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目的研究细粒棘球绦虫原头节可溶性抗原(protoscolex soluble antigens,PSA)对树突状细胞(dendritic cells,DC)表型分化及分泌炎症因子的影响,探索寄生虫对宿主免疫逃避的机制。方法运用免疫磁珠从健康BALB/c小鼠脾脏中分选DC,分别加入PSA(10μg/ml)、LPS(1μg/ml)、PSA(10μg/ml)+LPS(1μg/ml)以及等体积PBS进行刺激24h,收集细胞及培养上清,利用流式细胞术检测DC表面活化分子的表达水平,并采用流式液相多重蛋白定量技术检测细胞因子分泌情况。结果 PSA组DC表面表达CD40,CD80,CD86,MHCⅡ的比例为(42.33±0.59)%、(71.08±1.61)%、(73.63±5.31)%、(70.20±1.01)%,较PBS组的(29.1±8.14)%、(48.81±1.69)%、(37.37±2.57)%、(26.83.08±3.55)%均明显升高(P<0.05),较LPS组的(61.33±4.73)%、(81.23±2.03)%、(85.40±1.57)%、(86.86±2.01)%均显著降低(P<0.05)。PSA组培养上清中IL-6、TNF浓度为(53.67±6.23)pg/ml、(405.10±5.78)pg/ml,较PBS组的(9.44±4.06)pg/ml、(139.35±45.86)pg/ml、均显著升高(P<0.05),较LPS组的(48.23±7.85)pg/ml、(471.40±61.07)pg/ml均显著降低(P<0.05);且PSA+LPS组IL-6、TNF浓度为(73.42±8.00)pg/ml和(508.54±20.68),较PSA组、LPS组显著升高(P<0.05)。结论 PSA能促进DC活化及炎症因子释放,从而诱导机体产生正向免疫应答。
Objective To investigate the effect of protoscolex soluble antigens (PSA) on the phenotypic differentiation of dendritic cells (DCs) and the secretion of inflammatory cytokines in Echinococcus granulosus, and to explore the mechanism of parasite immunity to host immune evasion. Methods DCs were isolated from the spleens of healthy BALB / c mice by immunomagnetic beads. PSA (10μg / ml), LPS (1μg / ml), PSA The cells were stimulated with PBS for 24 h. The supernatants were collected and the expression of DCs was detected by flow cytometry. The secretion of cytokines was detected by flow cytometry and multiple protein quantification. Results The percentage of CD40, CD80, CD86 and MHCⅡ in the PSA group was (42.33 ± 0.59)%, (71.08 ± 1.61)%, (73.63 ± 5.31)% and (70.20 ± 1.01)%, respectively, (61.33 ± 4.73)%, (81.23)%, (48.81 ± 1.69)%, (37.37 ± 2.57)% and (26.83.08 ± 3.55)% in the LPS group (P <0.05) ± 2.03%, (85.40 ± 1.57)%, (86.86 ± 2.01)%, respectively (P <0.05). The levels of IL-6 and TNF in the supernatant of PSA group were (53.67 ± 6.23) pg / ml and (405.10 ± 5.78) pg / ml respectively, which were significantly higher than those in PBS group (9.44 ± 4.06 pg / ml and 139.35 ± 45.86 pg (P <0.05). Compared with LPS group (48.23 ± 7.85) pg / ml and (471.40 ± 61.07) pg / ml, the level of IL- 6 and TNF levels were (73.42 ± 8.00) pg / ml and (508.54 ± 20.68) respectively, which were significantly higher than those in PSA group and LPS group (P <0.05). Conclusion PSA can promote the activation of DCs and the release of inflammatory cytokines, thereby inducing the body to produce a positive immune response.