Strain-transcendent immune response to recombinant Var2CSA DBL5-ε domain block P. falciparum adhesion to placenta-derived BeWo cells under flow conditions.
Strain-transcendent immune response to recombinant Var2CSA DBL5-ε domain block P. falciparum adhesion to placenta-derived BeWo cells under flow conditions.
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DOI：
10.1371/journal.pone.0012558
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发表时间：
2010-09-03
期刊：
影响因子：
3.7
通讯作者：
Scherf A
中科院分区：
文献类型：
--
作者：
Fernandez P;Petres S;Mécheri S;Gysin J;Scherf A
妊娠相关性疟疾（PAM）是恶性疟原虫感染的红细胞（PE）粘附于胎盘受体硫酸软骨素A（CSA）的严重后果，所述红细胞表达大的富含半胱氨酸的多结构域蛋白var 2CSA。妇女对PAM产生耐药性，并对CSA结合寄生虫产生超越菌株的免疫力。鉴定能够引起广泛中和和粘附阻断抗体的var 2CSA区域是设计预防性疫苗策略的关键步骤。 大肠杆菌表达的var 2CSA DBL结构域在免疫小鼠和山羊之前被重折叠和纯化。使用流动细胞粘附抑制测定法测试蛋白G纯化的抗体阻断FCR 3CSA感染的红细胞与胎盘（BeWo）和猴脑内皮（ScC 2）细胞系结合的能力，所述测定法在0.05 Pa的剪切应力下密切模拟胎盘中存在的生理条件。DBL 5-ε、DBL 6-ε和DBL 5 -6-ε使用明矾和弗氏佐剂诱导交叉反应性抗体，其在0.5mg IgG/ml下以40至96%的值阻断细胞粘附。重要的是，针对来自3种不同寄生虫基因型（HB 3、Dd 2和7 G8）的重组DBL 5-ε产生的抗体显示出对异源寄生虫FCR 3CSA的38%至64%的菌株超越抑制。使用来自var 2CSA和明矾的单和双DBL结构域作为佐剂，我们鉴定了在实验动物中诱导免疫应答的重组亚基，其能够在流动细胞粘附测定中有效地阻断寄生虫粘附，所述流动细胞粘附测定密切模拟胎盘中的红细胞流动。这些亚基显示出包含在旨在保护免受PAM的疫苗中的有希望的特征。
Pregnancy-associated malaria (PAM) is a serious consequence of the adhesion to the placental receptor chondroitin sulfate A (CSA) of Plasmodium falciparum-infected erythrocytes (PE) expressing the large cysteine-rich multi-domain protein var2CSA. Women become resistant to PAM, and develop strain-transcending immunity against CSA-binding parasites. The identification of var2CSA regions that could elicit broadly neutralizing and adhesion-blocking antibodies is a key step for the design of prophylactic vaccine strategies. Escherichia coli expressed var2CSA DBL domains were refolded and purified prior to immunization of mice and a goat. Protein-G-purified antibodies were tested for their ability to block FCR3CSA-infected erythrocytes binding to placental (BeWo) and monkey brain endothelial (ScC2) cell lines using a flow cytoadhesion inhibition assay mimicking closely the physiological conditions present in the placenta at shear stress of 0.05 Pa. DBL5-ε, DBL6-ε and DBL5-6-ε induced cross-reactive antibodies using Alum and Freund as adjuvants, which blocked cytoadhesion at values ranging between 40 to 96% at 0.5 mg IgG per ml. Importantly, antibodies raised against recombinant DBL5-ε from 3 distinct parasites genotypes (HB3, Dd2 and 7G8) showed strain-transcending inhibition ranging from 38 to 64% for the heterologuous FCR3CSA. Using single and double DBL domains from var2CSA and Alum as adjuvant, we identified recombinant subunits inducing an immune response in experimental animals which is able to block efficiently parasite adhesion in a flow cytoadhesion assay that mimics closely the erythrocyte flow in the placenta. These subunits show promising features for inclusion into a vaccine aiming to protect against PAM.
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