【作者】 陈开森；

【导师】 傅颖媛；

【作者基本信息】 江西医学院 ， 病原生物学， 2005， 硕士

【摘要】 目的:探讨BALB/c小鼠系统感染白色念珠菌后免疫功能的动态变化。方法:BALB/c小鼠随机分成4组:正常对照组[常规条件下饲养];免疫功能低下组[小鼠背部皮下注射100mg/kg环磷酰胺(CP),隔天1次,共2次,第2次注射CP后24h后尾静脉注入0.2ml的无菌生理盐水];高剂量感染组[第2次注射CP后24h尾静脉感染白色念珠菌2×10~5CFU];低剂量感染组[第2次注入CP后24h尾静脉感染白色念珠菌2×10~4CFU]。分别于感染白色念珠菌1、4、7天后检测各组小鼠的免疫功能,指标如下:1.ELISA法测定感染1、4、7天后小鼠血清IL-18、IL-4、IFN-γ的水平。2.MTT法测定ConA诱导的小鼠脾脏T淋巴细胞增殖功能。3.RBC-C3bRR法观察各组小鼠红细胞的粘附功能。4.常规真菌培养,计算各组小鼠左肾白色念珠菌菌数(CFU)。结果:1. BALB/c小鼠系统感染白色念珠菌1天后,高剂量感染组和低剂量感染组的血清IL-18、IL-4、IFN-γ含量均低于正常对照组,但高于免疫功能低下组,差异有显著性。感染4天后,高剂量感染组的血清IL-18水平与正常对照组比较差异无显著性(P>0.05),但高于免疫功能低下对照组(P<0.05);低剂量感染组血清IL-18低于正常对照组(P<0.05),但与免疫功能低下对照组比较差异无显著性(P>0.05);高剂量感染组和低剂量感染组的血清IL-4、IFN-γ均高于正常对照组及免疫功能低下组。感染7天后,高剂量感染组的血清IL-18、IFN-γ均低于正常对照组及免疫功能低下组,差异有显著性,而血清IL-4高于正常对照组及免疫功能低下组;低剂量感染组的血清IL-18、IL-4与正常对照组及免疫功能低下组比较差异无显著性(P>0.05),但其血清IFN-γ水平明显高于正常对照组及免疫功能低下组,差异有显著性。2. ConA诱导的BLAB/c小鼠脾T淋巴细胞增殖功能:感染1天后,高剂量感染组和低剂量感染组的T淋巴细胞增殖功能均低于正常对照组但高于免疫功能低下组。感染4天后,低剂量感染组和高剂量感染组的T淋巴细胞增殖功能开始增强。感染七天后,低剂量感染组T淋巴细胞更多还原

【Abstract】 Objective: Exploring the kinetics of immunologic function in BALB/c mice systemically infected with Candida Albicans. Methods: The BALB/c mice are divided into 4 groups. Normal control group: routine feeding. Low immunologic function group: the mice injected s.c. with 100mg/kg CP(cyclophosphamide) every two days on the back of mice for two times, the mice were inoculated with 0.2ml aseptic N.S via the lateral tail vein within 24 hrs after treated with CP at twice. The high dose infecting group: The mice were inoculated with 2×105CFU of Candida Albicans via the lateral tail vein within 24 hrs after treated with CP at twice. The low dose infecting group: the mice were inoculated with 2×104CFU of Candida Albicans via the lateral tail vein within 24 hrs after treated with CP at twice. The indicators of immunological kinetics after infecting CA in mice of 4 groups were measured as follows: 1 ELISA method was used to detect the level of IL-18、IL-4 and IFN-γin the serum of the mice in each group at the 1、4、7 days after the mices infecting CA. 2 MTT method was used to measure spleen lymphocyte proliferation induced by ConA. 3 RBC-C3bRR test was used to observe the adhesive ability of erythrocyte. 4 The colony of Candida Albicans in the left kidney of these mice in each group were calculated and expressed in colony forming unit(CFU). Results: 1. On day 1 after systemically infected with Candida Albicans, the level of IL-18、IL-4、IFN-γin the serum from the BLAB/c mice of the high dose infecting group and the low dose infecting group elevated with statistically significant differen ce in comparison with low immunologic function group but decreased with normal co ntrol group. On day 4 after systemically infection, the level of IL-18 in the serum in th e high dose group have no statistically significant difference comparing with the nor mal control group(P>0.05), but decreased with statistically difference comparing wit h low immunologic function control group(P<0.05). the level of IL-18 in the serum in the low dose group have statistically significant difference comparing with the norm al control group(P<0.05), but have no statistically difference comparing with low imm unologic function control group(P>0.05). The level of IL-4 and IFN-γin the serum of the high dose group and the low dose group elevated with statistically significant difference in comparison with low immunologic function group and normal control gr oup. On day 7 post systemically infection, the level of IL-18、IFN-γin the serum in the high dose group decreased with significant difference in comparison with normal control group and low immunologic function group, however, the level of IL-4 in the serum increased with statistically significant difference in comparison with normal co ntrol group and low immunologic function group. The level of IL-18、IL-4 in the ser um in the low dose group increased with no statistically difference in comparison with the normal control group and the low immunologic function group(P>0.05), but the level of IFN-γsignificantly increased comparing with the normal control group and the low immunologic function group. 2. T lymphocyte of BLAB/c mice proliferation activity induced by ConA: On day 1 post infection, the high dose infecting group and the low dose infecting group was statistically significant difference comparing with normal control group and low immunologic function group. On the day 4 after infect ion, T lymphocyte proliferation activity in the high dose infecting group and the low dose infecting group have been increasing. On day 7 post infection, T lymphocyte proliferation activity in the low dose infecting group had no statistically significant difference comparing with normal control group (P> 0.05), but was higher than the level of the low immunologic function group(P<0.05), T lymphocyte proliferation activity in the high dose infecting group had no statistically significant difference comparing with the low immunologic function group (P> 0.05), but was lower than the level of normal control group(P<0.05) 3. On day 1 after infection, RBC-C3bRR activity in the high dose infecting group and the low dose infecting group had no statistically significant difference comparing with normal control group and low immuno logic function group(P>0.05),but increased comparing with the low immunologic fun ction group (P<0.0 5).On day 4 after infection, the RBC-C3bRR of the high dose infe cting group and the low dose infecting group increased in comparison with the normal control group and low immunologic function group(P<0. 05). On day 7 after infection, RB C-C3bRR of the low dose group elevated with statistically significant difference in comparison with low immunologic function group and normal control group. 4. The calculation of Candida Albicans in the left kidney after infection: On day 1 and 4 after infection, the CFU number of Candida Albicans in the left kidney of mice in the high dose infecting group was higher with statistically significant difference than the low dose infecting group (P<0. 05). On day 7 after infection, the left kidney of the high dose infecting group has Candida Albicans too, but the low dose infecting group have no Candida Albicans in the left kindey either. The colony of Candida Albicans on Sabouraud medium in which minced left kidney of mice from the normal control group and the low immunologic function group at the 1、4、7 days was cultivated res pectively had not been observed. Conclusion: 1. On day 1 after BLAB/c mice systemically infected with Candida Albicans, the cytokines of Th1/Th2 of high dose infecting group and low dose infecting group increased. On day 7 after systemically infected, the level of Th2 cytokines of the high dose infecting group was higher than the level of Th1 cytokines in the high dose infecting group, but the level of Th1 cytokines was higher than the level of Th2 cytokines in the low dose infecting group, which showed that the infection of Candida Albicans of BLAB/c mice have some stimulus to the immunologic function. At the infecting earlier stage, the cellular immunity and humoral immunity would enhance. At the infecting lateral stage, the cellular immunologic response become more importan at the low dose infecting, but the humoral immunologic response become more important in the high dose infecting. 2. When BLAB/c mice systemically infecting Candida Albicans, the T lymphocyteproliferation activity would be certain stimulus at the infecting earlier,but increased to the level of the normal group as time lasting(one week) at low dose infecting group, but do not recover to the normal level at high dose infecting group,which showed that BLAB/c mice infecting the low dose Candida Albicans reinforce T lymphocyte proliferation activity induced by ConA, however, BLAB/c mice infecting the high dose Candida Albicans enhance T lymphocyte proliferation activity induced by ConA at the infecting earlier stage, opposite at the lateral stage.3. The BLAB/c mice of infecting low dose Candida Albicans(one week) reinfore effect on the RBC-C3bRR function, but that have no effect to high dose infecting mice. 4. When BALB/c mice systemically infecting Candida Albicans, the ability of clearing Candida Albicans will enhance fellowing time going but the ability of clearance to Candida Albicans will be inhibited更多还原