摘要
目的 揭示红鹿抗结核杆菌感染的免疫反应途径。方法 共采用 2 0只红鹿 ,其中一组 10只对牛型结核杆菌(Mycobacteriumbovis,M bovis)有自然抵抗力 ,另一组 10只对M bovis敏感。两组各取 5只接种活的卡介苗 (BCG) ,余下 5只不接种。接种卡介苗 8周后所有的红鹿都注射有毒力的M bovis,在注射M bovis4周前以及 6周后分别收集外周血单核细胞 (Peripheralbloodmononuclearcells ,PBMC) ,PBMC在与美洲商陆原 (PokeweedMitogen ,PWM)共育后测定细胞因子Inter feron -γ(IFN -γ)和Interleukin - 4 (IL - 4 )mRNA的表达水平。mRNA的定量测定是在ABIPRISM 770 0序列检测系统上应用TaqManReal-time11PCR技术进行的。结果 在红鹿接受M bovis刺激前后PBMC中IFN -γ和IL - 4mRNA的表达水平有显著性差异。结论 红鹿抵抗结核杆菌感染的免疫反应是复杂的混合细胞反应型 ,Th1,Th2细胞反应均加强。
Aim This study was to elucidate the immune mechanisms operative in red deer infected with Mycobacterium bovis (M.bovis).Methods Twenty red deer,of which ten was resistant and ten was susceptible to M.bovis,were used in this study.Half of the resistant and half of the susceptible groups of red deer were vaccinated with live bacille Calmette-Guerin (BCG).All of animals had been challenged with virulent M.bovis eight weeks after vaccination.Peripheral blood mononuclear cells (PBMC) were collected four weeks before and six weeks after challenged with virulent M.bovis.The expression levels of cytokines mRNA were measured following stimulation of cells with Pokeweed Mitogen (PWM).Interferon-γ (IFN-γ) and interleukin-4(IL-4)mRNA expression in PBMC was quantified with TaqMan Real-time PCR using an ABI PRISM 7700 Sequence Detector System.Results It was found that the difference of expression levels of IFN-γ and IL-4 mRNA in PBMC from red deer had been statistically significant before and after challenge with virulent M.bovis.Conclusions The complex cellular activation patterns can be demonstrated in red deer infected with M.bovis.With a response compatible to mixed Th1,Th2 cell activation,Real-time PCR may be a method of choice for the quantification of mRNA.
出处
《中国人兽共患病杂志》
CAS
CSCD
北大核心
2003年第6期49-51,20,共4页
Chinese Journal of Zoonoses
基金
WHO奖学金资助
