestines. Mycobacterium marinum and Mycolicibacterium fortuitum were identified by culture of the hepatic granuloma, followed by PCR and rpoB gene sequencing. To our knowledge, this is the first description of M. marinum and M. fortuitum coinfection in this species. Although M. fortuitum has been isolated from reptiles, lesions associated with its presence in tissues have not been described previously. Interestingly, the mineralization within granulomas that we observed in our case is not reported in mycobacterial infection in reptiles, whereas this finding is common in mammals.
Subject(s)
Coinfection , Colubridae , Mycobacterium Infections, Nontuberculous , Mycobacterium marinum , Male , Animals , Mycobacterium Infections, Nontuberculous/veterinary , Mycobacterium Infections, Nontuberculous/microbiology , Coinfection/veterinary , Granuloma/veterinary , Granuloma/microbiology , MammalsABSTRACT
Approximately 27 million ducks are raised commercially in the US each year and Pekin ducks (Anas platyrhynchos domesticus) are the most commonly raised breed. Additionally, an increasing number of Americans are raising small flocks of poultry, which often include ducks, in their own backyards. This creates a need to better understand the imaging anatomy of ducks. However, in avian species, superimposition of organs limits evaluation of the coelomic cavity using the most common imaging modality, radiography. The use of cross-sectional imaging including computed tomography (CT) and magnetic resonance imaging (MRI) has increased in veterinary medicine research and clinical use. An anatomic atlas is essential for guiding research and diagnostic imaging. To date, no CT or MRI atlas exists for Pekin ducks. The purpose of the study is to generate a CT atlas of market-age Pekin ducks as well as CT attenuation values and measurements of coelomic organs with corresponding gross and MRI images. A total of 20 market-age, commercial Pekin ducks were imaged. Five male and five female ducks were imaged with CT; four of these ten ducks, two male and two female, were frozen and sliced in approximately 5 mm transverse sections for gross anatomic reference images. Six additional ducks, three male and three female ducks, were dissected to measure major organs. Four additional ducks were imaged using CT and MRI and necropsied for gross anatomical comparisons and landmarks. The atlas contained here may be used for future research projects and clinically to aid in diagnosis of disease in ducks.
Subject(s)
Ducks , Tomography, X-Ray Computed , Humans , Male , Female , Animals , Tomography, X-Ray Computed/veterinary , Magnetic Resonance Imaging/veterinaryABSTRACT
In collaboration with the American College of Veterinary Pathologists.
Subject(s)
Pathology, Veterinary , Veterinarians , Animals , Humans , United StatesABSTRACT
Neurologic diseases are common in domestic cats, and infectious agents are suspected to be the primary cause in 30-45% of cases. Among infectious etiologies, those of bacterial origin are only sporadically characterized in the literature, with few of these reports correlating gross and histologic findings with confirmatory bacteriologic identification. Here, we describe bacterial meningitis and meningoencephalomyelitis associated with Pasteurella multocida in 3 domestic cats. Purulent exudate expanding the cerebral meninges was grossly evident in 2 of the cases. In all 3 cases, histologic changes included multifocal suppurative-to-necrosuppurative meningitis and/or meningoencephalomyelitis of variable severity. Intralesional colonies of gram-negative, short rod-shaped to coccobacillary bacteria were evident histologically in only 1 case. P. multocida was confirmed by routine bacteriologic culture in all cases. Based on our cases, we hypothesize that the upper respiratory system serves as the main portal of entry for P. multocida, leading to invasion of the central nervous system and possible systemic hematogenous dissemination. A case series of meningoencephalomyelitis associated with P. multocida infection in cats has not been reported previously, to our knowledge. We also review briefly other causes of meningoencephalomyelitis in cats.
Subject(s)
Cat Diseases , Meningitis, Bacterial , Pasteurella Infections , Pasteurella multocida , Animals , Cats , Meningitis, Bacterial/veterinary , Pasteurella Infections/veterinaryABSTRACT
The objective of this pilot study was to examine the histologic effects associated with three known sclerosing agents and their ability to induce fibrosis in the subcutaneous space between the cervicocephalic air sac and skin. In the future, these drugs may prove useful in treating birds experiencing cervicocephalic diverticula rupture. The agents used were 1% polidocanol, absolute ethanol, and doxycycline hyclate. Twelve healthy adult chickens (Gallus gallus domesticus) were used in this study. The chickens were randomly allocated into three groups denoting day of euthanasia (day 4, 7, or 14). On day 0, all agents were injected (0.2 ml) subcutaneously, in a four-point grid fashion, in both the cervical and pectoral region of each bird. After euthanasia, the skin and subcutaneous tissues corresponding to the injection sites were harvested for histologic assessment. Tissue sections were assessed for fibrosis and lymphocytic and histiocytic inflammation. A scoring system was established to rank sclerosing agents by fibrosing and inflammatory ability. In the cervical region of chickens, 1% polidocanol induced the greatest inflammatory changes by day 7. Data suggest that doxycycline hyclate may produce the greatest cutaneous and subcutaneous fibrosis overall among all groups of birds. No adverse reactions were associated with any injection. Sterile saline produced the least amount of inflammation when assessed with the scoring system. Further investigation is needed to determine the safety of injections of larger volume with these chemicals and whether these findings can be extrapolated to birds with disease.
Subject(s)
Air Sacs/pathology , Chickens , Doxycycline/pharmacology , Ethanol/pharmacology , Polidocanol/pharmacology , Animals , Doxycycline/administration & dosage , Drug Therapy, Combination , Ethanol/administration & dosage , Fibrosis/chemically induced , Fibrosis/veterinary , Histiocytes , Inflammation/chemically induced , Inflammation/veterinary , Lymphocytes , Pilot Projects , Polidocanol/administration & dosage , Poultry Diseases/therapy , Rupture/therapy , Rupture/veterinary , Sclerosing Solutions/administration & dosage , Sclerosing Solutions/therapeutic use , Skin/drug effects , Skin/pathologyABSTRACT
Toxoplasma gondii is a zoonotic protozoan pathogen that infects many endothermic vertebrates, including humans; the domestic cat and other felids serve as the definitive host. Macropodids are considered highly susceptible to toxoplasmosis. Here, we describe the clinical, pathologic, and immunohistochemical findings of an outbreak of systemic toxoplasmosis in a mob of 11 red kangaroos (Macropus rufus), with high morbidity (73%) and mortality (100%) rates. Affected animals had either severe and rapidly deteriorating clinical conditions or sudden death, which was correlated with widespread necrotizing lesions in multiple organs and intralesional T. gondii organisms identified via MIC3-specific immunohistochemistry and confirmed by REP529-specific rtPCR. Quantification of parasites demonstrated the highest parasite density in pulmonary parenchyma compared with other tissues. Our study highlights the continued importance of this severe condition in Australian marsupials.
Subject(s)
Disease Outbreaks/veterinary , Macropodidae , Toxoplasma/isolation & purification , Toxoplasmosis, Animal/diagnosis , Acute Disease/epidemiology , Animals , Female , Immunohistochemistry/veterinary , Louisiana/epidemiology , Male , Toxoplasmosis, Animal/epidemiology , Toxoplasmosis, Animal/pathologyABSTRACT
OBJECTIVE: To assess the feasibility of a novel technique involving a vessel and tissue-sealing device (VTSD) for ovariectomy in chickens to evaluate the potential application of the procedure to other avian species. ANIMALS: 20 domestic laying hens (Gallus domesticus), of which 10 were immature (< 4 months old) and 10 were adults (> 18 months old). PROCEDURES: Ovariectomy was performed with a VTSD through a left lateral celiotomy. Birds were allowed to recover for 14 days after the procedure and then were euthanized for necropsy. A board-certified veterinary pathologist performed complete necropsies, with particular attention to identifying any remaining ovarian tissue. RESULTS: All birds survived the procedure. For the mature and juvenile birds, the mean ± SD durations of anesthesia (interval from intubation to extubation) were 67.2 ± 7.6 minutes and 50.5 ± 5.1 minutes, respectively, and mean durations of surgery were 45.3 ± 8.5 minutes and 31.6 ± 5.1 minutes, respectively. Three birds had severe hemorrhage during ovariectomy. At necropsy, ovarian tissue was present grossly in 2 mature birds and histologically in 6 additional birds (2 mature and 4 juvenile birds), indicating incomplete excision in 8 (40%) birds. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that the evaluated VTSD can be used to successfully perform ovariectomies in both juvenile and mature chickens, although the procedure was associated with major hemorrhage and incomplete excision of ovarian tissue in some cases. Use of this VTSD for ovariectomy in birds of other species, particularly birds with reproductive tract disease, should be investigated.
Subject(s)
Chickens , Veterinarians , Animals , Female , Humans , Laparotomy/veterinary , Ovariectomy/veterinaryABSTRACT
Bacterial pneumonia is a significant cause of morbidity, mortality, and health care expenditures. Optimum neutrophil recruitment and their function are critical defense mechanisms against respiratory pathogens. The nucleotide-binding oligomerization domain-like receptor (NLRP) 6 controls gut microbiota and immune response to systemic and enteric infections. However, the importance of NLRP6 in neutrophil homeostasis following lung infection remains elusive. To investigate the role of NLRs in neutrophil homeostasis, we used Nlrp6 gene-deficient (Nlrp6-/-) mice in a model of Klebsiella pneumoniae-induced pneumonia-derived sepsis. We demonstrated that NLRP6 is critical for host survival, bacterial clearance, neutrophil influx, and CXC-chemokine production. Kp-infected Nlrp6-/- mice have reduced numbers of hematopoietic stem cells and granulocyte-monocyte progenitors but increased retention of matured neutrophils in bone marrow. Neutrophil extracellular trap (NET) formation and NET-mediated bacterial killing were also impaired in Nlrp6-/- neutrophils in vitro. Furthermore, recombinant CXCL1 rescued the impaired host defense, granulopoietic response, and NETosis in Kp-infected Nlrp6-/- mice. Using A/J background mice and co-housing experiments, our findings revealed that the susceptible phenotype of Nlrp6-/- mice is not strain-specific and gut microbiota-dependent. Taken together, these data unveil NLRP6 as a central regulator of neutrophil recruitment, generation, and function during bacterial pneumonia followed by sepsis.
Subject(s)
Klebsiella Infections/metabolism , Klebsiella pneumoniae/physiology , Macrophages/immunology , Neutrophils/immunology , Pneumonia, Bacterial/metabolism , Receptors, Cell Surface/metabolism , Sepsis/metabolism , Animals , Chemokine CXCL1/genetics , Chemokine CXCL1/metabolism , Extracellular Traps/metabolism , Homeostasis , Humans , Klebsiella Infections/immunology , Mice , Mice, Inbred C57BL , Mice, Knockout , Neutrophil Activation , Pneumonia, Bacterial/immunology , Receptors, Cell Surface/genetics , Sepsis/immunology , THP-1 CellsABSTRACT
Cystic fibrosis is characterized by dehydration of the airway surface liquid layer with persistent mucus obstruction. Th2 immune responses are often manifested as increased mucous cell density (mucous cell metaplasia) associated with mucus obstruction. IL-33 is a known inducer of Th2 immune responses, but its roles in mucus obstruction and related phenotypes in a cystic fibrosis-like lung disease model (i.e., Scnn1b-Tg-positive [Tg+]) mouse, remain unclear. Accordingly, IL-33 knockout (IL-33KO) Tg+ mice were examined and compared with IL-33 heterozygous (IL-33HET) Tg+ mice. As compared with IL-33HET/Tg+ mice, IL-33KO/Tg+ mice had complete absence of bronchoalveolar lavage fluid eosinophilia, accompanied with significant reduction in bronchoalveolar lavage fluid concentration of IL-5, a cytokine associated with eosinophil differentiation and recruitment, and IL-4, a major Th2 cytokine. As compared with IL-33HET/Tg+ mice, IL-33KO/Tg+ mice had significantly reduced levels of Th2-associated gene signatures (Slc26a4, Clca1, Retnla, and Chi3l4), along with complete loss of intracellular mucopolysaccharide staining in the airway epithelium. As compared with IL-33HET/Tg+ mice, although the IL-33KO/Tg+ mice had significantly reduced levels of MUC5AC protein expression, they showed no reduction in the degree of mucus obstruction, MUC5B protein expression, bacterial burden, and neonatal mortality. Interestingly, the histological features, including subepithelial airway inflammation and alveolar space enlargement, were somewhat exaggerated in IL-33KO/Tg+ mice compared with IL-33HET/Tg+ mice. Taken together, our data indicate that although IL-33 modulates Th2 inflammatory responses and MUC5AC protein production, mucus obstruction is not dependent on IL-33.
Subject(s)
Cystic Fibrosis/immunology , Interleukin-33/metabolism , Lung/pathology , Mucin 5AC/metabolism , Th2 Cells/immunology , Animals , Bronchoalveolar Lavage Fluid/cytology , Bronchoalveolar Lavage Fluid/immunology , Cystic Fibrosis/diagnosis , Cystic Fibrosis/genetics , Cystic Fibrosis/pathology , Disease Models, Animal , Eosinophils/immunology , Eosinophils/metabolism , Epithelial Sodium Channels/genetics , Humans , Interleukin-33/genetics , Lung/cytology , Lung/immunology , Mice , Mice, Knockout , Mice, Transgenic , Mucus/immunology , Mucus/metabolism , Respiratory Mucosa/cytology , Respiratory Mucosa/immunology , Respiratory Mucosa/metabolism , Th2 Cells/metabolismABSTRACT
BACKGROUND: Liver-type fatty acid-binding protein (L-FABP) is a biomarker for early detection of renal disease in humans. Liver-type fatty acid-binding protein is cytotoxic oxidation products secreted from proximal tubules under ischemia and oxidative stress. OBJECTIVE: To examine renal expression and quantify urinary excretion of L-FABP in catswith renal disease. ANIMALS: One hundred and thirty-four client-owned cats including 34 cats with serum creatinine (sCre) values >1.6 mg/dL and 10 other cats that died in clinics. METHODS: Tissue expressions of L-FABP were examined by reverse transcription polymerase chain reaction and Western blotting. Urinary L-FABP (uL-FABP) and serum L-FABP (sL-FABP) levels were determined by enzyme-linked immunosorbent assay. Anti-liver-type fatty acid-binding protein antibody immunostained renal sections. RESULTS: Feline kidneys express L-FABP. Strong L-FABP signals were observed in the lumens of proximal tubular cells in 5 cats with high uL-FABP excretion, but not in 5 cats with low uL-FABP excretion. In 9 normal cats, uL-FABP index was <1.2 µg/g urinary creatinine (uCre). High uL-FABP indexes (>10.0 µg/g uCre) were detected in 7 of 100 cats with low sCre (<1.6 mg/dL) and 18 of 44 cats with high sCre (>1.6 mg/dL). There was a weak correlation between L-FABP index and sCre, serum symmetric dimethylarginine (SDMA), or blood urea nitrogen (BUN), and these correlation coefficients were increased by analyzing only data of cats with sCre >1.6 mg/dL. There was a weak correlation between u L-FABP index and sL-FABP in all tested cats, but not in cats with high sCre. CONCLUSIONS AND CLINICAL IMPORTANCE: This study demonstrates correlations between L-FABP and current renal biomarkers for chronic kidney disease in cats, such as sCre and SDMA. Liver-type fatty acid-binding protein may be a potential biomarker to predict early pathophysiological events in feline kidneys.
Subject(s)
Cat Diseases/urine , Fatty Acid-Binding Proteins/urine , Kidney Diseases/veterinary , Animals , Biomarkers/urine , Cat Diseases/blood , Cats , Female , Kidney Diseases/urine , Male , Urinalysis/veterinaryABSTRACT
SCOPE: Rheumatoid arthritis (RA) is an autoimmune disorder related to the inflammation of cartilage due to the infiltration of inflammatory cells. Sargassum serratifolium, a brown alga, possesses strong anti-inflammatory activities. METHODS AND RESULTS: The effect of meroterpenoid-rich fraction from the ethanol extract of S. serratifolium (MES) on RA and its underlying mechanisms on the inhibition of RA using a collagen-induced arthritis (CIA) mouse model are examined. The results show that MES ameliorates paw swelling and reduces the arthritis score. MES considerably decreases the secretion of pro-inflammatory cytokines in the serum and joint tissue of mice. Histopathological analysis demonstrates that MES strongly inhibited bone damage and inflammatory cell intrusion in the joint tissue. The expression of inflammatory enzymes and adhesion molecules is significantly inhibited in the serum and joint tissue of MES-fed mice. In addition, MES downregulates the nuclear factor κB (NF-κB) signaling pathway by suppressing the phosphorylation of protein kinase B, c-Jun N-terminal kinase, and p38 mitogen-activated protein kinases. CONCLUSIONS: MES supplementation remarkably reduces inflammatory response in CIA mouse model. These results indicate that MES can be used as a pharmaceutical agent against RA.
Subject(s)
Arthritis, Rheumatoid/drug therapy , Sargassum/chemistry , Terpenes/pharmacology , Alkenes/pharmacology , Animals , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/pharmacology , Arthritis, Experimental/drug therapy , Arthritis, Experimental/metabolism , Arthritis, Experimental/pathology , Arthritis, Rheumatoid/chemically induced , Arthritis, Rheumatoid/metabolism , Arthritis, Rheumatoid/pathology , Benzopyrans/pharmacology , Benzoquinones/pharmacology , Chondrocytes/drug effects , Chondrocytes/metabolism , Chondrocytes/pathology , Cyclooxygenase 2/metabolism , Cytokines/blood , Cytokines/metabolism , Ethanol/chemistry , Interleukin-6/metabolism , Joints/drug effects , Joints/pathology , Male , Mice, Inbred DBA , NF-kappa B/metabolism , Terpenes/chemistryABSTRACT
A 12 yr old spayed female domestic shorthair with a history of lethargy, anorexia, and a pendulous abdomen was referred after a cranial abdominal mass was palpated on physical examination. Thoracic radiographs and an abdominal ultrasound revealed a mass associated with the kidney and moderate hemoperitoneum. Exploratory laparotomy revealed abdominal hemorrhage originating from a right renal mass that was adhered to the caudal vena cava. Following a right nephrectomy, histopathology diagnosed the mass as a perirenal/renal myxosarcoma. Based upon thoracic radiographs and abdominal ultrasound, the patient remains disease free at 14 mo postoperatively.
Subject(s)
Cat Diseases/pathology , Kidney Neoplasms/veterinary , Myxosarcoma/veterinary , Animals , Cat Diseases/surgery , Cats , Female , Kidney Neoplasms/pathology , Kidney Neoplasms/surgery , Myxosarcoma/pathology , Myxosarcoma/surgeryABSTRACT
Melanoma is an immunogenic tumor that can metastasize quickly to proximal and distal sites, thus complicating the application of therapeutic modalities. Numerous mouse model systems have been used to gain understanding of the immunobiology and metastatic potential of melanoma. Here, we report the optimization of a syngeneic mouse melanoma model protocol using the mouse B16-derived melanoma cell line B16F10 that ensures the production of tumors on mice pinnae that are similar in size between animals and that enlarge in a time-dependent manner. In this model, B16F10 cells are first allowed to develop tumors after injection in the interscapular area or flank of C57BL/6J mice. Subsequently, the tumors are harvested, cells dissociated and injected into mouse pinnae. Dose-dependent studies revealed that injection of 2 × 105 cells allowed for slow tumor enlargement, producing tumors averaging 100 mm³ within 2 to 3 wk with a metastatic frequency of 100%. This experimental protocol will be useful in dissecting the immunobiology of melanoma tumor development and metastasis and the evaluation of immunotherapeutic antimelanoma therapies.