脑多头蚴重组蛋白rTm16和rTm-GST的免疫保护效果分析

doi:10.11843/j.issn.0366-6964.2018.12.020

摘要/Abstract

摘要：

为探究多头带绦虫16 kDa蛋白和GST蛋白对山羊脑多头蚴病的免疫保护效果，44只攀枝花本地黑山羊随机分为4组（rTm16疫苗组、rTm-GST疫苗组、rTm16+rTm-GST疫苗组以及对照组），皮下免疫三次，首免后4周进行2免，2免后6个月进行3免，定时采血并分离血清，使用间接ELISA方法检测血清中特异性IgG水平；末次免疫后2周，各组山羊每只口服攻击多头带绦虫的虫卵约5 500个，感染105 d后剖检记录脑多头蚴包囊数量，计算减囊率。结果显示：相较于对照组，rTm16组、rTm-GST组、rTm16+rTm-GST组分别获得50%、62.5%、87.5%的减囊率，rTm16+rTm-GST免疫组山羊保护效果最好（P=0.005）。ELISA检测结果显示，攻虫以前，rTm-GST组特异性抗体水平均显著低于rTm16组（P<0.05），而联合免疫组中rTm-GST所诱导的特异性抗体与rTm16所诱导的特异性抗体水平差异不显著（P>0.05）。rTm16与rTm-GST的配合使用能够诱导山羊产生较高抗体水平，从而获得较显著的免疫保护力。

Abstract:

To explore the immune protective effect of recombinant Tm16 and GST proteins against Coenurus cerebralis in goats, forty-four goats were divided into four groups, including rTm16 group, rTm-GST group, rTm16+rTm-GST group and control group, which were immunized 3 times by subcutaneous injection, respectively. Second vaccination was performed 4 weeks after the first vaccination, and 6 months later the last vaccination was performed. The serum was regularly collected from each goat, and the specific IgG response to each group was measured by indirect ELISA. Two weeks after the final vaccination, each goat was orally challenged with 5 500 eggs of Taenia multiceps. One hundred and five days post infection all goats were slaughtered, the cysts in brain and muscle were collected and counted. Result were as follows:Compared with the control group, the cysts of goat in group rTm16, rTm-GST and rTm16+rTm-GST reduced by 50%, 62.5% and 87.5%, respectively. Combined immunization of rTm16 with rTm-GST induced significant protection (P<0.05)against challenge of T. multiceps in goats. Result of iELISA showed that the specific IgG response against rTm-GST was significantly weaker than that against rTm16 (P < 0.05). However, the specific IgG response against rTm-GST was not significant compared with that against rTm16 in combined group (P>0.05). This revealed that the combined use of rTm16 and rTm-GST could induce higher antibody levels in goats, thus obtaining significant protective immunity.

中图分类号:

S852.734

郭承, 万洁, 杨应东, 文建国, 刘俞辰, 古小彬, 谢跃, 杨光友. 脑多头蚴重组蛋白rTm16和rTm-GST的免疫保护效果分析[J]. 畜牧兽医学报, 2018, 49(12): 2707-2714.

GUO Cheng, WAN Jie, YANG Ying-dong, WEN Jian-guo, LIU Yu-chen, GU Xiao-bin, XIE Yue, YANG Guang-you. Immune Protective Effect Analysis of Recombinant Protein rTm16 and rTm-GST from Coenurus cerebralis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(12): 2707-2714.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2018.12.020

https://www.xmsyxb.com/CN/Y2018/V49/I12/2707

参考文献

[1] CHRISTODOULOPOULOS G, DINKEL A, ROMIG T, et al. Cerebral and non-cerebral coenurosis:on the genotypic and phenotypic diversity of Taenia multiceps[J]. Parasitol Res, 2016, 115(12):4543-4558.
[2] SUN Y, WANG Y, HUANG X, et al. Characterization of glutathione S-transferase and its immunodiagnostic potential for detecting Taenia multiceps[J]. Vet Parasitol, 2017, 242:31-37.
[3] ABERA S, WUBIT T, NEJASH A. Cerebral coenurosis in small ruminants:a review[J]. J Anim Sci Adv, 2016, 6(3):1595-1608.
[4] AMER S, ELKHATAM A, FUKUDA Y, et al. Clinical, pathological, and molecular data concerning Coenurus cerebralis in sheep in Egypt[J]. Data Brief, 2018, 16:1-9.
[5] ALIYE J, DERESSA F B. Prevalence and economic importance of Coenurus Cerebralis in sheep and goats in and around "Legahida" district of bale zone, south eastern Ethiopia[J]. Comp Clin Pathol, 2017, 26(2):483-492.
[6] 牟静,杨应东,韦雷飞,等. 动物脑多头蚴病研究进展[J]. 动物医学进展, 2010, 31(2):96-99.
MU J, YANG Y D, WEI L F, et al. Progress on coenurosis cerebralis of animals[J]. Progress in Veterinary Medicine, 2010, 31(2):96-99. (in Chinese)
[7] ORYAN A, MOAZENI M, AMRABADI O, et al. Comparison of distribution pattern, pathogenesis and molecular characteristics of larval stages of Taenia multiceps in sheep and goats[J]. Small Rumin Res, 2015, 132:44-49.
[8] ANTONIOS S N, MINA S N. A case report of human coenurus cerebralis in Tanta, Egypt[J]. J Egypt Soc Parasitol, 2000, 30(3):959-960.
[9] COLLOMB J, MACHOUART M, BIAVA M F, et al. Contribution of nadh dehydrogenase subunit I and cytochrome C oxidase subunit I sequences toward identifying a case of human coenuriasis in france[J]. J Parasitol, 2007, 93(4):934-937.
[10] HUANG X, XU J, WANG Y, et al. GP50 as a promising early diagnostic antigen for Taenia multiceps infection in goats by indirect ELISA[J]. Parasit Vectors, 2016, 9:618.
[11] WANG Y, NIE H M, GU X B, et al. An ELISA using recombinant TmHSP70 for the diagnosis of Taenia multiceps infections in goats[J]. Vet Parasitol, 2015, 212(3-4):469-472.
[12] HUANG X, CHEN L, YANG Y D, et al. Expression, tissue localization and serodiagnostic potential of Taenia multiceps acidic ribosomal protein P2[J]. Parasit Vectors, 2015, 8:613.
[13] 安晓雪,杨光友,王颖旺,等. 多头带绦虫Tm7基因的表达及间接ELISA检测方法的建立[J]. 畜牧兽医学报, 2011, 42(9):1302-1308.
AN X X, YANG G Y, WANG Y W, et al. Prokaryotic expression of Tm7 gene of Taenia multiceps and establishment of indirect ELISA using the expressed protein[J]. Acta Veterinaria et Zootechnica Sinica, 2011, 42(9):1302-1308. (in Chinese)
[14] GUO C, WANG Y, HUANG X, et al. Molecular cloing and bioinformatics analysis of lactate dehydrogenase from Taenia multiceps[J]. Parasitol Res, 2017, 116(10):2845-2852.
[15] 张壮志,石保新,由弘,等. 多头绦虫膜蛋白(45M)对羊脑多头蚴病的免疫保护效果[J]. 中国寄生虫学与寄生虫病杂志, 2009, 27(4):336-339.
ZHANG Z Z, SHI B X, YOU H, et al. Immune protection of recombinant membrane protein against Taenia multiceps larvae in sheep[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2009, 27(4):336-339. (in Chinese)
[16] VERSTER A, TUSTIN R C. Preliminary report on the stimulation of immunity to the larval stage of Taenia multiceps[J]. J South Afr Vet Assoc, 1982, 53(3):175-176.
[17] GAUCI C, VURAL G, ÖNCEL T, et al. Vaccination with recombinant oncosphere antigens reduces the susceptibility of sheep to infection with Taenia multiceps[J]. Int J Parasitol, 2008, 38(8-9):1041-1050.
[18] NIE H M, XIE Y, FU Y, et al. Cloning and characterization of the fatty acid-binding protein gene from the protoscolex of Taenia multiceps[J]. Parasitol Res, 2013, 112(5):1833-1839.
[19] 牟静,杨应东,杨光友,等. 多头带绦虫六钩蚴Tm45W基因的克隆表达与免疫原性分析[J]. 中国兽医科学, 2011, 41(2):167-172.
MU J, YANG Y D, YANG G Y, et al. Cloning, prokaryotic expression and immunogenicity analysis of Tm45W gene from Taenia multiceps oncospheres[J]. Chinese Veterinary Science, 2011, 41(2):167-172. (in Chinese)
[20] 岳龙,李文卉,张念章,等. 多头带绦虫黏着蛋白TmAdh1基因的克隆及原核表达[J]. 中国兽医科学, 2015, 45(2):126-131.
YUE L, LI W H, ZHANG N Z, et al. Cloning and prokaryotic expression of TmAdh1 gene from Taenia multiceps[J]. Chinese Veterinary Science, 2015, 45(2):126-131. (in Chinese)
[21] 王颖旺,杨应东,杨光友,等. 多头带绦虫Tm18抗原基因的克隆、表达与免疫原性分析[J]. 中国人兽共患病学报, 2011, 27(4):320-324.
WANG Y W, YANG Y D, YANG G Y, et al. Cloning, expression and immunogenicity analysis of Tm18 gene of Taenia multiceps[J]. Chinese Journal of Zoonoses, 2011, 27(4):320-324. (in Chinese)
[22] 王颖旺,杨应东,杨光友,等. 多头带绦虫Tm16基因的克隆表达与免疫原性分析[J]. 中国兽医科学, 2010, 40(9):934-939.
WANG Y W, YANG Y D, YANG G Y, et al. Cloning and prokaryotic expression of Tm16 gene from Taenia multiceps oncospheres and immunogenicity analysis of its expressed product[J]. Chinese Veterinary Science, 2010, 40(9):934-939. (in Chinese)
[23] 张壮志,石保新,张文宝,等. 绵羊包虫病基因工程疫苗(Eg95)免疫试验[J]. 中国人兽共患病学报, 2008, 24(3):252-256.
ZHANG Z Z, SHI B X, ZHANG W B, et al. The prophylactic effect of the genetically engineering vaccine Eg95 on hydatid disease of sheep[J]. Chinese Journal of Zoonoses, 2008, 24(3):252-256. (in Chinese)
[24] EDWARDS G T, HERBERT I V. Observations on the course of Taenia multiceps infections in sheep:clinical signs and post-mortem findings[J]. Br Vet J, 1982, 138(6):489-500.
[25] 张文宝,哈江,蔡宏,等. 多头绦虫六钩蚴代谢物对绵羊脑包虫病的免疫效果观察[J]. 中国兽医科技, 1991, 21(12):35-36.
ZHANG W B, HA J, CAI H, et al. Immunity of Taenia multiceps oncosphere metabolites to coenurosis in sheep[J]. Chinese Journal of Veterinary Science and Technology, 1991, 21(12):35-36. (in Chinese)
[26] LIGHTOWLERS M W, GAUCI C G, CHOW C, et al. Molecular and genetic characterisation of the host-protective oncosphere antigens of taeniid cestode parasites[J]. Int J Parasitol, 2003, 33(11):1207-1217.
[27] HEIN W R, HARRISON G B L. Vaccines against veterinary helminths[J]. Vet Parasitol, 2005, 132(3-4):217-222.
[28] LIGHTOWLERS M W, GAUCI C G. Vaccines against cysticercosis and hydatidosis[J]. Vet Parasitol, 2001, 101(3-4):337-352.
[29] RICKARD M D, HARRISON G B L, HEATH D D, et al. Taenia ovis recombinant vaccine-'quo vadit'[J]. Parasitology, 1995, 110(S1):S5-S10.
[30] HARRISON G B L, HEATH D D, DEMPSTER R P, et al. Identification and cDNA cloning of two novel low molecular weight host-protective antigens from Taenia ovis oncospheres[J]. Int J Parasitol, 1996, 26(2):195-204.
[31] VEERAPATHRAN A, DAKSHINAMOORTHY G, GNANASEKAR M, et al. Evaluation of Wuchereria bancrofti GST as a vaccine candidate for lymphatic filariasis[J]. PLoS Negl Trop Dis, 2009, 3(6):e457.
[32] 马森. 谷胱甘肽过氧化物酶和谷胱甘肽转硫酶研究进展[J]. 动物医学进展, 2008, 29(10):53-56.
MA S. Progress on GSH-Px and GST[J]. Progress in Veterinary Medicine, 2008, 29(10):53-56. (in Chinese)
[33] 高鹏,雄英,杜娟,等. 细粒棘球绦虫(中国大陆株)谷胱甘肽s-转移酶重组蛋白(rEgGST)的免疫保护性研究[J]. 中国人兽共患病学报, 2011, 27(3):238-240.
GAO P, XIONG Y, DU J, et al. Immune protection of recombinant protein Glutathione s-transferase (rEgGST) of Echinococcus granulosus (Chinese mainland strain)[J]. Chinese Journal of Zoonoses, 2011, 27(3):238-240. (in Chinese)
[34] 施福恢,沈纬,田锷,等. 日本血吸虫重组抗原Sj GST Paramyosin和Sj 23对绵羊的保护性免疫试验[J]. 中国兽医科学, 1997, 27(11):3-6.
SHI F H, SHEN W, TIAN E, et al. Protective immunity test in sheep by recombinant antigen Sj GST Paramyosin and Sj 23 of Schistosoma japonicum[J]. Chinese Journal of Veterinary Science and Technology, 1997, 27(11):3-6. (in Chinese)
[35] MUTAPI F, BILLINGSLEY P F, SECOR W E. Infection and treatment immunizations for successful parasite vaccines[J]. Trends Parasitol, 2013, 29(3):135-141.
[36] JARIWALA A R, OLIVEIRA L M, DIEMERT D J, et al. Potency testing for the experimental Na-GST-1 hookworm vaccine[J]. Expert Rev Vaccines, 2010, 9(10):1219-1230.
[37] VARCASIA A, TOSCIRI G, COCCONE G N S, et al. Preliminary field trial of a vaccine against coenurosis caused by Taenia multiceps[J]. Vet Parasitol, 2009, 162(3-4):285-289.