IL-18对pVAX1/SjRPS4·CB质粒疫苗免疫小鼠抗日本血吸虫感染的免疫调节作用

中国寄生虫学与寄生虫病杂志 ›› 2018, Vol. 36 ›› Issue (4): 325-330.

IL-18对pVAX1/SjRPS4·CB质粒疫苗免疫小鼠抗日本血吸虫感染的免疫调节作用

程红兵^1,², 周云飞^1,^*(), 张树菊¹, 汪世平¹, 冯其梅¹, 刘益萍^1,³, 崔国艳^1,², 魏红^1,⁴, 李芬¹, 刘明社³

1 中南大学基础医学院医学寄生虫学系,长沙 410078
2 长治医学院微生物学教研室,长治 046000
3 长治医学院寄生虫学教研室,长治 046000
4 长治医学院附属和平医院感染管理科,长治 046000

收稿日期:2018-02-09 出版日期:2018-08-30 发布日期:2018-09-06
通讯作者: 周云飞
基金资助:
国家自然科学基金(No. 81672051,No. 81271862);长治医学院科研启动基金（普及项目）(No. QDZ201524)

The immunomodulatory effects of IL-18 against schistosome infection in mice with pVAX1/SjRPS4·CB plasmid vaccination

Hong-bing CHENG^1,², Yun-fei ZHOU^1,^*(), Shu-ju ZHANG¹, Shi-ping WANG¹, Qi-mei FENG¹, Yi-ping LIU^1,², Guo-yan CUI^1,², Hong WEI^1,⁴, Fen LI¹, Ming-she LIU³

1 Department of Parasitology, Xiangya Medical School, Central South University, Changsha 410078, China
2 Department of Microbiology, Changzhi Medical College, Changzhi 046000, China
3 Department of Parasitology, Changzhi Medical College, Changzhi 046000, China
4 Department of Infection Control, the Affiliated Heping Hospital of Changzhi Medical College, Changzhi 046000, China

Received:2018-02-09 Online:2018-08-30 Published:2018-09-06
Contact: Yun-fei ZHOU
Supported by:
Supported by the National Natural Science Foundation of China (No. 81672051, No. 81271862), and Research Starting Fund Project of Changzhi Medical College (No. QDZ201524).

摘要/Abstract

摘要：

目的探讨pVAX1/IL-18联合pVAX1/SjRPS4·CB质粒疫苗对小鼠抗日本血吸虫感染的免疫调节作用。方法 70只BALB/c雌鼠按随机数字表法均分为生理盐水组（NS组）、pVAX1空质粒组、pVAX1/IL-18组、pVAX1/SjRPS4·CB组和pVAX1/SjRPS4·CB + pVAX1/IL-18组等5组（每组14只）,每组小鼠在左后腿股四头肌注射相应质粒100 μg或等量生理盐水,每隔2周加强免疫1次,共免疫3次。小鼠感染前（免疫后3周）尾尖部取血,ELISA检测小鼠血清特异性抗体IgG以及抗体亚类IgG1、IgG2a水平。同时,各组小鼠经腹部贴片感染日本血吸虫尾蚴,（20 ± 1）条/鼠。感染后8周,门静脉灌注法收集日本血吸虫成虫,计算减虫率;取肝组织,显微镜下计数虫卵数,计算减卵率。无菌取脾脏,称重,计算小鼠脾脏指数,噻唑蓝（MTT）法测脾淋巴细胞体外增殖水平。分别于小鼠感染前（免疫后3周）以及感染后2、4、6、8周尾尖部取血,ELISA检测小鼠血清中γ干扰素（IFN-γ）、白细胞介素4（IL-4）水平。结果 5组小鼠免疫后3周,ELISA检测结果显示,pVAX1/SjRPS4·CB + pVAX1/IL-18组IgG、IgG1、IgG2的吸光度（A₄₅₀值）分别为1.03 ± 0.17、0.32 ± 0.08、0.78 ± 0.12,IgG2a/IgG1比值为2.44,均高于其他4组。pVAX1/SjRPS4·CB + pVAX1/IL-18组的减虫率、减卵率分别为52.0%、63.2%,均高于其他3组（P < 0.05）。脾脏指数结果显示,pVAX1/SjRPS4·CB + pVAX1/IL-18组小鼠脾脏指数为3.32 ± 0.37,高于其他4组（P < 0.05）。MTT法检测结果显示,pVAX1/SjRPS4·CB + pVAX1/IL-18组淋巴细胞体外增殖水平A₅₇₀值为4.45 ± 0.34,高于其他4组（P < 0.05）。小鼠感染前以及感染后2、4、6、8周,pVAX1/SjRPS4·CB + pVAX1/IL-18组IFN-γ含量分别为（569.07 ± 21.15）、（560.66 ± 30.84）、（577.46 ± 36.45）、（605.03 ± 36.91）、（636.33 ± 37.35）pg/ml,均高于其他4组（P < 0.05）;各组IL-4含量在感染前后差异无统计学意义（P > 0.05）。结论 IL-18可增强pVAX1/SjRPS4·CB疫苗抗日本血吸虫感染的免疫保护效果,且诱导小鼠产生较高的以Th1为主的免疫应答。

关键词: 日本血吸虫, 白细胞介素-18, pVAX/SjRPS4·CB, 疫苗

Abstract:

Objective To explore the immunoregulatory effect of pVAX1/IL-18 combined with pVAX1/SjRPS4·CB vaccine against schistosome infection in mice. Methods Seventy female BALB/c mice were divided into 5 groups: the normal saline (NS) group, pVAX1 plasmid group, pVAX1/IL-18 group, pVAX1/SjRPS4·CB group and pVAX1/SjRPS4·CB + pVAX1/IL-18 group. The mice were injected with 100 μg plasmid or an equal volume of NS at the quadriceps femoris of the left hind leg, once every other week, for a total of 3 vaccinations. Three weeks after the final vaccination, blood was collected from the tail, and the levels of IgG, IgG1 and IgG2a were determined by ELISA. The mice were then challenged with 20 ± 1 Schistosoma japonicum cercariae through abdominal patching and sacrificed 8 weeks later. Adult worms of S. japonicum were collected by portal vein perfusion to calculate the worm reduction rate. The liver tissue was collected to calculate the egg reduction rate in hepatic tissues under a microscope. Moreover, spleen was collected at a sterile condition, weighed, and the mouse spleen index was calculated. The splenic lymphocyte proliferation level in vitro was determined using MTT assay. Blood was collected from the tail before infection (at 3 weeks after the final vaccination) and at 2, 4, 6, and 8 weeks after infection, to assess the serum levels of IFN-γ and IL-4 by ELISA. Results At 3 weeks after the final vaccination, the A₄₅₀ values of IgG, IgG1 and IgG2 in the pVAX1/SjRPS4·CB + pVAX1/IL-18 group were 1.03 ± 0.17, 0.32 ± 0.08, and 0.78 ± 0.12, respectively, and the IgG2a/IgG1 ratio was 2.44, all of which were significantly higher than those in the other 4 groups. The worm reduction rate and the egg decrease rate in the pVAX1/SjRPS4·CB + pVAX1/IL-18 group were 52.0% and 63.2%, respectively, both significantly higher than those in the other 3 groups (P < 0.05). The spleen index in the pVAX1/SjRPS4·CB + pVAX1/IL-18 group (3.32 ± 0.37) was also significantly higher than those in the other 4 groups （P < 0.05）. MTT assay showed that the splenic lymphocyte proliferation level in vitro in the pVAX1/SjRPS4·CB + pVAX1/IL-18 group (A₅₇₀ value, 4.45 ± 0.34) was significantly higher than those in the other 4 groups （P < 0.05）. The IFN-γ levels in the pVAX1/SjRPS4·CB + pVAX1/IL-18 group before infection and at 2, 4, 6, and 8 weeks after infection were 569.07 ± 21.15 pg/ml, 560.66 ± 30.84 pg/ml, 577.46 ± 36.45 pg/ml, 605.03 ± 36.91 pg/ml and 636.33 ± 37.35 pg/ml, respectively, all significantly higher than those in the other 4 groups （P < 0.05）. However, the IL-4 level was not altered either before versus after infection, or among different groups （P > 0.05）. Conclusion IL-18 enhances the immunomodulatory effects of pVAX1/SjRPS4·CB against schistosome infection and induces Th1 type immune responses in mice.

Key words: Schistosoma japonica, IL-18, pVAX1/SjRPS4·CB, Vaccine

中图分类号:

R383.24

程红兵, 周云飞, 张树菊, 汪世平, 冯其梅, 刘益萍, 崔国艳, 魏红, 李芬, 刘明社. IL-18对pVAX1/SjRPS4·CB质粒疫苗免疫小鼠抗日本血吸虫感染的免疫调节作用[J]. 中国寄生虫学与寄生虫病杂志, 2018, 36(4): 325-330.

Hong-bing CHENG, Yun-fei ZHOU, Shu-ju ZHANG, Shi-ping WANG, Qi-mei FENG, Yi-ping LIU, Guo-yan CUI, Hong WEI, Fen LI, Ming-she LIU. The immunomodulatory effects of IL-18 against schistosome infection in mice with pVAX1/SjRPS4·CB plasmid vaccination[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2018, 36(4): 325-330.

图/表 2

参考文献 17

[1]	雷正龙, 周晓农. 血吸虫病“十二五”防治规划实施进展及面临的挑战[J]. 中国寄生虫学与寄生虫病杂志, 2014, 32(2): 91-85.
[2]	金嘉宁, 党辉, 张利娟, 等. 2016年全国血吸虫病监测点疫情分析[J]. 中国寄生虫学与寄生虫病杂志, 2017, 36(6): 542-548.
[3]	Tian Z, Wang XY, Zhou YF, et al. Schistosoma japonicum scFv-IL18 fusion DNA ameliorates hepatic fibrosis in schistos-omiasis-infected mice via improving local concentration of IL-18 in liver[J]. Exp Parasitol, 2013, 134(4): 447-454.
[4]	Chen J, Pan J, Wang J, et al. Soluble egg antigens of Schistosoma japonicum induce senescence in activated hepatic stellate cells by activation of the STAT3/p53/p21 pathway[J]. Sci Rep, 2016, 6: 30957.
[5]	彭微, 汪世平, 郑长黎, 等. 日本血吸虫pVAX1/SjRPS4·CB多价疫苗免疫对小鼠肝脏虫卵肉芽肿形成的影响[J]. 中国人兽共患病学报, 2012, 28(12): 1216-1219.
[6]	彭微, 汪世平, 郑长黎, 等. 日本血吸虫pVAX1/SjRPS4·CB多价疫苗免疫对小鼠肝脏虫卵肉芽肿形成中TNF-α表达的影响[J]. 中国人兽共患病学报, 2013, 28(6): 537-542.
[7]	Pearce EJ, MacDonald AS. The immunobiology of schistosomiasis[J]. Nat Rev Immunol, 2002, 2(7): 499-511.
[8]	Yoshimoto T, Takeda K, Tanaka T, et al. IL-12 upregulates IL-18 receptor expression on T cells, Thl cells and B cells: synergism with IL-l8 for IFN-γ production[J]. J Immunol, 1998, 161(7): 3400-3407.
[9]	张立煌, 姚航平. 吡喹酮联合IL-18基因治疗血吸虫感染小鼠对免疫功能的影响[J]. 中华微生物与免疫学杂志, 2001, 21(1): 87-90.
[10]	Men JT, Liu Q, Shang LM.IL-18 augments protective immunity of Sj23 plasmid DNA vaccine against Schistosoma japonicum in mice[J]. Chin J Vet Sci, 2009, 29(5): 610-613.
[11]	Huntley JF, Stabel JR, Paustian ML, et al. Expression library immunization confers protection against Mycobacterium avium subsp. paratuberculosis infection[J]. Infect Immun, 2005, 73(10): 6877-6884.
[12]	Bani-Hani AH, Leslie JA, Asanuma H, et al. IL-18 neutralization ameliorates obstruction-induced epithelial-mesenchymal transition and renal fibrosis[J]. Kidney Int, 2009, 76(5): 500-511.
[13]	Kockx M, Guo DL, Traini M, et al. Cyclosporin A decreases apolipoprotein E secretion from human macrophages via a protein phosphatase 2B-dependent and ATP-binding cassette transporter A1(ABCA1)-independent pathway[J]. J Biol Chem, 2009, 284(36): 24144-24154.
[14]	Hong JC, Kahan BD.Immunosuppressive agents in organ transplantation: past, present, and future[J]. Semin Nephrol, 2000, 20(2): 108-125.
[15]	李富荣, 石佑恩, 史大中, 等. 泡球蚴感染BALB/c小鼠IgG亚类和细胞因子的动态观察[J]. 中国寄生虫学与寄生虫病杂志, 2003, 21(6): 357-360.
[16]	Wei F, Liu Q, Zhai Y, et al. IL-18 enhances protective effect in mice immunized with a Schistosoma japonicum FABP DNA vaccine[J]. Acta Trop, 2009, 111(3): 284-288.
[17]	Wei F, Liu Q, Gao S, et al. Enhancement by IL-18 of the protective effect of a Schistosoma japonicum 26kDa GST plasmid DNA vaccine in mice[J]. Vaccine, 2008, 26(33): 4145-4149.