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Transcriptome analysis in chicken cecal epithelia upon infection by Eimeria tenella in vivo.

ABSTRACT: Coccidiosis, caused by various Eimeria species, is a major parasitic disease in chickens. However, our understanding on how chickens respond to coccidian infection is highly limited at both molecular and cellular levels. The present study employed the Affymetrix chicken genome array and performed transcriptome analysis on chicken cecal epithelia in response to infection for 4.5 days in vivo by the cecal-specific species E. tenella. By Significance Analysis of Microarrays (SAM), we have identified 7,099 probe sets with q-values at <0.05, in which 4,033 and 3,066 genes were found to be up- or down-regulated in response to parasite infection. The reliability of the microarray data were validated by real-time qRT-PCR of 20 genes with varied fold changes in expression (i.e., correlation coefficient between microarray and qRT-PCR datasets: R (2)?=?0.8773, p<0.0001). Gene ontology analysis, KEGG pathway mapping and manual annotations of regulated genes indicated that up-regulated genes were mainly involved in immunity/defense, responses to various stimuli, apoptosis/cell death and differentiation, signal transduction and extracellular matrix (ECM), whereas down-regulated genes were mainly encoding general metabolic enzymes, membrane components, and some transporters. Chickens mustered complex cecal eipthelia molecular and immunological responses in response to E. tenella infection, which included pathways involved in cytokine production and interactions, natural killer cell mediated cytotoxicity, and intestinal IgA production. In response to the pathogenesis and damage caused by infection, chicken cecal epithelia reduced general metabolism, DNA replication and repair, protein degradation, and mitochondrial functions.

SUBMITTER: Guo A

PROVIDER: S-EPMC3667848 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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Transcriptome analysis in chicken cecal epithelia upon infection by Eimeria tenella in vivo.

Guo Aijiang A Cai Jianping J Gong Wei W Yan Hongbin H Luo Xuenong X Tian Guangfu G Zhang Shaohua S Zhang Haili H Zhu Guan G Cai Xuepeng X

PloS one 20130530 5

Coccidiosis, caused by various Eimeria species, is a major parasitic disease in chickens. However, our understanding on how chickens respond to coccidian infection is highly limited at both molecular and cellular levels. The present study employed the Affymetrix chicken genome array and performed transcriptome analysis on chicken cecal epithelia in response to infection for 4.5 days in vivo by the cecal-specific species E. tenella. By Significance Analysis of Microarrays (SAM), we have identifie ...[more]

PMID: 23737974

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Project description:The T cell-mediated immune response plays an important role in coccidiosis. To reveal the host T cell immune response following Eimeria tenella (E. tenella) infection in chickens, we performed quantitative real-time PCR to analyze the dynamic expression of the Th1-related cytokines IFN-γ, IL-2, and IL-12; the Th17-related cytokines IL-17A, IL-17F, and IL-22; and the Treg-related cytokines IL-10, TGF-β, and CTLA-4 in the cecum and spleen at 0, 2, 4, 6, 8, and 10 d postinfection (dpi). In the cecal tissue, the expression of the Th1-related cytokine IFN-γ was significantly higher at 6 and 8 dpi than at other time points (11.97-fold and 39.78-fold, respectively, compared with 0 dpi, P < 0.05). IL-2 and IL-12 expression was significantly higher at 6 and 8 dpi than at 0, 2 and 10 dpi (P < 0.05). The expression of the Th17-related cytokines IL-17A and IL-17F at 2 and 4 dpi and IL-22 expression at 4 dpi were significantly higher than those at 0, 6, 8 and 10 dpi (P < 0.05). The expression of the Treg-related cytokines IL-10, TGF-β and CTLA-4 was significantly higher at 6 and 8 dpi than at 0, 2 and 4 dpi (P < 0.05). In the spleen, IFN-γ and IL-12 expression peaked at 4 dpi, while IL-2 expression peaked at 10 dpi. IL-17A, IL-17F and IL-22 expression was significantly higher at 2 and 4 dpi than at 0, 6, 8 and 10 dpi (P < 0.05). Treg-related cytokine TGF-β expression was almost unchanged and significantly decreased at only 4 dpi (P < 0.05), while CTLA-4 expression showed an overall decreasing trend from 0 to 8 dpi but increased significantly at 10 dpi (P < 0.05). The expression patterns of three T cell subset-related cytokines were different in the cecum and spleen. Furthermore, Th1 and Treg cells participate in the immune response mainly in the latter stage of coccidia infection (6 and 8 dpi), while Th17 cells play a role mainly in the early stages of infection (2 and 4 dpi). Our data will help to deepen the understanding of the complex T cell immune response after coccidia infection.

Project description:IL-6, IL-8, and C-C motif chemokine ligand 2 (CCLi2) are important factors in inflammatory and immune responses. To investigate their relationships in the spleen and cecum and between coccidiosis-infected and uninfected states, we performed quantitative real-time PCR to compare the relative expression difference of IL-6, IL-8, and CCLi2 in the same tissues between the infection and control groups. In addition, the correlations of the relative expression levels of these 3 genes were determined in the same and different tissues within the same group. The results showed that the expression levels of IL-6, IL-8, and CCLi2 in the spleen and cecum of the infected group were all higher than those of the uninfected group (P < 0.05). The correlation coefficients among the IL-6, IL-8, and CCLi2 expression levels in the spleen or cecum were all positive in both the infection and control groups. In the spleen tissues, CCLi2 expression was strongly correlated with IL-6 and IL-8 in the uninfected group (P < 0.01), and the correlation coefficients reached 0.853 (R2 = 0.728) and 0.996 (R2 = 0.992), respectively. The expression of CCLi2 was also strongly correlated with IL-8 (R reached 0.890, R2 = 0.792) in the infected group. In the cecal tissues, the expression levels of the 3 genes were all extremely significantly correlated in the uninfected group (P < 0.01), and the correlation coefficients ranged from 0.498 to 0.765, indicating moderate correlations. The expression of IL-6 was extremely significantly positively correlated with IL-8 and CCLi2 in the infected group (P < 0.01), with moderate correlations (R ranged from 0.469-0.639). In addition, the expression levels of the 3 genes were not significantly correlated (P > 0.05) between the spleen and cecum tissues in either the infection group or the control group. These results indicate that IL-6, IL-8, and CCLi2 were correlated and play an important role in coccidiosis infection of Jinghai yellow chicken. Our data also provide a basis for further exploring the role of these 3 genes in genetic breeding for coccidiosis resistance.

Dataset Information

Transcriptome analysis in chicken cecal epithelia upon infection by Eimeria tenella in vivo.

Publications

Transcriptome analysis in chicken cecal epithelia upon infection by Eimeria tenella in vivo.

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