马立克氏病易感鸡和抗性鸡脾脏组织转录组的特征差异分析

Analysis of characterization difference in the transcriptome of spleen tissues in Marek's disease susceptible and resistant chickens

为了探究马立克氏病(MD)易感鸡和抗性鸡脾脏组织转录组的特征差异,试验选择200只7日龄、健康的霞烟鸡(MD抗性品系核心群的后代),每只试验鸡腹腔注射含2500 pfu血清Ⅰ型马立克氏病病毒(MDV-1)超强毒株GX0101的血毒,攻毒210 d后,选取感染MDV-1超强毒株GX0101且脾脏明显具有MD肿瘤的鸡只作为易感组,未感染MDV-1超强毒株GX0101且无任何MD临床症状的鸡只作为MD抗性组,处死后提取脾脏组织总RNA,采用转录组测序技术进行差异表达基因(DEGs)的筛选,并进行基因本体(gene ontology,GO)功能富集分析、京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)信号通路分析和原发性免疫缺陷通路相关基因的表达分析。结果表明:在MD易感组与MD抗性组脾脏组织中共筛选得到152个DEGs,相对于MD易感组,MD抗性组脾脏组织有92个DEGs显著下调,60个DEGs显著上调(|lbFold Change|>1,FDR<0.05);共发现48个显著富集(P<0.05)GO条目,其中生物学过程共富集到23个条目,细胞组分共富集到16个条目,分子功能共富集到9个条目;筛选得到的DEGs在细胞周期、嘧啶代谢和原发性免疫缺陷通路等13个信号通路中显著富集。在原发性免疫缺陷通路中,与MD易感组比较,MD抗性组脾脏组织TNFRSF13B基因[编码钙调节因子(TACI)]显著下调(P<0.05),CD8α基因显著上调(P<0.05)。说明MD抗性鸡可通过下调体液免疫缺陷相关基因TACI和上调细胞免疫缺陷相关基因CD8α来提高机体对MD的抗性。

In order to investigate the characterization difference analysis on the transcriptome of spleen tissues in Marek's disease(MD)susceptible and resistant chickens,in this experiment,2007-day-old,healthy Xiayan chickens(the offspring of the core group of MD resistant breed)were injected intraperitoneally with the virus-containing blood of Marek's disease virus serotype Ⅰ(MDV-1)GXO101 of 2500 pfu very virulent strain.210 days after challenge,chickens infected with MDV-1 GX0101 very virulent strain and obviously MD tumors in the spleen were selected as the susceptible group,and chickens not infected with MDV-1 CXO101 very virulent strain and without any clinical symptoms of MD were selected as the MD-resistant group.Total RNA was extracted from spleen tissue after death,the differential expression genes(DEGs)were screened by transcriptome sequencing technology,and gene ontology(CO)function enrichment analysis,kyoto encyclopedia of genes and genomes(KEGG)signaling pathway analysis and primary immunodeficiency pathway-related gene expression analysis were performed.The results showed that a total of 152 DEGs were screened in spleen tissues between the MD-susceptible group and the MD-resistant group.Compared with the MD-susceptible group,92 DEGs were significantly downregulated,and 60 DEGs were significantly upregulated in spleen tissues of the MD-resistant group(Ilb Fold Changel>1,FDR<0.05).A total of 48 significant enrichment GO items(P<0.05)were found,including 23 enrichment items for biological processes,16 enrichment items for cellular components,and 9 enrichment items for molecular function.DEGs screened were significantly enriched in 13 signaling pathways including cell cycle,pyrimidine metabolism and primary immunodeficiency pathways.In the primary immunodeficiency pathway,TNFRSF13B gene[coding calcium regulatory factor(TACI)] was significantly down-regulated(P<0.05),and CD8αgene was significantly up-regulated(P<0.05)in the spleen of MD-resistant group compared with MD-susceptible group.These results suggest that MD-resistant chickens can improve their resistance to MD by down-regulating TACI related humoral immune deficiency and up-regulating CD8αrelated cellular immune deficiency.