RESUMO
Caseous lymphadenitis (CL) in sheep is a chronic contagious disease caused by Corynebacterium pseudotuberculosis, commonly characterized by abscess formation in peripheral lymph nodes and disseminated infections. Nonetheless, other microorganisms, including with zoonotic relevance, can be isolated from CL-resembling lymph nodes. Currently, mycobacteria have been reported in visceral granulomatous lesions in small ruminants, a fact that poses a public health issue, particularly in slaughtered sheep intended for human consumption. Cytology using fine needle aspiration and microbiological culturing are suitable tests for routine diagnostic, whereas present drawbacks and molecular methods have been confirmatory. Data about the occurrence of mycobacteria in both lymph nodes with aspect of CL and apparently healthy visceral nodes of sheep slaughtered for human consumption are scarce. In this study, 197 visceral lymph nodes of sheep showed lymphadenitis and 202 healthy visceral lymph nodes of slaughtered sheep intended for human consumption were submitted to conventional bacteriological diagnosis, mycobacteria culturing, and cytological evaluation. Compatible Corynebacterium isolates were subjected to multiplex PCR targeting 16S rRNA, rpoB, and pld genes to detect C. pseudotuberculosis. Based on microbiological identification, C. pseudotuberculosis (86/197; 43.7%), streptococci γ-hemolytic (17/197; 8.6%), and Trueperella pyogenes (12/197; 6.1%) were prevalent in lymph nodes with abscesses, as opposed to staphylococci (53/202; 26.2%) in apparently healthy lymph nodes. No mycobacteria were isolated. Cytology identified 49.2% (97/197) Gram-positive pleomorphic organisms (coryneform aspect). Multiplex PCR confirmed genetic material of C. pseudotuberculosis in 74.4% (64/86) of the samples with C. pseudotuberculosis isolation and 66% (64/97) samples with cytological coryneform aspect (κ = 86.78%; 95% CI = 79.87-93.68%). These findings emphasize the prevalence of C. pseudotuberculosis in abscess formation among peripheral lymph nodes of sheep. Other bacteria were also identified in lymph nodes sampled that resembling C. pseudotuberculosis-induced infections that may difficult the diagnosis. Multiplex PCR revealed a valuable assay to detect C. pseudotuberculosis, in addition to routine methods applied to CL-diagnosis. No mycobacteria were identified in lymph nodes sampled, with and without apparent lesions. Nonetheless, due to public health impacts, this pathogen should be considered as a differential diagnosis of C. pseudotuberculosis-induced infections during inspection procedures of slaughtered sheep intended for human consumption.
Assuntos
Bactérias/genética , Coinfecção/veterinária , Corynebacterium pseudotuberculosis/genética , Linfonodos/citologia , Linfonodos/microbiologia , Linfadenite/microbiologia , Linfadenite/veterinária , Mycobacterium/genética , Matadouros , Animais , Bactérias/classificação , Brasil/epidemiologia , Coinfecção/microbiologia , Estudos Transversais , Fazendas , Feminino , Masculino , Prevalência , RNA Ribossômico 16S/genética , Distribuição Aleatória , Ovinos/microbiologia , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/microbiologiaRESUMO
An association between increased susceptibility to infectious diseases and obesity has been described as a result of impaired immunity in obese individuals. It is not clear whether a similar linkage can be drawn between obesity and parasitic diseases. To evaluate the effect of obesity in the immune response to cutaneous Leishmania major infection, we studied the ability of C57BL/6 mice fed a hypercaloric diet (HSB) to control leishmaniasis. Mice with diet-induced obesity presented thicker lesions with higher parasite burden and a more intense inflammatory infiltrate in the infected ear after infection with L. major. There was no difference between control and obese mice in IFN-gamma or IL-4 production by auricular draining lymph node cells, but obese mice produced higher levels of IgG1 and IL-17. Peritoneal macrophages from obese mice were less efficient to kill L. major when infected in vitro than macrophages from control mice. In vitro stimulation of macrophages with IL-17 decreased their capacity to kill the parasite. Moreover, macrophages from obese mice presented higher arginase activity. To confirm the role of IL-17 in the context of obesity and infection, we studied lesion development in obese IL-17R-/- mice infected with L. major and found no difference in skin lesions and the leukocyte accumulation in the draining lymph node is redcuced in knockout mice compared between obese and lean animals. Our results indicate that diet-induced obesity impairs resistance to L. major in C57BL/6 mice and that IL-17 is involved in lesion development.
Assuntos
Leishmania major/patogenicidade , Leishmaniose Cutânea/imunologia , Obesidade , Animais , Dieta/efeitos adversos , Orelha/parasitologia , Feminino , Interferon gama , Interleucina-17 , Leishmaniose Cutânea/parasitologia , Linfonodos/citologia , Macrófagos Peritoneais/parasitologia , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fatores de RiscoRESUMO
It is well known that neutrophils are rapidly recruited to a site of injury or infection and perform a critical role in pathogen clearance and inflammation. However, they are also able to interact with and regulate innate and adaptive immune cells and some stimuli induce the migration of neutrophils to lymph nodes (LNs). Previously, we demonstrated that the immune complex (IC) generated by injecting OVA into the footpad of OVA/CFA immunized mice induced the migration of OVA+ neutrophils to draining LNs (dLNs). Here we investigate the effects of these neutrophils which reach dLNs on CD4+ T cell response. Our findings here strongly support a dual role for neutrophils in dLNs regarding CD4+ T cell response modulation. On the one hand, the CD4+ T cell population expands after the influx of OVA+ neutrophils to dLNs. These CD4+ T cells enlarge their proliferative response, activation markers and IL-17 and IFN-γ cytokine production. On the other hand, these neutrophils also restrict CD4+ T cell expansion. The neutrophils in the dLNs upregulate PD-L1 molecules and are capable of suppressing CD4+ T cell proliferation. These results indicate that neutrophils migration to dLNs have an important role in the homeostasis of adaptive immunity. This report describes for the first time that the influx of neutrophils to dLNs dependent on IC presence improves CD4+ T cell response, at the same time controlling CD4+ T cell proliferation through a PD-L1 dependent mechanism.
Assuntos
Antígeno B7-H1/metabolismo , Linfócitos T CD4-Positivos/imunologia , Movimento Celular/imunologia , Linfonodos/citologia , Neutrófilos/imunologia , Imunidade Adaptativa/imunologia , Transferência Adotiva , Animais , Complexo Antígeno-Anticorpo/efeitos dos fármacos , Complexo Antígeno-Anticorpo/imunologia , Antígeno B7-H1/genética , Proliferação de Células/efeitos dos fármacos , Técnicas de Inativação de Genes , Interferon gama/análise , Interleucina-17/análise , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Ovalbumina/farmacologiaRESUMO
Conventional dendritic cells (cDCs) are divided into the following different subtypes: cDC1, which promotes a Th1 response, and cDC2, which stimulates a Th2 and Th17 response. These cells have not been characterized in porcine lymphoid tissues. DEC205 is a receptor that increases antigen presentation and allows DCs to cross-present antigens. The objectives of this work were to characterize cDCs subsets in the tonsil, submaxillary and mesenteric lymph nodes and spleen lymphoid tissues and to determine their expression of DEC205 by flow cytometry. The cDC1 (MHCIIhighCADM1highCD172a-/low) and cDC2 (MHCIIhighCADM1highCD172a+) phenotypes were confirmed by the expression of characteristic cDC1 and cDC2 transcripts (FLT3, XCR1 and FCER1α). Among all lymphoid tissues, the spleen had the highest frequency of total cDCs. The cDC1:cDC2 ratio showed that all lymph tissues had higher levels of cDC1 than levels of cDC2. DEC205+ cDCs were found in all analyzed tissues, albeit with different frequencies. Our research will facilitate the study on the function of these cells and the investigation of the strategies for DEC205 targeting and functional studies.
Assuntos
Células Dendríticas/citologia , Linfonodos/citologia , Tonsila Palatina/citologia , Baço/citologia , Suínos/imunologia , Animais , Células Dendríticas/imunologia , Linfonodos/imunologia , Tonsila Palatina/imunologia , Baço/imunologiaRESUMO
O linfoma é uma forma de apresentação do distúrbio linfoproliferativo, em que o tumor se origina em um órgão hematopoiético sólido, como o linfonodo. É considerada uma neoplasia de maior incidência em cães e gatos. O trabalho tem como objetivo apresentar um relato de caso sobre um linfoma multicêntrico, com abordagem clínica e laboratorial. Um cão sem raça definida, foi atendido no Hospital Veterinário da Universidade Estadual do CEARÁ (UHV-UECE), apresentando hematúria e uma discreta linfadenomegalia no exame físico. Foram solicitados hemograma completo (HC), análise de bioquímica sérica, sumário de urina e exame ultrassonográfico (US). As alterações no hemograma foram anemia normocítica normocrômica e trombocitopenia. A análise dos exames de bioquímica sérica revelou um aumento na enzima aspartato amino-transferase (AST). No sumário de urina (SU), observaram-se alterações das características físicas e da sedimentoscopia. O exame ultrassonográfico mostrou alterações de ecogenicidade na bexiga, próstata, testículo e baço. Houve piora do quadro clínico do animal, com perda de peso e aumento evidente dos linfonodos. Um exame citológico foi realizado com aspirado de linfonodos, que apresentou configuração celular compatível com linfoma; sendo também encontradas células neoplásicas no esfregaço de sangue periférico, no hemograma. Diante dos achados e do agravamento do quadro clínico, o animal veio a óbito.
Lymphoma is a form of presentation of the lymphoproliferative disorder which the tumor originates from a solid hematopoietic organ such as the lymph node. It is considered one of the neoplasia of higher incidence in dogs and cats. This paper aims to present a case report on a multicenter lymphoma with a clinical and laboratory approach. A dog without defined breed, at the Hospital Veterinário da Universidade Estadual do CEARÁ (UHV-UECE) presenting hematuria and a discrete lymphadenomegaly on physical examination. Complete blood counts (HC), serum biochemical analyzes, urine summary and ultrasound examination were requested. Changes in the hemogram were normocyclic normocytic anemia and thrombocytopenia. Biochemical analysis revealed an increase in asparate amino transferase (AST). The urine summary (SU) showed changes in physical characteristics and sedimentation. Ultrasound examination (US) showed changes in the bladder, prostate, testis echogenicity and the spleens size. There was worsening of the animal's size over time with progression of weight loss and evident enlargement of the lymph nodes. A cytology was performed through a lymph node aspirate which had a lymphoma compatible cell configuration and neoplastic cells were also found in the hemogram peripheral blood smear. The animal died with deterioration of the clinical picture.
Assuntos
Animais , Cães , Linfoma/diagnóstico por imagem , Linfoma/veterinária , Linfonodos/citologia , Contagem de Células Sanguíneas/veterinária , Técnicas Citológicas/veterinária , Ultrassonografia/veterináriaRESUMO
O linfoma é uma forma de apresentação do distúrbio linfoproliferativo, em que o tumor se origina em um órgão hematopoiético sólido, como o linfonodo. É considerada uma neoplasia de maior incidência em cães e gatos. O trabalho tem como objetivo apresentar um relato de caso sobre um linfoma multicêntrico, com abordagem clínica e laboratorial. Um cão sem raça definida, foi atendido no Hospital Veterinário da Universidade Estadual do CEARÁ (UHV-UECE), apresentando hematúria e uma discreta linfadenomegalia no exame físico. Foram solicitados hemograma completo (HC), análise de bioquímica sérica, sumário de urina e exame ultrassonográfico (US). As alterações no hemograma foram anemia normocítica normocrômica e trombocitopenia. A análise dos exames de bioquímica sérica revelou um aumento na enzima aspartato amino-transferase (AST). No sumário de urina (SU), observaram-se alterações das características físicas e da sedimentoscopia. O exame ultrassonográfico mostrou alterações de ecogenicidade na bexiga, próstata, testículo e baço. Houve piora do quadro clínico do animal, com perda de peso e aumento evidente dos linfonodos. Um exame citológico foi realizado com aspirado de linfonodos, que apresentou configuração celular compatível com linfoma; sendo também encontradas células neoplásicas no esfregaço de sangue periférico, no hemograma. Diante dos achados e do agravamento do quadro clínico, o animal veio a óbito.(AU)
Lymphoma is a form of presentation of the lymphoproliferative disorder which the tumor originates from a solid hematopoietic organ such as the lymph node. It is considered one of the neoplasia of higher incidence in dogs and cats. This paper aims to present a case report on a multicenter lymphoma with a clinical and laboratory approach. A dog without defined breed, at the Hospital Veterinário da Universidade Estadual do CEARÁ (UHV-UECE) presenting hematuria and a discrete lymphadenomegaly on physical examination. Complete blood counts (HC), serum biochemical analyzes, urine summary and ultrasound examination were requested. Changes in the hemogram were normocyclic normocytic anemia and thrombocytopenia. Biochemical analysis revealed an increase in asparate amino transferase (AST). The urine summary (SU) showed changes in physical characteristics and sedimentation. Ultrasound examination (US) showed changes in the bladder, prostate, testis echogenicity and the spleens size. There was worsening of the animal's size over time with progression of weight loss and evident enlargement of the lymph nodes. A cytology was performed through a lymph node aspirate which had a lymphoma compatible cell configuration and neoplastic cells were also found in the hemogram peripheral blood smear. The animal died with deterioration of the clinical picture.(AU)
Assuntos
Animais , Cães , Linfoma/veterinária , Linfoma/diagnóstico por imagem , Linfonodos/citologia , Técnicas Citológicas/veterinária , Contagem de Células Sanguíneas/veterinária , Ultrassonografia/veterináriaRESUMO
Lymphoma is a malignant tumor characterized by cell proliferation of lymphoid origin and corresponds to 90% of all hematopoietic neoplasms of dogs. Regulatory T cells (Tregs) have been the target of many investigations in oncology due to their potential of down-regulating immune responses, as well as ensuring the maintenance of active mechanisms of tumor suppression. The aims of the present study were to compare the percentage of Tregs in peripheral blood between dogs with multicentric lymphoma and healthy animals, together with the percentage of Tregs in peripheral blood and lymph nodes of dogs with multicentric lymphoma. Twenty-six animals were enrolled in the study: 10 healthy dogs comprised the control group (CG) and 16 dogs with multicentric lymphoma comprised the Lymphoma Group (LG). We observed that dogs in the LG showed a significantly higher Tregs expression in peripheral blood compared to the CG. No significant difference was observed between Tregs expression in lymph nodes and peripheral blood of the LG, however. With these results, it is possible to conclude that multicentric lymphoma is a neoplasm with high Tregs expression, which poses this as a condition of interest when investigating treatments that can suppress Regulatory T cells.(AU)
O linfoma é uma neoplasia maligna caracterizada pela proliferação neoplásica de células originadas de tecido linfoide e corresponde a cerca de 90% das neoplasias hematopoiéticas em cães. Células T reguladoras (Tregs) têm sido alvo de diversas investigações na área da oncologia devido ao potencial de regulação negativa da resposta do sistema imune e à manutenção ativa do mecanismo de imunossupressão tumoral. O objetivo do presente estudo foi a comparação da porcentagem de Tregs no sangue periférico entre cães com linfoma multicêntrico e animais saudáveis e a porcentagem de Tregs no sangue periférico e nos linfonodos de cães com linfoma multicêntrico. Foram utilizados 26 animais: 10 cães saudáveis, como grupo controle (CG), e 16 cães com linfoma multicêntrico, como grupo linfoma (LG). Observou-se maior expressão de Tregs no sangue periférico de cães do LG em comparação ao CG. Entretanto, não foi observada diferença significativa entre as expressões de Treg nos linfonodos e no sangue periférico do LG. Com esses resultados, foi possível concluir que o linfoma multicêntrico apresenta alta expressão de Tregs, tornando-se condição interessante para o estudo de tratamentos capazes de suprimir as células T reguladoras.(AU)
Assuntos
Animais , Cães , Sangue , Linfonodos/citologia , Linfoma/veterinária , Linfócitos T Reguladores , Fatores de TranscriçãoRESUMO
Lymphoma is a malignant tumor characterized by cell proliferation of lymphoid origin and corresponds to 90% of all hematopoietic neoplasms of dogs. Regulatory T cells (Tregs) have been the target of many investigations in oncology due to their potential of down-regulating immune responses, as well as ensuring the maintenance of active mechanisms of tumor suppression. The aims of the present study were to compare the percentage of Tregs in peripheral blood between dogs with multicentric lymphoma and healthy animals, together with the percentage of Tregs in peripheral blood and lymph nodes of dogs with multicentric lymphoma. Twenty-six animals were enrolled in the study: 10 healthy dogs comprised the control group (CG) and 16 dogs with multicentric lymphoma comprised the Lymphoma Group (LG). We observed that dogs in the LG showed a significantly higher Tregs expression in peripheral blood compared to the CG. No significant difference was observed between Tregs expression in lymph nodes and peripheral blood of the LG, however. With these results, it is possible to conclude that multicentric lymphoma is a neoplasm with high Tregs expression, which poses this as a condition of interest when investigating treatments that can suppress Regulatory T cells.(AU)
O linfoma é uma neoplasia maligna caracterizada pela proliferação neoplásica de células originadas de tecido linfoide e corresponde a cerca de 90% das neoplasias hematopoiéticas em cães. Células T reguladoras (Tregs) têm sido alvo de diversas investigações na área da oncologia devido ao potencial de regulação negativa da resposta do sistema imune e à manutenção ativa do mecanismo de imunossupressão tumoral. O objetivo do presente estudo foi a comparação da porcentagem de Tregs no sangue periférico entre cães com linfoma multicêntrico e animais saudáveis e a porcentagem de Tregs no sangue periférico e nos linfonodos de cães com linfoma multicêntrico. Foram utilizados 26 animais: 10 cães saudáveis, como grupo controle (CG), e 16 cães com linfoma multicêntrico, como grupo linfoma (LG). Observou-se maior expressão de Tregs no sangue periférico de cães do LG em comparação ao CG. Entretanto, não foi observada diferença significativa entre as expressões de Treg nos linfonodos e no sangue periférico do LG. Com esses resultados, foi possível concluir que o linfoma multicêntrico apresenta alta expressão de Tregs, tornando-se condição interessante para o estudo de tratamentos capazes de suprimir as células T reguladoras.(AU)
Assuntos
Animais , Cães , Sangue , Linfonodos/citologia , Linfoma/veterinária , Linfócitos T Reguladores , Fatores de TranscriçãoRESUMO
Lamina propria dendritic cells (DCs) have a permanent turnover with constitutive migration to mesenteric lymph nodes and replenishment by progenitors. Luminal bacteria and dietary constituents provide key signals that endow DCs their unique properties in vivo. Taking into account that the intestinal immune system is greatly influenced by retinoids, we evaluated in B6 mice 3, 8, 16 and 24 h after feeding a single dose of vitamin A phenotype and function of cells present in mesenteric afferent lymph nodes as well as signals involved in migration. We studied the frequency of CD11c+MHC-II+CD103+CD86+ and RALDH+ DCs by flow cytometry, we determined CCL-21 and D6 levels in tissue homogenates by Western blot, and we co-cultured cells isolated from afferent lymphatics with sorted CD4+ lymphocytes to assess Foxp-3 induction and homing receptor expression. Sixteen hours after vitamin A administration, DCs isolated from afferent lymphatics were able to induce homing receptors and Foxp3 expression in CD4+ lymphocytes. Our results show that a single dose of vitamin A generated a stream of signals and amplified the tolerogenic activity of DCs migrating to lymphoid tissue.
Assuntos
Linfócitos T CD4-Positivos/metabolismo , Células Dendríticas/metabolismo , Suplementos Nutricionais , Fatores de Transcrição Forkhead/agonistas , Regulação da Expressão Gênica , Receptores de Retorno de Linfócitos/agonistas , Vitamina A/administração & dosagem , Animais , Antígenos CD/metabolismo , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD4-Positivos/imunologia , Movimento Celular , Células Cultivadas , Técnicas de Cocultura , Células Dendríticas/citologia , Células Dendríticas/imunologia , Feminino , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/metabolismo , Tolerância Imunológica , Linfa/citologia , Linfa/imunologia , Linfa/metabolismo , Linfonodos/citologia , Linfonodos/imunologia , Linfonodos/metabolismo , Mesentério , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Receptores de Retorno de Linfócitos/genética , Receptores de Retorno de Linfócitos/metabolismo , Organismos Livres de Patógenos EspecíficosRESUMO
The gastrointestinal immune system plays a pivotal role in the host relationship with food antigens, the homeostatic microbiome and enteric pathogens. Here, we describe how to collect and process liver and intestinal samples to efficiently isolate and analyse resident immune cells. Furthermore, we describe a step-by-step methodology showing how to high-dimensionally immunophenotype resident leucocytes using cytometry by time-of-flight, providing a well-characterized antibody platform that allows the identification of every leucocyte subset simultaneously. This protocol also includes instructions to purify and cultivate primary murine hepatocytes, a powerful tool to assess basic cell biology and toxicology assays. Gut and liver samples from the same mouse can be collected, processed and stained in less than 6 hr. This protocol enables the recovery of several populations of purified and viable immune cells from solid and fibrous organs, preventing unwanted loss of adherent cells during isolation.