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ABSTRACT: We evaluated differences in gene expression in pigs from the Porcine Reproductive and Respiratory Syndrome (PRRS) Host Genetics Consortium initiative showing a range of responses to PRRS virus infection. Pigs were allocated into four phenotypic groups according to their serum viral level and weight gain. RNA obtained from blood at 0, 4, 7, 11, 14, 28, and 42 days post-infection (DPI) was hybridized to the 70-mer 20K Pigoligoarray. We used a blocked reference design for the microarray experiment. This allowed us to account for individual biological variation in gene expression, and to assess baseline effects before infection (0 DPI). Additionally, this design has the flexibility of incorporating future data for differential expression analysis. We focused on evaluating transcripts showing significant interaction of weight gain and serum viral level. We identified 491 significant comparisons [false discovery rate (FDR) = 10%] across all DPI and phenotypic groups. We corroborated the overall trend in direction and level of expression (measured as fold change) at 4 DPI using qPCR (r = 0.91, p ≤ 0.0007). At 4 and 7 DPI, network and functional analyses were performed to assess if immune related gene sets were enriched for genes differentially expressed (DE) across four phenotypic groups. We identified cell death function as being significantly associated (FDR ≤ 5%) with several networks enriched for DE transcripts. We found the genes interferon-alpha 1(IFNA1), major histocompatibility complex, class II, DQ alpha 1 (SLA-DQA1), and major histocompatibility complex, class II, DR alpha (SLA-DRA) to be DE (p ≤ 0.05) between phenotypic groups. Finally, we performed a power analysis to estimate sample size and sampling time-points for future experiments. We concluded the best scenario for investigation of early response to PRRSV infection consists of sampling at 0, 4, and 7 DPI using about 30 pigs per phenotypic group.
Frontiers in genetics. 01/2012; 3:321.
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ABSTRACT: This report describes the cloning and characterization of expressed gene sequences of bovine, equine, and swine CXCL9 from RNA obtained from peripheral blood mononuclear cells (PBMC) and other tissues. The bovine coding region was 378 nucleotides in length, while the equine and swine coding regions were 381 nucleotides. Mapping showed that all three sequences were coded for in four exons in the genome, as are the human and mouse genes. The bovine, equine, and swine coding regions shared 83%, 86%, and 84% homology with human CXCL9, respectively, and all three were 74% homologous with mouse CXCL9. Cladogram comparison of the nucleotide sequences of CXCL9 showed that the bovine, equine and swine sequences were more closely related to one another than to either the human or the mouse sequences. However, the human sequence was more closely related to them than it was to the mouse sequence. These relationships were preserved when the deduced amino acid sequences were evaluated and all sequences showed conservation of the characteristic four cysteines. This work sets the stage for further work with these molecules; an integral goal of the U.S. Veterinary Immune Reagent Network is to develop reagents for investigating diseases in livestock species, poultry, and fish.
Veterinary Immunology and Immunopathology 06/2011; 141(3-4):317-21. · 2.08 Impact Factor
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ABSTRACT: The objective of this study was to determine cytokine and chemokine mRNA expression profiles in tracheobronchial lymph nodes from pigs singularly infected with porcine circovirus type 2 (PCV2), Mycoplasma hyopneumoniae (MHYO), or coinfected with both. Twenty-eight pigs were randomly assigned to one of four groups: (1) negative controls (NEG), (2) inoculated with MHYO (IMHYO), (3) inoculated with MHYO and PCV2 (CoI), and (4) inoculated with PCV2 (IPCV2). MHYO infection significantly (P<0.05) stimulated innate cytokines, IL1B and IL8. PCV2 infection significantly stimulated expression of IFNG, IL8, NOS2A and chemokines CCL2, CCL5, and CXCL10. IFNB, IL1B and IL12 were slightly increased with PCV2 infection and IFNA and IL4 were significantly downregulated. Compared to NEG pigs, coinfection resulted in a significant increase in expression of IFNG, IL1B, IL8, CCL5, CXCL10, and weak stimulation of IFNB, IL6 and IL10; IL13 and IFNA were significantly downregulated. Overall MHYO potentiated PCV2 infection by increasing IFNG and IL10 mRNA expression levels. The increase of IFNG and chemokines and decrease of IFNA in IPCV2 and CoI pigs were correlated with increased severity of lymphoid lesions and the presence of PCV2 antigen. In summary, this work provided evidence that the increased severity of lesions in PCV2 and MHYO coinfected pigs was associated mainly with the presence of PCV2 antigen and alterations of cytokine mRNA expression profiles.
Veterinary Immunology and Immunopathology 03/2011; 140(1-2):152-8. · 2.08 Impact Factor
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ABSTRACT: The 3rd Veterinary Immunology Committee (VIC) Toolkit Workshop took place at the 9th International Veterinary Immunology Symposium (IVIS) in Tokyo, Japan on 18th August 2010. The Workshop built on previous Toolkit Workshops and covered various aspects of reagent development, commercialization and provision to the veterinary immunology research community. The emphasis was on open communication about current progress and future plans to avoid duplication of effort and to update priorities for reagent development. There were presentations on the major reagent development and networking projects such as the BBSRC/RERAD Immunological Toolbox (2004-2009), US Veterinary Immune Reagent Network (VIRN 2006-2010) that has just received renewal funding for 2010-2014, and EU Network for Animal Diseases Infectiology Research Facilities project (NADIR 2009-2013). There were also presentations and discussions on the use of reagents for assay development, particularly multiplexing, and how these new technologies will underpin basic research developments. Mechanisms for improved information exchange, especially though websites with VIC playing a central role, were identified.
Veterinary Immunology and Immunopathology 03/2011; 148(1-2):197-201. · 2.08 Impact Factor
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ABSTRACT: The second International Symposium on Animal Genomics for Animal Health held in Paris, France 31 May-2 June, 2010, assembled more than 140 participants representing research organizations from 40 countries. The symposium included a roundtable discussion on critical needs, challenges and opportunities, and a forward look at the potential applications of animal genomics in animal health research. The aim of the roundtable discussion was to foster a dialogue between scientists working at the cutting edge of animal genomics research and animal health scientists. Importantly, stakeholders were included to provide input on priorities and the potential value of animal genomics to the animal health community. In an effort to facilitate the roundtable discussion, the organizers identified four priority areas to advance the use of genome-enabled technologies in animal health research. Contributions were obtained through open discussions and a questionnaire distributed at the start of the symposium. This report provides the outcome of the roundtable discussion for each of the four priority areas. For each priority, problems are identified, including potential solutions and recommendations. This report captures key points made by symposium participants during the roundtable discussion and serves as a roadmap to steer future research priorities in animal genomics research.
BMC proceedings 01/2011; 5 Suppl 4:S1.
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ABSTRACT: Understanding the role of host genetics in resistance to porcine reproductive and respiratory syndrome virus (PRRSV) infection, and the effects of PRRS on pig health and related growth, are goals of the PRRS Host Genetics Consortium (PHGC).
The project uses a nursery pig model to assess pig resistance/susceptibility to primary PRRSV infection. To date, 6 groups of 200 crossbred pigs from high health farms were donated by commercial sources. After acclimation, the pigs were infected with PRRSV in a biosecure facility and followed for 42 days post infection (dpi). Blood samples were collected at 0, 4, 7, 10, 14, 21, 28, 35 and 42 dpi for serum and whole blood RNA gene expression analyses; weekly weights were recorded for growth traits. All data have been entered into the PHGC relational database. Genomic DNAs from all PHGC1-6 pigs were prepared and genotyped with the Porcine SNP60 SNPchip.
Results have affirmed that all challenged pigs become PRRSV infected with peak viremia being observed between 4-21 dpi. Multivariate statistical analyses of viral load and weight data have identified PHGC pigs in different virus/weight categories. Sera are now being compared for factors involved in recovery from infection, including speed of response and levels of immune cytokines. Genome-wide association studies (GWAS) are underway to identify genes and chromosomal locations that identify PRRS resistant/susceptible pigs and pigs able to maintain growth while infected with PRRSV.
Overall, the PHGC project will enable researchers to discover and verify important genotypes and phenotypes that predict resistance/susceptibility to PRRSV infection. The availability of PHGC samples provides a unique opportunity to continue to develop deeper phenotypes on every PRRSV infected pig.
BMC proceedings 01/2011; 5 Suppl 4:S30.
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ABSTRACT: S100 calcium-binding protein A8 (S100A8) and S100 calcium-binding protein A9 (S100A9) are pivotal mediators of inflammatory and protective anti-infection responses for the mammalian host. In this study, we present the molecular cloning of porcine S100A8 (pS100A8) and porcine S100A9 (pS100A9). Both genes comprise 3 exons and 2 introns and are located on pig chromosome 4q21-q23 (closely linked to SW512). Homology comparison to other mammalian species affirmed that critical functional amino acids for post-transcriptional modification, inflammatory regulation, and formation of heterodimeric complexes exist in pS100A8 and pS100A9. Under normal conditions, both genes are preferentially expressed in porcine immune or immune-related organs, e.g., bone marrow, spleen, lymph nodes, and lung. Upon stimulation in porcine whole blood cultures with LPS or Poly(I:C), they are dramatically induced. Interestingly, the maximum increase of mRNA levels in blood cultures of Meishan pigs is significantly greater than that in Duroc pigs. We previously showed that pS100A8 and pS100A9 mRNA were up-regulated following Haemophilus parasuis (HPS) infection. We herein further confirm their up-regulation at the protein level in multiple HPS infected tissues (spleen, lung and liver). Functional cluster and network analysis based on our previous microarray data discovered that CEBPB may be one of the key transcription factors. A pS100A8/pS100A9-CASP3-SLC1A2 pathway regulating lipid metabolism was found. Both of their pro- and anti-inflammatory functions in response to HPS infection are highlighted.
Developmental and comparative immunology 12/2010; 35(4):490-500. · 3.29 Impact Factor
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ABSTRACT: This manuscript focuses on the advances made using genomic approaches to identify biomarkers that define genes and pathways that are correlated with swine resistance to infection with porcine reproductive and respiratory syndrome virus (PRRSV), the most economically important swine viral pathogen worldwide. International efforts are underway to assess resistance and susceptibility to infectious pathogens using tools such as gene arrays, single nucleotide polymorphisms (SNPs) chips, genome-wide association studies (GWAS), proteomics, and advanced bioinformatics. These studies should identify new candidate genes and biological pathways associated with host PRRS resistance and alternate viral disease processes and mechanisms; they may unveil biomarkers that account for genetic control of PRRS or, alternately, that reveal new targets for therapeutics or vaccines. Previous genomic approaches have expanded our understanding of quantitative trait loci (QTL) controlling traits of economic importance in pig production, e.g., feed efficiency, meat production, leanness; only recently have these included health traits and disease resistance. Genomic studies should have substantial impact for the pig industry since it is now possible to include the use of biomarkers for basic health traits alongside broader set of markers utilized for selection of pigs for improved performance and reproductive traits, as well as pork quality. Additionally these studies may reveal alternate PRRS control mechanisms that can be exploited for novel drugs, biotherapeutics and vaccine designs.
Virus Research 12/2010; 154(1-2):161-9. · 2.94 Impact Factor
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ABSTRACT: Recognized in the late 1980s in North America and Europe the syndrome that caused reproductive and respiratory problems in swine was initially called "mystery swine disease" and is now termed "porcine reproductive and respiratory syndrome (PRRS)". In the early 1990 s an arterivirus, referred to as PRRS virus (PRRSV), was determined to be the etiologic agent of this disease. Since then research has progressed substantially. Most recently "porcine high fever disease" was reported in China starting in 2006 with PRRSV being a critical virus associated with high morbidity and mortality (20%) associated with this syndrome which in 2010 is still causing severe pathology in pigs in China, with spread to Vietnam and Cambodia. This volume contains a series of reviews that highlight the virus, its pathogenesis, epidemiology, immunology, vaccinology and host genetic control. This paper provides a brief historical review of PRRS and the associated PRRSV. It presents areas of research gaps that inhibit current progress towards PRRS elimination through production of effective vaccines and current plans for PRRS elimination or eradication programs. It is hoped that this discussion will stimulate further collaboration between researchers and swine veterinarians throughout the world to provide answers that enhance our understanding of PRRS and PRRSV in an effort to eliminate this economically important disease.
Virus Research 10/2010; 154(1-2):1-6. · 2.94 Impact Factor
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ABSTRACT: Infection with porcine reproductive and respiratory syndrome virus (PRRSV) results in a weak antiviral immune response that leads to a persistent infection in a subset of pigs. We investigated the intensity and timing of the early cytokine responses to PRRSV infection to determine their utility as a predictor of persistence. As part of the "Big Pig" project, we evaluated cytokine gene expression in lymphoid tissues collected from pigs for up 202 days post-infection (dpi); serum samples were collected biweekly. Cytokine mRNA levels were compared between pigs that cleared the viral infection from serum and tissues (non-persistent [NP] pigs) to those of persistent (P) pigs, that had viral RNA in their serum for up to 126 dpi. The gene expression studies in the tracheobronchial lymph nodes (TBLN) of all the pigs showed upregulation of interferon-gamma (IFN-gamma)-associated T-helper 1 (Th-1) markers from 14-84 dpi, and of T-regulatory interleukin-10 (IL-10), but no upregulation of innate markers (IFN-A, IL-1B, and IL-8). At later time points (>112 dpi) these genes were no longer differentially expressed and thus were uninformative for persistence studies. Statistical analyses of serum cytokine levels indicated that innate cytokine (IL-1beta and IL-8) levels were upregulated early after infection. Interestingly, serum IL-8 levels in NP pigs were significantly higher than in P pigs at 14 dpi. When analyzed together, variations in all three of the serum cytokines tested (IL-8, IL-1beta, and IFN-gamma) was significantly correlated with virus level, accounting for approximately 84% of the variations observed. These results indicate that while each cytokine individually has minor effects on the length of virus replication, the combination of cytokine activities should be considered when understanding the role of immunity in persistence.
Viral immunology 04/2010; 23(2):127-34. · 1.78 Impact Factor
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ABSTRACT: This report describes the cloning and characterization of expressed gene sequences of the swine and bovine interferon-gamma inducible chemokine CXCL11, or I-TAC, associated with type 1 T-helper immune responses, and affirmation of bioactivity of their yeast-expressed protein products. The coding regions of both cDNA sequences were 303 nucleotides in length; each is coded for four exons in the genome. The bovine coding region shared 82% and 70% homology with human and mouse CXCL11, respectively, and the swine coding region 84% and 72% homology, respectively. As expected the swine and bovine CXCL11 sequences showed less homology with other human and mouse C-X-C motif chemokine sequences. Each cDNA was cloned into plasmids and transfected into Pichia pastoris (yeast) and the resultant expressed protein purified. Biological activity of each purified chemokine was affirmed by chemotaxis assays. Both swine and bovine CXCL11 were chemotactic for mitogen and IL-2 stimulated peripheral blood mononuclear cells. This is the first report for bioactivity of this chemokine in livestock species. This work provides valuable new reagents for investigating basic immunity as well as vaccine and disease responses in swine and cattle, goals of the U.S. Veterinary Immune Reagent Network which supported this effort.
Veterinary Immunology and Immunopathology 03/2010; 136(1-2):170-5. · 2.08 Impact Factor
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ABSTRACT: There are many diseases of fish, livestock and companion animals that impact negatively on animal health, welfare and productivity and for which there are no effective vaccines. The development of new vaccines is reliant on the availability of well-characterised immunological tools and reagents to understand host-pathogen interactions and identify protective immune responses. Veterinary immunology has always lagged behind mouse and human immunology in terms of development and availability of tools and reagents. However, several initiatives are underway to address this. The Veterinary Immunology Committee (VIC) Toolkit was initiated 6 years ago at the sixth International Veterinary Immunology Symposium (IVIS) in Uppsala and in the intervening period there have been several notable developments that have advanced reagent development and information exchange. This review will discuss advances in veterinary reagent development, networks, databases and commercial availability with particular reference to the second VIC Toolkit workshop held at the eighth IVIS in Ouro Preto, Brazil on the 15th of August 2007.
Veterinary Immunology and Immunopathology 11/2008; 128(1-3):24-9. · 2.08 Impact Factor
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ABSTRACT: To elucidate the host transcriptional response to Salmonella enterica serovar Typhimurium, Affymetrix porcine GeneChip analysis of pig mesenteric lymph nodes was used to identify 848 genes showing differential expression across different times after inoculation or when compared to non-inoculated controls. Annotation analyses showed that a high proportion of these differentially expressed (DE) genes are involved in immune and inflammatory responses. T helper 1, innate/inflammatory, and antigen-processing pathways were induced at 24 h post-inoculation (hpi) and/or 48 hpi, while apoptosis and antigen presentation/dendritic cell function pathways were downregulated at 8 hpi. Cluster analyses revealed that most DE genes annotated as NFkappaB targets were grouped into a specific induced subcluster, while many translation-related DE genes were found in a repressed subcluster. Quantitative polymerase chain reaction analyses confirmed the Affymetrix results, revealing transcriptional induction of NFkappaB target genes at 24 hpi and suppression of the NFkappaB pathway from 24 to 48 hpi. We propose that such NFkappaB suppression in antigen-presenting cells may be the mechanism by which S. Typhimurium eludes a strong inflammatory response to establish a carrier status in pigs.
Genomics 08/2007; 90(1):72-84. · 3.02 Impact Factor
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ABSTRACT: An investigation of the porcine response to gastrointestinal infection with Salmonella enterica serovars Choleraesuis (narrow host range) and Typhimurium (broad host range) revealed markedly different transcriptional profiles. Seven genes identified by suppression subtractive hybridization as up-regulated in the mesenteric lymph nodes at 24h (h) post-inoculation (p.i.) in serovar Choleraesuis-infected pigs (ARPC2, CCT7, HSPH1, LCP1, PTMA, SDCBP, VCP) and three genes in serovar Typhimurium-infected pigs (CD47/IAP, CXCL10, SCARB2) were analyzed by real-time PCR at 8h, 24 h, 48 h, 7 days (d) and 21 d p.i. A comparison between the two Salmonella infections revealed significant differences in transcriptional induction early in the infection (8-24h) for the serovar Typhimurium-infected pigs, whereas the serovar Choleraesuis-infected pigs exhibited significantly higher levels of gene expression at the later time points (48h-21 d), except for HSPH1. A similar gene expression trend was observed for immune-related genes involved in innate immunity and the inflammatory T helper 1 (Th1) response. Initial repression of gene expression in the serovar Choleraesuis-infected pigs from 8 to 48h p.i. (IFNG, IL12A, IL4, IL8, CSF2) coincided with extended transcriptional activation throughout the 21 d infection (IFNG, INDO, SOCS1, STAT1, IL1B, IL6, IL8, SLC11A1). The serovar Typhimurium-infected swine presented a more transient induction of immune-related genes (IFNG, INDO, IRF1, SOCS1, STAT1, IL1B, IL8, SLC11A1) early in the infection (24-48 h) followed by a significant repression of IL12A, IL12B, IL4, IL8 and CSF2. Collectively, these data reveal specific porcine genes with differences in gene expression kinetics that may be responsible for the variation in disease progression observed in swine infected with Typhimurium compared to Choleraesuis.
Molecular Immunology 05/2007; 44(11):2900-14. · 2.90 Impact Factor
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Joan K Lunney
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ABSTRACT: This review is a short update on the diversity of swine biomedical models and the importance of genomics in their continued development. The swine has been used as a major mammalian model for human studies because of the similarity in size and physiology, and in organ development and disease progression. The pig model allows for deliberately timed studies, imaging of internal vessels and organs using standard human technologies, and collection of repeated peripheral samples and, at kill, detailed mucosal tissues. The ability to use pigs from the same litter, or cloned or transgenic pigs, facilitates comparative analyses and genetic mapping. The availability of numerous well defined cell lines, representing a broad range of tissues, further facilitates testing of gene expression, drug susceptibility, etc. Thus the pig is an excellent biomedical model for humans. For genomic applications it is an asset that the pig genome has high sequence and chromosome structure homology with humans. With the swine genome sequence now well advanced there are improving genetic and proteomic tools for these comparative analyses. The review will discuss some of the genomic approaches used to probe these models. The review will highlight genomic studies of melanoma and of infectious disease resistance, discussing issues to consider in designing such studies. It will end with a short discussion of the potential for genomic approaches to develop new alternatives for control of the most economically important disease of pigs, porcine reproductive and respiratory syndrome (PRRS), and the potential for applying knowledge gained with this virus for human viral infectious disease studies.
International journal of biological sciences 02/2007; 3(3):179-84. · 2.70 Impact Factor
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ABSTRACT: The recent emergence of a unique group of North American type 1 porcine reproductive and respiratory syndrome virus (PRRSV) in the United States presents new disease control problems for a swine industry that has already been impacted seriously by North American type 2 PRRSV. In this study, a full-length cDNA infectious clone was generated from a low-virulence North American type 1 PRRSV isolate, SD01-08. In vitro studies demonstrated that the cloned virus maintained growth properties similar to those of the parental virus. Virological, pathological, and immunological observations from animals challenged with cloned viruses were similar to those from animals challenged with the parental virus and a modified live virus vaccine. To further explore the potential use as a viral backbone for expressing foreign genes, the green fluorescent protein (GFP) was inserted into a unique deletion site located at amino acid positions 348 and 349 of the predicted Nsp2 region in the virus, and expression of the Nsp2-GFP fusion protein was visualized by fluorescent microscopy. The availability of this North American type 1 infectious clone provides an important research tool for further study of the basic viral biology and pathogenic mechanisms of this group of type 1 PRRSV in the United States.
Journal of Virology 01/2007; 80(23):11447-55. · 5.40 Impact Factor
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ABSTRACT: Results from recent serological surveys and epidemiological studies show that pigs raised in a variety of management systems can be carriers of the tissue cyst stage of Toxoplasma gondi. This parasite can be transmitted to humans through the consumption of improperly prepared pork, making detection and removal of infected swine carcasses from the food chain an important food safety issue. Several methods are available for detection of T. gondii infected swine, including serological assays, polymerase chain reaction, and animal bioassays. The aim of the present study was to compare the detection sensitivities of six of these commonly used methods for detection of T. gondii infection in tissues from naturally and experimentally infected pigs. The results indicate that a serum-based ELISA is the most sensitive method, of those tested, for detection of T. gondii infected swine.
Veterinary Parasitology 11/2006; 141(1-2):9-17. · 2.58 Impact Factor
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ABSTRACT: Understanding the transcriptional response to pathogenic bacterial infection within food animals is of fundamental and applied interest. To determine the transcriptional response to Salmonella enterica serovar Choleraesuis (SC) infection, a 13,297-oligonucleotide swine array was used to analyze RNA from control, 24-h postinoculation (hpi), and 48-hpi porcine lung tissue from pigs infected with SC. In total, 57 genes showed differential expression (p < 0.001; false discovery rate = 12%). Quantitative real-time PCR (qRT-PCR) of 61 genes was used to confirm the microarray results and to identify pathways responding to infection. Of the 33 genes identified by microarray analysis as differentially expressed, 23 were confirmed by qRT-PCR results. A novel finding was that two transglutaminase family genes (TGM1 and TGM3) showed dramatic increases in expression postinoculation; combined with several other apoptotic genes, they indicated the induction of apoptotic pathways during SC infection. A predominant T helper 1-type immune response occurred during infection, with interferon gamma (IFNG) significantly increased at 48 hpi. Genes induced by IFNs (GBP1, GBP2, C1S, C1R, MHC2TA, PSMB8, TAP1, TAP2) showed increased expression during porcine lung infection. These data represent the first thorough investigation of gene regulation pathways that control an important porcine respiratory and foodborne bacterial infection.
Mammalian Genome 07/2006; 17(7):777-89. · 2.89 Impact Factor
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ABSTRACT: A first-generation porcine oligonucleotide set, representing 13,297 cDNAs and ESTs, has been designed by Qiagen-Operon for transcriptional profiling. To validate this set, microarrays containing each 70-mer oligonucleotide, referred to as the Qiagen-NRSP8 array, were hybridized with targets from porcine adult liver, lung, muscle, or small intestine. Transcriptome analyses showed that 11,328 of the oligonucleotides demonstrated expression in at least one tissue. Statistical analyses revealed that 1810 genes showed differential expression among tissues (Bonferroni adjusted p < 0.05). Biological pathways identified by DAVID/EASE analysis using a list of 423 tissue-selective genes matched archetypal pathways in the corresponding human or mouse tissue. Real-time quantitative PCR confirmed expression patterns for 9 of 11 genes tested. Our results demonstrate that this first-generation porcine oligonucleotide array is informative and the specificity is high. This is essential validation for investigators using the Qiagen-NRSP8 array for porcine functional genomics and for using the pig in modeling important physiological problems.
Genomics 11/2005; 86(5):618-25. · 3.02 Impact Factor
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Harry D Dawson,
Ethiopia Beshah,
Sandra Nishi,
Gloria Solano-Aguilar,
Motoko Morimoto,
Aiping Zhao,
Kathleen B Madden,
Tonya K Ledbetter,
J P Dubey,
Terez Shea-Donohue, Joan K Lunney,
Joseph F Urban
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ABSTRACT: Human infectious diseases have been studied in pigs because the two species have common microbial, parasitic, and zoonotic organisms, but there has been no systematic evaluation of cytokine gene expression in response to infectious agents in porcine species. In this study, pigs were inoculated with two clinically and economically important parasites, Toxoplasma gondii and Ascaris suum, and gene expression in 11 different tissues for 20 different swine Th1/Th2-related cytokines, cytokine receptors, and markers of immune activation were evaluated by real-time PCR. A generalized Th1-like pattern of gene expression was evident in pigs infected with T. gondii, along with an increased anti-inflammatory gene expression pattern during the recovery phase of the infection. In contrast, an elevated Th2-like pattern was expressed during the period of expulsion of A. suum fourth-stage larvae from the small intestine of pigs, along with low-level Th1-like and anti-inflammatory cytokine gene expression. Prototypical immune and physiological markers of infection were observed in bronchial alveolar lavage cells, small intestinal smooth muscle, and epithelial cells. This study validated the use of a robust quantitative gene expression assay to detect immune and inflammatory markers at multiple host tissue sites, enhanced the definition of two important swine diseases, and supported the use of swine as an experimental model for the study of immunity to infectious agents relevant to humans.
Infection and Immunity 03/2005; 73(2):1116-28. · 4.16 Impact Factor
