Temporal microRNA expression profile of pig peripheral blood during postnatal development.

Gene expression profiles of blood can reflect the physiopathologic status of the immune system. The dynamic microRNA (miRNA) expression profiles of peripheral blood from pigs at different developmental stages, and how differential expression of miRNAs might relate to immune system development, are unknown. In this study, peripheral blood samples taken at five developmental stages were used to construct 15 miRNA libraries (three biological replicates/stage): 0 days (newborn), 30 days (weaning), 60 days (weaned), and 180 and 360 days (puberty). We identified 295 known mature miRNAs. Hierarchical clustering of the miRNA expression profile showed significant differences between individuals at the neonatal and postnatal stages. Functional enrichment analysis revealed that miRNAs differentially expressed between pairwise comparisons of the developmental stages were over-represented in immune-related pathways such as toll-like receptor signaling. The time-course of expression of the over-representated miRNAs exhibited a pattern of steady decline over time, for both the complete miRNA compendium and immune-related miRNAs. We identified six marker miRNAs that were highly negatively correlated with chronologic age and enriched for genes involved in immune-related pathways. This study of a peripheral blood miRNA transcriptome offers insight into immune system development in swine and provides a resource for pig genome annotation.

Silence of TGF-ß1 gene expression reduces prrsv replication and potentiates immunity of immune cells of tibetan pig.

Transforming growth factor beta 1 (TGF-ß1) was of importance in the pathogenesis of porcine reproductive and respiratory syndrome virus (PRRSV). To determine whether knockdown of TGF-ß1 gene expression could facilitate the control of PRRSV infection, specific sequences for expressing shRNA targeted to porcine TGF-ß1 gene were synthesized and cloned into pSilencer 3.1-H1 neovector. Then they were used to transfect peripheral blood mononuclear cells of Tibetan pig (Tp-PBMCs) followed by PRRSV inoculation. The positive recombinant plasmids were screened for inhibition of TGF-ß1 gene expression by real-time quantitative RT-PCR. Conversely, the mRNA level of PRRSV in shRNA treated Tp-PBMCs dramatically decreased, and there were significant increases of the transcription of immune genes, such as interleukin-2 (IL-2), interleukin-4 (IL-4), interferon-alpha (IFN-α), interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), toll-like receptor 3 (TLR3), toll-like receptor 7 (TLR7), Myeloid differentiation primary response gene (88) (MyD88), and interleukin-27p28 (IL-27p28). However, the expressions of IL-8 and IL-10 genes significantly reduced in comparison to the control infected cells. In addition, transfection with the shRNA plasmids significantly elevated the viability of immune cells. Therefore the knockdown of TGF-ß1 gene expression by shRNA not only inhibits the replication of PRRSV but also improves immune responsiveness following viral infection, suggesting a novel way to facilitate the control of PRRSV infection in pigs.

Knockdown expression of IL-10Rα gene inhibits PRRSV replication and elevates immune responses in PBMCs of Tibetan pig in vitro.

Increase of interleukin-10 (IL-10) induced by Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection has been intensely studied to inhibit the anti-viral responses of host pigs. Blockade of expression of IL-10 receptor (IL-10R) by RNA interference (RNAi) may relieve the immunosuppression caused by excessive IL-10 in PRRSV infection. The recombinant short hairpin expressing plasmid targeted to pig IL-10Rα was transfected into peripheral blood mononuclear cells of Tibetan pig (Tp-PBMCs) prior to PRRSV inoculation, then the replication of PRRSV and immune responses in Tp-PBMCs were evaluated. The recombinant interfering plasmid greatly decreased PRRSV yield. The transcriptional level of IL-10Rαwas obviously inhibited by recombinant interfering plasmid; the expression of IL-10 was also down-regulated, while that of TGF-ß1 was not affected. Furthermore, the recombinant plasmid notably up-regulated the mRNA levels of TLR3, TLR7, IFN-α, IFN-γ, IL-2, IL-4, IL-12p40 and MyD88, while that of IL-8 was apparently decreased; In addition, cell viability of Tp-PBMCs was clearly enhanced by the interfering recombinant plasmid. Our results suggest that knockdown the expression of pig IL-10Rα can evidently inhibit the PRRSV infection and enhance the anti-viral immune responses of pig immune cells, which may be a promising way for preventing virus infection and developing new effective immune-regulator to strengthen the host immunity against PRRS.

Transcriptomic profiling in muscle and adipose tissue identifies genes related to growth and lipid deposition.

Growth performance and meat quality are important traits for the pig industry and consumers. Adipose tissue is the main site at which fat storage and fatty acid synthesis occur. Therefore, we combined high-throughput transcriptomic sequencing in adipose and muscle tissues with the quantification of corresponding phenotypic features using seven Chinese indigenous pig breeds and one Western commercial breed (Yorkshire). We obtained data on 101 phenotypic traits, from which principal component analysis distinguished two groups: one associated with the Chinese breeds and one with Yorkshire. The numbers of differentially expressed genes between all Chinese breeds and Yorkshire were shown to be 673 and 1056 in adipose and muscle tissues, respectively. Functional enrichment analysis revealed that these genes are associated with biological functions and canonical pathways related to oxidoreductase activity, immune response, and metabolic process. Weighted gene coexpression network analysis found more coexpression modules significantly correlated with the measured phenotypic traits in adipose than in muscle, indicating that adipose regulates meat and carcass quality. Using the combination of differential expression, QTL information, gene significance, and module hub genes, we identified a large number of candidate genes potentially related to economically important traits in pig, which should help us improve meat production and quality.

A mouse protein interactome through combined literature mining with multiple sources of interaction evidence.

Protein-protein interactions (PPIs) play crucial roles in a number of biological processes. Recently, protein interaction networks (PINs) for several model organisms and humans have been generated, but few large-scale researches for mice have ever been made neither experimentally nor computationally. In the work, we undertook an effort to map a mouse PIN, in which protein interactions are hidden in enormous amount of biomedical literatures. Following a co-occurrence-based text-mining approach, a probabilistic model--naïve Bayesian was used to filter false-positive interactions by integrating heterogeneous kinds of evidence from genomic and proteomic datasets. A support vector machine algorithm was further used to choose protein pairs with physical interactions. By comparing with the currently available PPI datasets from several model organisms and humans, it showed that the derived mouse PINs have similar topological properties at the global level, but a high local divergence. The mouse protein interaction dataset is stored in the Mouse protein-protein interaction DataBase (MppDB) that is useful source of information for system-level understanding of gene function and biological processes in mammals. Access to the MppDB database is public available at http://bio.scu.edu.cn/mppi.

Regulating effects of novel CpG chitosan-nanoparticles on immune responses of mice to porcine paratyphoid vaccines.

OBJECTIVE: To study the regulating effects of a novel CpG oligodeoxynuleotide and the synergistic effect of chitosan-nanoparticles (CNP) with CpG on immune responses of mice, which were used to develop a novel immunoadjuvant to boost immune response to conventional vaccines. METHODS: A novel CpG ODN containing 11 CpG motifs was synthesized and its bioactivities to stimulate the proliferation of lymphocytes of pig in vitro were detected. Then it was entrapped with CNP prepared in our laboratory by the method of ionic cross linkage, and immunized Kunming mice were co-inoculated with paratyphoid vaccine. The peripheral blood was collected weekly from the tail vein of inoculated mice to detect the contents of IgG, IgA, IgM, and specific antibody against salmonella as well as the levels of interleukin-2 (IL2), IL-4, and IL-6 by SABC-ELISA assay. The numbers of leucocytes, monocytes, granuloytes, and lymphocytes were calculated separately using the routine method. The experimental mice were orally challenged with virulent salmonella 35 days after inoculation. RESULTS: This CpG ODN could remarkably provoke the proliferation of lymphocytes of pig in vitro in contrast with the control (P < 0.05). Compared with those of the control, immunoglobulins, including IgG, IgA, IgM, and specific antibodies to paratyphoid vaccine, increased significantly in sera from the CpG or CpG-CNP-vaccinated mice (P < 0.05). IL-2, IL-4, and IL-6 increased remarkably in sera from immunized mice (P < 0.05). The leucocytes, monocytes, granuloytes, and lymphocytes of the mice immunized with CpG or CpG-CNP were also increased in number (P < 0.05). After the challenge, these immunity values were elevated in the mice vaccinated with CpG or CpG-CNP. The immunized mice all survived, while the control mice fell ill with evident lesions with diffuse hemorrhage in stomach, small intestine, and peritoneum. CONCLUSIONS: CpG ODN entrapped with CNP is a promising effective immunoadjuvant for vaccination, which promotes humoral and cellular immune responses, enhances immunity and resistance against salmonella by co-administration with paratyphoid vaccine.