Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 20
Filtrar
1.
Anim. Reprod. (Online) ; 18(2): e20210009, 2021. tab, graf
Artigo em Inglês | LILACS-Express | VETINDEX | ID: biblio-1285137

Resumo

Abstract Toll-like receptors (TLRs) are involved to the maternal immune tolerance. The spleen is essential for adaptive immune reactions. However, it is unclear that early pregnancy regulates TLR-mediated signalings in the maternal spleen. The purpose of this study was to investigate the effects of early pregnancy on expression of TLR signaling members in the ovine spleen. Ovine spleens were collected at day 16 of the estrous cycle, and at days 13, 16 and 25 of pregnancy (n = 6 for each group). Real-time quantitative PCR, western blot and immunohistochemistry analysis were used to detect TLR signaling members, including TLR2, TLR3, TLR4, TLR5, TLR7, TLR9, myeloid differentiation primary-response protein 88 (MyD88), tumor necrosis factor receptor associated factor 6 (TRAF6) and interleukin-1-receptor-associated kinase 1 (IRAK1). The results showed that expression levels of TLR2, TLR4 and IRAK1 were downregulated, but expression levels of TLR3, TLR5, TLR7, TLR9, TRAF6 and MyD88 were increased during early pregnancy. In addition, MyD88 protein was located in the capsule, trabeculae and splenic cords of the maternal spleen. This paper reports for the first time that early pregnancy has effects on TLR signaling pathways in the ovine spleen, which is beneficial for understanding the maternal immune tolerance during early pregnancy.

2.
Anim. Reprod. ; 18(2): e20210009, 2021. graf, tab, ilus
Artigo em Inglês | VETINDEX | ID: vti-31908

Resumo

Toll-like receptors (TLRs) are involved to the maternal immune tolerance. The spleen is essential for adaptive immune reactions. However, it is unclear that early pregnancy regulates TLR-mediated signalings in the maternal spleen. The purpose of this study was to investigate the effects of early pregnancy on expression of TLR signaling members in the ovine spleen. Ovine spleens were collected at day 16 of the estrous cycle, and at days 13, 16 and 25 of pregnancy (n = 6 for each group). Real-time quantitative PCR, western blot and immunohistochemistry analysis were used to detect TLR signaling members, including TLR2, TLR3, TLR4, TLR5, TLR7, TLR9, myeloid differentiation primary-response protein 88 (MyD88), tumor necrosis factor receptor associated factor 6 (TRAF6) and interleukin-1-receptor-associated kinase 1 (IRAK1). The results showed that expression levels of TLR2, TLR4 and IRAK1 were downregulated, but expression levels of TLR3, TLR5, TLR7, TLR9, TRAF6 and MyD88 were increased during early pregnancy. In addition, MyD88 protein was located in the capsule, trabeculae and splenic cords of the maternal spleen. This paper reports for the first time that early pregnancy has effects on TLR signaling pathways in the ovine spleen, which is beneficial for understanding the maternal immune tolerance during early pregnancy.(AU)


Assuntos
Animais , Feminino , Gravidez , Ovinos/embriologia , Prenhez , Receptores Toll-Like , Baço
3.
Anim. Reprod. ; 18(1): e20200048, fev. 2021. ilus
Artigo em Inglês | VETINDEX | ID: vti-31083

Resumo

This review focuses on the innate immune events modulated by conceptus signaling during early pregnancy in ruminants. Interferon-tau (IFN-) plays a role in the recognition of pregnancy in ruminants, which involves more than the inhibition of luteolytic pulses of PGF2 to maintain corpus luteum function. For successful pregnancy establishment, the allogenic conceptus needs to prevent rejection by the female. Therefore, IFN- exerts paracrine and endocrine actions to regulate the innate immune system and prevent conceptus rejection. Additionally, other immune regulators work in parallel with IFN-, such as the pattern recognition receptors (PRR). These receptors are activated during viral and bacterial infections and in early pregnancy, but it remains unknown whether PPR expression and function are controlled by IFN-. Therefore, this review focuses on the main components of the innate immune response that are involved with early pregnancy and their importance to avoid conceptus rejection.(AU)


Assuntos
Animais , Feminino , Gravidez , Ruminantes/embriologia , Ruminantes/imunologia , Prenhez , Luteólise , Sistema Imunitário , Interleucinas , Dinoprosta
4.
Anim. Reprod. (Online) ; 18(1): e20200048, 2021. graf
Artigo em Inglês | LILACS-Express | VETINDEX | ID: biblio-1285122

Resumo

Abstract This review focuses on the innate immune events modulated by conceptus signaling during early pregnancy in ruminants. Interferon-tau (IFN-τ) plays a role in the recognition of pregnancy in ruminants, which involves more than the inhibition of luteolytic pulses of PGF2α to maintain corpus luteum function. For successful pregnancy establishment, the allogenic conceptus needs to prevent rejection by the female. Therefore, IFN-τ exerts paracrine and endocrine actions to regulate the innate immune system and prevent conceptus rejection. Additionally, other immune regulators work in parallel with IFN-τ, such as the pattern recognition receptors (PRR). These receptors are activated during viral and bacterial infections and in early pregnancy, but it remains unknown whether PPR expression and function are controlled by IFN-τ. Therefore, this review focuses on the main components of the innate immune response that are involved with early pregnancy and their importance to avoid conceptus rejection.

5.
Anim. Reprod. (Online) ; 14(supl. 1): 1271-1277, 2017. ilus, graf, tab
Artigo em Inglês | VETINDEX | ID: biblio-1461323

Resumo

The aim of this study was to elucidate the presence of components of the innate immune system inthe bovine corpus luteum (CL) by detecting the expression and cell-specific localization of TLR2 and TLR4 during different stages of the estrous cycle in a control study design. Bovine CL samples were collected from a local slaughterhouse and assigned to three groups as follows: developing CL (dCL; n = 6, approx. days 3–6), mature CL (mCL; n = 5, approx. days 8–12), and regressing CL (rCL; n = 5, approx. days 17–19). An upregulation of TLR2 mRNA was detected only in rCL(P < 0.05). Localization of the TLR2 protein was particularly apparent in luteal cells and a prominent immunofluorescent signal corresponding to TLR2 was detected only in rCL. TLR4 mRNA were higher in mCLand rCL compared to dCL(P < 0.05). The presence of the TLR4 protein in bovine CL was clearly detected in the luteal cells of both mCL and rCL. The results of this study suggest a role for TLRs in the development, maintenance, and regression of bovine CL. TLR signaling mediated pathway in luteal cells may involve in the regression of CL via regulation of TLR2 and TLR4.


Assuntos
Feminino , Animais , Bovinos , Bovinos/fisiologia , Ciclo Estral , Corpo Lúteo/anormalidades , /análise , /análise
6.
Anim. Reprod. ; 14(supl. 1): 1271-1277, 2017. ilus, graf, tab
Artigo em Inglês | VETINDEX | ID: vti-728533

Resumo

The aim of this study was to elucidate the presence of components of the innate immune system inthe bovine corpus luteum (CL) by detecting the expression and cell-specific localization of TLR2 and TLR4 during different stages of the estrous cycle in a control study design. Bovine CL samples were collected from a local slaughterhouse and assigned to three groups as follows: developing CL (dCL; n = 6, approx. days 3–6), mature CL (mCL; n = 5, approx. days 8–12), and regressing CL (rCL; n = 5, approx. days 17–19). An upregulation of TLR2 mRNA was detected only in rCL(P < 0.05). Localization of the TLR2 protein was particularly apparent in luteal cells and a prominent immunofluorescent signal corresponding to TLR2 was detected only in rCL. TLR4 mRNA were higher in mCLand rCL compared to dCL(P < 0.05). The presence of the TLR4 protein in bovine CL was clearly detected in the luteal cells of both mCL and rCL. The results of this study suggest a role for TLRs in the development, maintenance, and regression of bovine CL. TLR signaling mediated pathway in luteal cells may involve in the regression of CL via regulation of TLR2 and TLR4.(AU)


Assuntos
Animais , Feminino , Bovinos , Bovinos/fisiologia , Corpo Lúteo/anormalidades , Ciclo Estral , Receptor 2 Toll-Like/análise , Receptor 4 Toll-Like/análise
7.
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-954846

Resumo

Agaricus brasiliensis é um cogumelo medicinal com atividades imunomoduladoras e antitumorais atribuídas aos ß-glucanos presentes na fração polissacarídica de seu corpo de frutificação. Uma vez que os ß-glucanos aumentam a imunorresponsividade celular, neste estudo objetivamos avaliar o efeito de uma fração rica em polissacarídeos tratados com ácido (ATF) de A. brasiliensis sobre a capacidade de monócitos humanos de aderir / fagocitar células de levedura C. albicans . expressão de receptores de reconhecimento de padrões e sua capacidade de produzir citocinas. Métodos: A adesão / fagocitose de C. albicans marcada com FITC foi avaliada por citometria de fluxo. As células foram incubadas com anticorpos marcados com fluorocromo específicos para TLR2 e 4, ßGR e MR e também avaliadas por citometria de fluxo. Os monócitos foram cultivados com ATF, e os sobrenadantes da cultura foram coletados para análise da produção de citocinas in vitro por ELISA (TNF-α, IL-1ß, IL-12 e IL-10). Resultados: ATF aumentou significativamente a aderência / fagocitose de C. albicans por monócitos e isso foi associado com expressão aumentada de TLR2 e TLR4, enquanto nenhum efeito foi observado em ßGR ou MR. Além disso, a expressão de TLR4 e TLR2 foi associada a níveis mais elevados de produção in vitro de TNF-α e IL-1, respectivamente. A produção de IL-10 também foi aumentada pelo tratamento com ATF, mas não encontramos associação entre sua produção e a expressão de receptores Toll-like. Conclusão: Nossos resultados nos forneceram evidências de que polissacarídeos de A. brasiliensis afetam monócitos humanos provavelmente através da modulação de receptores Toll-like.(AU)


Assuntos
Polissacarídeos , Técnicas In Vitro , Agaricus , Candida albicans , Citocinas , Receptores Toll-Like
8.
R. bras. Ci. avíc. ; 19(1): 87-94, jan.-mar. 2017. tab, graf
Artigo em Inglês | VETINDEX | ID: vti-688269

Resumo

Toll-like receptors (TLRs) are well-characterized in mice and rats, but little is known on their role in broiler chickens during acute heat stress (AHS). The aim of the present study was to estimate the change in TLR4 mRNA expression in the liver, kidney's pleen, heart, and small intestine of broiler chickens under AHS. A total of 240 healthy Arbor Acres (AA) broiler chickens were randomly divided into four groups: control group (22±1°C; 0h), HS2, HS5 and HS10 (38±1°C; 2h, 5h and 10h of heat stress, respectively). As AHS duration increased, TLR4 mRNA expression slightly decreased at HS2 and HS5, but was dramatically elevated in HS10 (p 0.001) compared with the control group in the small intestine, as well as in the spleen at HS2 (p=0.001) and HS10 (p 0.001). The mRNA expression levels of TLR4 significantly increased in the liver, heart, and kidneys (p 0.001) at HS10, and in the kidneys at HS5 (p=0.003). It is found that TLR4 mRNA expression at HS10 in different organs was significantly higher (p 0.001) compared with HS2 and HS5. The results of the present study suggest that AHS may modulate the functional responses of the liver, kidney, spleen, heart, and small intestine of broilers by regulating TLR4 mRNA expression.(AU)


Assuntos
Animais , Transtornos de Estresse por Calor/complicações , Transtornos de Estresse por Calor/veterinária , Receptor 4 Toll-Like/administração & dosagem , Receptor 4 Toll-Like/análise , Galinhas/anormalidades , Galinhas/fisiologia , Galinhas
9.
Artigo em Inglês | VETINDEX | ID: vti-33405

Resumo

Agaricus brasiliensis é um cogumelo medicinal com atividades imunomoduladoras e antitumorais atribuídas aos ß-glucanos presentes na fração polissacarídica de seu corpo de frutificação. Uma vez que os ß-glucanos aumentam a imunorresponsividade celular, neste estudo objetivamos avaliar o efeito de uma fração rica em polissacarídeos tratados com ácido (ATF) de A. brasiliensis sobre a capacidade de monócitos humanos de aderir / fagocitar células de levedura C. albicans . expressão de receptores de reconhecimento de padrões e sua capacidade de produzir citocinas. Métodos: A adesão / fagocitose de C. albicans marcada com FITC foi avaliada por citometria de fluxo. As células foram incubadas com anticorpos marcados com fluorocromo específicos para TLR2 e 4, ßGR e MR e também avaliadas por citometria de fluxo. Os monócitos foram cultivados com ATF, e os sobrenadantes da cultura foram coletados para análise da produção de citocinas in vitro por ELISA (TNF-α, IL-1ß, IL-12 e IL-10). Resultados: ATF aumentou significativamente a aderência / fagocitose de C. albicans por monócitos e isso foi associado com expressão aumentada de TLR2 e TLR4, enquanto nenhum efeito foi observado em ßGR ou MR. Além disso, a expressão de TLR4 e TLR2 foi associada a níveis mais elevados de produção in vitro de TNF-α e IL-1, respectivamente. A produção de IL-10 também foi aumentada pelo tratamento com ATF, mas não encontramos associação entre sua produção e a expressão de receptores Toll-like. Conclusão: Nossos resultados nos forneceram evidências de que polissacarídeos de A. brasiliensis afetam monócitos humanos provavelmente através da modulação de receptores Toll-like.(AU)


Assuntos
Polissacarídeos , Técnicas In Vitro , Agaricus , Candida albicans , Citocinas , Receptores Toll-Like
10.
Rev. bras. ciênc. avic ; 19(1): 87-94, jan.-mar. 2017. tab, graf
Artigo em Inglês | VETINDEX | ID: biblio-1490363

Resumo

Toll-like receptors (TLRs) are well-characterized in mice and rats, but little is known on their role in broiler chickens during acute heat stress (AHS). The aim of the present study was to estimate the change in TLR4 mRNA expression in the liver, kidney's pleen, heart, and small intestine of broiler chickens under AHS. A total of 240 healthy Arbor Acres (AA) broiler chickens were randomly divided into four groups: control group (22±1°C; 0h), HS2, HS5 and HS10 (38±1°C; 2h, 5h and 10h of heat stress, respectively). As AHS duration increased, TLR4 mRNA expression slightly decreased at HS2 and HS5, but was dramatically elevated in HS10 (p 0.001) compared with the control group in the small intestine, as well as in the spleen at HS2 (p=0.001) and HS10 (p 0.001). The mRNA expression levels of TLR4 significantly increased in the liver, heart, and kidneys (p 0.001) at HS10, and in the kidneys at HS5 (p=0.003). It is found that TLR4 mRNA expression at HS10 in different organs was significantly higher (p 0.001) compared with HS2 and HS5. The results of the present study suggest that AHS may modulate the functional responses of the liver, kidney, spleen, heart, and small intestine of broilers by regulating TLR4 mRNA expression.


Assuntos
Animais , Galinhas/anormalidades , Galinhas/fisiologia , /administração & dosagem , /análise , Transtornos de Estresse por Calor/complicações , Transtornos de Estresse por Calor/veterinária , Galinhas
11.
Artigo em Inglês | LILACS-Express | LILACS, VETINDEX | ID: biblio-1484708

Resumo

Abstract Background Agaricus brasiliensis is a medicinal mushroom with immunomodulatory and antitumor activities attributed to the -glucans presented in the polysaccharide fraction of its fruiting body. Since -glucans enhance cellular immunoresponsiveness, in this study we aimed to evaluate the effect of an acid-treated polysaccharide-rich fraction (ATF) of A. brasiliensis on the ability of human monocytes to adhere/phagocyte C. albicans yeast cells, their expression of pattern recognition receptors and their ability to produce cytokines. Methods Adhesion/phagocytosis of FITC-labeled C. albicans was evaluated by flow cytometry. Cells were incubated with specific fluorochrome-labeled antibodies for TLR2 and 4, GR and MR and also evaluated by flow cytometry. Monocytes were cultured with ATF, and culture supernatants were collected for analysis of in vitro cytokine production by ELISA (TNF-, IL-1, IL-12 and IL-10). Results ATF significantly increased the adherence/phagocytosis of C. albicans by monocytes and this was associated with enhanced expression of TLR2 and TLR4, while no effect was observed on GR or MR. Moreover, expression of TLR4 and TLR2 was associated with higher levels of in vitro production of TNF- and IL-1, respectively. Production of IL-10 was also increased by ATF treatment, but we found no association between its production and the expression of Toll-like receptors. Conclusion Our results provided us with evidence that A. brasiliensis polysaccharides affect human monocytes probably through the modulation of Toll-like receptors.

12.
Arq. bras. med. vet. zootec. (Online) ; 69(4): 835-842, jul.-ago. 2017. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-876594

Resumo

Zebu and Holstein x Zebu crossbred have low incidence of uterine infection when compared to Holstein cows. Resistance to uterine infections may be associated with the ability to recognize invading microorganisms. Endometrial transcription of microbial molecular patterns receptors has been investigated in the postpartum period of Holstein cows, but it is completely unknown in Zebu or Holstein x Zebu cows. In this study, 9 Gyr and 12 F1 Holstein x Gyr cows were submitted to endometrial biopsies at the first and seventh days postpartum, with the objective to measure transcription levels of toll-like receptors (TLRs) 1/6, 2, 4, 5, and 9; nucleotide-binding oligomerization domain (NOD)-like receptors 1 and 2; and coreceptors cluster of differentiation 14 (CD14) and myeloid differentiation protein-2 (MD-2). There was a significant (P<0.05) decrease in transcription of TLR5 in Gyr, and an increase in transcription of TLR9 in F1 cows, between the first and seventh day postpartum. Both groups had low incidences of uterine infections up to 42 days postpartum. Uterine involution completed at 27.7 ± 10.1 and 25.1 ± 4.7 days postpartum for Gyr and F1 cows, respectively. In Gyr cows, higher transcription levels of TLR1/6 and NOD1 correlated to a longer period required for uterine involution. In F1 cows, lower levels of TLR1/6, TLR2 and NOD2 correlated to a longer period required for uterine involution. In conclusion, some pathogen recognition receptors associated significantly with the time required for uterine involution in Gyr and F1 cows.(AU)


Vacas Zebu e mestiças Holandês x Zebu apresentam baixas incidências de infecções uterinas quando comparadas às Holandesas. A resistência às infecções uterinas pode estar relacionada com a capacidade de reconhecimento dos microrganismos invasores. A transcrição endometrial de receptores de padrões moleculares microbianos tem sido investigada em vacas Holandesas recém-paridas, porém ainda é desconhecida em vacas Zebu e mestiças Holandês x Zebu. No presente estudo, nove vacas Gir e 12 F1 Holandês x Gir foram submetidas a biópsias endometriais no primeiro e no sétimo dia após o parto, com o objetivo de mensurar os níveis de transcrição gênica dos receptores tipo Toll (TLRs) 1/6, 2, 4, 5 e 9; receptores tipo NOD 1 e 2; e dos coreceptores CD14 e MD-2. Houve diminuição significativa (P < 0,05) do nível de transcrição de TLR5 em vacas Gir e aumento da transcrição de TLR9 em vacas F1, entre o primeiro e o sétimo dia após o parto. Os dois grupos apresentaram baixas incidências de infecções uterinas até 42 dias pós-parto. O período de involução uterina foi de 27,7 ± 10,1 e 25,1 ± 4,7 dias pós-parto, para vacas Gir e F1, respectivamente. No grupo de vacas Gir, altos níveis de transcrição de TLR1/6 e NOD1 tiveram correlação significativa com o prolongamento do período de involução uterina. No grupo de vacas F1, baixos níveis de transcrição de TLR1/6, TLR2 e NOD2 foram associados a maiores períodos de involução uterina. Portanto, os níveis de transcrição endometrial de alguns receptores de padrões moleculares microbianos na primeira semana após o parto podem estar relacionados com o tempo requerido para ocorrência da involução uterina em vacas Gir e F1.(AU)


Assuntos
Animais , Feminino , Bovinos , Biópsia/veterinária , Endométrio/ultraestrutura , Receptor Toll-Like 9/análise , Imunidade Inata , Transcrição Gênica
13.
Arq. bras. med. vet. zootec. (Online) ; 69(4): 835-842, jul.-ago. 2017. tab, graf
Artigo em Inglês | VETINDEX | ID: vti-18045

Resumo

Zebu and Holstein x Zebu crossbred have low incidence of uterine infection when compared to Holstein cows. Resistance to uterine infections may be associated with the ability to recognize invading microorganisms. Endometrial transcription of microbial molecular patterns receptors has been investigated in the postpartum period of Holstein cows, but it is completely unknown in Zebu or Holstein x Zebu cows. In this study, 9 Gyr and 12 F1 Holstein x Gyr cows were submitted to endometrial biopsies at the first and seventh days postpartum, with the objective to measure transcription levels of toll-like receptors (TLRs) 1/6, 2, 4, 5, and 9; nucleotide-binding oligomerization domain (NOD)-like receptors 1 and 2; and coreceptors cluster of differentiation 14 (CD14) and myeloid differentiation protein-2 (MD-2). There was a significant (P<0.05) decrease in transcription of TLR5 in Gyr, and an increase in transcription of TLR9 in F1 cows, between the first and seventh day postpartum. Both groups had low incidences of uterine infections up to 42 days postpartum. Uterine involution completed at 27.7 ± 10.1 and 25.1 ± 4.7 days postpartum for Gyr and F1 cows, respectively. In Gyr cows, higher transcription levels of TLR1/6 and NOD1 correlated to a longer period required for uterine involution. In F1 cows, lower levels of TLR1/6, TLR2 and NOD2 correlated to a longer period required for uterine involution. In conclusion, some pathogen recognition receptors associated significantly with the time required for uterine involution in Gyr and F1 cows.(AU)


Vacas Zebu e mestiças Holandês x Zebu apresentam baixas incidências de infecções uterinas quando comparadas às Holandesas. A resistência às infecções uterinas pode estar relacionada com a capacidade de reconhecimento dos microrganismos invasores. A transcrição endometrial de receptores de padrões moleculares microbianos tem sido investigada em vacas Holandesas recém-paridas, porém ainda é desconhecida em vacas Zebu e mestiças Holandês x Zebu. No presente estudo, nove vacas Gir e 12 F1 Holandês x Gir foram submetidas a biópsias endometriais no primeiro e no sétimo dia após o parto, com o objetivo de mensurar os níveis de transcrição gênica dos receptores tipo Toll (TLRs) 1/6, 2, 4, 5 e 9; receptores tipo NOD 1 e 2; e dos coreceptores CD14 e MD-2. Houve diminuição significativa (P < 0,05) do nível de transcrição de TLR5 em vacas Gir e aumento da transcrição de TLR9 em vacas F1, entre o primeiro e o sétimo dia após o parto. Os dois grupos apresentaram baixas incidências de infecções uterinas até 42 dias pós-parto. O período de involução uterina foi de 27,7 ± 10,1 e 25,1 ± 4,7 dias pós-parto, para vacas Gir e F1, respectivamente. No grupo de vacas Gir, altos níveis de transcrição de TLR1/6 e NOD1 tiveram correlação significativa com o prolongamento do período de involução uterina. No grupo de vacas F1, baixos níveis de transcrição de TLR1/6, TLR2 e NOD2 foram associados a maiores períodos de involução uterina. Portanto, os níveis de transcrição endometrial de alguns receptores de padrões moleculares microbianos na primeira semana após o parto podem estar relacionados com o tempo requerido para ocorrência da involução uterina em vacas Gir e F1.(AU)


Assuntos
Animais , Feminino , Bovinos , Receptor Toll-Like 9/análise , Endométrio/ultraestrutura , Biópsia/veterinária , Imunidade Inata , Transcrição Gênica
14.
Tese em Português | VETTESES | ID: vtt-220237

Resumo

A bubalinocultura vem apresentando elevado desempenho nos últimos anos, especialmente na Amazônia Brasileira, embora esta apresente condições ambientais adversas que influenciam no surgimento de doenças, incluindo infecções oculares. Assim, a determinação de genes relacionados à imunidade inata ocular é necessária em populações bubalinas. Buscamos determinar os perfis de expressão de genes relacionados ao sistema imune inato na membrana conjuntival de búfalos da Amazônia e caracterizar a região 3-UTR do gene Interferon gama determinando o seu perfil de expressão nessa população. Para isso, amostras de tecido conjuntival ocular e sangue foram coletadas de búfalos abatidos no Estado do Amapá. Os animais foram classificados em três grupos (G1, G2 e G3) de acordo com o grau quantitativo de tecido linfoide associado à conjuntiva por técnicas histológicas das amostras conjuntivais as quais foram submetidas a quantificação da expressão de citocinas inflamatórias, receptores tolllike 4 e -defensina-110 relativa ao gene endógeno. Amostras de sangue foram submetidas a PCR convencional usando iniciadores da região 3 UTR do gene Interferon gama. Os produtos foram purificados, sequenciados e as sequências editadas com outras correspondentes de búfalos e de espécies mamíferas. As mesmas foram alinhadas e comparadas a partir da análise de similaridade genética e análise filogenética. Na conjuntiva dos animais do G1 houve expressão apenas para IFNG e TLR4. Enquanto o G2 houve alta expressão para os genes IL6, IL10, IFNG, TLR4 e os animais do G3 apresentaram elevada expressão para todos os genes estudados. O grau de similaridade genética formou os grupos dos ruminantes, primatas, carnívoros, quirópteros, cetáceos e indivíduos isolados como suiformes e rinocerontes. Ruminantes e cetáceos apresentam alta relação filogenética enquanto houveram diferentes distâncias evolutivas de búfalos com as demais espécies de mamíferos. Nossos estudos serviram para definir os processos imunológicos inatos que ocorrem na membrana conjuntival de uma população de búfalos e determinar o perfil evolutivo em relação a outras espécies mamíferas.


Buffalo production has shown high performance in recent years, especially in the Brazilian Amazon, although it presents adverse environmental conditions that influence the emergence of diseases, including eye infections. Thus, the determination of genes related to ocular innate immunity is necessary in buffalo populations. We sought to determine the expression profiles of genes related to the innate immune system in the conjunctival membrane of Amazon buffaloes and to characterize the 3-UTR region of the Interferon gamma gene, determining its expression profile in this population. For this purpose, samples of ocular conjunctival tissue and blood were collected from buffaloes slaughtered in the State of Amapá. The animals were classified into three groups (G1, G2 and G3) according to the quantitative degree of lymphoid tissue associated with the conjunctiva by histological techniques of the conjunctival samples which were also subjected to quantification of the expression of inflammatory cytokines, tolllike receptors 4 and -defensin-110 relative to the endogenous gene. Blood samples were subjected to conventional PCR using primers from the 3' UTR region of the Interferon gamma gene. The products were purified, sequenced and the sequences edited with corresponding ones from buffalo and mammalian species. They were aligned and compared based on genetic similarity and phylogenetic analysis. In the conjunctiva of animals from G1 there was expression only for IFNG and TLR4. While G2 there was high expression for genes IL6, IL10, IFNG, TLR4 and animals from G3 showed high expression for all genes studied. The degree of genetic similarity formed the groups of ruminants, primates, carnivores, bats, cetaceans and isolated individuals such as suiformes and rhinos. Ruminants and cetaceans have a high phylogenetic relationship while there were different evolutionary distances between buffaloes and other mammal species. Our studies served to define the innate immunological processes that occur in the conjunctival membrane of a buffalo population and to determine the evolutionary profile in relation to other mammalian species.

15.
R. bras. Reprod. Anim. ; 38(4): 214-219, Out-Dez. 2014. tab
Artigo em Português | VETINDEX | ID: vti-28640

Resumo

A imunidade inata é o principal mecanismo de defesa responsável pelo controle da contaminaçãouterina em vacas recém-paridas. O reconhecimento inicial de determinados componentes estruturais demicrorganismos invasores do ambiente uterino é realizado por receptores específicos, localizados nas célulasendometriais. Estudos sobre a atuação desses receptores são importantes para elucidar os mecanismos queregulam a resposta endometrial contra os patógenos, a qual pode evoluir para a proteção imunológica ou para oestabelecimento de infecções uterinas nos casos de falhas na ativação da resposta inflamatória. O objetivo destarevisão é relatar as principais informações disponíveis sobre imunidade inata uterina em vacas após o parto.(AU)


Innate immunity is the main defense mechanism responsible for controlling uterine contamination inpostpartum cows. The initial recognition of certain structural components of microorganisms that invade theuterine environment is mediated by specific receptors located in endometrial cells. Studies about the role ofthese receptors are important for elucidating the mechanisms that regulate endometrial response againstpathogens, which can progress to immune protection or establishment of uterine infections in cases of failures inthe activation of the inflammatory response. The objective of this review is to describe the most relevantknowledge about uterine innate immunity in postpartum cows.(AU)


Assuntos
Animais , Feminino , Bovinos , Imunidade Inata , Infecções , Bovinos/fisiologia , Período Pós-Parto
16.
Rev. bras. reprod. anim ; 38(4): 214-219, Out-Dez. 2014. tab
Artigo em Português | VETINDEX | ID: biblio-1492128

Resumo

A imunidade inata é o principal mecanismo de defesa responsável pelo controle da contaminaçãouterina em vacas recém-paridas. O reconhecimento inicial de determinados componentes estruturais demicrorganismos invasores do ambiente uterino é realizado por receptores específicos, localizados nas célulasendometriais. Estudos sobre a atuação desses receptores são importantes para elucidar os mecanismos queregulam a resposta endometrial contra os patógenos, a qual pode evoluir para a proteção imunológica ou para oestabelecimento de infecções uterinas nos casos de falhas na ativação da resposta inflamatória. O objetivo destarevisão é relatar as principais informações disponíveis sobre imunidade inata uterina em vacas após o parto.


Innate immunity is the main defense mechanism responsible for controlling uterine contamination inpostpartum cows. The initial recognition of certain structural components of microorganisms that invade theuterine environment is mediated by specific receptors located in endometrial cells. Studies about the role ofthese receptors are important for elucidating the mechanisms that regulate endometrial response againstpathogens, which can progress to immune protection or establishment of uterine infections in cases of failures inthe activation of the inflammatory response. The objective of this review is to describe the most relevantknowledge about uterine innate immunity in postpartum cows.


Assuntos
Feminino , Animais , Bovinos , Bovinos/fisiologia , Imunidade Inata , Infecções , Período Pós-Parto
17.
Acta cir. bras. ; 29(supl.3): 33-38, 2014. graf
Artigo em Inglês | VETINDEX | ID: vti-11222

Resumo

To evaluate the expression profile of genes related to Toll Like Receptors (TLR) pathways of human Primary Epidermal keratinocytes of patients with severe burns. After obtaining viable fragments of skin with and without burning, culture hKEP was initiated by the enzymatic method using Dispase (Sigma-Aldrich). These cells were treated with Trizol(r) (Life Technologies) for extraction of total RNA. This was quantified and analyzed for purity for obtaining cDNA for the analysis of gene expression using specific TLR pathways PCR Arrays plates (SA Biosciences). After the analysis of gene expression we found that 21% of these genes were differentially expressed, of which 100% were repressed or hyporegulated. Among these, the following genes (fold decrease): HSPA1A (-58), HRAS (-36), MAP2K3 (-23), TOLLIP (-23), RELA (-18), FOS (-16), and TLR1 (-6.0). This study contributes to the understanding of the molecular mechanisms related to TLR pathways and underlying wound infection caused by the burn. Furthermore, it may provide new strategies to restore normal expression of these genes and thereby change the healing process and improve clinical outcome.(AU)


Assuntos
Animais , Queimaduras , Ferimentos e Lesões/complicações , Queratinócitos/fisiologia
18.
Acta cir. bras. ; 28(3): 216-220, 2013. ilus
Artigo em Inglês | VETINDEX | ID: vti-8960

Resumo

PURPOSE: To describe a method to characterize the gelatinase activity of cultured human periodontal fibroblasts stimulated with Pam3Cys and E. coli LPS, ligands of TLR2 and TLR4 respectively, and by centrifugation of the cultures, simulating an orthodontic force. METHODS: To study MMP-2 activity, primary cultures of human periodontal fibroblasts were stimulated with the addition of TLRs 2 and 4 ligands and the application of mechanical force by centrifugation at 141 x g for 30 min. Supernatant media was collected 24 hours later to perform protein quantification and zymography. RESULTS: MMP-2 activity suffered an increase in cultures co-stimulated with TLRs 2 and 4 ligands alone or with the presence of mechanical force application compared to basal levels. CONCLUSION: Zymography, one of the several methods to study MMPs activities, is a simple, qualitative and efficient method based on electrophoresis of bis-acrylamide gels copolymerized with a protein substrate.(AU)


Assuntos
Humanos , Animais , Fibroblastos/citologia , Ligamento Periodontal/anatomia & histologia , Proteínas , Eletroforese
19.
Tese em Português | VETTESES | ID: vtt-207885

Resumo

A resposta imune aviária pode ser dividida em resposta imune inata e adquirida. A imunidade inata é a primeira linha de defesa contra patógenos microbianos e possui como um de seus componentes moleculares os receptores de reconhecimento de padrão (PRRs). Três famílias dos PRRs têm sido maior alvo de pesquisas nas aves: os receptores semelhantes ao Toll (TLR), receptores semelhantes ao NOD (NLR) e os receptores semelhante a RIG-1 (RLR). Estruturas denominadas Padrão Molecular Associado ao Patógeno (PAMP) e Padrão Molecular Associado ao Dano (DAMP) são reconhecidos por esses receptores, induzem respostas imunes inatas e podem modular a imunidade adquirida. O objetivo deste trabalho foi selecionar PAMPs e DAMPs agonistas de PRRS das aves,analisar in silico sequências genéticas do PAMP flagelina e DAMP HSP70 das aves, clonar e expressar de de forma recombinante esses agonistas. O modelo de clonagem e expressão utilizado para a geração do PAMP e DAMP alvos desta pesquisa, pET100-D/TOPO, clonados em E. coli TOP10 e expressos em E. coli C4 (DE3) e C43 (DE3) mostrou ser eficiente, e ambas as proteínas foram expressas e confirmadas por Western Blotting. Os resultados da caracterização in silico realizadas para as proteínas expressas justificaram a adoção do fragmento genético das sequências da flagelina e HSP70 devido a seleção de uma região de cada agonista que foi traduzida em uma proteína com características físico-químicas e imunológicas viáveis para sua adoção como imunobiógicos das aves. Os resultados obtidos nesta tese aumenta a possibilidade de obter novos ensaios in vitro e in vivo que visam analisar PAMPs e DAMPs recombinantes como estimulantes dos componentes do sistema imune inato das aves e desta forma atuarem como possíveis adjuvantes vacinais.


The immune responses of birds are carried out in coordination by the innate and acquired immune systems. The innate immunity is the first line of defense against microbial pathogens, and is structured with cells equipped with receptors for pattern recognition (PRR). Three families of PRR have been the subject of research in birds: Toll-like receptors (TLR), NOD-like receptors (NLR), and RIG-1-like receptors (RLR). Structures known as Pathogen Associated Molecular Pattern (PAMP), present in pathogens, and Damage Associated Molecular Standard (DAMP), present in the damaged tissue, are recognized by cells with these receptors, and induce innate immune responses which will modulate the acquired immunity. The objective of this work was to perform a review that addresses the intrinsic characteristics of the innate immune system of birds, pointing out the mechanisms of action of PRRs in avian species, and consider the perspective of studies aimed at improving the vaccine responses through the adoption of PAMPs and DAMPs as adjuvants. In addition, we aimed at selecting and analyzing in silico the genetic sequences of the flagellin (PAMP) and HSP70 (DAMP) from birds and to produce these PRR agonists through genetic engineering. The in silico expressed proteins characterization results justified the adoption of the genetic fragment of the fljB (flagellin encoding) and HSP70 gene sequences. The selected region was chosen considering the ideal physico-chemical and immunological characteristics of the translated segment and considering the immunogenicity to birds. The expression system pET100-D / TOPO was used for the generation of flagellin and HSP70 and subsequently cloned in E. coli TOP10. Using E. coli C4 (DE3) and C43 (DE3) for expression, the system showed to be efficient, enabling both proteins be correctly expressed and confirmed by western blotting. The results obtained indicate the possibility of using the recombinant proteins for the immunomodulation in birds. In vitro and In vivo studies are needed aiming at analyzing recombinant PAMPs and DAMPs as stimulants of the innate immune system of birds and their potential use as vaccine adjuvants

20.
Tese em Português | VETTESES | ID: vtt-203025

Resumo

A leishmaniose visceral canina é uma importante zoonose causada pelo protozoário Leishmania infantum, transmitida pela picada de insetos flebotomíneos. Apesar do avanço no conhecimento dos mecanismos imunológicos frente à infecção por Leishmania nos últimos anos, há poucos estudos sobre o estabelecimento da resposta imune em cães, especialmente sobre os mecanismos de imunidade inata. Nesse contexto, visando medidas futuras de controle dos casos caninos e humanos, a imunoterapia desponta como uma opção viável. Desta forma, o objetivo desse trabalho foi determinar o perfil de expressão gênica dos receptores TLR9, NOD1 e NOD2 e avaliar sua correlação com a sobrevivência intracelular de L. infantum em macrófagos caninos. Após exames de triagem e confirmação de diagnóstico negativo para a doença, foram selecionados 43 animais clinicamente saudáveis para coleta de sangue para cultivo celular. Após diferenciação de monócitos em macrófagos, parte do sobrenadante da cultura foi coletado e armazenado a -80ºC para posterior extração de RNA e realização de PCR em tempo real, para avaliação da expressão dos genes de imunidade inata. A outra parte foi experimentalmente infectada por L. infantum, com posterior avaliação da sobrevivência intracelular do parasito. Observou-se que a expressão desses genes é baixa, além de correlação positiva entre a expressão de NOD2 e a sobrevivência intracelular de L. infantum. Esses resultados evidenciam a importância de estudos que visem descrever, de forma mais aprofundada, a função destes e de outros receptores, tanto na infecção experimental como em animais naturalmente infectados.


Canine visceral leishmaniasis is an important zoonosis caused by Leishmania infantum, transmitted by sand flies. Despite the advance in the understanding of the immune mechanisms against Leishmania infection in recent years, there are few studies about the establishment of the immune response in dogs, especially on the mechanisms of innate immunity. In this context, aiming future control measures of canine and human cases, immunotherapy has emerged as a viable option. This way, the aim of this study was to determine the gene expression of the receptors TLR9, NOD1 and NOD2 in canine macrophages and evaluate their correlation with the intracellular survival of L. infantum. After screening tests and confirmation of negative diagnosis for the disease, 43 clinically healthy animals were selected for blood collection for cell culture. After monocytes differentiation into macrophages, part of the culture supernatant was collected and stored at -80°C for subsequent RNA extraction and real-time PCR, for evaluation of the expression of innate immunity genes. The other part was experimentally infected with L. infantum, with further evaluation of the intracellular survival of the parasite. It was observed the low expression of these receptors and a positive correlation between the expression of NOD2 and intracellular survival of L. infantum. These results highlight the importance of studies that aim to describe, in a more detailed manner, the function of these and other receptors, both in experimental infection as in naturally infected animals.

SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA