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Objective. To evaluate the Plasmodium falciparum CTL epitope vaccines in HLA class I allele specific human cell lines that have high frequency among Chinese population. Methods. Synthesized oligonucleotides encoding for P.f. CTL epitope genes, constructed eukaryotic expression plasmids, transfected the minigenes into HLA class I allele specific human cell lines and identified endogenous expressing of the minigenes by RT-PCR and HLA stabilization assay. Results. Two mini-genes encoding Plasmodium falciparum CTL epitopes were designed and cloned, respectively, into an eukaryotic expressing vector to form TR26 which was restricted to HLA-B51, SH6 which was restricted to HLA-A2.1, and TS, which had the two aforementioned mini-genes fused in tandem. All of these CTL epitope genes were transfected and endogenously expressed in respective cell lines containing appropriate HLA molecules. The obviously increased expressions of HLA class I molecules were detected in the transfected cell lines. It was demonstrated that the two discrete Plasmodium falciparum epitope genes were effectively processed and presented, and the close proximity of the two epitope genes in one chain as in mini-gene TS did not interfere with the processing and presenting of each epitope gene in corresponding cell line. Conclusion. A successful expression and presentation of multiple CTL epitope mini-gene in MHC class I allele specific human cell lines were demonstrated by an in vitro assay, which could be corresponding to the vaccination of CTL vaccines in people with different MHC I molecules. This work also suggested the possibility of constructing a multiple CTL epitope plasmodium falciparum DNA vaccine that could cover most of Chinese population.
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两种恶性疟快速诊断方法的比较
随着疟疾发病率和感染度的下降,采用常规镜检方法诊断疟疾费时、费力且检出率偏低。为此,国内外有关专家一直努力寻求新的疟疾快速诊断方法,如QBC、Dipstik、ICT、LDH-P、PCR等方法,应用诸方法进行恶性疟检测的报道甚多[1,2],但有关应用DEAE-纤维素薄层色谱法(DEAE thin-layer chromatography,DEAE-TLC)通过检测LDH-P诊断恶性疟的报道尚未见到。为找到一种更适合在我国推广应用的疟疾快速诊断方法,作者选择ICT及DEAE-TLC两种方法对恶性疟病人血样进行检测,并对两种检测方法及结果作了比较。1 材料与方法1.1 ICT试剂盒为澳大利亚Austcard公司产品。
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恶性疟BCG多价疫苗及DNA疫苗的实验研制
本项目在恶性疟疫苗候选分子裂殖子表面蛋白-2(MSA2)及环子孢子蛋白(CSP)的分子生物学特征和免疫原性研究的基础上,构建了CSP、MSA2之DNA疫苗,同时率先在国内外构建CSP、MSA2之重组BCG活疫苗,并对疫苗的免疫保护机制、免疫应答及疫苗安全性进行了研究.
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疟原虫乳酸脱氢酶活性检测法和有限稀释法在筛选和建立恶性疟原虫克隆株时的应用
建立恶性疟原虫克隆株存在着两个技术难点:一是怎样对原虫感染率、原虫绝对数均小的标本进行疟原虫检测;二是怎样从一个疟原虫分离株中选择一个疟原虫.针对这两个问题,我所在建立恶性疟原虫克隆株过程中,检测微量疟原虫时采用乳酸脱氢酶活性检测法.在分离疟原虫个体时采用有限稀释法,对疟原虫高度且有限地稀释,以实现对疟原虫个体的分离,获得成功,为建立疟原虫克隆株奠定了基础.现报告如下: