Li Xiao-Hong, Cao Jian-Ping, Tang Lin-Hua, Xu Yu-Xin, Liu Shu-Xian, Wang Sheng-Jun, Cheng Jing
National Institue of Parasitic Diseases, Chinese Center for Disease Control and Prevention, WHO Collaborating Centre for Malaria, Schistsosomiasis and Filariasis, Shanghai 200025, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2010 Jun 30;28(3):185-9.
To study the protective effects on the infection of Schistosoma japonicum in C57BL/6 mice induced by dendritic cells DCs pulsed with GST in combination with CpG oligodeoxynucleotide.
GST was purified and used to stimulate DC2.4 cell line. Antigen loading was analyzed by immunofluorescence method. Thirty-five C57BL/6 mice were divided into seven groups(5 mice per group). Mice in groups A, B, C, D and E were immunized subcutaneously with DCs, DCs treated with PSA, DCs pulsed with GST, DCs stimulated with GST+CpG ODN, DCs stimulated with CpG ODN, respectively. For the above five groups, each mouse received 100 microl cell suspension at the density of 10(7)/ml subcutaneously for three times at 2-week intervals. Each mouse of group F was immunized subcutaneously with 50 microg GST formulated in complete Freund's adjuvant first, and 50 microg, 10 microg GST respectively in incomplete Freund's adjuvant for the last two doses. Group G received PBS and served as control. Serum samples were collected 10 days after the final immunization, and were analyzed for specific antibodies by ELISA. At two weeks after the final immunization, each mouse were challenged by 30 +/- 1 cercariae of S. japonicum. Six weeks after infection the mice were sacrificed, and number of worms was counted.
Light green fluorescence was observed in dendritic cells under the fluoroscope after pulsing with GST which indicated the protein loaded dendritic cells. The IgG level in groups C, D and F was 0.555 2 +/- 0.078 9, 0.715 0 +/- 0.052 3, and 2.127 0 +/- 0.411 5, respectively, all higher than that of group G (P < 0.05). The worm reduction rate of group D was 53.3%, followed by group F (24.0%) and group C (21.3%). There was no significantly difference in the worm reduction rate between group D and groups F and C (P > 0.05).
Dendritic cells pulsed with GST in combination with CpG oligodeoxynucleotide induce significant immunoprotection against the infection of Schistosoma japonicum.
研究用谷胱甘肽 S-转移酶（GST）与 CpG 寡脱氧核苷酸共脉冲的树突状细胞（DCs）对 C57BL/6 小鼠日本血吸虫感染的保护作用。
纯化 GST 并用其刺激 DC2.4 细胞系。采用免疫荧光法分析抗原负载情况。将 35 只 C57BL/6 小鼠分为 7 组（每组 5 只）。A、B、C、D 和 E 组小鼠分别皮下注射 DCs、经前列腺特异性抗原（PSA）处理的 DCs、用 GST 脉冲的 DCs、用 GST + CpG ODN 刺激的 DCs、用 CpG ODN 刺激的 DCs。对于上述 5 组，每只小鼠皮下注射密度为 10(7)/ml 的 100 μl 细胞悬液，每隔 2 周注射 3 次。F 组每只小鼠首先皮下注射用完全弗氏佐剂配制的 50 μg GST，后两次剂量分别用不完全弗氏佐剂配制的 50 μg、10 μg GST。G 组接受 PBS 作为对照。末次免疫后 10 天采集血清样本，用酶联免疫吸附测定（ELISA）法分析特异性抗体。末次免疫后 2 周，每只小鼠用 30±1 条日本血吸虫尾蚴攻击。感染 6 周后处死小鼠，计数虫体数量。
用 GST 脉冲后，在荧光显微镜下观察到树突状细胞呈浅绿色荧光，表明蛋白质负载到了树突状细胞上。C、D 和 F 组的 IgG 水平分别为 0.555 2±0.078 9、0.715 0±0.052 3 和 2.127 0±0.411 5，均高于 G 组（P < 0.05）。D 组的减虫率为 53.3%，其次是 F 组（24.0%）和 C 组（21.3%）。D 组与 F 组和 C 组的减虫率差异无统计学意义（P > 0.05）。
用 GST 与 CpG 寡脱氧核苷酸共脉冲的树突状细胞可诱导对日本血吸虫感染的显著免疫保护作用。
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