An Opsonic Phagocytosis Assay for Plasmodium falciparum Sporozoites.
An Opsonic Phagocytosis Assay for Plasmodium falciparum Sporozoites.
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恶性疟原虫子孢子的调理吞噬作用测定。
DOI：
10.1128/cvi.00445-16
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发表时间：
2017
期刊：
影响因子：
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通讯作者：
Kappe,StefanHI
中科院分区：
文献类型：
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作者：
Steel,RyanWJ;Sack,BrandonK;Tsuji,Moriya;Navarro,MaryJaneL;Betz,Will;Fishbaugher,MattE;Flannery,ErikaL;Kappe,StefanHI
恶性疟原虫疟疾仍然是全世界最致命的寄生虫病。靶向红细胞前期子孢子和肝脏阶段的疫苗具有完全预防血液阶段感染和疾病以及向前传播的潜力。子孢子表面和分泌的蛋白质是包含在红细胞前期阶段特异性的基于抗体的疫苗中的主要候选物。临床前功能测定，以确定体液相关的保护在vitro和验证新的子孢子蛋白靶点，包括在多亚单位疫苗目前不考虑子孢子靶向抗体与免疫系统的其他组件的相互作用。在这里，我们描述了一个简单的流式细胞术测定的发展，以定量评估针对恶性疟原虫子孢子的抗体，以促进其吞噬的能力。我们证明，这种子孢子调理吞噬试验（SOPA）与单克隆抗体和人免疫血清相容，并可使用冷冻保存的恶性疟原虫子孢子进行。这种简单、方便的检测方法将有助于评估疟原虫抗原疫苗接种后的抗体反应及其与免疫系统吞噬细胞的相互作用。
Plasmodium falciparum malaria remains the deadliest parasitic disease worldwide. Vaccines targeting the preerythrocytic sporozoite and liver stages have the potential to entirely prevent blood-stage infection and disease, as well as onward transmission. Sporozoite surface and secreted proteins are leading candidates for inclusion in a preerythrocytic stage-specific, antibody-based vaccine. Preclinical functional assays to identify humoral correlates of protectionin vitroand to validate novel sporozoite protein targets for inclusion in multisubunit vaccines currently do not consider the interaction of sporozoite-targeting antibodies with other components of the immune system. Here, we describe the development of a simple flow cytometric assay to quantitatively assess the ability of antibodies directed against P. falciparum sporozoites to facilitate their phagocytosis. We demonstrate that thissporozoiteopsonicphagocytosisassay (SOPA) is compatible with both monoclonal antibodies and human immune serum and can be performed using cryopreserved P. falciparum sporozoites. This simple, accessible assay will aid with the assessment of antibody responses to vaccination with Plasmodium antigens and their interaction with phagocytic cells of the immune system.