Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 342
Filter
1.
Front Immunol ; 15: 1361240, 2024.
Article in English | MEDLINE | ID: mdl-38698868

ABSTRACT

N-glycosylation influences the effectiveness of immune globulin G (IgG) and thus the immunological downstream responses of immune cells. This impact arises from the presence of N-glycans within the Fc region, which not only alters the conformation of IgG but also influences its steric hindrance. Consequently, these modifications affect the interaction between IgG and its binding partners within the immune system. Moreover, this posttranslational modification vary according to the physiological condition of each individual. In this study, we examined the N-glycosylation of IgG in pigs from birth to five months of age. Our analysis identified a total of 48 distinct N-glycan structures. Remarkably, we observed defined changes in the composition of these N-glycans during postnatal development. The presence of agalactosylated and sialylated structures increases in relation to the number of N-glycans terminated by galactose residues during the first months of life. This shift may indicate a transition from passively transferred antibodies from the colostrum of the sow to the active production of endogenous IgG by the pig's own immune system.


Subject(s)
Glycosylation , Immunoglobulin G , Protein Processing, Post-Translational , Sus scrofa , Female , Male , Immunoglobulin G/analysis , Immunoglobulin G/isolation & purification , Immunoglobulin G/metabolism , Polysaccharides/analysis , Sialic Acids/analysis , Sus scrofa/growth & development , Sus scrofa/immunology , Sus scrofa/metabolism , Weaning , Animals
2.
Front Immunol ; 12: 784978, 2021.
Article in English | MEDLINE | ID: mdl-34899750

ABSTRACT

In recent years, the increase in awareness of antimicrobial resistance together with the societal demand of healthier meat products have driven attention to health-related traits in livestock production. Previous studies have reported medium to high heritabilities for these traits and described genomic regions associated with them. Despite its genetic component, health- and immunity-related traits are complex and its study by association analysis with genomic markers may be missing some information. To analyse multiple phenotypes and gene-by-gene interactions, systems biology approaches, such as the association weight matrix (AWM), allows combining genome wide association study results with network inference algorithms. The present study aimed to identify gene networks, key regulators and candidate genes associated to immunocompetence in pigs by integrating multiple health-related traits, enriched for innate immune phenotypes, using the AWM approach. The co-association network analysis unveiled a network comprised of 3,636 nodes (genes) and 451,407 edges (interactions), including a total of 246 regulators. From these, five genes (ARNT2, BRMS1L, MED12L, SUPT3H and TRIM25) were selected as key regulators as they were associated with the maximum number of genes with the minimum overlapping (1,827 genes in total). The five regulators were involved in pathways related to immunity such as lymphocyte differentiation and activation, platelet activation and degranulation, megakaryocyte differentiation, FcγR-mediated phagocytosis and response to nitric oxide, among others, but also in immunometabolism. Furthermore, we identified genes co-associated with the key regulators previously reported as candidate genes (e.g., ANGPT1, CD4, CD36, DOCK1, PDE4B, PRKCE, PTPRC and SH2B3) for immunity traits in humans and pigs, but also new candidate ones (e.g., ACSL3, CXADR, HBB, MMP12, PTPN6, WLS) that were not previously described. The co-association analysis revealed new regulators associated with health-related traits in pigs. This approach also identified gene-by-gene interactions and candidate genes involved in pathways related to cell fate and metabolic and immune functions. Our results shed new light in the regulatory mechanisms involved in pig immunity and reinforce the use of the pig as biomedical model.


Subject(s)
Gene Regulatory Networks/immunology , Immunity, Innate/genetics , Quantitative Trait Loci/immunology , Sus scrofa/genetics , Animal Husbandry , Animals , Female , Genome-Wide Association Study , Male , Models, Animal , Sus scrofa/immunology
3.
BMC Genomics ; 22(1): 717, 2021 Oct 05.
Article in English | MEDLINE | ID: mdl-34610786

ABSTRACT

BACKGROUND: In recent years, animal welfare and health has become more and more important in pig breeding. So far, numerous parameters have been considered as important biomarkers, especially in the immune reaction and inflammation. Previous studies have shown moderate to high heritabilities in most of these traits. However, the genetic background of health and robustness of pigs needs to be extensively clarified. The objective of this study was to identify genomic regions with a biological relevance for the immunocompetence of piglets. Genome-wide Association Studies (GWAS) in 535 Landrace (LR) and 461 Large White (LW) piglets were performed, investigating 20 immune relevant traits. Besides the health indicators of the complete and differential blood count, eight different cytokines and haptoglobin were recorded in all piglets and their biological dams to capture mediating processes and acute phase reactions. Additionally, all animals were genotyped using the Illumina PorcineSNP60v2 BeadChip. RESULTS: In summary, GWAS detected 25 genome-wide and 452 chromosome-wide significant SNPs associated with 17 immune relevant traits in the two maternal pig lines LR and LW. Only small differences were observed considering the maternal immune records as covariate within the statistical model. Furthermore, the study identified across- and within-breed differences as well as relevant candidate genes. In LR more significant associations and related candidate genes were detected, compared with LW. The results detected in LR and LW are partly in accordance with previously identified quantitative trait loci (QTL) regions. In addition, promising novel genomic regions were identified which might be of interest for further detailed analysis. Especially putative pleiotropic regions on SSC5, SSC12, SSC15, SSC16 and SSC17 are of major interest with regard to the interacting structure of the immune system. The comparison with already identified QTL gives indications on interactions with traits affecting piglet survival and also production traits. CONCLUSION: In conclusion, results suggest a polygenic and breed-specific background of immune relevant traits. The current study provides knowledge about regions with biological relevance for health and immune traits. Identified markers and putative pleiotropic regions provide first indications in the context of balancing a breeding-based modification of the porcine immune system.


Subject(s)
Immune System , Sus scrofa , Animals , Genetic Association Studies/veterinary , Genotype , Phenotype , Quantitative Trait Loci , Sus scrofa/genetics , Sus scrofa/immunology , Swine
4.
Arq. bras. med. vet. zootec. (Online) ; 73(5): 1047-1057, Sept.-Oct. 2021. tab, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1345276

ABSTRACT

Colostrum is the main source of immunoglobulins (Ig) for neonate piglets and plays a crucial role within the health and growth of the piglet. Currently in pig farming, there are still no widespread practical methods for measuring the Ig concentration in colostrum at herd level. We evaluated sows' colostrum IgG concentration using an optical and a digital Brix refractometer and their performance was correlated to an IgG ELISA test, and flow cytometry. Colostrum concentrations of IgG and IgA averaged 74.05 ± 21.37mg/mL and 20.2 ± 5.32mg/mL respectively. The mean value of the Brix percentages for optical refractometer was 26.32%, and for digital was 28.32%. The Brix refractometer measurements of colostrum samples presented high correlation for IgG content analyzed by ELISA (Optical = 0.74, Digital = 0.87; P <0.001). Considering the immunophenotyping, the values for IgG and IgA lymphoblasts indicated a highly significant relationship to ELISA (IgG=0.77, IgA=0.84; P<0.001). The Brix refractometer can be considered a useful tool to be included in a colostrum monitoring program to improve potentially neonatal health. In addition, we demonstrated that flow cytometry can be an important tool to analyze and characterize the immunological potential of sow colostrum.(AU)


O colostro é a principal fonte de imunoglobulinas (Ig) para leitões recém-nascidos e desempenha um papel crucial na saúde e no crescimento dos leitões. Atualmente, na suinocultura, ainda não existem métodos amplamente utilizados na prática de produção para medir a concentração de imunoglobulinas no colostro suíno. Avaliou-se a concentração de IgG no colostro de porcas usando refratômetros Brix óptico e digital, e o desempenho foi comparado com ELISA e citometria de fluxo. As concentrações de IgG e IgA no colostro foram 74,05 ± 21,37mg/mL e 20,2 ± 5,32mg/mL, respectivamente. A percentagem de Brix média das amostras de colostro para o refratômetro óptico foi 26,32%, e para o digital foi 28,32%. As medições dos refratômetros de Brix apresentaram elevada correlação com a concentrações de IgG medidas por ELISA (óptico=0,74, digital=0,87; P<0,001). Considerando a imunofenotipagem, os valores dos linfoblastos IgG e IgA apresentaram alta correlação com o ELISA (IgG=0,77, IgA=0,84; P<0,001). O refratômetro Brix pode ser considerado uma ferramenta útil para ser incluída em um programa de monitoramento de colostro para melhorar a saúde neonatal. Além disso, foi demonstrado que a citometria de fluxo pode ser uma ferramenta importante para analisar e caracterizar o potencial imunológico do colostro de porcas.(AU)


Subject(s)
Animals , Female , Pregnancy , Immunoglobulin G , Colostrum , Sus scrofa/immunology , Immunoglobulin A , Flow Cytometry/veterinary
6.
Genes (Basel) ; 12(9)2021 08 31.
Article in English | MEDLINE | ID: mdl-34573362

ABSTRACT

Enhancing resistance and tolerance to pathogens remains an important selection objective in the production of livestock animals. Single nucleotide polymorphisms (SNPs) vary gene expression at the transcriptional level, influencing an individual's immune regulation and susceptibility to diseases. In this study, we investigated the distribution of SNP sites in immune-related genes and their correlations with cell surface markers of immune cells within purebred (Taiwan black, Duroc, Landrace and Yorkshire) and crossbred (Landrace-Yorkshire) pigs. Thirty-nine SNPs of immune-related genes, including 11 cytokines, 5 chemokines and 23 Toll-like receptors (TLRs) (interferon-α and γ (IFN-α, γ), tumor necrosis factor-α (TNF-α), granulocyte-macrophage colony-stimulating factor (GM-CSF), Monocyte chemoattractant protein-1 (MCP-1) and TLR3, TLR4, TLR7, TLR8, and TLR9) were selected, and the percentages of positive cells with five cell surface markers of CD4, CD8, CD80/86, MHCI, and MHCII were analyzed. There were 28 SNPs that were significantly different among breeds, particularly between Landrace and Taiwan black. For instance, the frequency of SNP1 IFN-α -235A/G in Taiwan black and Landrace was 11.11% and 96.15%, respectively. In addition, 18 SNPs significantly correlated with the expression of cell surface markers, including CD4, CD8, CD80/86, and MHCII. The percentage of CD4+ (39.27%) in SNP33 TLR-8 543C/C was significantly higher than those in A/C (24.34%), at p < 0.05. Together, our findings show that Taiwan black pigs had a unique genotype distribution, whereas Landrace and Yorkshire had a more similar genotype distribution. Thus, an understanding of the genetic uniqueness of each breed could help to identify functionally important SNPs in immunoregulation.


Subject(s)
Disease Resistance/genetics , Genetic Predisposition to Disease , Sus scrofa/genetics , Animals , Biomarkers , Immunophenotyping , Polymorphism, Single Nucleotide , Selective Breeding , Sus scrofa/blood , Sus scrofa/immunology
7.
BMC Vet Res ; 17(1): 166, 2021 Apr 15.
Article in English | MEDLINE | ID: mdl-33858408

ABSTRACT

BACKGROUND: Health and growth of pigs are affected by the hygiene of housing. Lower growth performance observed in poor hygiene of housing conditions is explained by reduced feed intake and metabolic changes caused by the activation of body defences. In a previous experiment, we reported contrasted average values of body weight gain, concentrations of circulating metabolites, redox and immune indicators in blood of pigs housed in good or poor hygiene conditions during the growing period. This study addressed inter-individual variability in these responses to determine whether a particular blood profile explains average daily gain (ADG) of the pig. RESULTS: The data originated from 160 growing pigs, half of which subjected to a hygiene challenge for 6 weeks (W0 to W6) and the others housed in good hygiene conditions. Pigs originated from two lines divergently selected for residual feed intake (RFI). Individual body weights were recorded during this period, and relative ADG (rADGW0-W6) was calculated as the ADG corrected by the initial body weight measured at W0. Blood samples were taken before (W0) and 3 weeks (W3) after the beginning of the challenge. The analysed dataset consisted of 51 metabolites and indicators of immune and inflammatory responses measured on 136 pigs having no missing value for any variables, when calculated as the differences W3 minus W0 in circulating concentrations. An algorithm tested all possible linear regression models and then selected the best ones to explain rADGW0-W6. Six variables were identified across the best models and correlated with rADGW0-W6 with a goodness of fit (adjusted R2) of about 67%. They were changes in haptoglobin, global antioxidant capacity of plasma (Biological Antioxidant Power or BAP), free fatty acids, and 3 amino acids: leucine, tryptophan, and 1-methylhistidine. The effects of housing conditions and RFI lines were comprised in the variables of the selected models and none of these conditions improved accuracy of the predictive models, leading to genericity of the pinpointed metabolic changes in relation to variability of ADG. CONCLUSIONS: This approach allows us to identify blood variables, whose changes in blood concentrations correlated to ADG under contrasted sanitary conditions.


Subject(s)
Housing, Animal/standards , Hygiene , Sus scrofa/growth & development , Animal Husbandry/methods , Animals , Body Weight/physiology , Eating/genetics , Eating/physiology , Female , Male , Sus scrofa/blood , Sus scrofa/immunology , Sus scrofa/metabolism
8.
Vet Parasitol Reg Stud Reports ; 23: 100522, 2021 01.
Article in English | MEDLINE | ID: mdl-33678377

ABSTRACT

The protozoan Neospora caninum is an important cause of abortion in cattle worldwide, with domestic dogs serving as the definitive hosts. Although hunting activities have been indicated as a potential risk factor for N. caninum infection in dogs, no serological evidence has so far been reported in hunting dogs, wild boars (Sus scrofa), and hunters. A total of 98 wild boars, 168 hunting dogs, and 15 hunters from three Brazilian regions were sampled and tested for anti-N. caninum antibodies by indirect immunofluorescence antibody test, resulting in 15/168 (9%) seropositive hunting dogs, and 0/98 wild boars, and 0/15 hunters seropositive. The absence of N. caninum antibodies in wild boars may suggest low exposure to oocysts shed by hunting dogs and wild canids in three different Brazilian regions. Finally, the absence of hunter seropositivity supports the current lack of evidence that N. caninum is a zoonotic parasite.


Subject(s)
Antibodies, Protozoan/blood , Neospora , Sus scrofa/blood , Working Dogs , Animals , Brazil/epidemiology , Dogs , Seroepidemiologic Studies , Sus scrofa/immunology , Working Dogs/blood , Working Dogs/immunology
9.
Arch Anim Nutr ; 75(2): 121-136, 2021 Apr.
Article in English | MEDLINE | ID: mdl-33557604

ABSTRACT

The objective of this study was to determine whether dietary tea tree oil (TTO) supplementation could effectively replace the antibiotics through modulating the antioxidant capacity and intestinal microbiota profile, and then decreasing the diarrhoea incidence and improving the growth performance of weaned pigs. A total of 216 weaned pigs with initial body weights (BW) of 9.19 ± 1.86 kg were randomly allocated to three dietary treatments in a completely randomised design. The dietary treatments included a corn-soybean meal basal diet (CON) without any antibiotics, and two experimental diets formulated by adding 75 mg/kg aureomycin (AGP) or 100 mg/kg TTO into the basal diet, respectively. Pigs fed the TTO diet showed greater gain to feed ratio (p < 0.05) than those fed CON and AGP diets during d 0-14 and d 14-28. Both dietary TTO and AGP supplementation tended to increase the average daily gain of weaned pigs during d 14-28 (p = 0.06) and the overall 28-d period (p = 0.07), and significantly reduced (p < 0.05) the diarrhoea incidence during d 0-14 compared with the CON treatment. In addition, dietary TTO supplementation improved the apparent total tract digestibility of dry matter and ether extract (p < 0.05), and increased (p < 0.05) the propionate and butyrate concentrations in faecal samples of weaned pigs. Moreover, pigs fed the TTO diet showed greater total antioxidant capacity, greater superoxide dismutase and interleukin-10 concentrations, and lower malondialdehyde concentration in serum than those fed the CON diet (p < 0.05). Furthermore, pigs fed the TTO diet demonstrated greater relative abundance of Clostridiaceae_1, while those fed the AGP diet exhibited greater relative abundance of Lactobacillaceae at family level. In conclusion, dietary TTO supplementation could improve growth performance in weaned pigs, which could be mainly attributed to the benefits on nutrient digestibility, antioxidative capacity and microbial community profile.


Subject(s)
Gastrointestinal Microbiome/physiology , Sus scrofa , Tea Tree Oil/metabolism , Animal Feed/analysis , Animal Nutritional Physiological Phenomena , Animals , Diet/veterinary , Dietary Supplements/analysis , Female , Gastrointestinal Microbiome/drug effects , Male , Random Allocation , Sus scrofa/growth & development , Sus scrofa/immunology , Sus scrofa/metabolism , Sus scrofa/microbiology , Tea Tree Oil/administration & dosage
10.
Res Vet Sci ; 136: 127-137, 2021 May.
Article in English | MEDLINE | ID: mdl-33609970

ABSTRACT

Reproductive cycling in fattening gilts can be associated with undesirable effects, such as estrus-related aggressive behavior, reduced feed intake and, in production systems where gilts are co-housed with entire males, unwanted pregnancy. Immunization against Gonadotrophin Releasing Factor (IM) can temporarily suppress ovarian activity, including related negative consequences on animal welfare and productivity. Feed intake has been shown to be higher after IM, resulting in both increased growth and increased carcass fat. A series of studies was conducted to confirm these effects on production and look at their dynamics over time. Three trials were performed to a similar design, each involving 240 gilts divided into 4 experimental groups at 12 weeks of age. One group remained untreated while the others had the two dose, IM course completed 8, 6 or 4 weeks before harvest, which was on a single day at 24, 25 or 26 weeks of age depending on the study. Feed intake was measured daily and bodyweight weekly, allowing growth parameters to be calculated on a weekly basis and for specific longer periods. Carcass weight, backfat depth and lean meat percentage were recorded at harvest. No effects were observed before the second application of the immunological product (V2) and completion of the IM course. Starting in the second week after V2 all IM groups showed a marked and consistent increase in Average Daily Feed Intake (ADFI), typically peaking at over 120% of the control group 3 to 4 weeks after V2 and then slowly declining, but still remaining elevated at 8 weeks. Weekly Average Daily Gain (ADG) showed a similar pattern but with a faster decline, resulting in the initially favorable impact on feed efficiency becoming less favorable as the V2 to harvest interval (V2H) progressed. Carcass weights were higher in IM gilts and backfat depths were greater, with the effects increasing with increasing V2H. Correspondingly, carcass lean meat percentage tended to decrease, although the higher carcass weights meant that the absolute weight of lean meat remained similar or higher. Carcass yield was generally unaffected by IM, but some between-group differences were statistically significant, and it is possible that different factors predominated at different times after V2, creating a complex relationship with V2H duration. The optimum IM protocol will depend on local conditions and production objectives but, as a generalization and assuming ad libitum feeding, a shorter V2H will favor efficient growth, while a longer duration will maximize carcass changes, such as increased fat coverage. It is suggested that the growth performance changes seen after IM in gilts might be viewed as a process of adjustment to a heavier and fatter target body type.


Subject(s)
Gonadotropin-Releasing Hormone/metabolism , Immunization/veterinary , Meat/analysis , Sus scrofa/physiology , Animals , Female , Sus scrofa/growth & development , Sus scrofa/immunology , Time Factors
11.
Vet Med Sci ; 7(4): 1347-1358, 2021 07.
Article in English | MEDLINE | ID: mdl-33620158

ABSTRACT

The purpose of this study was to investigate the effects of adding Pennisetum purpureum (P. purpureum, also known as Napier grass or elephant grass) to the diets of late gestation on the antioxidant indexes, immune indexes and faecal microbiota of sows. At the 90 days of gestation, 300 healthy sows were randomly divided into three groups, and they received the basic commercial diet or added 5% P. purpureum and 10% P. purpureum, respectively. The experiment started from 90 days of gestation to parturition. The results showed that the total antioxidant capacity, immunoglobulins and serum equol concentrations of sows on 100 days of gestation and at parturition increased linearly (p < .05) with the increase of the content of P. purpureum in the gestation diet. The 5% P. purpureum increased the relative abundance of Bacteroidetes (p = .027) and Actinobacteria (p < .001) at phylum level, Coriobacteriaceae (p < .001) at family level and Prevotellaceae_UCG_001 (p = .004) at genus level, and decreased the relative abundance of Escherichia_Shigella (p < .001) at genus level. In summary, this study shows that the additive of P. purpureum can increase the concentration of serum equol, improve the antioxidant capacity and immune function of sow in late gestation. In addition, the additive of 5% P. purpureum in the diet might change the composition of intestinal microbiota of sows, particularly the relative abundance of Coriobacteriaceae (p < .001) increased.


Subject(s)
Antioxidants/metabolism , Dietary Supplements/analysis , Feces/microbiology , Immunity, Innate , Microbiota , Pennisetum/chemistry , Pregnancy, Animal/physiology , Sus scrofa/immunology , Animal Feed/analysis , Animals , Diet/veterinary , Female , Immunity, Innate/drug effects , Microbiota/drug effects , Pregnancy , Pregnancy, Animal/drug effects
12.
Rapid Commun Mass Spectrom ; 35(9): e9063, 2021 May 15.
Article in English | MEDLINE | ID: mdl-33538041

ABSTRACT

RATIONALE: While high-throughput proteomic methods have been widely applied to monoclonal antibodies and human immunoglobulin gamma (IgG) samples, less information is available on porcine IgG. As pigs are considered one of the most suitable species for xenotransplantation, it is important to characterize IgG amino acid sequences and glycosylation profiles, which is the focus of this study. METHODS: Three different purified porcine IgG samples, including wild-type and knockout species, were digested with trypsin and enriched for glycopeptides. Digestion mixtures were spiked with a mixture of six standard peptides. Analysis was performed using electrospray ionization liquid chromatography-tandem mass spectrometry (MS/MS) in standard MS/MS data-dependent acquisition mode on a hybrid triple quadrupole time-of-flight mass spectrometer. RESULTS: To facilitate the classification of subtypes detected experimentally, UniprotKB database entries were organized using comparative alignment scores. Sequences were grouped based on 11 different subtypes as translated from GenBank entries. Proteomic searches were accomplished automatically using specialized software, whereas glycoprotein searches were performed manually by monitoring the extracted chromatograms of diagnostic MS/MS glycan fragments and studying their corresponding mass spectra; 40-50 non-glycosylated peptides and 4-5 glycosylated peptides were detected in each sample, with several glycoforms per sequence. CONCLUSIONS: Proteomic analysis of porcine IgG is complicated by factors such as the presence of several subtypes, redundant heavy chain (HC) sequences in protein databases, and the lack of consistent cross-referencing between databases. Aligning and comparing HC sequences were necessary to eliminate redundancy. This study highlights the complexity of pig IgG and shows the importance of MS in proteomics and glycoproteomics.


Subject(s)
Chromatography, Liquid/veterinary , Glycoproteins/analysis , Immunoglobulin G/chemistry , Proteomics/methods , Sus scrofa/immunology , Tandem Mass Spectrometry/veterinary , Amino Acid Sequence , Animals , Chromatography, Liquid/methods , Gene Knockout Techniques , Glycopeptides/analysis , Glycopeptides/chemistry , Glycoproteins/metabolism , Glycosylation , Immunoglobulin G/genetics , Immunoglobulin G/metabolism , Immunoglobulin Heavy Chains/chemistry , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Tandem Mass Spectrometry/methods
13.
Transplantation ; 105(2): 300-307, 2021 02 01.
Article in English | MEDLINE | ID: mdl-32433239

ABSTRACT

Advances in genetic engineering, particularly CRISPR/Cas9, have resulted in the development of a triple glycan-knockout (TKO) pig. There is minimal human antipig antibody binding to TKO pig cells. The TKO background has decreased antibody binding to a sufficiently low level that any additional xenoantigens expressed on the cells can now be more easily detected. One of these xenoantigens is the swine major histocompatibility complex, termed swine leukocyte antigens (SLA). SLA are the homolog to HLAs, a protein complex expressed on human tissue capable of stimulating the development of new antibodies in allotransplantation. These antibodies can result in graft failure through hyperacute, acute, or chronic rejection. Our knowledge of SLA, particularly in the last 5 years, has grown considerably. The presence, cause, and methods to detect anti-SLA antibodies will need to be carefully considered for the first clinical trial of xenotransplantation. The focus of this review is to summarize the role of SLA in xenotransplantation and consider whether it will prove to be a major barrier. Techniques are now available to mutate target SLA amino acids to ensure that cross-reactive anti-HLA antibodies no longer bind to SLA on the cells of the organ-source pigs. While deletion of SLA expression is possible, it would render the pig at risk for infectious complications. The ideal organ-source pig for HLA highly sensitized recipients may therefore be 1 with site-specific mutations to eliminate cross-reactive binding.


Subject(s)
Antibodies, Heterophile/blood , Antigens, Heterophile/immunology , Graft Rejection/immunology , Histocompatibility Antigens Class I/immunology , Sus scrofa/immunology , Transplantation, Heterologous , Animals , Animals, Genetically Modified , Antibody Specificity , Antigens, Heterophile/genetics , Graft Rejection/blood , Graft Rejection/prevention & control , Graft Survival , Histocompatibility Antigens Class I/genetics , Humans , Species Specificity , Sus scrofa/genetics , Transplantation Tolerance , Transplantation, Heterologous/adverse effects
14.
Nat Biomed Eng ; 5(2): 134-143, 2021 02.
Article in English | MEDLINE | ID: mdl-32958897

ABSTRACT

The clinical applicability of porcine xenotransplantation-a long-investigated alternative to the scarce availability of human organs for patients with organ failure-is limited by molecular incompatibilities between the immune systems of pigs and humans as well as by the risk of transmitting porcine endogenous retroviruses (PERVs). We recently showed the production of pigs with genomically inactivated PERVs. Here, using a combination of CRISPR-Cas9 and transposon technologies, we show that pigs with all PERVs inactivated can also be genetically engineered to eliminate three xenoantigens and to express nine human transgenes that enhance the pigs' immunological compatibility and blood-coagulation compatibility with humans. The engineered pigs exhibit normal physiology, fertility and germline transmission of the 13 genes and 42 alleles edited. Using in vitro assays, we show that cells from the engineered pigs are resistant to human humoral rejection, cell-mediated damage and pathogenesis associated with dysregulated coagulation. The extensive genome engineering of pigs for greater compatibility with the human immune system may eventually enable safe and effective porcine xenotransplantation.


Subject(s)
CRISPR-Cas Systems , Genetic Engineering/methods , Germ Cells/metabolism , Sus scrofa/genetics , Sus scrofa/virology , Transplantation, Heterologous , Animals , CRISPR-Associated Protein 9/genetics , Cells, Cultured , Galactosyltransferases/genetics , Gene Knockout Techniques , Mixed Function Oxygenases/genetics , N-Acetylgalactosaminyltransferases/genetics , Sus scrofa/immunology
15.
Transbound Emerg Dis ; 68(1): 183-193, 2021 Jan.
Article in English | MEDLINE | ID: mdl-32652870

ABSTRACT

Although vaccination is the main measure to control influenza A virus (IAV) in swine, there is limited information on the efficacy of sow vaccination on reducing IAV infections in pigs at weaning. We assessed the effect of sow vaccination on IAV infection in pigs at weaning in a cohort of 52 breeding herds studied prospectively. Herds were voluntarily enrolled according to their IAV history, sow vaccination protocol and monitored during six months (prospective longitudinal study). On each herd, nasal swabs were collected monthly from 30 pigs at weaning and tested for IAV by RT-PCR. IAV was detected in 25% (75/305) of sampling events. Of 9,150 nasal swab pools (3 individual nasal swabs/pool), 15% (458/3050) of pools tested IAV positive. IAV infections in pigs at weaning were lower in vaccinated herds compared to non-vaccinated ones. Moreover, no significant differences were seen between prefarrow and whole herd protocols, or the use of commercial versus autogenous IAV vaccines. Prefarrow and whole herd vaccination protocols reduced the odds of groups testing IAV positive at weaning in comparison with no vaccination. Our results are relevant when considering implementation of sow vaccination to control influenza infections in pigs at weaning and, hence, minimize transmission to growing pigs and other farms.


Subject(s)
Influenza A virus/immunology , Influenza Vaccines/administration & dosage , Orthomyxoviridae Infections/veterinary , Swine Diseases/prevention & control , Vaccination/veterinary , Weaning , Animals , Female , Longitudinal Studies , Orthomyxoviridae Infections/prevention & control , Prospective Studies , Sus scrofa/immunology , Sus scrofa/physiology , Swine
16.
Transplantation ; 105(2): 318-327, 2021 02 01.
Article in English | MEDLINE | ID: mdl-32796494

ABSTRACT

BACKGROUND: Natural preformed anti-pig IgM/IgG antibodies in primates play an important role in xenograft rejection. As it is not clear how IgE and IgA engage in the immune system in xenotransplantation, we investigated natural preformed and elicited anti-pig IgE/IgA in naive primates and after xenotransplantation in nonhuman primates. METHODS: The binding of IgM/IgG/IgE/IgA antibodies to red blood cells (RBCs) from wild-type (WT), α1,3-galactosyltransferase gene-knockout (GTKO), and GTKO/cytidine monophospho-N-acetylneuraminic acid hydroxylase gene-knockout/ß-1,4 N-acetylgalactosaminyltransferase 2 gene-knockout (ie, triple-knockout pigs) pigs were measured by flow cytometry in naive human (n = 50) and baboon (n = 14) sera. Antibody binding to WT and GTKO pig RBCs (pRBCs) was also measured in the sera of baboons (nonsensitized n = 7, sensitized n = 2) and rhesus monkeys (nonsensitized n = 2, sensitized n = 11) following WT or GTKO pig organ/tissue xenotransplantation. Deposition of IgM/IgG/IgE/IgA in the grafts was detected by immunohistochemistry. RESULTS: The majority of humans had natural preformed IgM/IgG/IgE/IgA to WT and GTKO pRBCs. In contrast, IgM/IgG/IgE/IgA to triple-knockout pRBCs were present at lower levels and frequency (P < 0.01). Baboons also had IgM/IgG/IgE/IgA antibodies against WT pRBCs, but fewer to GTKO and triple-knockout (P < 0.01). After xenotransplantation into nonhuman primates, when IgM/IgG increased, IgE/IgA also increased, but to a lesser extent. In addition to IgM/IgG, IgE or IgA deposition was observed in rejected pig xenografts. CONCLUSIONS: Primates develop serum anti-pig IgE/IgA antibodies both naturally and during xenograft rejection. The pathophysiological role, if any, of anti-pig IgE/IgA antibodies remains unknown.


Subject(s)
Antibodies, Heterophile/blood , Erythrocytes/immunology , Graft Rejection/immunology , Immunoglobulin A/blood , Immunoglobulin E/blood , Transplantation, Heterologous/adverse effects , Animals , Animals, Genetically Modified , Galactosyltransferases/genetics , Galactosyltransferases/immunology , Graft Rejection/blood , Graft Rejection/pathology , Humans , Macaca mulatta , Mixed Function Oxygenases/genetics , Mixed Function Oxygenases/immunology , N-Acetylgalactosaminyltransferases/genetics , N-Acetylgalactosaminyltransferases/immunology , Papio , Species Specificity , Sus scrofa/genetics , Sus scrofa/immunology
17.
J Appl Anim Welf Sci ; 24(3): 246-259, 2021.
Article in English | MEDLINE | ID: mdl-32894989

ABSTRACT

Single- and group-housed pregnant sows were assessed during 8 weeks (4th and 8th to 14th) with regard to physiological, reproductive, welfare indicators and gene expression profiling. Compared to single-housed sows, group-housed sows had decreased non-return to estrus at 56 days after artificial insemination (AI) (83% vs 92%) and farrowing rate (78% vs 88%), respectively. Furthermore, group-housed sows showed a higher degree (p < 0.01) of advantageous physiological indicators, such as albumin (odds ratio [OR] = 4.4), alkaline phosphatase (OR = 1.5), bactericidal (OR = 3.2) and complement (OR = 24.3), and disadvantageous for alanine amino transferase (OR = 0.5), bilirubin (OR = 0.4), lysozyme (OR = 0.3) and C-reactive protein compared to single-housed. Eighty-seven genes related to immune response were underexpressed (log fold change ≤ 1.5; p < 0.05) during the 8th to 14th weeks in the group compared to single-housed sows, which in turn showed an immunomodulatory reduction on the expression of 43 genes during the 11th to 14th compared to the 4th week. Overall, the results were interpreted as indicative of greater comfort state of the group compared to single-housed sows.


Subject(s)
Animal Welfare , Housing, Animal , Sus scrofa/physiology , Animal Husbandry/methods , Animals , Female , Fertility , Immunity/genetics , Pregnancy , Sus scrofa/immunology , Transcriptome
18.
Trop Anim Health Prod ; 53(1): 43, 2020 Nov 24.
Article in English | MEDLINE | ID: mdl-33231781

ABSTRACT

This research investigated the impact of dietary beta-glucan-rich molasses yeast powder (MYP) supplementation on the antibody response to swine fever virus (Titer) and hematology of starter-grower pig. Sixteen cross pigs (30 kg body weight) were equally split into four groups; each group with four replicates and fed four dietary treatments that consisted of basal diets (control) and the basal diets added with 2.5, 5.0, and 7.5% MYP. Feed and water were consumed ad libitum for 44 days. Feed intake (FI), MYP intake (MYPI), beta-glucan intake (BGI), and Mannan-oligosaccharide intake (MOSI) were recorded daily. Titer was evaluated after 15 (Titer15) and 30 (Titer30) days after vaccination, while hematology was analyzed at the end of the experiment. The results indicated that it was unchangeable for ADFI (P > 0.05). No impacts were observed on hematological variables and Titer15 in MYP fed pigs (P > 0.05). However, supplementation with 7.5% MYP increased platelet count (PC) and Titer30 (P < 0.01), but decreased hematocrit (Hct) (P < 0.05). Titer 30 and titer 15 were linked to MYPI, BGI, and MOSI (P < 0.05). Based on the study, feeding starter-grower pigs diets supplemented with 7.5% MYP might enhance the antibody response to swine fever virus 30 days after vaccination, and it has a potential role in the application in prevention of swine fever virus disease.


Subject(s)
Antibody Formation/drug effects , Classical Swine Fever Virus/immunology , Molasses/analysis , Sus scrofa/immunology , Yeast, Dried/metabolism , beta-Glucans/metabolism , Animal Feed/analysis , Animals , Diet/veterinary , Dietary Supplements/analysis , Dose-Response Relationship, Drug , Female , Hematologic Tests/veterinary , Male , Sus scrofa/blood , Yeast, Dried/administration & dosage , beta-Glucans/administration & dosage
19.
Toxins (Basel) ; 12(10)2020 10 09.
Article in English | MEDLINE | ID: mdl-33050248

ABSTRACT

Effects of folic acid and protein levels on growth and serum chemistry in pigs fed aflatoxin were determined in two experiments. Increasing aflatoxin (250 to 800 ppb) decreased (P < 0.05) weight gain and feed intake for both of the 35-day trials. In Experiment 1, increasing aflatoxin (0, 250, 500 ppb), increased linearly (P < 0.05) aspartate aminotransferase (AST), alkaline phosphatase (ALKP) and ɣ-glutamyl transferase (GGT). Folic acid (0, 2.0, 5.0, 12.5 ppm) increased linearly (P < 0.05) serum K, Ca, P, Mg, and AST with the largest effect observed at 12.5 ppm. Folic acid decreased (P < 0.05) blood urea nitrogen (BUN): creatinine and Na:K. In Experiment 2, aflatoxin (800 ppb) increased (P < 0.05) glucose and GGT, and decreased (P < 0.05) Na:K and albumin:globulin. Increasing protein from 15 to 18% elevated BUN: creatinine (P < 0.05), albumin: globulin (P < 0.05), albumin (P < 0.05) and ALKP (P < 0.05). Folic acid (2 ppm) elevated (P < 0.05) BUN, and interacted with both aflatoxin (P < 0.10) and protein (P < 0.05) on BUN. Adding folic acid to aflatoxin contaminated diets improved some measures of clinical chemistry in Experiment 1 but not traditional growth performance measures. The higher protein level reduced the effects of aflatoxicosis on growth.


Subject(s)
Aflatoxins/toxicity , Animal Feed/microbiology , Dietary Proteins/administration & dosage , Dietary Supplements , Folic Acid/administration & dosage , Mycotoxicosis/veterinary , Sus scrofa/growth & development , Swine Diseases/prevention & control , Animals , Biomarkers/blood , Dietary Proteins/metabolism , Mycotoxicosis/immunology , Mycotoxicosis/metabolism , Mycotoxicosis/prevention & control , Sus scrofa/immunology , Sus scrofa/metabolism , Swine , Swine Diseases/metabolism , Weaning
20.
Elife ; 92020 10 07.
Article in English | MEDLINE | ID: mdl-33026343

ABSTRACT

Submucosal glands (SMGs) are a prominent structure that lines human cartilaginous airways. Although it has been assumed that SMGs contribute to respiratory defense, that hypothesis has gone without a direct test. Therefore, we studied pigs, which have lungs like humans, and disrupted the gene for ectodysplasin (EDA-KO), which initiates SMG development. EDA-KO pigs lacked SMGs throughout the airways. Their airway surface liquid had a reduced ability to kill bacteria, consistent with SMG production of antimicrobials. In wild-type pigs, SMGs secrete mucus that emerges onto the airway surface as strands. Lack of SMGs and mucus strands disrupted mucociliary transport in EDA-KO pigs. Consequently, EDA-KO pigs failed to eradicate a bacterial challenge in lung regions normally populated by SMGs. These in vivo and ex vivo results indicate that SMGs are required for normal antimicrobial activity and mucociliary transport, two key host defenses that protect the lung.


Subject(s)
Ectodysplasins/genetics , Exocrine Glands/immunology , Respiratory Mucosa/immunology , Staphylococcus aureus/physiology , Sus scrofa/immunology , Animals , Ectodysplasins/immunology , Female , Gene Knockout Techniques , Male , Sus scrofa/genetics
SELECTION OF CITATIONS
SEARCH DETAIL