沙门氏菌和聚肌胞处理条件下鸡免疫相关候选基因mRNA表达特性

doi:10.3864/j.issn.0578-1752.2014.17.016

摘要/Abstract

摘要： 【目的】利用高密度SNP芯片完成了对北京油鸡血清免疫球蛋白Y含量等9个免疫性状的关联分析，筛选得到了与性状显著关联位点和候选基因。基于该研究结果，以集中分布在16号染色体的候选基因CD1b、 BMA1（B locus M alpha chain 1）、TRIM27（tripartite motif-containing 27）和ZNF692（zinc finger protein 692）作为研究对象，以北京油鸡为实验材料，在聚肌胞（Polyinosinic acid-polycytidylic acid, Poly I:C）和细菌的处理下进一步对候选基因表达特性进行分析和鉴定。【方法】随机选取80只12日龄北京油鸡分为3组：空白对照组、聚肌胞处理组和肠炎沙门氏菌（Salmonella enteritidis, SE）处理组，分别饲养在独立的隔离器内。两处理组分别胸肌注射聚肌胞注射液和SE菌液，对照组注射生理盐水，于处理后12 h、24 h、3 d和6 d（days post infection, DPI）检测血清炎症因子的变化水平和候选基因表达规律。【结果】经过聚肌胞和SE处理后，体重在24 h之后显著低于空白组（P＜0.05），体温24 h以内显著升高；血清IFN-α、IL-4和IL-6水平先升高后降低，在第24小时或第3天达到峰值，TNF-α水平持续升高，3 d后极显著高于空白组（P＜0.01）。两种处理条件下CD1b的mRNA表达没有组织特异性，其他3个基因在胸腺和法氏囊中高表达。在胸腺组织中，CD1b在两处理间12和24 h的表达量存在显著差异（P＜0.01），在聚肌胞处理组整个感染阶段没有显著性变化，在SE组是先升高后降低的趋势，感染后24 h时表达量最高（P＜0.01）；BMA1在12 h和3 d时两处理组间差异显著，聚肌胞处理组在12 h时表达量低于SE处理组，而在3 d时显著高于SE处理组（P＜0.01）；TRIM27在聚肌胞感染6 d时表达量显著高于空白组（P＜0.05），ZNF692的表达量在3组间没有差异。在法氏囊中，CD1b表达量在两个处理组中存在差异，在SE组12 h时表达量高；BMA1和TRIM27的表达量在两组间没有差异，TRIM27在3 d时表达量最高（P＜0.01），ZNF692在SE组感染24 h时相对表达量高于聚肌胞处理组，在两组中都在3 d时表达最高。【结论】CD1b、BMA1、TRIM27和ZNF692参与了聚肌胞和肠炎沙门氏菌引起的免疫反应过程，是与免疫相关的功能基因；CD1b主要在肠炎沙门氏菌感染的前期发挥作用；BMA1和ZNF692分别参与胸腺和法氏囊的免疫反应。

关键词: CD1b , BMA1 , TRIM27 , ZNF692 , mRNA表达 , 鸡

Abstract: 【Objective】 Genome-wide association studies were performed by using the chicken 60k high density SNP array for nine immune traits in Beijing-You chicken and identification of candidate genes and loci responsible for these traits were identified. Here, by treating Beijing-You chicken with polyinosinic acid-polycytidylic acid, Poly(I:C) and Salmonella enteritidis (SE), several candidate genes were further studied based on authors’ GWAS result, including CD1b, BMA1(B locus M alpha chain 1), TRIM27 (tripartite motif-containing 27) and ZNF692(zinc finger protein 692). 【Method】 Eighty 12-d-old Beijing-You Chickens were divided into three groups: the control group, Poly(I:C) treatment and SE treatment groups. All the experimental birds were reared in isolated facility. The treatment groups were, respectively, injected intramuscularly into the breast with 0.5 mL of Poly(I:C) and SE bacterial suspension containing 108 CFU and the control group was given 0.5 mL saline. Chickens were sacrificed at 12 h, 24 h, 3 and 6 days of post infection (DPI). Blood samples were collected and serum was stored. The bursa of Fabricius, thymus and spleen were rapidly removed to test inflammatory factors and gene expression. 【Result】 The weight was significantly lower than control group 24 h post infection (P＜0.01) and temperature of chickens was significantly changed in 24 h after treatments. In serum, concentrations of IFN-α, IL-4 and IL-6 were significantly higher than the control and reached a peak at 24 h or 3 d (P＜0.01). TNF-α increased in all periods and significantly higher than the control group after 3 days. The expression of candidate gene CD1b was not tissue-specific, but BMA1, TRIM27 and ZNF692 were highly expressed in thymus and bursa fabricius. In thymus, the expression of CD1b was significantly different between two treatments at 12 h and 24 h post infection. When treated with SE, the expression of CD1b was significantly increased and reached a peak at 24 h (P＜0.01). The mRNA expression of BMA1 was significantly different between two treatment groups in 12 h and 3 d post infection, when treated with Poly(I:C), the expression at 12 h post infection was lower than the SE group, but it was significantly higher than SE group (P＜0.01). The expression of TRIM27 on 6 DPI in Poly(I:C) treatment was significantly higher than control group, expression of ZNF692 was not different among three groups. In bursa of Fabricius, the expression of CD1b was the highest at 12 h after infection. There were no differences in the expression of genes BMA1 and TRIM27 between two treatments. The expression of ZNF692 was higher on 1 DPI in SE treatment than Poly(I:C), and also the highest on 3 DPI in both groups. 【Conclusion】 CD1b, BMA1,TRIM27 and ZNF692 might be involved in the immune response after treating with S. enteritidis and Poly(I:C). CD1b gene had an important role in the early stage of SE infections. BMA1 and ZNF692 were involved in immune response of thymus and bursa.

Key words: CD1b , BMA1 , TRIM27 , ZNF692 , mRNA expression , chickens

孙艳，李建超，李鹏，郑麦青，刘冉冉，李庆贺，文杰，赵桂苹. 沙门氏菌和聚肌胞处理条件下鸡免疫相关候选基因mRNA表达特性[J]. 中国农业科学, 2014, 47(17): 3483-3491.

SUN Yan, LI Jian-chao, LI Peng, ZHENG Mai-qing, LIU Ran-ran, LI Qing-he, WEN Jie, ZHAO Gui-ping. Transcription Profiles of Immune-Related Genes in Chickens Infected by Salmonella Enteritidis and Poly(I:C)[J]. Scientia Agricultura Sinica, 2014, 47(17): 3483-3491.

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