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1.
Anim Sci J ; 91(1): e13450, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32881233

RESUMO

Mycoplasma pneumonia of swine (MPS) is caused by Mycoplasma hyopneumoniae (M.hp) and is a common chronic respiratory disease of pigs. Recently, a genetically selected variant of the Landrace pig (Miyagino L2) has a lower incidence of pulmonary MPS lesions. We investigated the pathological and immunological characteristics of MPS resistance in these pigs (n = 24) by comparing with the normal landrace pig (control: n = 24). The pathological MPS lung lesion score in MPS-selected landrace pigs was significantly lower than in the control. The gene expression of interleukin (IL)-12p40, which acts as a chemoattractant and a component of the bioactive cytokines IL-12 and IL-23, was significantly higher at the hilar lymph nodes, lung, and spleen in MPS-selected landrace pigs than in control landrace pigs, and these were negatively correlated with the macroscopic MPS lung lesion score. In summary, we demonstrate that resistance against MPS in Miyagino L2 pigs is associated with IL-12p40 up-regulation, in comparison with normal landrace pigs without the MPS vaccine. In addition, a comparative study of macroscopic MPS lung lesions and IL-12p40 gene expression in lung and hilar lymph nodes may lead to beneficial selection traits for the genetic selection for MPS resistance in pigs.


Assuntos
Subunidade p40 da Interleucina-12/genética , Subunidade p40 da Interleucina-12/metabolismo , Pulmão/imunologia , Linfonodos/imunologia , Pneumonia Suína Micoplasmática/genética , Pneumonia Suína Micoplasmática/imunologia , Suínos/genética , Suínos/imunologia , Animais , Expressão Gênica , Predisposição Genética para Doença , Masculino , Característica Quantitativa Herdável , Seleção Genética , Regulação para Cima/genética
2.
Transplantation ; 104(8): 1566-1573, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32732833

RESUMO

BACKGROUND: Xenogeneic organ transplantation has been proposed as a potential approach to fundamentally solve organ shortage problem. Xenogeneic immune responses across species is one of the major obstacles for clinic application of xeno-organ transplantation. The generation of glycoprotein galactosyltransferase α 1, 3 (GGTA1) knockout pigs has greatly contributed to the reduction of hyperacute xenograft rejection. However, severe xenograft rejection can still be induced by xenoimmune responses to the porcine major histocompatibility complex antigens swine leukocyte antigen class I and class II. METHODS: We simultaneously depleted GGTA1, ß2-microglobulin (ß2M), and major histocompatibility complex class II transactivator (CIITA) genes using clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins technology in Bamma pig fibroblast cells, which were further used to generate GGTA1ß2MCIITA triple knockout (GBC-3KO) pigs by nuclear transfer. RESULTS: The genotype of GBC-3KO pigs was confirmed by polymerase chain reaction and Sanger sequencing, and the loss of expression of α-1,3-galactose, SLA-I, and SLA-II was demonstrated by flow cytometric analysis using fluorescent-conjugated lectin from bandeiraea simplicifolia, anti-ß2-microglobulin, and swine leukocyte antigen class II DR antibodies. Furthermore, mixed lymphocyte reaction assay revealed that peripheral blood mononuclear cells from GBC-3KO pigs were significantly less effective than (WT) pig peripheral blood mononuclear cells in inducing human CD3CD4 and CD3CD8 T-cell activation and proliferation. In addition, GBC-3KO pig skin grafts showed a significantly prolonged survival in immunocompetent C57BL/6 mice, when compared with wild-type pig skin grafts. CONCLUSIONS: Taken together, these results demonstrate that elimination of GGTA1, ß2M, and CIITA genes in pigs can effectively alleviate xenogeneic immune responses and prolong pig organ survival in xenogenesis. We believe that this work will facilitate future research in xenotransplantation.


Assuntos
Rejeição de Enxerto/prevenção & controle , Xenoenxertos/imunologia , Transplante de Órgãos/métodos , Transplante Heterólogo/métodos , Aloenxertos/provisão & distribução , Animais , Animais Geneticamente Modificados/imunologia , Sistemas CRISPR-Cas/genética , Modelos Animais de Doenças , Feminino , Galactosiltransferases/genética , Galactosiltransferases/imunologia , Técnicas de Inativação de Genes/métodos , Genes MHC da Classe II/genética , Genes MHC da Classe II/imunologia , Rejeição de Enxerto/imunologia , Sobrevivência de Enxerto/genética , Sobrevivência de Enxerto/imunologia , Xenoenxertos/transplante , Humanos , Masculino , Camundongos , Transplante de Órgãos/efeitos adversos , Suínos/genética , Suínos/imunologia , Transplante Heterólogo/efeitos adversos , Microglobulina beta-2/genética , Microglobulina beta-2/imunologia
3.
Anim Sci J ; 91(1): e13429, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32696533

RESUMO

To evaluate the effects of crude protein (CP) and lactose (LAC) for weaned piglets on performance, intestinal morphology, and expression of genes related to intestinal integrity and immune system, 144 piglets with initial weight 7.17 ± 0.97 kg were allotted in a randomized design, in a 2 × 3 factorial arrangement (20.0% and 24.0% CP and 8.0%, 12.0%, and 16.0% LAC) with eight replicates. Piglets fed 20.0% CP had greater weight gain and feed intake. Including 12.0% LAC in the 20.0% CP diet provided higher villous height in the duodenum than 8.0% LAC, and 12.0% or 16.0% LAC in the 24.0% CP diet resulted in higher villous height in the jejunum and ileum, and higher villi/crypt ratio in the ileum than 8.0% LAC. No effects of CP and LAC on interleukin-1ß and tumor necrosis factor-α mRNA were observed. The 16.0% LAC diet provided higher gene expression of transforming-ß1 growth factor. Feeding 20.0% CP resulted in better performance than 24.0% CP. The 12.0% LAC diet promoted greater genetic expression of occludin and zonula occludens. Including 12.0% LAC in the diet may improve intestinal epithelial morphology and integrity, and these improvements are more evident when piglets are fed diets with 24.0% CP.


Assuntos
Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Dieta/veterinária , Proteínas na Dieta/análise , Expressão Gênica , Mucosa Intestinal/anatomia & histologia , Mucosa Intestinal/imunologia , Lactose/análise , Suínos/crescimento & desenvolvimento , Suínos/genética , Fator de Crescimento Transformador beta1/genética , Fator de Crescimento Transformador beta1/metabolismo , Animais , Ingestão de Alimentos , Feminino , Masculino , Ocludina/genética , Ocludina/metabolismo , Suínos/anatomia & histologia , Suínos/imunologia , Junções Íntimas/genética , Junções Íntimas/metabolismo , Ganho de Peso
4.
PLoS One ; 15(6): e0233773, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32559198

RESUMO

In early life and around weaning, pigs are at risk of developing infectious diseases which compromise animal welfare and have major economic consequences for the pig industry. A promising strategy to enhance resistance against infectious diseases is immunomodulation by feed additives. To assess the immune stimulating potential of feed additives in vitro, bone marrow-derived dendritic cells were used. These cells play a central role in the innate and adaptive immune system and are the first cells encountered by antigens that pass the epithelial barrier. Two different feed additives were tested on dendritic cells cultured from fresh and cryopreserved bone marrow cells; a widely used commercial feed additive based on yeast-derived ß-glucans and the gram-negative probiotic strain E. coli Nissle 1917. E. coli Nissle 1917, but not ß-glucans, induced a dose-dependent upregulation of the cell maturation marker CD80/86, whereas both feed additives induced a dose-dependent production of pro- and anti-inflammatory cytokines, including TNFα, IL-1ß, IL-6 and IL-10. Furthermore, E. coli Nissle 1917 consistently induced higher levels of cytokine production than ß-glucans. These immunomodulatory responses could be assessed by fresh as well as cryopreserved in vitro cultured porcine bone marrow-derived dendritic cells. Taken together, these results demonstrate that both ß-glucans and E. coli Nissle 1917 are able to enhance dendritic cell maturation, but in a differential manner. A more mature dendritic cell phenotype could contribute to a more efficient response to infections. Moreover, both fresh and cryopreserved bone marrow-derived dendritic cells can be used as in vitro pre-screening tools which enable an evidence based prediction of the potential immune stimulating effects of different feed additives.


Assuntos
Células da Medula Óssea/imunologia , Células Dendríticas/imunologia , Fatores Imunológicos/farmacologia , Probióticos/farmacologia , Suínos/imunologia , beta-Glucanas/farmacologia , Ração Animal , Animais , Antígeno B7-1/genética , Antígeno B7-1/metabolismo , Antígeno B7-2/genética , Antígeno B7-2/metabolismo , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos dos fármacos , Diferenciação Celular , Células Cultivadas , Células Dendríticas/citologia , Células Dendríticas/efeitos dos fármacos , Escherichia coli , Fatores Imunológicos/administração & dosagem , Interleucinas/genética , Interleucinas/metabolismo , Probióticos/administração & dosagem , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , beta-Glucanas/administração & dosagem
5.
PLoS One ; 15(5): e0232782, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32369529

RESUMO

The efficacy of foot-and-mouth disease virus (FMDV) inactivated vaccines is mainly dependent on the integrity of the whole (146S) viral particles. If the intact capsids disassemble to 12S subunits, antibodies against internal-not protective epitopes, may be induced. Serological correlates with protection may be hampered if antibodies against internal epitopes are measured. Here we compared the performance of different ELISAs with the virus-neutralization test (VNT) that measures antibodies against exposed epitopes. Sera from pigs immunized with one dose of an expired commercial FMDV vaccine were used. This vaccine contained about 50% of O1/Campos and over 90% of A24/Cruzeiro strains total antigen as whole 146S particles. Specific-total antibodies were measured with the standard liquid-phase blocking ELISA (LPBE). We also developed an indirect ELISA (IE) using sucrose gradient purified 146S particles as capture antigen to titrate total antibodies, IgM, IgG1 and IgG2. A good correlation was found between VNT titers and IgG-ELISAs for A24/Cruzeiro, with the lowest correlation coefficient estimated for IgG2 titers. For O1/Campos, however, the presence of antibodies against epitopes different from those of the whole capsid, elicited by the presence of 12S particles in the vaccine, hampered the correlation between LPBE and VNT, which was improved by using purified O1/Campos 146S-particles for the liquid-phase of the LPBE. Interestingly, 146S particles but not 12S were efficiently bound to the ELISA plates, confirming the efficiency of the IE to detect antibodies against exposed epitopes. Our results indicate that any serological test assessing total antibodies or IgG1 against epitopes exposed in intact 146S-particles correlate with the levels of serum neutralizing antibodies in vaccinated pigs, and might potentially replace the VNT, upon validation. We recommend that antigen used for serological assays aimed to measure protective antibodies against FMDV should be controlled to ensure the preservation of 146S viral particles.


Assuntos
Vírus da Febre Aftosa/imunologia , Febre Aftosa/terapia , Doenças dos Suínos/terapia , Suínos/virologia , Vacinas Virais/uso terapêutico , Animais , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Febre Aftosa/imunologia , Testes de Neutralização , Suínos/imunologia , Doenças dos Suínos/imunologia , Vacinas Virais/imunologia
6.
J Anim Sci ; 98(5)2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-32307528

RESUMO

The aim of this study was to investigate the effects of diet complexity and l-Thr supplementation level on the growth performance, immune response, intestinal barrier function, and microbial metabolites in nursery pigs. Thirty-two weaned pigs (body weight 7.23 ± 0.48 kg) were randomly assigned to dietary treatments in a 2 × 2 factorial arrangement based on diet complexity (complex or simple) and dietary Thr content. The complex diet contained fish meal, plasma protein, and dried whey to mimic a conventional nursery diet. The simple diet was formulated with corn, wheat, and soybean meal and did not contain any animal products. l-Thr was supplemented to each diet to supply either 100% (STD Thr) or 115% (SUP Thr) of the NRC (2012) requirement for standardized ileal digestible Thr. Pigs were individually housed and fed experimental diets ad libitum for 14 d. Diet complexity, dietary Thr content, and their interactions were considered the main effects. Pigs fed the simple diet had greater (P < 0.05) plasma interleukin (IL)-10 and IL-6 concentrations compared with those fed the complex diet on days 7 and 14, respectively. Simple diet-fed pigs tended to show greater (P < 0.10) expression of genes encoding for tumor necrosis factor-α, claudin-1, and zonula occludens-1 in the jejunum compared with complex diet-fed pigs. The simple diet-fed pigs had greater (P < 0.05) concentrations of NH3-N in the jejunum digesta than did complex diet-fed pigs. The SUP Thr increased (P < 0.05) villus height and goblet cell (GC) density in villi and crypts in the jejunum and deepened (P < 0.05) crypts in the proximal colon. The SUP Thr resulted in the upregulation (P < 0.05) of occludin gene expression and a tendency toward the downregulation (P = 0.10) of IL-6 gene expression in the jejunum. Interactions (P < 0.05) between diet complexity and l-Thr supplementation level were observed in GC density in the crypt, NH3-N concentration in the jejunum, and the contents of acetate, propionate, and total volatile fatty acids in the colon. In conclusion, feeding a simple diet to nursery pigs resulted in systemic and intestinal inflammation. The SUP Thr diet did not normalize the simple diet-induced inflammation but improved gut integrity. SUP Thr seems to have greater benefits with a simple diet than with a complex diet. Therefore, SUP Thr in a simple diet could be a beneficial nutritional strategy for enhancing gut health.


Assuntos
Suplementos Nutricionais/análise , Metabolismo Energético/efeitos dos fármacos , Microbioma Gastrointestinal/fisiologia , Imunidade Inata/efeitos dos fármacos , Suínos/fisiologia , Treonina/administração & dosagem , Ração Animal/análise , Animais , Composição Corporal , Peso Corporal/efeitos dos fármacos , Dieta/veterinária , Ácidos Graxos Voláteis/análise , Microbioma Gastrointestinal/efeitos dos fármacos , Interleucina-10/análise , Mucosa Intestinal/efeitos dos fármacos , Masculino , Nitrogênio/metabolismo , Soja , Suínos/crescimento & desenvolvimento , Suínos/imunologia , Desmame , Zea mays
7.
Vet Microbiol ; 242: 108599, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32122603

RESUMO

Streptococcus suis is an emerging swine and human pathogen causing severe infections and sudden death. During infection, complement C3a and C5a were reported to induce immune cells towards infection and injury sites via their corresponding receptors C3aR and C5aR. However, how S. suis evade immune surveillance mediated by C3aR and C5aR remains unclear. In this study, we analyze and construct an S. suis bacterial two-hybrid prey library containing 39 LPXTG motif anchored proteins and 18 secreted proteins. Two highly possible C3aR-binding proteins: thiol-activated toxin Suilysin, putative RTX family exoprotein A gene and three highly possible C5aR-binding proteins: thiol-activated toxin Suilysin, putative 5'-nucleotidase and subtilisin-like serine protease are identified through bacterial two-hybrid assay. Far-western blot assay confirms that a cholesterol-binding cytolysin Suilysin can interact with both C3aR and C5aR. Chemotaxis assays demonstrate that recombinant and natural Suilysin can inhibit monocyte chemotaxis mediated by C3a and C5a. These findings enlarge our knowledge of suilysin biological significance and provide a new perspective on S. suis complement evasion.


Assuntos
Proteínas de Bactérias/imunologia , Quimiotaxia , Proteínas Hemolisinas/imunologia , Monócitos/imunologia , Receptor da Anafilatoxina C5a/imunologia , Receptores de Complemento/imunologia , Animais , Células Cultivadas , Humanos , Infecções Estreptocócicas/microbiologia , Streptococcus suis/química , Streptococcus suis/imunologia , Suínos/imunologia , Suínos/microbiologia , Células THP-1
8.
J Anim Sci ; 98(3)2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-32191809

RESUMO

Hermetia illucens larvae meal (HILM) are rich in proteins and chitin, and represent an innovative feed ingredient for animals. However, little is known about the intestinal bacteria and immune homeostasis response of HILM as a fishmeal replacement on weanling piglets. Thus, this study aimed to investigate the changes in specific ileal and cecal bacterial populations and their metabolic profiles, and ileal immune indexes in weanling piglets fed with a diet containing HILM. A total of 128 weanling piglets were fed either a basal diet or 1 of 3 diets with 1%, 2%, and 4% HILM (HI0, HI1, HI2, and HI4, respectively). Each group consisted of 8 pens (replicates), with 4 pigs per pen. After 28 d of feeding, 8 barrows per treatment were euthanized, the ileal and cecal digesta, and ileal mucosa were collected for analyzing bacterial population and metabolic profiles, and immune indexes, respectively. Results showed that HILM increased (P < 0.05, maximum in HI2) the number of Lactobacillus and Bifidobacterium in the ileum and cecum, but quadratically decreased (P < 0.05, minimum in HI2) the number of Escherichia coli. In the cecum, the number of Firmicutes, Ruminococcus, Clostridium cluster IV, and Prevotella showed a quadratic response to increasing (P < 0.05, maximum in HI2) HILM levels. Lactate and butyrate concentrations in the ileum and cecum were quadratically increased (P < 0.05, maximum in HI2) with increasing HILM levels. In the cecum, the amines, phenol, and indole compounds concentrations were quadratically decreased (P < 0.05, minimum in HI2) with increasing HILM levels, while total short-chain fatty acids and acetate concentrations were quadratically increased (P < 0.05, maximum in HI2). In the ileum, the TLR4, NF-κB, MyD88, and TNF-α mRNA expressions were quadratically decreased (P < 0.05, minimum in HI2) with increasing HILM levels, while the mRNA expression of IL-10, barrier function (MUC1, ZO-1, Occludin, and Claudin-2), and development-related genes (IGF-1, GLP-2, and EGF) was quadratically increased (P < 0.05, maximum in HI2). Furthermore, the changes in the mucosal gene expression were associated with changes in the bacterial populations and their metabolites. Collectively, these results showed that a diet supplemented with 2% HILM affected specific bacterial populations and metabolic profiles, and maintained ileal immune status. These findings provide new insights into the use of insect meal as a suitable alternative protein source for swine feeding.


Assuntos
Ração Animal/análise , Bactérias/classificação , Bifidobacterium/isolamento & purificação , Dípteros , Intestinos/microbiologia , Suínos/fisiologia , Animais , Bactérias/metabolismo , Dieta/veterinária , Suplementos Nutricionais , Escherichia coli/metabolismo , Ácidos Graxos Voláteis/metabolismo , Homeostase , Lactobacillus/isolamento & purificação , Larva , Suínos/imunologia
9.
J Anim Sci ; 98(1)2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31909792

RESUMO

An isotope tracer study was conducted to evaluate the effects of immune system stimulation (ISS) on the irreversible loss of cysteine (Cys) to taurine (Tau) and sulfate (SO4), as well as glutathione (GSH) synthesis, during the fed state in pigs. We previously have reported that ISS increases plasma Cys flux and the GSH synthesis rate at the tissue and whole-body levels in growing pigs. Thus, the current article presents the data on the irreversible loss of Cys during ISS in pigs. Ten gilts (BW: 7.0 ±â€…0.12 kg) were feed restricted a sulfur amino acids (SAA) limiting diet and injected twice with either saline (n = 4) or increasing amounts of E. coli lipopolysaccharide (n = 6). The day after the second injection, a 5-h primed continuous intravenous infusion of 35S-Cys was conducted. ISS reduced plasma Cys and total SAA concentrations (16% and 21%, respectively; P < 0.05). However, ISS had no effect on the plasma concentrations of Tau and SO4, nor did it affect the appearance of 35S in plasma Tau, plasma SO4, urinary Tau, or urinary SO4 (P > 0.19). On a whole-body basis and including urinary excretion, ISS increased the appearance of 35S in Tau by 67% (P < 0.05), but tended to decrease the appearance of 35S in SO4 by 22% (P < 0.09). Overall, the current findings indicate that during ISS, decreased plasma SAA concentrations and increased plasma Cys flux are attributed in part to increased rates of Cys conversion to Tau, but not Cys catabolism to SO4. Thus, increased utilization of Cys for the synthesis of immune system metabolites, such as GSH and Tau, is likely the main contributor to increased Cys flux during ISS in pigs. In addition, the irreversible loss of Cys during ISS is small and has a minimal impact on the daily SAA requirements of starter pigs.


Assuntos
Cisteína/metabolismo , Sistema Imunitário/efeitos dos fármacos , Suínos/fisiologia , Taurina/metabolismo , Ração Animal , Animais , Dieta/veterinária , Escherichia coli/química , Feminino , Glutationa/metabolismo , Lipopolissacarídeos/administração & dosagem , Sulfatos/metabolismo , Suínos/imunologia
10.
Gene ; 731: 144356, 2020 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-31935504

RESUMO

The genetic basis of differential host immune response vis-à-vis transcriptome profile was explored in PBMCs of indigenous (Ghurrah) and crossbred pigs after classical swine fever vaccination and in monocyte derived macrophages (MDMs) challenged with virulent classical swine fever (CSF) virus. The humoral immune response (E2 antibody) was higher (74.87%) in crossbred than indigenous pigs (58.20%) at 21st days post vaccination (21dpv). The rate of reduction of ratio of CD4+/CD8+ was higher in crossbred pigs than indigenous pigs at 7th days post vaccination (7dpv). The immune genes IFIT1, IFIT5, RELA, NFKB2, TNF and LAT2 were up regulated at 7dpv in RNA seq data set and was in concordance during qRT-PCR validation. The Laminin Subunit Beta 1 (LAMB1) was significantly (p ≤ 0.05) down-regulated in MDMs of indigenous pigs and consequently a significantly (p ≤ 0.01) higher copy number of virulent CSF virus was evidenced in macrophages of crossbred pigs than indigenous pigs. Activation of LXR:RXR pathway at 60 h post infection (60hpi) in MDMs of indigenous versus crossbred pigs inhibited nuclear translocation of NF-κB, resulted into transrepression of proinflammatory genes. But it helped in maintenance of HDL level by lowering down cholesterol/LDL level in MDMs of indigenous pigs. The key immune genes (TLR2, TLR4, IL10, IL8, CD86, CD54, CASP1) of TREM1 signaling pathway were upregulated at 7dpv in PBMCs but those genes were downregulated at 60hpi in MDMs indigenous pigs. Using qRT-PCR, the validation of differentially expressed, immunologically important genes (LAMB1, OAS1, TLR 4, TLR8 and CD86) in MDMs revealed that expression of these genes were in concordance with RNA-seq data.


Assuntos
Peste Suína Clássica/genética , Leucócitos Mononucleares/metabolismo , Macrófagos/metabolismo , Suínos , Transcriptoma , Animais , Células Cultivadas , Peste Suína Clássica/sangue , Peste Suína Clássica/imunologia , Peste Suína Clássica/prevenção & controle , Vírus da Febre Suína Clássica/imunologia , Vírus da Febre Suína Clássica/fisiologia , Estudo de Associação Genômica Ampla/veterinária , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/imunologia , Hibridização Genética/fisiologia , Imunidade Celular/genética , Imunidade Humoral/genética , Leucócitos Mononucleares/patologia , Leucócitos Mononucleares/virologia , Macrófagos/patologia , Macrófagos/virologia , Suínos/genética , Suínos/imunologia , Suínos/metabolismo , Suínos/virologia
11.
Sci Rep ; 10(1): 743, 2020 01 20.
Artigo em Inglês | MEDLINE | ID: mdl-31959823

RESUMO

Swine leukocyte antigens play indispensable roles in immune responses by recognizing a large number of foreign antigens and thus, their genetic diversity plays a critical role in their functions. In this study, we developed a new high-resolution typing method for pig SLA-1 and successfully typed 307 individuals from diverse genetic backgrounds including 11 pure breeds, 1 cross bred, and 12 cell lines. We identified a total of 52 alleles including 18 novel alleles and 9 SLA-1 duplication haplotypes, including 4 new haplotypes. We observed significant differences in the distribution of SLA-1 alleles among the different pig breeds, including the breed specific alleles. SLA-1 duplication was observed in 33% of the chromosomes and was especially high in the biomedical model breeds such as SNU (100%) and NIH (76%) miniature pigs. Our analysis showed that SLA-1 duplication is associated with the increased level of SLA-1 mRNA expression in porcine cells compared to that of the single copy haplotype. Therefore, we provide here the results of the most extensive genetic analysis on pig SLA-1.


Assuntos
Cruzamento , Variações do Número de Cópias de DNA , Variação Genética , Heterozigoto , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Antígenos de Histocompatibilidade Classe I/genética , Teste de Histocompatibilidade/métodos , Suínos/genética , Suínos/imunologia , Alelos , Animais , Linhagem Celular , Expressão Gênica , Haplótipos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
12.
Annu Rev Anim Biosci ; 8: 171-198, 2020 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-31846353

RESUMO

In pigs, the major histocompatibility complex (MHC), or swine leukocyte antigen (SLA) complex, maps to Sus scrofa chromosome 7. It consists of three regions, the class I and class III regions mapping to 7p1.1 and the class II region mapping to 7q1.1. The swine MHC is divided by the centromere, which is unique among mammals studied to date. The SLA complexspans between 2.4 and 2.7 Mb, depending on haplotype, and encodes approximately 150 loci, with at least 120 genes predicted to be functional. Here we update the whole SLA complex based on the Sscrofa11.1 build and annotate the organization for all recognized SLA genes and their allelic sequences. We present SLA nomenclature and typing methods and discuss the expression of SLA proteins, as well as their role in antigen presentation and immune, disease, and vaccine responses. Finally, we explore the role of SLA genes in transplantation and xenotransplantation and their importance in swine biomedical models.


Assuntos
Antígenos de Histocompatibilidade Classe I/genética , Antígenos de Histocompatibilidade Classe I/imunologia , Suínos/imunologia , Animais , Regulação da Expressão Gênica , Modelos Animais , Suínos/genética , Doenças dos Suínos/imunologia , Transplantes/imunologia
13.
Animal ; 14(4): 824-833, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31650940

RESUMO

The modern hyper-prolific sow gives birth to more piglets than she has functional teats (in the following called supernumerary piglets). The aim of the present study was (1) to investigate the production consequences of hyper-prolific sows rearing supernumerary piglets equal to the mean live-born litter size, and (2) investigate whether potential negative effects on survival and growth could be alleviated by providing access to milk replacer and/or providing easier access to the udder (by loose housing). At day 1 (D1) postpartum (pp), 93 litters were standardised to 14 or 17 piglets (LS14/LS17) after which no piglets were moved between sows leading to decreased litter size if piglets died. Litters were provided with or without milk replacer in milk cups (+MILK/-MILK), and sows were either crated or loose housed (CRATE/LOOSE) in a 2 × 2 × 2 factorial design. Piglet mortality was higher in LS17 compared to LS14 (P < 0.01; OR = 2.0), higher in -MILK compared to +MILK (P = 0.01; OR = 1.2) and higher in LOOSE compared to CRATE (P = 0.02; OR = 1.8). This study showed that sow rearing of supernumerary piglets while supplying with milk replacer can increase piglet survival. It also showed that early mortality before piglets learned to drink milk replacer posed a challenge using this automatic milk replacer system. An interaction between access to milk replacer and the standardised litter size D1 affected litter weight (P < 0.01) and piglet weight day 28 (D28) (P = 0.03). The highest litter weight D28 was found in LS17 +MILK (P < 0.01) but with a lower individual piglet weight than in LS14 -MILK. Piglet weight D28 was higher in LS14 -MILK compared to LS17 regardless of access to milk replacer. Heterogeneity in piglet weight within litters D28 was larger in LS17 (P = 0.03) but could be reduced with +MILK in CRATE (P < 0.01). No effects were found on sow weight loss and feed intake (P > 0.05). In conclusion, the results showed that sows cannot rear the supernumerary piglets without further management interventions to reduce mortality. Supplying supernumerary piglets equal to the mean live-born litter size of hyper-prolific sows with milk replacer can from results of this study be an alternative strategy to the use of nurse sows.


Assuntos
Abrigo para Animais , Tamanho da Ninhada de Vivíparos , Substitutos do Leite , Suínos/imunologia , Animais , Peso Corporal , Feminino , Lactação , Parto , Período Pós-Parto , Gravidez , Suínos/crescimento & desenvolvimento , Suínos/fisiologia
14.
Rapid Commun Mass Spectrom ; 34(7): e8639, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-31659824

RESUMO

RATIONALE: There are no approved animal drugs for management of inflammation in swine due to lack of validated animal models. To assess efficacy, it was essential to develop proteomics approaches to identify suitable biomarkers of inflammation as presented in this study. METHODS: Serum samples were collected from a group of four pigs prior to (baseline) and 24 and 48 h following lipopolysaccharide (LPS) stimulation to reveal proteomic changes during inflammation. Two other pigs served as untreated controls. Proteins were separated by either one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) or two-dimensional (2D) gel electrophoresis (2DE) prior to analysis by nano-flow liquid chromatography (nLC) coupled to tandem mass spectrometry (MS/MS). RESULTS: We identified 165 proteins using SDS-PAGE, of which 47 proteins were also detected by 2DE prior to nLC/MS/MS. More than half (72%) of all characterized proteins were modulated as a result of LPS stimulation, many of which are known to be involved with innate and adaptive immunity. Pig serum samples obtained 24 h after LPS initiation of inflammation showed protein modulations of serum albumin, serotransferrin, light and heavy immunoglobulin chains (IGs), and major acute phase proteins including haptoglobin (HPT), serum amyloid A2 (SAA2), C-reactive protein (CRP), ß-2-glycoprotein 1 (B-2GP1), alpha-2-HS-glycoprotein (A2HS), α-1-antitrypsin (A1AT), and α-1-acid glycoprotein (A1AG). SAA2 was distinguished from the other SAA isoforms by the unique peptide sequence of SAA2. CONCLUSIONS: The results provided proteomics analysis of swine serum due to LPS stimulation and indicated the importance of SAA2, which appears to be unique and may be regarded as a potential clinical diagnostic biomarker of inflammation.


Assuntos
Proteínas Sanguíneas/análise , Inflamação/sangue , Lipopolissacarídeos/imunologia , Suínos/sangue , Imunidade Adaptativa , Animais , Biomarcadores/sangue , Proteínas Sanguíneas/imunologia , Cromatografia Líquida , Eletroforese em Gel Bidimensional , Eletroforese em Gel de Poliacrilamida , Imunidade Inata , Inflamação/imunologia , Lipopolissacarídeos/sangue , Proteômica , Suínos/imunologia , Espectrometria de Massas em Tandem
15.
Dev Comp Immunol ; 103: 103503, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31563460

RESUMO

Chemokines refer to chemoattractant cytokines, which have crucial functions in inflammation and immune responses in multiple cellular processes. In the present study, we described the potential role of porcine CCL5 in embryo implantation and fetal-maternal environment during early pregnancy. We first carried out phylogenetic analysis of porcine CCL5, and analyzed the cell specific localization of CCL5 and its receptor CCR3 in a kinetic approach within porcine estrous cycles and early gestation stage. In addition, CCL5 stimulated porcine uterine luminal epithelial (pLE) and porcine trophectoderm (pTr) cell proliferations, and cell cycle progressions via AKT and MAPK intracellular signaling tractions. Furthermore, CCL5 attenuated tunicamycin-induced endoplasmic reticulum (ER) stress signaling, and lipopolysaccharides-triggered inflammatory responses in pLE and pTr cells. Taken together, our study showed that CCL5 is involved in the placental development or promotes the placental development.


Assuntos
Quimiocina CCL5/metabolismo , Placenta/imunologia , Prenhez , Suínos/imunologia , Sequência de Aminoácidos , Animais , Ciclo Celular , Proliferação de Células , Quimiocina CCL5/genética , Implantação do Embrião/imunologia , Endométrio/citologia , Endométrio/metabolismo , Estresse do Retículo Endoplasmático , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Ciclo Estral/imunologia , Feminino , Inflamação , Filogenia , Placenta/metabolismo , Placentação , Gravidez , Receptores CCR3/metabolismo , Alinhamento de Sequência , Transdução de Sinais , Suínos/classificação , Suínos/genética , Trofoblastos/citologia , Trofoblastos/metabolismo , Regulação para Cima
16.
Food Funct ; 10(12): 7844-7854, 2019 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-31793606

RESUMO

We investigated the effects of Clostridium butyricum and Enterococcus faecalis (probiotics) in a piglet model. Weaned piglets (180) were randomly divided into three treatment groups and fed basal diet and basal diet supplemented with 6 × 109 CFU C. butyricum per kg and 2 × 1010 CFU E. faecalis per kg, respectively. The probiotics improved the final body weight, average daily gain, and feed conversion rate, while they reduced the diarrhea rate. The serum aspartate aminotransferase and alanine aminotransferase activities in probiotic-supplemented piglets were decreased on days 14 and 28. Piglets supplemented with probiotics presented an increased serum immunoglobulin (Ig)M level on day 14 and IgA, IgG, and IgM levels on day 28 compared with control piglets, respectively. Moreover, the probiotics increased the jejunal villus length and jejunal villus height to crypt depth ratio, while they decreased the jejunal crypt depth compared with those of the control. Similarly, an increase in inflammation-related pathway factor expression was observed after probiotic administration. Piglets supplemented with probiotics had a higher concentration of volatile fatty acids in the colonic contents than that in the control. High-throughput sequencing indicated that the probiotics modulated the colon bacterial diversity. Species richness and the alpha diversity index of bacterial samples in probiotic-supplemented piglets were higher than those in the control. Piglets supplemented with C. butyricum presented a considerably high relative abundance of C. butyricum compared with that in the control. Overall, C. butyricum and E. faecalis can promote growth performance, protect the intestinal villi morphology, improve immunity, and optimize the intestinal flora in weaned piglets.


Assuntos
Clostridium butyricum/fisiologia , Diarreia/tratamento farmacológico , Enterococcus faecalis/fisiologia , Ácidos Graxos Voláteis/metabolismo , Microbioma Gastrointestinal/efeitos dos fármacos , Intestinos/anatomia & histologia , Probióticos/administração & dosagem , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/metabolismo , Diarreia/imunologia , Diarreia/microbiologia , Modelos Animais de Doenças , Feminino , Humanos , Intestinos/crescimento & desenvolvimento , Intestinos/imunologia , Intestinos/microbiologia , Masculino , Suínos/crescimento & desenvolvimento , Suínos/imunologia , Suínos/microbiologia
17.
Int J Mol Sci ; 20(24)2019 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-31847111

RESUMO

High mobility group box 1 (HMGB1) is a DNA-binding nuclear protein that can be actively secreted by immune cells after different immune stimuli or passively released from cells undergoing necrosis. HMGB1 amplifies inflammation, and its hypersecretion contributes to multiple organ dysfunction syndrome and death. We tested possible immunomodulatory effect of commensal Lactobacillus amylovorus (LA), Lactobacillus mucosae (LM) or probiotic Escherichia coli Nissle 1917 (EcN) in infection of gnotobiotic piglets with Salmonella Typhimurium (ST). Transcription of HMGB1 and Toll-like receptors (TLR) 2, 4, and 9 and receptor for advanced glycation end products (RAGE), TLR4-related molecules (MD-2, CD14, and LBP), and adaptor proteins (MyD88 and TRIF) in the ileum and colon were measured by RT-qPCR. Expression of TLR4 and its related molecules were highly upregulated in the ST-infected intestine, which was suppressed by EcN, but not LA nor LM. In contrast, HMGB1 expression was unaffected by ST infection or commensal/probiotic administration. HMGB1 protein levels in the intestine measured by ELISA were increased in ST-infected piglets, but they were decreased by previous colonization with E. coli Nissle 1917 only. We conclude that the stability of HMGB1 mRNA expression in all piglet groups could show its importance for DNA transcription and physiological cell functions. The presence of HMGB1 protein in the intestinal lumen probably indicates cellular damage.


Assuntos
Escherichia coli/imunologia , Vida Livre de Germes/imunologia , Proteína HMGB1/imunologia , Lactobacillus acidophilus/imunologia , Probióticos , Salmonella typhimurium/imunologia , Transdução de Sinais/imunologia , Suínos , Receptor 4 Toll-Like/imunologia , Animais , Intestinos/imunologia , Intestinos/microbiologia , Suínos/imunologia , Suínos/microbiologia
18.
BMB Rep ; 52(11): 625-634, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31722780

RESUMO

Severe combined immunodeficiency (SCID) is a group of inherited disorders characterized by compromised T lymphocyte differentiation related to abnormal development of other lymphocytes [i.e., B and/or natural killer (NK) cells], leading to death early in life unless treated immediately with hematopoietic stem cell transplant. Functional NK cells may impact engraftment success of life-saving procedures such as bone marrow transplantation in human SCID patients. Therefore, in animal models, a T cell-/B cell-/NK cell+ environment provides a valuable tool for understanding the function of the innate immune system and for developing targeted NK therapies against human immune diseases. In this review, we focus on underlying mechanisms of human SCID, recent progress in the development of SCID animal models, and utilization of SCID pig model in biomedical sciences. Numerous physiologies in pig are comparable to those in human such as immune system, X-linked heritability, typical T-B+NK- cellular phenotype, and anatomy. Due to analogous features of pig to those of human, studies have found that immunodeficient pig is the most appropriate model for human SCID. [BMB Reports 2019; 52(11): 625-634].


Assuntos
Modelos Animais de Doenças , Imunodeficiência Combinada Severa/imunologia , Suínos/imunologia , Animais , Humanos , Medicina Regenerativa , Transplante Heterólogo/métodos
19.
Sci Immunol ; 4(41)2019 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-31676497

RESUMO

The success of organ transplantation is limited by the complications of immunosuppression, by chronic rejection, and by the insufficient organ supply, and thousands of patients die every year while waiting for a transplant. With recent progress in xenotransplantation permitting porcine organ graft survival of months or even years in nonhuman primates, there is renewed interest in its potential to alleviate the organ shortage. Many of these advances are the result of our heightened capacity to modify pigs genetically, particularly with the development of CRISPR-Cas9-based gene editing methodologies. Although this approach allows the engineering of pig organs that are less prone to rejection, the clinical application of xenotransplantation will require the ability to avoid the ravages of a multifaceted attack on the immune system while preserving the capacity to protect both the recipient and the graft from infectious microorganisms. In this review, we will discuss the potential and limitations of these modifications and how the engineering of the graft can be leveraged to alter the host immune response so that all types of immune attack are avoided.


Assuntos
Sobrevivência de Enxerto/imunologia , Transplante de Órgãos , Suínos , Animais , Humanos , Suínos/imunologia , Engenharia Tecidual
20.
Front Immunol ; 10: 2561, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31736976

RESUMO

Natural killer (NK) cells are cells of the innate immunity and play an important role in the defense against viral infections and cancer, but also contribute to shaping adaptive immune responses. Long-lived tissue-resident NK cells have been described in man and mouse, particularly in the liver, contributing to the idea that the functional palette of NK cells may be broader than originally thought, and may include memory-like responses and maintaining tissue homeostasis. Remarkably, liver resident (lr)NK cells in man and mouse show substantial species-specific differences, in particular reverse expression patterns of the T-box transcription factors Eomesodermin (Eomes) and T-bet (EomeshighT-betlow in man and vice versa in mouse). In pig, compared to blood NK cells which are CD3-CD8αhigh cells, the porcine liver contains an abundant additional CD3-CD8αdim NK cell subpopulation. In the current study, we show that this porcine CD3-CD8αdim liver NK population is highly similar to its human lrNK counterpart and therefore different from mouse lrNK cells. Like human lrNK cells, this porcine NK cell population shows an EomeshighT-betlow expression pattern. In addition, like its human counterpart, the porcine liver NK population is CD49e- and CXCR6+. Furthermore, the porcine EomeshighT-betlow liver NK cell population is able to produce IFN-γ upon IL-2/12/18 stimulation but lacks the ability to kill K562 or pseudorabies virus-infected target cells, although limited degranulation could be observed upon incubation with K562 cells or upon CD16 crosslinking. All together, these results show that porcine EomeshighT-betlow NK cells in the liver strongly resemble human lrNK cells, and therefore indicate that the pig may represent a unique model to study the function of these lrNK cells in health and disease.


Assuntos
Citocinas/imunologia , Células Matadoras Naturais/imunologia , Fígado/imunologia , Suínos/imunologia , Animais , Encefalomielite/imunologia , Encefalomielite/patologia , Encefalomielite/veterinária , Herpesvirus Suídeo 1/imunologia , Humanos , Células K562 , Células Matadoras Naturais/virologia , Fígado/patologia , Fígado/virologia , Suínos/virologia , Doenças dos Suínos/imunologia , Doenças dos Suínos/patologia
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