【作者】 田野；

【导师】 崔言顺；

【作者基本信息】 山东农业大学 ， 预防兽医， 2007， 硕士

【摘要】 胶体金(Colloidal gold)是由氯金酸(HAuCl4)在还原剂的作用下聚合成的带负电的疏水胶溶液,具有高电子密度、介电特性和催化作用,能与多种生物大分子结合,且不影响其生物活性。1971年胶体金(CG)被Faulk正式引入免疫化学中后,在免疫金标记方面被广泛应用。近年来,科技工作者对胶体金的研究扩展到了免疫原性方面,但是目前国内外对胶体金的研究报导仅限于CG和其他物质(半抗原和抗原)结合后对试验动物免疫功能的影响方面,而对单独使用CG对试验动物免疫功能影响的研究,尚未见报导。为了明确不同胶体金直径和剂量对动物机体免疫功能的影响,为胶体金在免疫原性方面的应用寻求理论依据。本研究在参考Dykman等人研究资料的基础上,通过对试验小鼠机体注射不同直径、不同剂量的CG,采取免疫学研究的测试方法,对试验小鼠机体的T淋巴细胞增殖活性、血清IL-2含量、腹腔巨噬细胞活性及脾脏NK细胞杀伤活性等能够衡量机体特异和非特异性免疫的重要指标进行了检测,研究结果如下:实验一胶体金对小鼠脾脏和外周血T淋巴细胞增殖反应的影响。通过直径、剂量、时间三个因素对小鼠特异性细胞免疫水平的综合影响的研究,结果表明:与对照相比,不同直径的胶体金对小鼠脾脏和外周血T淋巴细胞增殖反应均有一定影响,但不同直径和剂量组合对其影响不同,其中5nm胶体金影响不显著, 10nm和15nm胶体金影响显著,而且剂量越大,效果越明显;以15nm,0.3 mL的胶体金对T淋巴细胞的影响最为显著。从时间上看,在14d时各处理组胶体金的效果最为明显。实验二胶体金对小鼠外周血血清和脾脏淋巴细胞诱生白介素2(IL-2)含量的影响。采用免疫放射法,对试验小鼠外周血血清和脾脏淋巴细胞诱生IL-2含量的进行了测试,结果显示:脾脏淋巴细胞诱生的IL-2的规律性比血清中的规律明显,但是总体趋势相同。10nm和15nm组的CG在各个剂量组都可以使血清及其淋巴细胞诱导的IL-2分泌增多,且与剂量呈明显的正相关,但是5nm和20nm组的效果均不明显。实验三免疫胶体金对小鼠腹腔巨噬细胞功能和脾脏NK细胞活性的影响。试验结果表明,与对照组相比,各直径组中的低、中、高剂量组均可明显增强巨嗜和NK细胞活性,并以15nm 0.2mL组最为显著,但是同直径组CG低、中、高三个剂量组之间,高剂量组优于中、低剂量组。综合这三个试验的结果分析,可以说明,不同直径和剂量的CG对机体免疫活性有不同程度的影响,其中直径15nm、10nmCG的0.3mL和0.2mL剂量组不仅对T淋巴细胞增殖活性和血清中IL-2含量有显著促进作用,而且可显著提高腹腔巨噬细胞活性及脾脏NK细胞杀伤活性,而0.1mL剂量组虽有提高但不显著;直径5nm、20nm各个剂量组对机体免疫活性的影响效果波动较大,其中中、高剂量组对四项指标均有一定的促进作用,但效果不显著,低剂量组则没有促进作用。表明15nm和10nm中、高剂量的胶体金可显著提高小鼠机体特异性和非特异性细胞免疫功能。更多还原

【Abstract】 Colloidal gold (CG) is an kind of hydrophobe solution gathered by the hydrochloro-auric acid under the electronegative circumstance, It has high electron density, electricity medium ability and the catalytic effect. It can combine many kinds of biological macro-molecule and does not affect its biological activity. After the colloidal gold (CG) was introduced into immunochemistry by the Faulk in 1971, it is widely applied in the area of immunity mark. In recent years, the researchers developed the immunity colloid gold and began to research the immunity original application. Rencently the reports of domestic and foreign research are only restricted in influence of combination of CG and other matter (half antigen and antigen) on the experimental animal immunity function, but it has no reported about influence of single CG on the experimental animal immunity function.In view of this, from the reference to the former work of Dykman’s studies, in this paper we have injected different diameter, different dosage CG to the mice organism , using the test method of the immunology research. We tested the mice organism’s T-lymphocyte multiplication activeness, IL-2 content in the blood serum, abdominal cavity macrophage activeness and spleen NK cell activeness and so on, which are important indexes that can judge of the special and the non- specificity immunity of the organism. The examination has been carried on for the purpose of definitude the influence of CG on the animal organism immunity function , through the comparison between the different experimental group results and the comparison group results, and then we can carry on the preliminary discussion for the application of colloid gold in the immunity original area. The main results of this paper include:Results 1: The influence of colloid gold on the mice spleen and serum T lymphocyte proliferating response. We synthetically studied the influence of colloid gold on the mice body fluid immunity level and the cellular immunity level through three factors that is the diameter, the dosage, the time. Result shows that the influence of colloid gold on the mice spleen and serum T lymphocyte proliferating response is different because of different diameter, the influence of 5nm colloid gold is not remarkable, 10nm and the 15nm is remarkable (p<0.05), and when the dosage is bigger, the effect is more remarkable. The influence of colloid gold which is 15nm, 0.3 mL on the T-lymphocyte is most remarkable. Measured by days, the influence of colloid gold every diameter which are14 days are most remarkable.Result 2: The influence of colloid gold on content of IL-2 in the mice serum and spleen lymphocyte.This experimentation adopts RIA assays to mensurate the content of IL-2 in the mice serum and spleen lymphocyte.The spleen lymphocyte IL-2 has a better orderliness than the mice serum IL-2 ,but they have the same direction . Compared with the comparison group, content of IL-2 in the blood serum and lymphocyte of all kinds of 10 and 15nm groups are in the majority, and assume obvious direct ratio relations with the dosage.Result 3: The influence of immune colloids gold on the activity of abdominal cavity macrophage and NK cell.The experiment shows that, compared with the comparison group, all the low, middle, the high doses team in various diameters group can notably strengthen the activity of abdominal cavity macrophage and NK cell, and the 15nm 0.2mL group is most remarkable. And in the same diameter group, the high doses team surpasses the low and middle monitoring team.All the results shows that different effects on mice which injects CG for mice.0.3mL、0.2mL of 15 nm、10 nm CG can enhance the activity of lymphocyte proliferating and increase content of IL-2 in serum and induction in spleen lymphocyte, meanwhile they can also enhance the activity of abdominal cavity macrophage and NK cell notably; 0.1 mL doses can improve also but unconspicuous. The effects on the doses of 5nm、20nmCG is fluctuate,the group of middle and high doses can improve the four criterions but unconspicuous; the group of low doses have no effect. We can learn that the middle and high doses of 15 nm and 10 nm colloidal gold can enhancing specific and nonspecific immune response in laboratory mice.更多还原