Response of swine lung to H1N1 swine influenza virus infection revealed by transcription analysis
ABSTRACT: As a mild, highly contagious, respiratory disease, swine influenza always damages the innate immune systems, and increases susceptibility to secondary infections which results in considerable morbidity and mortality in pigs. Nevertheless, the systematical host response of pigs to swine influenza virus infection remains largely unknown. To explore these, a time-course gene expression profiling was performed to detect comprehensive analysis of the global host response induced by H1N1 swine influenza virus in pigs. At the age of day 35, 15 pigs were randomly allocated to the non-infected group and 15 to the infected group. Each piglet of the infected group was intranasaly challenged with A/swine/Hubei/101/2009(H1N1) strain and Each piglet of the non-infected group was treated similarly with an identical volume of PBS as control.
Project description:Swine H1N1 influenza virus and streptococcus suis serotype 2 (SS2) are two important contributors to the porcine respiratory disease complex, which have significant economic impacts. Clinically, swine influenza virus and swine streptococcus suis co-infection is common, which will increase the mortality. However, the pathogenesis of the co-infection remains largely unkown. To explore it, gene expression profiling was to performed to detect comprehensive analysis of the global host response induced by H1N1 virus infection alone, SS2 infection alone, H1N1-SS2 co-infection and PBS control. Overall design: A total of 12 healthy pigs were randomly allocated to 4 groups with 3 pigs each: H1N1 group, SS2 group, H1N1-SS2 group and control group. At the age of 35 days, pigs in H1N1 and H1N1-SS2 groups were intranasaly challenged with A/swine/Hubei/101/2009(H1N1) strain and others were treated similarly with PBS. 3 days after H1N1 virus challenge, pigs in SS2 and H1N1-SS2 groups were inoculated intranasally with PBS containing SS2 strain ZY05, others were treated similarly with PBS.
Project description:This SuperSeries is composed of the following subset Series: GSE35738: 2009 pandemic H1N1 virus causes disease and upregulation of genes related to inflammatory and immune response, cell death, and lipid metabolism in pigs GSE40088: Comparative transcriptomic analysis of acute host responses during 2009 pandemic H1N1 influenza infection in mouse, macaque, and swine (macaque dataset) GSE40091: Comparative transcriptomic analysis of acute host responses during 2009 pandemic H1N1 influenza infection in mouse, macaque, and swine (mouse dataset) Refer to individual Series
Project description:In order to identify the swine genes which play roles in the regulation of swine influenza A virus replication, the gene microarray was performed to explore the systematical host response to the swine H1N1/2009 influenza A virus infection in porcine cells. Overall design: Two pools of total RNA were extracted from the swine H1N1/2009 influenza A virus infected or uninfected NPTr cells and hybrided on Affymetrix Porcine genome array Array.
Project description:In recent years, the roles of microRNAs playing in the regulation of influenza viruses replication caused researchers' much attenion. However, much work focused on the interactions between human, mice or chicken microRNAs with human or avian influenza viruses rather than the interactions of swine microRNAs and swine influenza viruses. To investigate the roles of swine microRNAs playing in the regulation of swine influenza A virus replication, the microRNA microarray was performed to identify which swine microRNAs were involved in swine H1N1/2009 influenza A virus infection. Overall design: Two pools of total RNA were extracted from the swine H1N1/2009 influenza A virus infected or uninfected NPTr cells and hybrided on Affymetrix GeneChip miRNA 2.0 Array.
Project description:Background: Swine influenza is a highly contagious viral infection in pigs affecting the respiratory tract that can have significant economic impacts. Streptococcus suis serotype 2 is one of the most important post-weaning bacterial pathogens in swine causing different infections, including pneumonia. Both pathogens are important contributors to the porcine respiratory disease complex. Outbreaks of swine influenza virus with a significant level of co-infections due to S. suis have lately been reported. In order to analyze a global response to the dual infection, we carried out a comprehensive gene expression profiling using a microarray approach to study the swine tracheal epithelial (NPTr) cell response to a co-infection with H1N1 swine influenza virus (swH1N1) and S. suis serotype 2. Results: Gene clustering showed that the swH1N1 and swH1N1/S. suis infections modified the expression of genes in a similar manner. Additionally, infection of NPTr cells by S. suis alone did not result in many differentially expressed genes compared to mock-infected cells. However, some important genes coding for inflammatory mediators, such as chemokines, interleukins, cell adhesion molecules and eicosanoids, were significantly upregulated in the presence of both pathogens comparing to infection with each pathogen taken individually. This synergy may also be the consequence of an increased adhesion/invasion of epithelial cells previously infected by swH1N1, as recently reported. Conclusion: In a co-infection situation, influenza virus would replicate in the respiratory epithelium inducing an inflammatory infiltrate comprised of mononuclear cells and neutrophils. Despite that these cells are unable to phagocyte and kill S. suis, they are highly activated by this pathogen. S. suis is not considered a primary pulmonary pathogen, but an exacerbated production of pro-inflammatory mediators during a co-infection with influenza virus may be of critical importance in the pathogenesis and outcome of this respiratory disease complex. Total RNA obtained from NPTr cells infected with S. suis, H1N1, or S. suis & H1N1. Four replicates in both groups.
Project description:The determinants of influenza transmission remain poorly understood. Swine influenza viruses preferentially attach to receptors found in the upper airways; however, most swine influenza viruses fail to transmit efficiently from swine to humans, and from human-to-human. The pandemic 2009 H1N1 (H1N1pdm) virus was a rare exception of a swine virus that acquired efficient transmissibility from human-to-human, and is reflected in efficient respiratory droplet transmission in ferrets. We hypothesize that virus-induced host responses in the upper airways correlate with airborne transmission in ferrets. To address this question, we used the H1N1pdm virus and swine influenza A/swine/Hong Kong/201/2010 (HK201) virus that has comparable titre in the ferret nasopharynx, but it exhibits differential transmissibility in ferrets via respiratory droplet route. We performed a transcriptomic analysis of tissues from the upper and lower respiratory tract from ferrets infected with either H1N1pdm or HK201 viruses using ferret-specific Agilent oligonucleotide arrays. We found differences in the kinetics of the innate immune response elicited by these two viruses that varied across tissues. Overall design: Ferrets were mock inoculated with 0.5mL media or inoculated with 105 TCID50 of the A/CA/04/09 (H1N1) or A/Sw/HK/ 210/10 (H1N1) influenza viruses intra-nasally. There are a total of 9 ferrets per virus group and six ferrets for the mock group. Ferrets were euthanized at day 1 (n = 4 per virus group) and day 2 (n = 5 per virus group) post-inoculation (p.i.). Three mock inoculated ferrets were euthanized at days 1 and 2 p.i. Tissues from nasal turbinate, soft palate, pharynx, trachea, bronchus, and lungs were collected at necropsy, placed in RNAlater overnight at 4°C and conserved at -80°C. RNA was extracted and gene expression was analyzed by custom 4x180K ferret-specific oligonucleotide arrays (Agilent Cat# G4862A, design ID 48472)
Project description:Purpose: The goals of this study is to analyze differentially expressed microRNAs in pulmonary alveolar macrophages from pigs infected with H1N1 swine influenza A virus at different time points. Methods: miRNA profiles in PAMs from pigs infected with H1N1 SwIV at three time points were generated by deep sequencing. Differentially expressed miRNAs were then identified and their targets were predicted. For these targets, GO and KEGG analyses were performed and protein-protein interaction networks regulated by DE miRNAs were constructed. Results: Comparing to control group, 70 and 16 DE miRNAs were respectively identified on post-infection day (PID) 4 and PID 7. 56 DE miRNAs were identified between PID4 and PID7. GO and KEGG analyses indicated that most of targets for these DE miRNAs were related to immune and inflammatory responses. Conclusions: Our study represents the first integrative analysis of DE miRNAs in PAMs infected with H1N1 SwIV at different time points. The results in this study would enable us to better understand the underlying pathogenesis of H1N1 SwIV infection in pigs. Furthermore, these data would be very useful for investigating the functions and regulatory mechanisms of miRNAs in human inﬂuenza because pig serves as an excellent animal model to study human diseases. miRNA profiles in PAMs from pigs infected with H1N1 SwIV at three time points were generated by deep sequencing.
Project description:Background: The 2009 pandemic H1N1 influenza virus emerged in swine and quickly became a major global health threat. In mouse, non-human primate, and swine infection models, the pH1N1 virus efficiently replicates in the lung and induces pro-inflammatory host responses; however, whether similar or different cellular pathways were impacted by pH1N1 virus across independent infection models remains to be further defined. To address this, we have performed a comparative transcriptomic analysis of acute host responses to a single pH1N1 influenza virus, A/California/04/2009 (CA04), in the lung of mice, macaques and swine. Results: Despite similarities in the clinical course, we observed differences in inflammatory molecules elicited, and the kinetics of their gene expression changes across all three species. The retinoid X receptor (RXR) signaling pathway controlling pro-inflammatory and metabolic processes was differentially regulated during infection in each species, though the heterodimeric RXR partner, pathway associated signaling molecules, and gene expression patterns differed in each species. Conclusions: By comparing transcriptional changes in the context of clinical and virological measures, we identified differences in the host transcriptional response to pH1N1 virus across independent models of acute infection. Antiviral resistance and the emergence of new influenza viruses have placed more focus on developing drugs that target the immune system. Underlying overt clinical disease are molecular events that suggest therapeutic targets identified in one host may not be appropriate in another. The goal of this experiment was to use global gene expression profiling to understand swine lung host responses to pandemic H1N1 influenza A/Californica/04/2009 (CA04) virus infection and compare acute host responses across independent species. Four-week-old crossbred pigs (Sus Scrofa) were inoculated intratracheally with either 10^6 TCID50/pig egg-derived 2009 pandemic influenza A/California/04/2009 virus (n = 5) or mock inoculated with non-infectious cell culture supernatant (control; n = 4). Animals were euthanized on day 7 post-infection and lung samples were used for microarray.
Project description:Swine influenza virus (SIV) is a prevalent respiratory pathogen in pigs that has deleterious consequences to animal health, production, and public health. Pigs represent an important reservoir for influenza as well as a potential mixing vessel for novel gene reassortments. Despite the central role of the pig in the 2009 pandemic and 2012 variant H3N2 outbreak, much remains unknown about the impact of swine immunity on SIV transmission, pathogenesis, and evolution. An incomplete understanding of interactions between the porcine immune system and SIV has hindered the development of new diagnostic tools and CD8+ T cell influenza epitope based vaccines. In order to address this gap in knowledge, we identified swine leukocyte antigen (SLA) restricted influenza virus peptides presented by swine respiratory epithelial cells using an immunoproteomics approach. The majority of MHC associated peptides belonged to matrix 1, nucleocapsid, and nonstructural 1 proteins. Specific epitopes, such as M1229-242, NS177-89, and NP417-426, may have value in epitope based vaccines. Future investigations examining the potential cross-reactive nature of these peptides are needed to confirm antigen recognition by cytotoxic T lymphocytes and utility as vaccine candidates.
Project description:The aim of this study was to acquire a better understanding of porcine reproductive and respiratory syndrome (PRRS) disease through a deeper knowledge of gene expression changes that occur in pulmonary lymph nodes by comparing PRRS virus (PRRSV), porcine circovirus type 2 (PCV-2), and swine influenza virus (IAV-S) infections. The PRRSV, IAV-S and PCV-2 viral infections followed a clinical course in these domestic pigs typical of experimental infection of young pigs with these viruses. PRRSV isolate SDSU-73 was pathogenic in this study inducing fever, anorexia, listlessness, and dyspnea. Overall design: Differentially expressed tags (with respect to control) at all time points were ascertained. The experimental results were integrated with previous studies to develop a robust model of swine respiratory virus infection. Here we identified and compared gene expression changes in tracheobronchial lymph nodes (TBLN) following viral infection using Digital Gene Expression Tag Profiling (DGETP). Pigs were infected with 1 x 10^5 cell culture infectious dose 50% per pig of PRRSV (strain SDSU73), porcine circovirus type 2 (PCV2; group 2), or swine influenza A virus (IAV-S; A/SW/OH/511445/2007). Pigs were allotted to one of 4 treatment groups: sham inoculated control, PRRSV-challenge, PCV2-challenge, or IAV-S-challenge.