Difference of T cell and B cell activation in two proteins with similar antigenicity but great distinct immunogenicity

大肠杆菌表达的戊型肝炎病毒(HEV)衣壳蛋白ORF2的aa394-606片段NE2可以体外形成同源多聚体,其N端延伸突变体HEV239蛋白（ORF2aa368-606）在体外可以自发组装成类病毒颗粒。二者具有近乎完全一致的抗原性，但HEV239的免疫原性比可溶的非颗粒蛋白NE2强200倍左右。有研究表明，颗粒蛋白抗原比可溶性抗原能够更加有效的被抗原递呈细胞捕获从而具有更强的免疫原性，亦有文献报道认为，这是由于颗粒抗原能更有效的激活B细胞。因此对HEV239与NE2的研究将能使我们更好的了解颗粒抗原与可溶性抗原免疫原性差别产生的原因，并为将来合理的设计开发基因工程亚单位疫苗提供新的思路。 本文...An E. coli expressed recombinant antigen NE2(HEV ORF2 aa394-606) was able to aggregate into homo-oligomer, and one of it’s N terminal extension mutant which was expressed in E. coli and named HEV 239 （HEV ORF2 aa368-606） was found to aggregate into particle. The antigenicity of HEV 239 is virtually identical to E2, which is not particulate but soluble. However, HEV 239 is over 200 times more immun...学位：理学硕士院系专业：生命科学学院生物医学科学系_生物化学与分子生物学学号：2005130212

一个包含HCVIRES的高效真核双顺反子表达载体的构建

In this paper, a new eukaryotic bi-cistronic expression vector containing Hepatitis C Virus(HCV) internal ribosome entry site (IRES) expressing two foreign genes from one mRNA was constructed. The sequence starting from the 5' untranslated region of 18nt to 32nt in HCV core coding region was cloned and then the encephalomyocarditis virus (ECMV) IRES sequence in the commercial vector pIRES was substituted to construct a new vector pCVIR. Green fluorescent protein (GFP) and Hepatitis B virus surface antigen (HBsAg) coding genes were inserted up-stream and down-stream of IRES sequence. The fluorescence intensity of GFP and HBsAg were determined by flow cytometry and ELISA respectively., thus, the expression efficiency of the two vectors, pCVIR and pIRES could be compared. The experimental results showed that the vector pCVIR could translate the GFP and HBsAg genes down-stream of its HCV IRES sequence more efficiently without impairing the expression of genes up-stream of IRES sequence than the vector pIRES.It is..

不同HBsAg疫苗联合免疫后诱导小鼠细胞和体液免疫应答的研究

目的:了解HBsAg的蛋白疫苗(P)、痘苗病毒疫苗(V)、DNA疫苗(D)联合免疫小鼠诱导的特异性体液和细胞免疫应答。方法:以P、V或D疫苗中的一种疫苗初次免疫BALB/c小鼠后,于第2、5、8、11周再用另一种疫苗加强,共产生9种免疫组合:即PP、PV、PD、VP、VV、VD、DP、DV及DD。于初免后第2、5、8、11周采血检测血清中抗HBsAgIgG的总滴度及其IgG1和IgG2a亚类,并于每次加强免疫后第7天,检测小鼠脾脏的CTL对P815S细胞的特异性杀伤率。结果:在P、V、D3种疫苗中,V疫苗诱导产生抗HBsAg抗体的速度最快,P疫苗诱导的体液免疫回忆反应最强,D疫苗诱导产生的抗体最弱。除PP疫苗组合诱导的抗体明显倾向于IgG1外,其他均无明显的倾向性。各种免疫组合中,VD和DV疫苗组诱导的CTL应答最强,对P815S的特异性杀伤率分别为71%和64%。结论:在各种联合免疫组合中,PV、PD、VP和VD疫苗组的抗体应答较好;而DV和VD疫苗组诱导的CTL杀伤效应最强

戊型肝炎病毒重组颗粒性蛋白疫苗在小鼠体内诱导的免疫应答研究

HEV 239是福建省医学分子病毒学研究中心实验室研制的一种戊型肝炎病毒(HEV)重组颗粒性蛋白疫苗,该文旨在研究HEV239蛋白疫苗在小鼠体内诱导产生特异性免疫应答的情况。将5μg HEV 239蛋白疫苗(239-Pro)、加铝佐剂疫苗(239-Vac)或加弗氏佐剂疫苗(239-CFA)肌肉注射免疫BALB/c鼠3次,第8周检测鼠血清抗HEV抗体及其亚类,同时用ELISPOT方法检测细胞毒性T细胞(CTL)应答。结果显示:239-Vac诱导的抗体滴度与239-CFA相当,高于无佐剂的239-Pro。239-Vac诱导的抗体中,IgG1/IgG2a比值显著高于239-CFA和239-Pro,主要为Th2型应答。除239-CFA之外,239-Vac和239-Pro也可诱导出一定的HEV抗原特异性I型Tc应答。提示:重组抗原HEV 239能诱导良好的抗体应答及一定的Tc1应答

戊型肝炎病毒颗粒性蛋白疫苗H-2~d限制性Th表位的筛选

戊型肝炎病毒衣壳蛋白重组抗原HEV 239能形成类病毒颗粒,具备演变成多价疫苗的载体的潜力,此文旨在筛选、鉴定其内包含的H-2d限制性Th表位。以50μg HEV 239蛋白与完全弗式佐剂混合后皮下免疫BALB/c鼠,以覆盖HEV 239蛋白全长的15氨基酸肽库体外刺激其脾细胞,用IFN--γELISPOT方法检测其细胞免疫应答,并通过磁珠剔除脾细胞中CD4+T细胞或CD8+T细胞以分析筛选得到的T细胞表位的特性。结果显示:HEV 239中包含优势的T细胞表位P34(HEV PORF2 AA533~AA547,HSKTF FVLPL RGKLS)及数个较弱的T细胞表位,P34对HEV 239免疫的BALB/c鼠脾细胞的刺激效果与HEV 239蛋白相当,剔除实验表明该表位为CD4+T细胞表位,即Th表位

戊肝病毒Th表位肽免疫可增强其载体蛋白的体液免疫应答

戊型肝炎病毒衣壳蛋白内包含一个强H-2d限制性Th表位P34。以该表位肽免疫BALB/c鼠,其脾细胞能够在体外识别重组戊型肝炎病毒衣壳蛋白,剔除实验表明应答细胞几乎完全是CD4+T细胞,证明P34表位肽能有效诱导产生特异性Th细胞。以P34肽初免小鼠,再以包含该表位的重组戊型肝炎病毒抗原(E2)免疫,结果表明,10μg、20μgE2免疫组在免疫后第1周即有部分小鼠产生抗体,到第3周所有小鼠均能够产生抗体;而对照肽P18初免的小鼠,以20μgE2加强免疫亦无法诱导小鼠产生抗体。这表明,Th表位肽P34初免诱导产生的Th细胞能够有效促进小鼠对携带该表位的载体蛋白的体液免疫应答

戊型肝炎重组蛋白聚乳酸羟基乙酸微球疫苗的研究

目的:研究戊型肝炎重组蛋白(NE2)聚乳酸羟基乙酸(PLGA)微球疫苗诱导免疫应答的情况。方法:复乳法制备微球,考察粒径分布等特性。通过间接ELISA法检测其诱导BALB/c小鼠体内IgG、IgG2a和IgG1水平,并通过IFN--ELISPOT方法检测其诱导BALB/c小鼠体内抗原特异性免疫应答情况。结果:微球的平均粒径为7.1 m。注射小鼠6周后(第4周加强免疫1次),微球疫苗诱导产生的抗戊型肝炎病毒IgG抗体水平较同剂量铝佐剂组明显升高(间接ELISA:OD450/620 10.09 vs.5.32)。IgG2a抗体量略高于铝佐剂组,OD450/620分别为0.17、0.04。IgG1抗体量明显高于铝佐剂组,OD450/620分别为20.48、15.00。IFN--ELISPOT结果显示,微球疫苗能很好的诱导NE2或P34肽抗原特异性免疫应答。结论:戊型肝炎重组蛋白聚乳酸羟基乙酸微球作为疫苗输送体系能明显的提高抗原的免疫原性,有很好的应用前景

福祉職場におけるStaff diversityの研究

There was a major change in 2000 in the national policy on social welfare. The Long-term Care Insurance System started in April 2000. In addition, in June 2000, drastic amendments were made to the Social Welfare Service, after an interval of fifty years. Such changes are having an influence on the management of welfare service business. One influence is that management must have a clearer market mechanism for welfare service business. Today welfare services mainly involve manual labor, and thus, management of personnel expenditures has become the main problem. Although most private companies are diligently carrying out systematic research in HRM (human resource management), in the public welfare service business field study HRM is lagging. In order to deal with this problem, we analyzed data that we collected at 251 facilities of the social welfare service association for the elderly in Hyogo prefecture. In this paper, we would like to present an HRM solution

21世紀地域福祉への提言 : 延岡からの発信

First: How to share the welfare services for the elderly in Nobeoka-City between the public and private sectors.Second: How to promote the social welfare based on the cooperation among business, governmental and academic fields.Third: How to make a good use of local dialects at the place of social welfare