草鱼干扰素的体外诱生及其组织细胞的来源  

IN VITRO INDUCTION AND TISSUE AND CELL ORIGIN OF INTERFERON IN GRASS CARP (CTENOPHARYNGODON IDELLUS)

作　　者：邵健忠[1] 项黎新[1] 

机构地区：[1]浙江大学生命科学学院,杭州310012

出　　处：《动物学报》2001年第6期677-683,共7页ACTA ZOOLOGICA SINICA

基　　金：国家自然科学基金资助项目 (No .3940 0 10 0 ;39970 5 89);浙江省自然科学基金资助项目 (No .39730 0 )~~

摘　　要：草鱼九种组织干扰素 (IFN)体外诱生试验结果表明 ,从头肾、脾脏、外周血和胸腺组织中均能诱生出IFN。进一步对四种组织的细胞组分进行分离和IFN诱导 ,证实IFN由其中的白细胞所合成。理化性质、中和试验、Dot ELISA和Western blot等研究表明 ,体外诱生的白细胞IFN与体内诱生的血清IFN是同一种物质。在此基础上 ,又对白细胞组分进行分离和IFN诱导 ,并结合免疫荧光标记技术 ,证实IFN由白细胞中的淋巴细胞所合成。进而采用灭活增殖细胞法 ,对T、B淋巴细胞进行了功能性分离和IFN诱导 。Interferon is an important cytokine that has been shown to have many functions such as anti virus and immunomodulation. The tissue and cell origin of interferon had been well studied in humans and higher vertebrates, but less known in fishes. This paper reported the tissue and cell origin of Grass carp ( Ctenopharyngodon idellus ) interferon for the first time in order to investigate and understand the biological role of interferon in fish. The in vitro induction experiment of Grass carp interferon from nine tissues of peripheral blood,head kidney,kidney,thymus,spleen,liver,intestines,heart and muscle were undertaken. The results showed that the Grass carp interferon was mainly produced by the immune related tissues of head kidney, spleen, peripheral blood and thymus which were full of leucocytes. Therefore, the leucocytes were further isolated from these tissues by Percoll gradient and density centrifugation and induced the interferon in vitro . The results confirmed that leucocytes were interferon producing cells. Biochemical and biophysical characteristic studies indicated that the leucocyte interferon induced in vitro had same character with serum interferon induced in vivo . Furthermore, the antigenic characteristic analyses of neutralization test, dot ELISA and Western blot were performed between two kinds of interferon, and the results also showed that the leucocyte interferon was the same as that of serum interferon. In order to find out what kind of cell in leucocytes was the interferon producing cell, lymphocytes were purified from leucocytes by Ficoll Urografin gradient and density centrifugation for the further induction of interferon, and the results showed that the lymphocytes were the interferon producing cells. Finally, T and B lymphocyte was selectively separated by genetic incorporation of BudR and ultraviolet illumination inactivation technique to induce interferon respectively, the results indicated that the Grass carp interferon was produced by T lymphocyte.

关 键 词：草鱼 干扰素 体外诱生 组织来源 形成细胞 

分 类 号：Q959.4[生物学—动物学] S942.1[农业科学—水产养殖]