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目的观察BCG-CpG-DNA重复给药对小鼠免疫器官和免疫功能的影响,评价BCG-CpG-DNA的免疫毒性。方法将BALB/c小鼠随机分为4组,对照组注射生理盐水,实验组分别注射低(100μg)、中(250μg)、高(500μg)剂量的BCG-CpG-DNA,于28d内分别经背部皮下免疫7次,分别于初次免疫后14d、末次免疫后3d和3周,检测各组小鼠免疫器官脏器系数、淋巴细胞转化功能、细胞因子释放(ELISPOT法)、T细胞亚群变化(免疫荧光法)和NK细胞杀伤活性(乳酸脱氢酶法)。结果 BCG-CpG-DNA重复给药7次,主要影响小鼠初级免疫器官,表现为脾肿大,对胸腺、肝脏和肾脏无影响;对细胞免疫功能的影响是增强淋巴细胞转化功能;降低中、高剂量组CD3+T细胞亚群含量;提高体内分泌IFNγ和IL-4的细胞数二者的比例;增强NK细胞的杀伤能力。结论 BCG-CpG-DNA重复给药累计达3.5mg,小鼠表现为免疫功能增强,对免疫器官和免疫功能无不良影响,未见免疫毒性副作用。
Objective To observe the effects of repeated administration of BCG-CpG-DNA on immune organs and immune function in mice and evaluate the immunotoxicity of BCG-CpG-DNA. Methods BALB / c mice were randomly divided into four groups. The control group was injected with normal saline. The experimental group was given BCG-CpG-DNA of low (100μg), moderate (250μg) and high The mice were immunized subcutaneously 7 times in the back, respectively. The organ function, lymphocyte transformation, ELISPOT and T cell subsets of immune organs in mice were detected 14 d after the first immunization and 3d and 3 weeks after the last immunization respectively. (Immunofluorescence) and NK cell killing activity (lactate dehydrogenase method). Results BCG-CpG-DNA was administered repeatedly seven times, mainly affecting the primary immune organs in mice, showing splenomegaly, no effect on the thymus, liver and kidney; the effect on cellular immune function was enhanced lymphocyte transformation; , High-dose group of CD3 + T cell subsets; increase the proportion of cells secreting IFNγ and IL-4 in vivo; and enhance the cytotoxicity of NK cells. CONCLUSIONS: BCG-CpG-DNA was administered repeatedly in a total amount of 3.5mg. Mice showed enhanced immune function and no adverse effects on immune organs and immune function, with no immunotoxic side effects.