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首页> 外文期刊>Infection and immunity >Protective Immunization with a Novel Membrane Protein of Plasmodium yoelii-Infected Erythrocytes
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Protective Immunization with a Novel Membrane Protein of Plasmodium yoelii-Infected Erythrocytes

机译：新型膜蛋白的约氏疟原虫感染的红细胞的保护性免疫。

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Immunization with a particulate fraction of blood-stage antigens was shown previously to protect mice against Plasmodium yoelii malaria. To identify antigens inducing the protective response, sera from immunized mice were used to screen a P. yoelii cDNA expression library. Sequence analysis of one 2.6-kb cDNA clone indicated that the identified gene,pypag-1, encoded a novel plasmodial antigen. Two nonoverlapping regions of pypag-1 were expressed inEscherichia coli. The first recombinant antigen, pAg-1N, contained the N-terminal 337 residues, which included a putative transmembrane domain and a region relatively rich in tryptophan residues. The second recombinant antigen, pAg-1C, contained the remaining C-terminal 211 residues, which included 31 copies of a 5-amino-acid degenerative repeat. Immunoblot studies using rabbit antiserum raised against recombinant pAg-1N showed that the native pypAg-1 protein migrated at approximately 98 kDa, considerably slower than its predicted molecular mass of 66 kDa. Immunofluorescence studies localized the expression of the native pypAg-1 protein both to the cytoplasm and at the surface of P. yoelii-infected erythrocytes. Immunization with either pAg-1N or pAg-1C induced a four- to sevenfold reduction in P. yoeliiblood-stage parasitemia. As such, pypAg-1 appears to contain at least two distinct protective epitopes. To our knowledge, this is the first characterization of a protective antigen of P. yoelii that is associated with the erythrocyte membrane.
机译：以前显示过使用微粒级的血液阶段抗原进行免疫可以保护小鼠免于约氏疟原虫疟疾。为了鉴定诱导保护性应答的抗原，使用来自免疫小鼠的血清筛选 P。 yoelii cDNA表达文库。对一个2.6-kb cDNA克隆的序列分析表明，鉴定出的基因 pypag-1 编码一种新的疟原虫抗原。 pypag-1 的两个非重叠区域在大肠杆菌中表达。第一个重组抗原pAg-1N包含N末端337个残基，其中包括一个推定的跨膜结构域和一个相对较丰富的色氨酸残基的区域。第二种重组抗原pAg-1C包含剩余的C末端211残基，其中包括31个拷贝的5个氨基酸的变性重复序列。使用针对重组pAg-1N的兔抗血清进行的免疫印迹研究表明，天然pypAg-1蛋白的迁移速率约为98 kDa，大大低于其预测的66 kDa的分子量。免疫荧光研究将天然pypAg-1蛋白的表达定位于细胞质和 P表面。 yoelii 感染的红细胞。用pAg-1N或pAg-1C免疫可导致 P降低4到7倍。 yoelii 血期寄生虫病。这样，pypAg-1似乎包含至少两个不同的保护性表位。据我们所知，这是 P保护性抗原的第一个特征。与红细胞膜有关的yoelii 。

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