Shen Ding-Wen, Luo Jin-Ping, Li Yong-Long, Liu Wen-Qi, Long Xiao-Chun
Medical College of Xianning College, Xianning 437100, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2007 Feb 28;25(1):17-21.
To explore the mechanism of protective immunity against Schistosoma japonicum infection induced by Sj26 gene transfected dendritic cell (DC).
48 BALB/c mice were divided randomly into 4 groups with 12 each. The mice were injected through auricle for three times with Sj26 gene transfected DC (Group A), pcDNA3 transfected DC (Group B), untreated DC (Group C) and RPMI-1640 (Group D) respectively, and challenged with 40+/-2 cercariae of S. japonicum per mouse 2 weeks after the last immunization. Sera from mice were examined for IgG antibody, IFN-gamma and IL-4 by ELISA. Western blot was used for detecting specific anti-Sj26 IgG antibody. The production of IFN-gamma and IL-4 in the supernatant of spleen cells stimulated with soluble egg antigen (SEA) and ConA was quantified by sandwich ABC-ELISA. The proliferation of spleen cells were measured with MTT method.
IgG antibody increased significantly in the mice of group A at 2 weeks after the last immunization (absorbency A491=0.117), higher than that of group B (A491=0.061) and group C (A491=0.058) (P<0.05). The Mr 26000 antigen of S. japonicum was strongly recognized by sera from group A by Western blot. The level of IL-4 in mice of each group showed no significant difference before and after immunization. The level of IFN-gamma in group A (101.4+/-4.9 pg/ml) was significantly higher than that before immunization (15.0+/-1.9 pg/ml) and that of group B (40.1+/-3.1 pg/ml) and group C (35.6+/-1.2 pg/ml) (P<0.01). The level of IFN-gamma in spleen cells from group A in response to ConA and SEA (171.2 and 70.8 pg/ml, respectively) was higher than that of group D (91 and 49.7 pg/ml, respectively) (P<0.01). The level of IL-4 in spleen cells from group A in response to ConA and SEA (79.7 and 50.7 pg/ml, respectively) was lower than that of group D (125.2 and 70.5 pg/ml, respectively) (P<0.01). The stimulating index of spleen cells from group A was 4.1 and 2.82 in response to ConA and SEA respectively, higher than that of other groups (compared with group D, P<0.05).
Sj26 gene transfected dendritic cell induces predominant Th1 type immune response which might play a role in protection against S. japonicum infection.
探讨日本血吸虫Sj26基因转染树突状细胞(DC)诱导抗日本血吸虫感染保护性免疫的机制。
48只BALB/c小鼠随机分为4组,每组12只。分别经耳缘静脉注射Sj26基因转染的DC(A组)、pcDNA3转染的DC(B组)、未处理的DC(C组)和RPMI-1640(D组),共3次,末次免疫后2周,每只小鼠经腹部皮肤感染40±2条日本血吸虫尾蚴。用ELISA检测小鼠血清IgG抗体、IFN-γ和IL-4水平。用Western blot检测特异性抗Sj26 IgG抗体。用夹心ABC-ELISA定量检测可溶性虫卵抗原(SEA)和ConA刺激的脾细胞上清中IFN-γ和IL-4的产生。用MTT法检测脾细胞增殖。
末次免疫后2周,A组小鼠IgG抗体显著升高(吸光度A491=0.117),高于B组(A491=0.061)和C组(A491=0.058)(P<0.05)。Western blot显示A组血清能强烈识别日本血吸虫Mr 26000抗原。各组小鼠免疫前后IL-4水平无显著差异。A组IFN-γ水平(101.4±4.9 pg/ml)显著高于免疫前(15.0±1.9 pg/ml)及B组(40.1±3.1 pg/ml)和C组(35.6±1.2 pg/ml)(P<0.01)。A组脾细胞对ConA和SEA刺激产生的IFN-γ水平(分别为171.2和70.8 pg/ml)高于D组(分别为91和49.7 pg/ml)(P<0.01)。A组脾细胞对ConA和SEA刺激产生的IL-4水平(分别为79.7和50.7 pg/ml)低于D组(分别为125.2和70.5 pg/ml)(P<0.01)。A组脾细胞对ConA和SEA的刺激指数分别为4.1和2.82,高于其他组(与D组比较,P<0.05)。
Sj26基因转染的树突状细胞诱导以Th1型为主的免疫反应,可能在抗日本血吸虫感染中起保护作用。