Full-length recombinant Plasmodium falciparum VAR2CSA binds specifically to CSPG and induces potent parasite adhesion-blocking antibodies.
Full-length recombinant Plasmodium falciparum VAR2CSA binds specifically to CSPG and induces potent parasite adhesion-blocking antibodies.
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DOI：
10.1016/j.jmb.2010.01.040
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发表时间：
2010-04-02
影响因子：
5.6
通讯作者：
Salanti A
中科院分区：
文献类型：
--
作者：
Khunrae P;Dahlbäck M;Nielsen MA;Andersen G;Ditlev SB;Resende M;Pinto VV;Theander TG;Higgins MK;Salanti A
恶性疟原虫疟疾仍然是世界上造成人类痛苦和贫穷的主要原因之一。每年，这种疾病夺去100万至300万人的生命，主要是在撒哈拉以南非洲。感染红细胞粘附于血管内皮或胎盘是恶性疟原虫严重感染发病机制中的关键事件。在孕妇中，寄生虫表达一种名为VAR2CSA的恶性疟原虫红细胞膜蛋白1（PfEMP 1）家族的单一和独特成员，这与IE特异性粘附胎盘中硫酸软骨素A（CSA）的能力有关。来自VAR2CSA分子的几个Duffy结合样结构域已经在体外显示出与CSA结合，但是也已经证明来自除VAR2CSA之外的PfEMP 1蛋白的Duffy结合样结构域可以结合CSA。此外，VAR2CSA结构域与糖胺聚糖结合的特异性与表达VAR2CSA的IE的特异性不匹配。这导致推测天然VAR2CSA的结构域需要聚集在一起以形成特异性结合位点，或者VAR2CSA可能通过桥接分子与CSA结合。在这里，我们描述的表达和纯化的完整的细胞外区域的VAR2CSA分泌在高产量的昆虫细胞。使用表面等离子体共振，我们证明，VAR2CSA单独结合与纳摩尔亲和力的人硫酸软骨素蛋白聚糖和显着较弱的亲和力，其他糖胺聚糖，显示出类似的特异性观察到的IE。针对全长VAR2CSA产生的抗体完全抑制重组VAR2CSA结合，以及寄生虫与硫酸软骨素蛋白聚糖的结合。这是第一个描述全长PfEMP 1的成功生产和功能的研究。诱导IgG的结合和抗粘附效力的特异性，以及高产率生产，鼓励在疫苗开发策略中使用全长PfEMP 1。
Plasmodium falciparum malaria remains one of the world's leading causes of human suffering and poverty. Each year, the disease takes 1–3 million lives, mainly in sub-Saharan Africa. The adhesion of infected erythrocytes (IEs) to vascular endothelium or placenta is the key event in the pathogenesis of severe P. falciparum infection. In pregnant women, the parasites express a single and unique member of the P. falciparum erythrocyte membrane protein 1 (PfEMP1) family named VAR2CSA, which is associated with the ability of the IEs to adhere specifically to chondroitin sulphate A (CSA) in the placenta. Several Duffy-binding-like domains from VAR2CSA molecules have been shown in vitro to bind to CSA, but it has also been demonstrated that Duffy-binding-like domains from PfEMP1 proteins other than VAR2CSA can bind CSA. In addition, the specificity of the binding of VAR2CSA domains to glycosaminoglycans does not match that of VAR2CSA-expressing IEs. This has led to speculation that the domains of native VAR2CSA need to come together to form a specific binding site or that VAR2CSA might bind to CSA through a bridging molecule. Here, we describe the expression and purification of the complete extracellular region of VAR2CSA secreted at high yields from insect cells. Using surface plasmon resonance, we demonstrate that VAR2CSA alone binds with nanomolar affinity to human chondroitin sulphate proteoglycan and with significantly weaker affinity to other glycosaminoglycans, showing a specificity similar to that observed for IEs. Antibodies raised against full-length VAR2CSA completely inhibit recombinant VAR2CSA binding, as well as parasite binding to chondroitin sulphate proteoglycan. This is the first study to describe the successful production and functionality of a full-length PfEMP1. The specificity of the binding and anti-adhesion potency of induced IgG, together with high-yield production, encourages the use of full-length PfEMP1 in vaccine development strategies.
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影响因子：
4
作者：
Resende, Mafalda;Ditlev, Sisse B.;Dahlback, Madeleine
通讯作者：
Dahlback, Madeleine
影响因子：
5.6
作者：
Khunrae, Pongsak;Philip, Judith M. D.;Bull, Duncan R.;Higgins, Matthew K.
通讯作者：
Higgins, Matthew K.
影响因子：
64.5
作者：
BARUCH, DI;PASLOSKE, BL;HOWARD, RJ
通讯作者：
HOWARD, RJ
影响因子：
3.1
作者：
Bir, Nivedita;Yazdani, Syed Shams;Chitnis, Chetan E.
通讯作者：
Chitnis, Chetan E.
影响因子：
3.1
作者：
Avril, Marion;Kulasekara, Bridget R.;Smith, Joseph D.
通讯作者：
Smith, Joseph D.