为研究主要组织相容性抗原(MHC)不同的G2系和G5系鸡在感染马立克氏病毒(MDV)后立即早期病毒复制及细胞因子转录水平的动态变化,本实验对14日龄G2和G5系SPF鸡滴鼻接种超强毒(vv MDV)Md5株,以28S r RNA基因为内参,利用双重荧光定量RT-PCR(dq RT-PCR)方法对接种后4 h、24 h、48 h和96 h肺淋巴细胞、脾脏和外周血淋巴细胞(PBL)中MDV meq、IFN-γ、IL-18、IL-4和IL-10的m RNA含量进行检测。结果显示,接种后96 h仅在两品系感染鸡的肺淋巴细胞和脾脏中检测到了meq m RNA,G2系鸡群肺淋巴细胞中病毒m RNA含量高于G5系,但脾脏中较低。96 h肺淋巴细胞和脾脏中IFN-γ、IL-4和IL-10转录水平大幅上调,并且G2系鸡群均略高于G5。本研究结果提示G2系对MDV的抵抗能力强于G5,这两种品系鸡感染MDV后可能主要启动以Th2型为主的细胞免疫反应,为研究MDV对不同MHC单倍型鸡的致病性提供了实验依据。
To compare the transcription levels of Marek's disease virus (MDV) gene and four cytokine genes during immediate-early stage after MDV-infected MHC-B haplotype chickens, twenty-four of 14-days old B haplotype chicken lines of each G2 and G5 were nasal tripped with very virulent MDV strain Md5. Then, MDV meq, IFN-γ IL-18, IL-4 and IL-10 mRNA from lung leukocytes, spleens and peripheral blood lymphocyte (PBL) of the chickens were quantified by duplex quantitative PCR (dqRT-PCR) at 4, 24, 48 and 96 hrs post infection. The result showed that meq mRNA could be detected in lung leukocytes and spleens of both infected-groups at 96 hrs, higher in lung leukocytes than in spleens, higher of G2 than that of G5 in lung, but lower of G2 than G5 in spleens. IFN-γ IL-4 and IL-10 were significantly up regulated in lung leukocytes and spleens of both lines at 96 hrs. IFN-γ IL-18, IL-4 and IL-10 in G2 lung leukocytes and spleens were higher than in those of G5. The study indicated that G2 line is more resistant to MDV than G5, and the disease course will lead to Th2 type immune response. The data will give experimental evidence to MDV research.