大肠杆菌调节小鼠DC细胞表面CD40及B7分子表达
发布时间：2019-06-21 05:01
【摘要】： 目的:研究大肠杆菌刺激小鼠DC细胞对其表面分子CD40、B7-1、B7-2表达的影响。 方法:1、用不同浓度(2×107、2×106)大肠杆菌刺激小鼠DC细胞(DC2.4,2×105个/孔),LPS处理组作为阳性对照,PBS处理组作为阴性对照,在刺激4h及24h后收集细胞,提取RNA,采用RT-PCR法检测DC细胞CD40 mRNA的表达。2、扩大培养(DC细胞为1×106个/孔,大肠杆菌及LPS亦相应扩大5倍),感染4h及24h后收集细胞,用流式细胞术检测其细胞表面分子CD40、B7-1、B7-2的表达。 结果:1、与阴性对照组比较,小鼠DC细胞在大肠杆菌刺激后,其表面CD40分子显著增加,两组比较有统计学意义(均P 0. 05)。其中,刺激4h增加水平低于刺激24h[(6714.5±1367.74)copies/μl vs (13105.0±801.06)copies/μl;(10101.0±797.69)copies/μl vs (15785.0±676.23)copies/μl],两组比较有显著统计学意义(均P 0. 01) ,较低浓度(1×107)刺激增加水平高于较高浓度刺激(1×108)([10101.0±797.69)copies/μl v(s6714.5±1367.74)copies/μl;(15785.0±676.23)copies/μl vs (13105.0±801.06)copies/μl],两组比较有显著统计学意义(均P 0. 01)。2、用大肠杆菌刺激小鼠DC细胞,与PBS阴性对照和LPS阳性对照比较,其表面分子B7-1、B7-2均无明显变化,无统计学意义。 结论:1、培养的小鼠DC细胞有CD40分子表达,大肠杆菌刺激后表达上调,其上调程度与刺激时间成正相关,与刺激浓度成负相关,初步证明Th细胞极化受不同抗原负载量的影响。2、培养的小鼠DC细胞大量表达B7-1、B7-2分子,大肠杆菌刺激后表达未见明显变化。
[Abstract]:Objective: to study the effect of Escherichia coli on the expression of CD40,B7-1,B7-2 in mouse DC cells. Methods: 1. Mouse DC cells were stimulated with different concentrations of E. coli (2 脳 107,2 脳 106) (DC2.4,2 脳 10 ~ 5 / well), LPS treatment group as positive control and PBS treatment group as negative control). The cells were collected after stimulation for 4 h and 24 h. RNA, was extracted to detect the expression of CD40 mRNA in DC cells by RT-PCR assay. 2, expanded culture (DC cells were 1 脳 10 ~ 6 / well, Escherichia coli and LPS were also expanded 5 times), and cells were collected 4 h and 24 h after infection. The expression of CD40,B7-1,B7-2 on the cell surface was detected by flow cytometry. Results: 1. Compared with the negative control group, the surface CD40 molecules of mouse DC cells stimulated by E. coli increased significantly, and there was significant difference between the two groups (all P 0. 0). ) Among them, the increase level of stimulation for 4 h was lower than that of stimulation for 24 h [(6714.5 卤1367.74) copies/ 渭 l vs (13105.0 卤801.06) copies/ 渭 l; (10101.0 卤797.69) copies/ 渭 l vs (15785.0 卤676.23) copies/ 渭 l], and there was significant difference between the two groups (all P 0). 01), the increased level of lower concentration (1 脳 107) stimulation was significantly higher than that of higher concentration stimulation (1 脳 108) ([10101.0 卤797.69) copies/ 渭 l v (s6714.5 卤1367.74) copies/ 渭 l; (15785.0 卤676.23) copies/ 渭 l vs (13105.0 卤801.06) copies/ 渭 l]. 01). 2. Compared with PBS negative control and LPS positive control, the surface molecules B7 and B7 of DC cells stimulated by E. coli had no significant change, and there was no significant difference between B7 and B7. Conclusion: 1. The expression of CD40 molecule in cultured mouse DC cells is up-regulated after stimulation with Escherichia coli. The up-regulation degree is positively correlated with stimulation time and negatively correlated with stimulation concentration. It is preliminarily proved that the polarization of Th cells is affected by different antigen loading. 2, the expression of B7 鈮，
本文编号：2503807
[Abstract]:Objective: to study the effect of Escherichia coli on the expression of CD40,B7-1,B7-2 in mouse DC cells. Methods: 1. Mouse DC cells were stimulated with different concentrations of E. coli (2 脳 107,2 脳 106) (DC2.4,2 脳 10 ~ 5 / well), LPS treatment group as positive control and PBS treatment group as negative control). The cells were collected after stimulation for 4 h and 24 h. RNA, was extracted to detect the expression of CD40 mRNA in DC cells by RT-PCR assay. 2, expanded culture (DC cells were 1 脳 10 ~ 6 / well, Escherichia coli and LPS were also expanded 5 times), and cells were collected 4 h and 24 h after infection. The expression of CD40,B7-1,B7-2 on the cell surface was detected by flow cytometry. Results: 1. Compared with the negative control group, the surface CD40 molecules of mouse DC cells stimulated by E. coli increased significantly, and there was significant difference between the two groups (all P 0. 0). ) Among them, the increase level of stimulation for 4 h was lower than that of stimulation for 24 h [(6714.5 卤1367.74) copies/ 渭 l vs (13105.0 卤801.06) copies/ 渭 l; (10101.0 卤797.69) copies/ 渭 l vs (15785.0 卤676.23) copies/ 渭 l], and there was significant difference between the two groups (all P 0). 01), the increased level of lower concentration (1 脳 107) stimulation was significantly higher than that of higher concentration stimulation (1 脳 108) ([10101.0 卤797.69) copies/ 渭 l v (s6714.5 卤1367.74) copies/ 渭 l; (15785.0 卤676.23) copies/ 渭 l vs (13105.0 卤801.06) copies/ 渭 l]. 01). 2. Compared with PBS negative control and LPS positive control, the surface molecules B7 and B7 of DC cells stimulated by E. coli had no significant change, and there was no significant difference between B7 and B7. Conclusion: 1. The expression of CD40 molecule in cultured mouse DC cells is up-regulated after stimulation with Escherichia coli. The up-regulation degree is positively correlated with stimulation time and negatively correlated with stimulation concentration. It is preliminarily proved that the polarization of Th cells is affected by different antigen loading. 2, the expression of B7 鈮，
本文编号：2503807
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