分支杆菌菌体裂解物增强鸡体免疫力及其作用机理研究

发布时间：2018-05-14 11:54

本文选题：菌体蛋白 + 多肽　；参考：《南京农业大学》2006年硕士论文

【摘要】：接种卡介苗是当今各国预防结核病最有效的手段之一。分支杆菌属的成员如草分支杆菌、母牛分支杆菌等也被广泛的应用于疫苗和佐剂的制备。但是，由于是全菌制剂，，菌体中对免疫功能无作用、甚至起抑制作用的成分同时存在在疫苗中，卡介苗使结核菌素的皮试出现阳性反应，因而限制了它们的应用。本文从结核分支杆菌中提取菌体蛋白和多肽，通过生物化学的方法对其进行定量检测。经过动物试验，探讨分支杆菌菌体蛋白和多肽对鸡特异性免疫和非特异性免疫的效应，最终筛选出一种低毒、高效的新型免疫增强剂并确定了临床最佳用药剂量。利用超声波低温破碎技术，提取、制备结核分支杆菌中的菌体蛋白和多肽，应用生物化学方法(lowry法)测定菌体蛋白和多肽的含量；Tricine-SDS-PAGE电泳分析菌体蛋白组分。试验结果表明：不同批次提取的菌体蛋白含量相对稳定，没有明显差异；组分分析结果表明，菌体蛋白由多种组分构成，分子量由66kD到6.5kD不等。按人常用剂量，根据人／鸡体表面积比，计算鸡用药量。利用所提取的结核分支杆菌菌体蛋白和多肽投服试验鸡。210只鸡被均分为分支杆菌菌体蛋白高剂量组(0.9mg／只)、分支杆菌菌体蛋白中剂量组(0.45mg／只)、分支杆菌菌体蛋白低剂量组(0.225mg／只)、透析多肽组(0.3mg／只)、未破碎全菌体组(0.3g／只)、分支杆菌菌体蛋白注射组(0.45mg／只)和对照组(生理盐水)七个组。除注射组外，其余六组均采用口服给药。各组试验鸡于给药后7d、14d、21d、28d分四次扑杀，分别检测鸡外周血淋巴细胞转化率、鸡新城疫抗体滴度、红细胞免疫水平、外周血淋巴细胞CD_3~+亚群和淋巴器官的组织发育情况。试验结果表明，分支杆菌菌体蛋白投服试验组均能提高鸡体的特异性免疫和非特异性免疫功能，尤以菌体蛋白注射组和菌体蛋白中剂量口服组诱导机体免疫效果为好。
[Abstract]:Bacillus Calmette Guerin is one of the most effective means of preventing tuberculosis in various countries. Members of the genus Mycobacterium, such as Mycobacterium tumefaciens and Mycobacterium cow, are also widely used in the preparation of vaccines and adjuvant. However, because of the whole bacteria preparation, there is no effect on the immune function in the bacteria, and even the components of inhibition are present in the vaccine. BCG showed positive reactions to the skin test of tuberculin, thus limiting their application.
In this paper, bacterial protein and polypeptide were extracted from Mycobacterium tuberculosis and tested by biochemical method. After animal experiments, the effect of Mycobacterium protein and polypeptide on specific immunity and non specific immunity of chicken was investigated. Finally, a new type of low toxic and efficient immunosuppressive agent was selected and the clinical best was determined. A good dose of medicament.
The bacterial protein and polypeptide in Mycobacterium tuberculosis were extracted and prepared by ultrasonic low temperature crushing technology. The contents of bacterial protein and polypeptide were measured by the biochemical method (Lowry method). The bacterial protein components were analyzed by Tricine-SDS-PAGE electrophoresis. The results showed that the protein content of different batch of bacterial proteins was relatively stable and no obvious. The results of component analysis showed that the bacterial protein was composed of many components, with molecular weights ranging from 66kD to 6.5kD.
According to the common dose of human / chicken body surface area ratio, the amount of chicken medicine was calculated. The.210 chickens were divided into high dose group (0.9mg / only), the dose group of Mycobacterium body protein (0.45mg / only), and the low dose of Mycobacterium body protein in the Mycobacterium tumefaciens. Group (0.225mg / only), dialysis polypeptide group (0.3mg / only), unbroken whole bacteria group (0.3g / only), Mycobacterium tumefaciens protein injection group (0.45mg / only) and control group (physiological saline) seven groups. Except for the injection group, the other six groups were taken oral administration. Each group of test chickens was given 7d, 14d, 21d, 28d, four times after administration, respectively, to detect the peripheral blood drenching of chickens. The rate of Ba cell transformation, the titer of the antibody to the Newcastle disease, the level of the red cell immunity, the CD_3~+ subgroup of the peripheral blood lymphocyte and the tissue development of the lymphoid organs. The experimental results showed that the specific immune and non specific immune function of the chicken body could be improved by the experimental group of Mycobacterium tumefaciens, especially in the mycelial protein injection group and the mycelial egg. The white medium dose group had good immune effect.

【学位授予单位】：南京农业大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392

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