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当前位置: 首页 > 文章 > 海豚链球菌重组C5a肽酶对斑点叉尾的免疫保护性研究 上海海洋大学学报 2015,24 (3) 334-340
Position: Home > Articles > The immunoprotectivity study of the pSCPI of Streptococcus iniae against channel catfish,Ictalurus punctatus Journal of Shanghai Ocean University 2015,24 (3) 334-340

海豚链球菌重组C5a肽酶对斑点叉尾的免疫保护性研究

作  者：
郑宗林;汪开毓;肖孟玮;王均;李岚敏
单  位：
四川农业大学动物医学院;四川农业大学鱼病研究中心;四川农业大学鱼病研究中心动物疫病与人类康四川省重点实验室四川农业大学;西南大学荣昌校区水产系
关键词：
海豚链球菌;斑点叉尾;IgM;免疫保护率
摘  要：
采用饱和硫酸铵法对斑点叉尾IgM进行粗提,用亲硫树脂对IgM进行纯化,将纯化的IgM免疫家兔,并用ELISA检测兔抗斑点叉尾IgM抗体效价;制备重组C5a肽酶(pSCPI),分别用1、2和3μg/g的抗原免疫斑点叉尾,以PBS作为对照;测定了第21、28、35、42和49天斑点叉尾产生的特异性抗体的水平,并于免疫后第4周用海豚链球菌DGX07株进行攻毒。结果显示:通过亲硫树脂亲和层析后得到了较纯的IgM重链和轻链,分别约70ku和27ku,浓度达到6.4mg/mL,免疫家兔后的兔抗斑点叉尾IgM抗体效价达1∶51200;重组蛋白pSCPI免疫斑点叉尾后第3周抗体水平开始增加,第4周达到峰值,第5周后开始下降,且3μg/g浓度组第4周抗体水平最高,但与2μg/g浓度组第4周抗体水平差异不显著;攻毒实验显示,3个组14d平均累计死亡率分别为47.5%、45%和47.5%,平均相对保护率分别为40.63%、43.75%和40.63%,且死亡鱼体内分离到唯一海豚链球菌。以上结果表明重组pSCPI蛋白具有较好的免疫保护作用,能够作为海豚链球菌亚单位疫苗的候选之一。
译  名：
The immunoprotectivity study of the pSCPI of Streptococcus iniae against channel catfish,Ictalurus punctatus
作  者：
ZHENG Zonglin;WANG Kaiyu;XIAO Mengwei;WANG Jun;LI Lanmin;HE Yang;CHEN Defang;HUANG Lingyuan;Fisheries Department of Sichuau Agricultural University;Fisheries Department of Southwest University Rongchang Campus;Key Laboratory of Animal Disease and Human Health of Sichuan Province,Sichuan Agricultural University;
关键词：
Streptococcus iniae;;Ictalurus punctatus;;IgM;;immunoprotectivity
摘  要：
The saturated ammonium sulfate method was used to crude-extract the IgM of channel catfish,then purifying the IgM using sulfur resin affinity chromatography,followed by immunizing the rabbit using the purified IgM and detecting the antibody titer of the rabbit anti-IgM serum using the enzymelinked immunosorbent assay(ELISA).We prepared the recombinant protein p SCPI based on the method used in Chapter 2,vaccinating the healthy catfish with the concentration of the antigen protein 1 μg / g fish,2 μg / g and 3 μg / g fish,respectively and with PBS as a control.We detected the antibody level of the specific antibody produced by the vaccinated catfish at 21,28,35,42 and 49 days post-vaccination,and challenged with the S.iniae DGX07 at 4 weeks post-vaccination.The results showed that: the purified IgM heavy chain and light chain by using sulfur resin affinity chromatography were about 70 ku and 27 ku,separately.And the concentration of the IgM was up to 6.4 mg / m L,the antibody titer of the rabbit anti-IgM serum was 1∶ 51 200.Also we observed that the antibody level began to increase at 3 weeks post-vaccination and with the highest antibody level occurring at 4-week p.v.,but it began to decline at 5-week p.v..The antibody level of the antigen concentration with 3μg / g fish-vaccinated fish was highest at each examined time point,which had no significant difference compared with the 2 μg / g fish-vaccinated fish.In 1 μg / g,2 μg / g and 3 μg / g fishvaccinated fish,S.iniae DGX07 challenge caused accumulated mortality rates of 47.5%,45% and 47.5%respectively,which corresponded to RPS rates of 40.63%,43.75% and 40.63% respectively with PBS as a control.For all vaccination trials,examination of moribund fish indicated that S.iniae was the only type of bacterial strains isolated from S.iniae-challenged fish,suggesting that mortalities were caused by the challenging bacteria.All these results indicated that the recombinant protein p SCPI was effective and had a better immunoprotectivity,which could be used as a candidate subunit vaccine against S.iniae.

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