Dataset Information

Concordant expression pattern across multiple immune tissues of commercial broilers in response to avian pathogenic Escherichia coli (APEC).

ABSTRACT: In this study, RNA-seq gene expression data were obtained for three different immune tissues (bone marrow, thymus, and bursa) at two time points from birds with three phenotypes (non-infected, resistant, and susceptible birds). A total of 380 significant differentially expressed genes (DEGs) were commonly expressed in each of the three tissues in the contrast of susceptible vs. non-infected birds at 5 days post-infection (dpi) and 106 significant DEGs were shared in susceptible vs. resistant birds at 5 dpi. For the co-expressed DEGs, a relatively high consistency in expression pattern was identified in the three tissues for the two contrasts. These co-expressed DEGs were involved in the biological process of response to stimulus, regulation of cell proliferation, signal transduction, and immune system. Moreover, three gene networks were identified for the co-expressed DEGs, showing the activation of the humoral immune response, cell survival, growth and death were the concordant response for the three lymphocyte tissues. Several potential biomarker genes, HPGD, PROX1, FOSL2, E2F1, and ANP32A were found, which may have critical functions in resistant birds to resist APEC infection. Taken together, this study provides a novel insight that may elucidate the molecular mechanisms underlying host response to systemic APEC infection, as well as enhances the probability of advancement in biomedical research.

SUBMITTER: Sun H

PROVIDER: S-EPMC6682839 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature

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Concordant expression pattern across multiple immune tissues of commercial broilers in response to avian pathogenic <i>Escherichia coli</i> (APEC).

Sun Hongyan H Li Huan H

3 Biotech 20190805 9

In this study, RNA-seq gene expression data were obtained for three different immune tissues (bone marrow, thymus, and bursa) at two time points from birds with three phenotypes (non-infected, resistant, and susceptible birds). A total of 380 significant differentially expressed genes (DEGs) were commonly expressed in each of the three tissues in the contrast of susceptible vs. non-infected birds at 5 days post-infection (dpi) and 106 significant DEGs were shared in susceptible vs. resistant bir ...[more]

PMID: 31406642

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Project description:Many reports show an association between the Pst system, the Pho regulon related genes and bacterial virulence. Our previous results showed that a functional Pst system is required for full virulence, resistance to serum, polymyxin B and acid shock However, the interplay between the Pst system and virulence has an unknown molecular basis. To understand global APEC virulent strain responses to Pho regulon activation, we conducted transcriptome profiling experiments comparing the isogenic Pst mutant with its wild type strain chi7122 using the Affymetrix GeneChipÂ® E. coli Genome 2.0 Array. To understand the global responses to Pho regulon activation of APEC, we use the Affymetrix technology. We conducted transcriptome profiling experiments comparing the APEC chi7122 strain and its isogenic Pst mutant grown in rich phosphate medium using the Affymetrix GeneChipÂ® E. coli Genome 2.0 Array. The Affymetrix GeneChipÂ® E. coli Genome 2.0 Array contains the genome of the E. coli MG1655 and three pathogenic E. coli strain (EDL933, Sakai and CFT073) representing 20,366 genes. While comparing genes expression between Pst mutant and the wild type chi7122 strain, 471 genes are either up- (254) or down-regulated (217) of at least 1.5-fold, with a p-value inferior or equal to 0.05 and a false discovery rate of 2.71%. Experiment Overall Design: RNA extraction was performed onto the chi7122 and K3 strains on four biological replicates. An overnight culture grown at 37Â°C was diluted 100-fold into 5 ml of LB broth and was allowed to grow to mid-log phase (OD600 0,6). Cultures were centrifuged and RNAs were isolated by using the RiboPureâ¢-Bacteria Kit (Ambion, Austin, TX), according to the manufacturerâs recommendations, with the exception that the DNAse 1 treatment was performed twice. RNA extractions were performed on four different days (the chi7122 and K3 RNA were extracted at the same time) and a total of eight hybridizations were performed. Hybridization was carried out at McGill University & Genome Quebec Innovation Centre according to the Affymetrix recommendation (Affymetrix Expression Manual Section 3 701029 rev. 4). GeneChip scan was carried out at McGill University & Genome Quebec Innovation Centre according to the Affymetrix recommendation (Affymetrix Expression Manual Section 3 701029 rev. 4), and data were processed using the FlexArray software (Michal Blazejczyk, Mathieu Miron, Robert Nadon (2007). FlexArray: A statistical data analysis software for gene expression microarrays. Genome Quebec, Montreal, Canada). Raw data were normalized using the RMA algorithm and log2 were generated. The expression value was generated by subtracting the mean value of each replicate of the mutant strain by wild-type strain.

Project description:Many studies have examined avian pathogenic Escherichia coli (APEC) from commercial broilers but few have examined isolates from native chickens. This study compared APEC isolates from commercial broilers and native chickens in regard to the phylogenetic group and the phenotypic and genotypic antimicrobial resistance profiles. From 100 suspect colibacillosis cases in both commercial broilers and native chickens, a total of 90 broiler isolates and 42 native chicken isolates were identified as E. coli by biochemical tests. Phylogenetic grouping revealed that 90 broiler APEC isolates belonged to A group (5.56%), B1 group (22.22%), B2 group (31.11%), and D group (41.11%). The 42 native chicken APEC isolates belonged to A group (35.71%), B1 group (26.19%), B2 group (30.95%), and D group (7.14%). The difference in the allocation to groups A and D of the 2 isolate types was significant (P < 0.05). The APEC broiler isolates had a significantly higher multidrug-resistant (MDR) rate (80%) than the native chicken isolates (14.29%) (P < 0.05). The APEC broiler isolates demonstrated significantly higher resistance rates than the native chicken isolates for amoxicillin (98.89%; 78.57% respectively), chloramphenicol (42.2%; 9.5%), enrofloxacin (68.9%; 7.1%), gentamicin (11.1%; 0%), nalidixic acid (72.2%; 7.1%), sulfamethoxazole + trimethoprim (45.6%; 2.4%), and tetracycline (88.9%; 76.2%) (P < 0.05). The APEC broiler isolates had a significantly higher presence compared with the native chicken isolates of the following resistance genes:- by blaTEM (43.3%; 21.4%, respectively), cml-A (34.4%; 2.4%), tetA (76.7%; 40.5%), tetB (26.7%; 0%), sul2 (23.3%; 14.3%), and dhfrI (13.3%; 0%) (P < 0.05). The qnrB and qnrS genes were detected (12.16%; 72.97% respectively), in the APEC broiler isolates resistant to nalidixic acid and/or enrofloxacin while only qnrS genes was detected in all 3 APEC native chicken isolates. Regarding the point mutations of gyrA and parC, all isolates were positive to gyrA83S, gyrA87D, gyrA87L, gyrA87NY, parC80S and parC80I except that gyrA83S was not present in 20 APEC broiler isolates. Antimicrobial stewardship programs should be targeted at the backyard poultry sector as well as the commercial poultry sector.

Project description:Purpose: This study is to more comprehensively understand the genome-wide host response to avian pathogen E. coli (APEC). Methods: Male broiler chicks were challenged with APEC (or mock-challenged as controls), and bone marrow was harvested at 1 and 5 days post-infection (dpi). Based upon necropsy-scored lesions on liver, pericardium, and air sacs, the challenged birds were assigned to mild or severe pathology categories, representing resistant and susceptible phenotypes, respectively. RNA sequence data were first analyzed using the R package EdgeR to identify differentially expressed genes.GO and pathway analysis using the R package GOseq identified many immune-related pathways. Results: RNA sequencing resulted in 11 to 40 million single-end raw reads of 100 bp per sample. After alignment, an average of 80 % of the reads, with 5 % representing multiple mapping, were mapped to the chicken reference transcriptome. Among these detected unique transcripts, there were 2,404 novel genes mainly distributed on chromosomes 1, 2, 4, 3, Z, 5 in decreasing number. Genes were detected as differentially expressed (DE) between treatments (susceptible, resistant, and mock-challenged) at a given time point, or DE between 1 and 5 dpi within the same treatment group. Compared to mock-challenged birds on 5 dpi, susceptible birds exhibited extensive enhancement of their T cell and B cell development, as well as innate and adaptive immune response. However, for the 1 dpi group, susceptible birds exhibited a decreased gene expression of T and B cell development. The differences between the two phenotypes, susceptible and resistant birds, at 5 dpi showed that susceptible birds had increased gene expression of B-cell development and adaptive immune response. Conclusions: This is the first report to our knowledge examining the role of bone marrow cells in responding to APEC infection in broilers. This transcriptome study provides insight and global overview into the response of genes involved in the earliest phases of the immune response to APEC infection. Our data indicate a dynamic interaction between the innate and adaptive immune responses to APEC infection in susceptible birds, providing flexibility and redundancy in the host’s induction of cytokines and chemokines. Additionally, B cell and T cell development are also extensively affected by APEC infection for susceptible birds, resulting in drastic host impairment in early response to infection. The findings of present study shed light on the underlying chicken immune modulation against APEC infection. Also, this study will build a solid foundation for identifying host genetic variation that may be manipulated to enhance resistance to infection and may be useful as colibacillosis control targets.

Project description:BackgroundAvian pathogenic Escherichia coli (APEC) is a major cause of disease impacting animal health. The bone marrow is the reservoir of immature immune cells; however, it has not been examined to date for gene expression related to developmental changes (cell differentiation, maturation, programming) after APEC infection. Here, we study gene expression in the bone marrow between infected and non-infected animals, and between infected animals with mild (resistant) versus severe (susceptible) pathology, at two times post-infection.ResultsWe sequenced 24 bone marrow RNA libraries generated from the six different treatment groups with four replicates each, and obtained an average of 22 million single-end, 100-bp reads per library. Genes were detected as differentially expressed (DE) between APEC treatments (mild pathology, severe pathology, and mock-challenged) at a given time point, or DE between 1 and 5 days post-infection (dpi) within the same treatment group. Results demonstrate that many immune cells, genes and related pathways are key contributors to the different responses to APEC infection between susceptible and resistant birds and between susceptible and non-challenged birds, at both times post-infection. In susceptible birds, lymphocyte differentiation, proliferation, and maturation were greatly impaired, while the innate and adaptive immune responses, including dendritic cells, monocytes and killer cell activity, TLR- and NOD-like receptor signaling, as well as T helper cells and many cytokine activities, were markedly enhanced. The resistant birds' immune system, however, was similar to that of non-challenged birds.ConclusionThe DE genes in the immune cells and identified signaling models are representative of activation and resolution of infection in susceptible birds at both post-infection days. These novel results characterizing transcriptomic response to APEC infection reveal that there is combinatorial activity of multiple genes controlling myeloid cells, and B and T cell lymphopoiesis, as well as immune responses occurring in the bone marrow in these early stages of response to infection.

Dataset Information

Concordant expression pattern across multiple immune tissues of commercial broilers in response to avian pathogenic Escherichia coli (APEC).

Publications

Concordant expression pattern across multiple immune tissues of commercial broilers in response to avian pathogenic <i>Escherichia coli</i> (APEC).

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