ALV-J感染对鸡脾脏巨噬细胞天然免疫因子的影响  

Innate Immune Factor Induced by Avian Leukosis Virus Subgroup J in Chicken Spleen Macrophages

作　　者：张曦 谢婷婷 冯敏 张细权[1,2] ZHANG Xi;XIE Tingting;FENG Min;ZHANG Xiquan(College of Animal Science,South China Agricultural University,Guangzhou,Guangdong 510642;Key Lab of Chicken Genetics,Breeding and Reproduction,Ministry of Agriculture and Rural Affair,Guangzhou,Guangdong 510642)

机构地区：[1]华南农业大学动物科学学院,广东广州510642 [2]农业农村部鸡遗传育种与繁殖重点试验室,广东广州510642

出　　处：《中国家禽》2024年第3期44-50,共7页China Poultry

基　　金：现代农业产业技术体系专项资金(CARS-41-G22);国家自然科学基金项目(31970540)。

摘　　要：为探究J亚群禽白血病病毒(Avian leukosis virus subgroup J,ALV-J)对鸡脾脏巨噬细胞天然免疫反应的影响,试验利用密度梯度离心法和贴壁法,从鸡脾脏组织中分离得到单核细胞,并通过显微镜观察培养至24 h、72 h、120 h单核细胞的分化过程;通过PCR扩增ALV-J特异性引物H5/H7,验证脾脏巨噬细胞对ALV-J的易感情况;同时,通过qPCR验证感染ALV-J后鸡原代脾脏巨噬细胞中炎性因子、干扰素及肿瘤抑制因子的表达。结果显示:单核细胞分化至第120 h形成形态正常的巨噬细胞,通过流式细胞技术检测鸡巨噬细胞表面特异性标记蛋白发现,84.9%的细胞为KUL01阳性细胞;鸡原代脾脏巨噬细胞对ALV-J易感,并且ALV-J可整合进鸡原代脾脏巨噬细胞基因组中;对感染ALV-J的鸡脾脏巨噬细胞进行形态观察,发现ALV-J感染导致鸡脾脏巨噬细胞形态逐渐萎缩直至死亡;ALV-J感染6 h、12 h、24 h及48 h可诱导鸡原代脾脏巨噬细胞炎性因子(IL-1β、IL-6)的表达显著上调,在ALV-J感染24 h、48 h诱导肿瘤抑制因子(TNF-α)的表达显著上调(P<0.05);并且在早期(感染3 h)诱导干扰素(IFN-β)及干扰素刺激基因(CH25H、STAT1、PKR)表达,后期(感染48 h)抑制部分干扰素刺激基因(OAS、STAT1、ZAP、Mx、PKR)的表达。研究表明ALV-J感染鸡原代脾脏巨噬细胞早期可诱发强烈的天然免疫反应,后期可能通过抑制IFN-β及部分干扰素刺激基因的表达实现免疫逃逸。The aim of this study was to investigate the effect of avian leukosis virus subgroup J(ALV-J)on natural immune response of chicken splenic macrophages.Monocytes were isolated from chicken splenic tissues using density gradient centrifuga⁃tion and adherent culture method,and the differentiation process of monocytes was observed by microscopy on 24 h,72 h and 120 h;The susceptibility of spleen macrophages to ALV-J infection was verified by PCR amplification by ALV-J-specific primers H5/H7;The expression of inflammatory factors,interferons and tumor suppressors in primary splenic macrophages after infection with ALV-J was also detected by qPCR.The results showed that monocytes differentiate to the 120 h to form morphologically nor⁃mal macrophages,and the flow cytometry detected chicken macrophage surface-specific marker proteins showed that 84.9%of cells were KUL01-positive cells.The chicken primary splenic macrophages were susceptible to ALV-J and ALV-J could be inte⁃grated into the genome of chicken primary splenic macrophages.The chicken primary splenic macrophages after infection with ALV-J revealed that infection with ALV-J resulted in gradual morphological atrophy of chicken spleen macrophages until death.ALV-J infection induced significant up-regulation of inflammatory factors(IL-1β,IL-6)at 6 h,12 h,24 h,48 h after infection,induced significant up-regulation of interferon(IFN-β)and tumor suppressor(TNF-α)at 24 h,48 h after infection in chicken pri⁃mary splenic macrophages(P<0.05),and induced up-regulation of expression of some interferon stimulated genes(CH25H,STAT1,PKR)in the early stage(3 h after infection)but suppressed the expression of some interferon stimulated genes(OAS,STAT1,ZAP,Mx,PKR)in the later stage(48 h after infection).The results indicated that ALV-J infection of chicken primary splenic macrophages induced the innate immune response at an early stage,and immune escape might be achieved in the later stage by suppressing the expression of IFN-βas well as some interferon-stimulated ge

关 键 词：鸡 鸡原代脾脏巨噬细胞 ALV-J感染 天然免疫因子 

分 类 号：S855.3[农业科学—临床兽医学]