In recent years, the immunomodulatory role of adipose tissue (AT) has gained attention, yet the metabolic basis for immune homeostasis in AT remains unclear. Catecholamines (CAs) activate adipocyte β3-adrenergic receptors (β3-AR) to promote lipid metabolism, while cholesterol metabolism has bidirectional immunoregulatory properties. Whether the interaction between CAs and cholesterol regulates immune function in AT is unknown. Using chickens as models, we induced distinct immune states via dexamethasone (Dex) treatment and Newcastle disease virus (NDV) vaccination. Through qRT-PCR and metabolomics, we analyzed dynamic changes in lymphocytes, CAs, and cholesterol metabolism in AT. The results indicated that B cells were present in chicken AT and positively responded to different immune states through up-regulation. AT positively responded to Dex-induced immunosuppression (DIIS) and NDV-induced immune responses by altering cholesterol and CAs metabolisms. Moreover, the differences in cholesterol and CAs metabolisms were the key ways by which Dex affected immune response in AT. AT could synthesize bile acids and steroid derivatives, and secondary immunization possible was the key stage with active changes in cholesterol derivatives in AT. We identified a circuit pathway: “lymphocyte-catecholamine-adipocyte-cholesterol-cholesterol derivatives-lymphocyte,” which may underpin immune microenvironment homeostasis in AT. Additionally, miR-206 was actively involved in the processes of DIIS, and the miR-206/MSMO1 pathway potentially regulated cholesterol metabolism in AT. This study provides a perspective for in-depth understanding of the immune regulation mechanism of AT and offer a direction for developing cholesterol derivatives for immune regulation.

The bursa of Fabricius (BF), a central immune organ of birds, plays a critical role in immune regulation. Cholesterol metabolism is an important pathway for regulating lymphocyte proliferation and activation. However, the mechanisms by which cholesterol metabolism influences immune regulation in avian BF remain unclear. In this study, dexamethasone (Dex) and Newcastle disease virus (NDV) attenuated vaccine were used to simulate stress and induce immune response in chickens, respectively. The dynamic expression patterns and interrelationships of B cell receptor signaling subunit β (CD79B), sphingosine 1-phosphate receptor 1 (S1PR1), and 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) genes in chicken BF were analyzed, and the miRNA regulatory mechanisms of cholesterol metabolism were further identified. The results showed that, under stress condition, the BF actively responded to stress processes by circulating B cells migrating into BF, and exhibited similar changes in the primary and secondary stresses; under immune response condition, the BF participated in immune responses primarily through the migration of B cells from BF into circulatory system, which showed differences between the primary and secondary immune response. BF responded to stress mainly through up-regulating cholesterol metabolism, and the change in cholesterol metabolism was one of potential key pathways by which stress inhibited the immune function of BF. Moreover, the dynamic migrations of B cells and changes of cholesterol metabolism in BF possibly constituted the two-way regulatory modes to maintain the immune homeostasis of BF. The miR-29a/c-3p/HMGCR pathway served as a potential mechanism for cholesterol metabolism in chicken BF under different immune conditions. This study can provide innovative references for further exploring the mechanisms of stress affecting the immune function of BF.

In poultry production, stress-induced immunosuppression (SIIS) often leads to poor immune response to vaccines, causing immunized chickens to still develop diseases. If the molecular mechanisms by which SIIS affects immune response can be comprehensively elucidated, it can provide new insights and strategies for improving and addressing this issue. However, the detailed molecular regulatory mechanisms remain unclear. In this study, dexamethasone-induced immunosuppression (DIIS) was used to simulate SIIS in chickens, and the Newcastle disease virus (NDV) attenuated vaccine was used for vaccination. The study aimed to explore the expression characteristics and functions of immune checkpoint gene PD-L1 (programmed cell death ligand 1), the regulatory mechanism of PD-L1 gene mediated by miR-17-5p, and the potential application of circulating miR-17-5p in the process of SIIS inhibiting the immune response to the NDV vaccine. Quantitative real-time PCR (qRT-PCR) results showed that PD-L1 gene was differentially expressed in nine candidate tissues (P < 0.05), and actively involved in the SIIS mediated immunosuppression of NDV vaccine response. There were significant regulatory relationships between miR-17-5p and PD-L1 gene in multiple tissues, suggesting that the miR-17-5p-PD-L1 pathway was a key regulatory mechanism of SIIS inhibiting NDV immune response. Furthermore, expression analysis demonstrated that SIIS significantly altered circulating miR-17-5p levels in serum at several time points, such as 2 days post immunization (dpi), 4 dpi and 5 dpi, indicating the potential of serum circulating miR-17-5p as a molecular marker. This study provides valuable references for detecting, preventing, and treating the effects of stress affecting immune response.

Immunosuppression induced by infectious bursal disease virus (IBDV) and its subsequent secondary infections remain serious problems that urgently need to be addressed in the poultry industry. Of even greater concern, the molecular mechanism of IBDV-induced immunosuppression is not fully understood. In this study, expression characteristics of the immune checkpoint programmed cell death-ligand 1 (PD-L1) gene were explored during the chicken immune response induced by IBDV-attenuated vaccine, and the competing endogenous RNA (ceRNA) regulatory mechanism of PD-L1 gene in vivo was identified by quantitative real-time PCR (qRT-PCR). The results showed that PD-L1 gene expression was closely related to the immune response to IBDV, and played important regulatory roles in the immune-related tissues at different stages of the immune response. Significant game relationships in expression levels between miR-17 family members (miR-17-5p, miR-20a-5p, and miR-20b-5p), circITSN2, and PD-L1 gene were identified in vivo, so the circITSN2-miR-17-5p/20a-5p/20b-5p-PD-L1 network was a potential molecular regulatory mechanism of PD-L1 in the immune response to IBDV vaccine, and the heart (5 dpi), proventriculus (5 dpi), and lung (21 dpi) were the key tissues. This study can provide valuable references for further investigation of the molecular mechanisms of immunosuppression induced by IBDV.RESEARCH HIGHLIGHTSPD-L1 gene is correlated with IBDV immune response in chickens.PD-L1 is a key gene regulating the immune functions of the heart, lung, and proventriculus.CircITSN2-miR-17-5p/20a-5p/20b-5p-PD-L1 is a potential mechanism in IBDV immunity.

The mammalian adipose tissue (AT) plays a key role in regulating immune function and anti-infective protection to maintain tissue regional homeostasis. However, it is still unclear whether there are differences in the participation of AT in primary and secondary immune response, and whether avian AT has the similar immune function characteristics to mammals. In this study, we used Newcastle disease virus (NDV) attenuated vaccine to induce primary and secondary immune response in chickens, and the changes of the key regulatory gene NR4A3 (nuclear receptor subfamily 4 group A member 3) of T cells activation and its targeted miR-20a-5p were detected by quantitative real-time PCR (qRT-PCR). The results showed that NR4A3 actively participated in immune response of AT, and showed significant differences in expression activities between the two immune processes. "MiR-20a-5p/NR4A3" pathway was a potential molecular mechanism involved in the regulation of immune function in AT. Moreover, AT responded differently to the primary and secondary immune response possibly through the different patterns of source, apoptosis and migration for lymphocytes (such as CD8β+ T cells). This study can provide directional guidance for further studying immune functions of avian AT.

Stress-induced immunosuppression (SIIS) is one of the most common problems in intensive poultry production, which can cause immunized chickens to still develop diseases and bring huge losses to production. Recently, adipose tissue, as an immunomodulatory organ, has become a hot topic of attention. However, the function and mechanism of adipose tissue involved in SIIS and its influence on the immune response are still unclear. In this study, we dynamically analyzed the correlations between the T cells migration and change of sphingosine-1-phosphate receptor 1 (S1PR1) gene in adipose tissue using chicken models with different immune states, and further explored the regulatory mechanisms and application. The results showed that SIIS could significantly change the expressions of lymphocytes migration related S1PR1 gene, and SIIS could inhibit the Newcastle disease virus (NDV) immune response partially by affecting the migration and proliferation of TCRα+ T cells in adipose tissue. Moreover, the miR-145-5p/S1PR1 pathway was a potential key mechanism to regulate T cells migration in adipose tissue, and circulating miR-145-5p had potential value as a molecular marker. This research can provide innovative reference for in-depth studying the immunoregulatory function and mechanism of adipose tissue.

Stress-induced immunosuppression (SIIS) is one of common problems in the intensive poultry industry, affecting the effect of vaccine immunization and leading to high incidences of diseases. In this study, the expression characteristics and regulatory mechanisms of miR-214 in the processes of SIIS and its influence on the immune response to avian influenza virus (AIV) vaccine in chicken were explored. The qRT-PCR results showed that serum circulating miR-214 was significantly differentially expressed (especially on 2, 5, and 28 days post immunization (dpi)) in the processes, so had the potential as a molecular marker. MiR-214 expressions from multiple tissues were closely associated with the changes in circulating miR-214 expression levels. MiR-214-PTEN regulatory network was a potential key regulatory mechanism for the heart, bursa of Fabricius, and glandular stomach to participate in the process of SIIS affecting AIV immune response. This study can provide references for further understanding of stress affecting immune response.

Stress-induced immunosuppression (SIIS) can weaken the immune response effect of poultry vaccination, and bring huge hidden dangers and economic losses to the poultry industry. However, the detailed molecular mechanisms are still not fully understood. Unveiling the common mechanism of SIIS affecting the immune response to different vaccines is critical for detecting and minimizing the losses caused by SIIS. This study used glucocorticoid dexamethasone (Dex) to simulate SIIS, and three classic avian vaccines (including avian influenza virus (AIV), Newcastle disease virus (NDV), and infectious bursal disease virus (IBDV)) were used to induce immune responses in chicken. Quantitative real-time PCR (qRT-PCR) revealed the expression characteristics and functions of circMYO1B and miR-155 in the processes of SIIS affecting the immune response to the aforementioned avian vaccines, as well as their targeted regulatory relationship. Subsequent bioinformatics analysis predicted FOS, one of the potential target genes of miR-155. The results showed that circMYO1B/miR-155 pathway served as a key common mechanism by which SIIS affected the immune response to the three vaccines. Both heart and proventriculus appeared to be the crucial tissues for this process, with five days post immunization (dpi) emerging as the primary time of interest. Moreover, mitogen-activated protein kinase (MAPK) signaling system played a key role in modulating the immune response subsequent to SIIS administration. Our findings provide new insights into the immune function of competitive endogenous RNA (ceRNA), which have important function in the detection and treatment of SIIS affecting vaccine immunity.

Lipid metabolism plays an important role in maintaining lipid homeostasis and regulating immune functions. However, the regulations and mechanisms of lipid metabolism on the regional immune function of avian adipose tissue (AT) have not been reported. In this study, qRT-PCR was used to investigate the changes and relationships of different lipid metabolism pathways in chicken AT during stress-induced immunosuppression (SIIS) inhibiting immune response to Newcastle disease virus vaccine, then the miRNA regulation patterns of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) gene and its potential applications were further identified. The results showed that AT actively responded to SIIS, and ATGL, CPT1A and HMGCR were all the key genes involved in the processes of SIIS inhibiting the immune responses. SIIS significantly inhibited the natural and specific immune phases of the primary immune response and the initiation phase of the secondary immune response in AT by suppressing T cells by up-regulating steroid anabolism. Moreover, steroid metabolism could play dual roles in regulating the regional immune functions of AT. The miR-29a/c-3p-HMGCR network was a potential regulation mechanism of steroid metabolism in AT, and serum circulating miR-29a/c-3p had the potential as molecular markers. The study can provide valuable references for an in-depth investigation of the regional immune functions regulated by lipid metabolism in AT.

生物工程下游技术课程是生物技术专业的核心专业课,为了提升生物工程下游技术实验课程的教学效能,提高对生物技术人才培养质量的支撑度,适应生物经济发展的需要,我们构建了生物工程下游技术进阶式实验教学模式.该文从师生双向对生物工程下游技术进阶式实验教学模式的教学效果进行了评价,为持续改进教学质量提供依据.

Stress-induced immunosuppression (SIIS) is one of the common problems in intensive poultry production, which often reduces the prevention and control effects of various vaccines, including infectious bursal disease virus (IBDV) vaccine, and brings enormous economic losses to the poultry industry. However, the molecular mechanisms of SIIS inhibiting immune response to IBDV vaccine remain unclear. In this study, suppressor of cytokine signaling 3 (SOCS3) gene was selected and stress-induced immunosuppressed chickens were simulated using dexamethasone (Dex). Quantitative real-time PCR (qRT-PCR) was conducted to analyze its expression characteristics and game relationships between SOCS3 gene and miR-27b-3p (it could target SOCS3 gene) in the process of SIIS inhibiting immune response to IBDV vaccine in chicken, and the potential application value of circulating miR-27b-3p as a biomarker was also identified. The results showed that SOCS3 gene and miR-27b-3p were significantly differentially expressed in the candidate tissues during SIIS inhibiting the immune response to IBDV (P < 0.05), respectively, which were key factors involved in the process. Moreover, miR-27b-3p and SOCS3 gene showed game regulation relationships in several tissues during the process, so the miR-27b-3p/SOCS3 regulatory network was one of the key mechanisms of SOCS3 gene participating in the process. Circulating miR-27b-3p was differentially expressed in serum at 10 time points (1, 2, 3, 4, 5, 7, 14, 21, 28, and 35 days postimmunization (dpi)) in the process (P < 0.05), showing that circulating miR-27b-3p was a valid candidate target as a molecular marker for detecting SIIS inhibiting the IBDV immune response. This study can provide references for further studying molecular mechanisms of stress affecting immune response.

At present, stress-induced immunosuppression is still a hidden threat that leads to immunization failure and outbreaks of poultry diseases, and causes huge economic losses to the modern poultry industry. However, the molecular mechanisms of stress-induced immunosuppression affecting viral vaccine immunity are still poorly understood. Here, we identified circAKIRIN2 as a conserved circular transcript in chicken, and explored its expression patterns in different immune states by quantitative real-time PCR (qRT-PCR), then conducted bio-informatics analysis. The results showed that circAKIRIN2 actively participated in the process of stress-induced immunosuppression affecting the immune response to infectious bursal disease virus (IBDV) vaccine. The key time points for circAKIRIN2 involving in the process were 2 day post immunization (dpi), 5 dpi, and 28 dpi, especially at the acquired immune stage. The important tissues that responded to the process included the heart, liver, and lung, all of which changed significantly. In addition, circAKIRIN2 as a competing endogenous RNA (ceRNA) sponging zinc finger and BTB domain containing 20 (ZBTB20) was a potential molecular mechanism for regulating immune functions in the process. In conclusion, circAKIRIN2 is a key regulatory factor for stress-induced immunosuppression affecting the IBDV vaccine immune response, and this study can provide a new perspective for exploring the molecular regulatory mechanisms of stress-induced immunosuppression affecting immune response.

Adipose tissue (AT) is considered as a regional immune organ and plays an important role in the anti-infection immune response. However, the function and mechanism of chicken AT in response to secondary immune response remain poorly understood. Here, we used mRNA and microRNA (miRNA) sequencing technology to survey the transcriptomic landscape of chicken abdominal adipose tissue (AAT) during the first and second immunization with Newcastle disease virus (NDV) vaccine, and carried out bioinformatics analysis, such as Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis, protein-protein interaction (PPI) analysis, and miRNA-mRNA integrated analysis. The results indicated that chicken AAT actively responded to the secondary immune response. DNA replication and cytoskeleton regulation as the regulatory functions of immune activation changed significantly, and weakened lipid metabolism was an effective strategy for the secondary immunity. Mechanically, the regulatory network between the differentially expressed miRNAs (DEMs) and their targeted differentially expressed genes (DEGs), such as miR-206/miR-499-5p-nuclear receptor subfamily 4 group A member 3 (NR4A3)/methylsterol monooxygenase 1 (MSMO1) pathway, was one of the potential key mechanisms by which AAT responded to the secondary immune response. In conclusion, regional immunity of chicken AT responds to secondary immunity by promoting immune activation and weakening lipid metabolism, and this study can instruct future research on antiviral strategy.

MiR-155 and CTLA-4 are important factors involved in the regulation of immune function. However, there is no report about their involvement in function regulation of stress-induced immunosuppression affecting immune response. In this study, the chicken model of stress-induced immunosuppression affecting immune response (simulation with dexamethasone and immunization with Newcastle disease virus (NDV) attenuated vaccine) was established, then the expression characteristics of miR-155 and CTLA-4 gene were analyzed at several key time points during the processes of stress-induced immunosuppression affecting NDV vaccine immune response at serum and tissue levels. The results showed that miR-155 and CTLA-4 were the key factors involved in stress-induced immunosuppression and NDV immune response, whose functions involved in the regulation of immune function were different in different tissues and time points, and 2 day post immunization (dpi), 5dpi and 21dpi were the possible key regulatory time points. CTLA-4 , the target gene of miR-155, had significant game regulation relationships between them in various tissues, such as bursa of Fabricius , thymus and liver, indicating that miR-155-CTLA-4 pathway was one of the main mechanisms of their involvement in the regulations of stress-induced immunosuppression affecting NDV immune response. This study can lay the foundation for in-depth exploration of miR-155-CTLA-4 pathway involved in the regulation of immune function.

Maternal undernutrition is highly prevalent in developing countries, leading to severe fetus/infant mortality, intrauterine growth restriction, stunting, and severe wasting. However, the potential impairments of maternal undernutrition to metabolic pathways in offspring are not defined completely. In this study, 2 groups of pregnant domestic pigs received nutritionally balanced gestation diets with or without 50% feed intake restriction from 0 to 35 gestation days and 70% from 35 to 114 gestation days. Full-term fetuses were collected via C-section on day 113/114 of gestation. MicroRNA and mRNA deep sequencing were analyzed using the Illumina GAIIx system on fetal liver samples. The mRNA-miRNA correlation and associated signaling pathways were analyzed via CLC Genomics Workbench and Ingenuity Pathway Analysis Software. A total of 1189 and 34 differentially expressed mRNA and miRNAs were identified between full-nutrition (F) and restricted-nutrition (R) groups. The correlation analyses showed that metabolic and signaling pathways such as oxidative phosphorylation, death receptor signaling, neuroinflammation signaling pathway, and estrogen receptor signaling pathways were significantly modified, and the gene modifications in these pathways were associated with the miRNA changes induced by the maternal undernutrition. For example, the upregulated (P<.05) oxidative phosphorylation pathway in R group was validated using RT-qPCR, and the correlational analysis indicated that miR-221, 103, 107, 184, and 4497 correlate with their target genes NDUFA1, NDUFA11, NDUFB10 and NDUFS7 in this pathway. These results provide the framework for further understanding maternal malnutrition's negative impacts on hepatic metabolic pathways via miRNA-mRNA interactions in full-term fetal pigs.

Stress-induced immunosuppression is one of the most common hazards in poultry intensive production, which often leads to vaccination failure and severe economic losses. At present, there is no report about the function and mechanism of circulating miRNA on stress-induced immunosuppression affecting immune response. In this study, the changes of circulating miR-20a-5p under stress-induced immunosuppressive condition were analyzed by qRT-PCR, and the key time points, tissues and mechanisms for functional regulation of miR-20a-5p in the process of stress-induced immunosuppression affecting avian influenza virus (AIV) vaccine immune response were identified. The results showed that stress-induced immunosuppression down-regulated miR-20a-5p and further affected AIV vaccine immune response, in which 5 day post immunization (dpi) was a key time point, and the heart, lung, and proventriculus were the important tissues. The game relationship analysis between miR-20a-5p and its target nuclear receptor subfamily 4 group A member 3 (NR4A3) gene showed that "miR-20a-5p/ NR4A3" pathway was the potential key mechanism of this process, especially for heart and lung. This study provides insights into the molecular mechanisms of stress-induced immunosuppression affecting immune response.

Studies have shown that circulating microRNAs (miRNAs) are important players in the immune response and stress-induced immunosuppression. However, the function and mechanism of stress-induced immunosuppression affecting the immune response to the Newcastle disease virus (NDV) vaccine remain largely unknown. This study analyzed the changes of 15 NDV-related circulating miRNAs at different immune stages by qRT-PCR, aiming to explore the key timepoints, potential biomarkers, and mechanisms for the functional regulation of candidate circulating miRNAs under immunosuppressed conditions. The results showed that stress-induced immunosuppression induced differential expressions of the candidate circulating miRNAs, especially at 2 days post immunization (dpi), 14 dpi, and 28 dpi. In addition, stress-induced immunosuppression significantly affected the immune response to NDV vaccine, which was manifested by significant changes in candidate circulating miRNAs at 2 dpi, 5 dpi, and 21 dpi. The featured expressions of candidate circulating miRNAs indicated their potential application as biomarkers in immunity and immunosuppression. Bioinformatics analysis revealed that the candidate circulating miRNAs possibly regulated immune function through key targeted genes, such as Mg2+/Mn2+-dependent 1A (PPM1A) and Nemo-like kinase (NLK), in the MAPK signaling pathway. This study provides a theoretical reference for studying the function and mechanism of circulating miRNAs in immune regulation.

Vaccination is one of effective means to prevent viral infectious diseases in poultry. However, the functions of circulating miRNAs in immune response remain unknown. In this study, miR-155, a key factor in the regulation of immune function, was selected to study its expression, potential function and mechanism in the 12-day-old chicken immune responses to three vaccines (avian influenza virus inactivated vaccine, Newcastle disease virus attenuated vaccine and infectious bursal disease virus attenuated vaccine), respectively. The experiment aimed to explore the relationships between the expression levels of serum circulating miR-155 and immune responses. The results showed that the expression levels of serum circulating miR-155 were significantly different during the three immune responses, but had similarities at several time points post inoculation. 2 day post inoculation (dpi), 5dpi, and 21dpi were the possible common key time points of the three immune responses. Moreover, spleen (2dpi), bursa of Fabricius and cecal tonsil (5dpi), and liver (21dpi) were the possible key tissues associated with the differential expression levels of serum circulating miR-155. Bioinformatics analysis showed that several key target genes (such as KRAS, RAP1B, and RPS6KA3) of miR-155 possibly played a key role in immune function regulation through MAPK and mTOR signaling pathways. The study can lay the foundation for further studying the function and application of circulating miR-155 in chicken immune responses.

Stress-induced immunosuppression can affect the immune effect of vaccine. However, the mechanism of stress-induced immunosuppression affecting immune response to infectious bursal disease virus (IBDV) vaccine in chicken is still unclear. In this study, thirteen IBDV related circulating miRNAs were selected to study their expressions, possible functions and mechanisms in dexamethasone (Dex)-induced immunosuppressed chicken vaccinated with IBDV attenuated vaccine. The experiment aimed to explore the relationship between the expressions of IBDV related circulating miRNAs and stress-induced immunosuppression. The quantitative real-time PCR (qRT-PCR) results showed that Dex-induced immunosuppression could induce the differential expressions of the candidate serum circulating miRNAs, especially on the 2nd, 5th, 7th and 28th day after dexamethasone treatment. Dex-induced immunosuppression could affect the immune response to the IBDV vaccine, which was possibly achieved by partially regulating the differential expressions of the IBDV related circulating miRNAs. Bioinformatics analysis showed that the candidate miRNAs could regulate the immune function mainly through targeting genes (such as CREB1 and MAPK1) in TGF-β and MAPK signaling pathways. This study can provide a preliminary reference for further studying the function and mechanism of circulating miRNAs in immune regulation.

MicroRNAs are a class of evolutionary conserved non-coding small RNAs that play key regulatory roles at the post-transcriptional level.In recent years, studies have shown that miR-214 plays an important role in regulating several biological processes such as cell proliferation and differentiation, tumorigenesis, inflammation and immunity, and has become a hotspot in the miRNA field.In this review, the regulatory functions of miR-214 in the proliferation, differentiation and functional activities of immune-related cells, such as dendritic cells, T cells and NK cells, were briefly reviewed.Also, the mechanisms of miR-214 involved in tumor immunity, inflammatory regulation and antivirus were discussed.Finally, the value and application prospects of miR-214 as a molecular marker in inflammation and tumor related diseases were analyzed briefly.We hope it can provide reference for further study on the mechanism and application of miR-214.