Development of a new technology using synthetic malarial peptides for immunological analysis of an endemic population

开发利用合成疟疾肽对流行人群进行免疫分析的新技术

• 批准号: 08557021
• 负责人: SUZUKI Mamoru
• 金额: $ 9.92万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08557021/
• 关键词: malaria; synthetic antigen; immunology; diagnosis; sporozoite; enolase; 診断

In this research, we synthesized the following two peptides first, Boc-GIGNPNAGIG-Oet and cyclo[(NPNAGAG)ィイD22ィエD2] with regard to the basic structure of the circum sporozoite protein, (NANP)ィイD2nィエD2 Structure of the peptides were analyzed by Nuclear Magnetic Resonance, Fourier-transform Infraredspectrum, and Circular Dichroism Spectroscopy, and it was revealed that those peptides had β-structure within the polypeptides which would work as an appropriate antigenic structure for the detection of the malarial antibody. When we examined their antigenicity by fluorescence-ELISA by applying malaria patients' sera from Thailand, quite a nice reaction was observed suggesting their potential importance as diagnostic materials.Another synthetic polypeptides studied in this research was a part of enolase, a glycolytic enzyme. We analyzed the tertially structure of enolase with reference to the crystal structure of yeast enolase. The identity of the peptides were calculated as 61% and we concluded that we could built up the structure of Plasmodial enolase by computer graphics. Then we chose a part of the polypeptides which consisted of Asn with β-turn structure: GFAPNILNANEALDLL. The synthetic polypeptides of this helical structure was also proved to be a good antigenic molecule for the detection of malarial antibody. It is also proved that this molecule was highly reactive against the sera from complicated malaria patients, but less reactive against those from mild malaria patients. We, thus, regarded this molecule as a good material for us to develop appropriate diagnostic technology not only for individuals, but also for the evaluation of epidemiological situation of endemic areas.

在这项研究中,我们合成了以下两个肽首先,Boc-GIGNPNAGIG-Oet和三轮车[(NPNAGAG)ィイD22摊位ィエD2)关于环孢子体的基本结构蛋白(NANP)ィイD2nィエD2肽的结构被核磁共振分析,傅里叶变换Infraredspectrum,和圆二色性光谱,结果表明，这些多肽具有β-结构，可作为检测疟疾抗体的合适抗原结构。当我们应用泰国疟疾患者的血清，用荧光- elisa检测它们的抗原性时，观察到相当好的反应，表明它们作为诊断材料的潜在重要性。本研究研究的另一种合成多肽是烯醇化酶的一部分，一种糖酵解酶。我们参照酵母烯醇化酶的晶体结构分析了烯醇化酶的结构。经计算，这些肽段的同源性为61%，可以用计算机图形学的方法建立疟原虫烯醇化酶的结构。然后我们选择了一部分由Asn组成的β-turn结构的多肽：GFAPNILNANEALDLL。合成的这种螺旋结构的多肽也被证明是检测疟疾抗体的良好抗原分子。结果表明，该分子对复杂疟疾患者血清具有较强的抗疟活性，而对轻度疟疾患者血清的抗疟活性较弱。因此，我们认为该分子不仅可以为个体提供合适的诊断技术，而且可以为流行地区的流行病学情况评估提供良好的材料。

项目成果

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