柔嫩艾美耳球虫卵囊免疫后鸡盲肠扁桃体GM-CSF基因表达的动态变化

Expressing Profiling of GM-CSF Gene in Cecal Tonsil of Chickens Vaccinated with Eimeria tenella Oocysts

为检测和评价鸡粒细胞-巨噬细胞集落刺激因子(ChGM-CSF)在抗球虫免疫过程中的作用,以RT-PCR技术从鸡盲肠扁桃体中克隆获得了ChGM-CSF编码基因的开放阅读框(ORF),其长为435 bp。将试验鸡随机分为对照组和试验组,试验组于12、22日龄时用E.tenella孢子化卵囊进行免疫,分别于设定时间每次取5只鸡采集盲肠扁桃体,以鸡β-actin为参照建立半定量RT-PCR方法检测ChGM-CSF的表达动态。结果表明,在第1次免疫后的第1天(13日龄,P<0.05)和第2次免疫后的第7天(29日龄,P<0.01),试验组ChGM-CSF的表达量显著高于对照组;在第1次免疫后的第9天(21日龄)和第2次免疫后的第3天(25日龄)ChGM-CSF的表达量显著低于对照组(P<0.01)。本研究提示ChGM-CSF参与了鸡体的抗球虫感染过程,为进一步探讨ChGM-CSF的生物学活性及作为鸡球虫病基因工程疫苗佐剂的研究奠定基础。

In order to evaluate the effects of chicken granulocyte macrophage colony stimulating factor（GM-CSF） on the anticoccidial immunity, the ORF of ChGM-CSF were cloned from chicken cecal tonsil using RT-PCR with a pair of specific primers designed from the reported sequence in GenBank. The full-length of ChGM-CSF ORF is 435 bp. The semi-quantitative RT-PCR using chicken β-actin as inner-reference for detecting the expression of ChGM-CSF was further developed and then was used for detecting the expressing patterns of ChGM-CSF in chickens vaccinated with sporulated oocysts of E. tenella at 12 and 22 day-old. Our results indicated that the expression level of ChGM-CSF in experimental group significantly increased at day 1 post-primary（P〈0.05） and day 7 post-boost（P〈0.01） infection, decreased significantly（P〈0.01）at day 9 post-primary and at day 3 post-boost infection. It is suggested that ChGM-CSF can play important roles in the immune responses against E. tenella infection in chickens.