Dissection-independent production of Plasmodium sporozoites from whole mosquitoes.
Dissection-independent production of Plasmodium sporozoites from whole mosquitoes.
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DOI：
10.26508/lsa.202101094
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发表时间：
2021-07
影响因子：
4.4
通讯作者：
Baum J
中科院分区：
文献类型：
--
作者：
Blight J;Sala KA;Atcheson E;Kramer H;El-Turabi A;Real E;Dahalan FA;Bettencourt P;Dickinson-Craig E;Alves E;Salman AM;Janse CJ;Ashcroft FM;Hill AV;Reyes-Sandoval A;Blagborough AM;Baum J
克服了传统的分离方法的局限性，疟疾子孢子从蚊子，一个新的解剖的方法，产生纯的，感染性的子孢子。研制疟疾保护性疫苗的进展仍然缓慢。到目前为止，只有有限的保护已经实现了常规免疫接种后，无论是整个寄生虫（孢子）或亚基为基础的疫苗。疫苗进展和红细胞前寄生虫生物学的一个主要障碍是继续依赖于人工唾液腺解剖来从受感染的蚊子中分离子孢子。在这里，我们报告了一个多步骤的方法，基于批量处理的均质化的整个蚊子，浆料，和密度梯度过滤，结合自由流动电泳快速产生一个纯的，感染性的子孢子接种物。在体外肝细胞感染测定中，以这种方式产生的人感染性恶性疟原虫和啮齿动物感染性伯氏疟原虫子孢子的感染性是唾液腺解剖子孢子的2至3倍。在体内啮齿动物疟疾模型中，当静脉内递送免疫时，相同的伯氏疟原虫子孢子赋予针对蚊子叮咬攻击的无菌保护，或者当肌内递送免疫时，赋予60-70%的保护。通过提高纯度、感染性和免疫原性，这种方法代表了生产研究级子孢子能力的关键进步，这将影响未来大规模提供基于全寄生虫的疟疾疫苗。
Overcoming limitations of traditional dissection-dependent methods for malarial sporozoite isolation from mosquitoes, a new dissection-free method is presented that yields pure, infective sporozoites. Progress towards a protective vaccine against malaria remains slow. To date, only limited protection has been routinely achieved following immunisation with either whole-parasite (sporozoite) or subunit-based vaccines. One major roadblock to vaccine progress, and to pre-erythrocytic parasite biology in general, is the continued reliance on manual salivary gland dissection for sporozoite isolation from infected mosquitoes. Here, we report development of a multi-step method, based on batch processing of homogenised whole mosquitoes, slurry, and density-gradient filtration, which combined with free-flow electrophoresis rapidly produces a pure, infective sporozoite inoculum. Human-infective Plasmodium falciparum and rodent-infective Plasmodium berghei sporozoites produced in this way are two- to threefold more infective than salivary gland dissection sporozoites in in vitro hepatocyte infection assays. In an in vivo rodent malaria model, the same P. berghei sporozoites confer sterile protection from mosquito-bite challenge when immunisation is delivered intravenously or 60–70% protection when delivered intramuscularly. By improving purity, infectivity, and immunogenicity, this method represents a key advancement in capacity to produce research-grade sporozoites, which should impact delivery of a whole-parasite based malaria vaccine at scale in the future.
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影响因子：
2.1
作者：
BEAUDOIN, RL;STROME, CPA;TUBERGEN, TA
通讯作者：
TUBERGEN, TA
影响因子：
3.7
作者：
Angrisano, Fiona;Riglar, David T.;Baum, Jake
通讯作者：
Baum, Jake
影响因子：
4.7
作者：
Hegge, Stephan;Kudryashev, Mikhail;Frischknecht, Friedrich
通讯作者：
Frischknecht, Friedrich
DOI：
10.1084/jem.20101488
发表时间：
2011-02-14
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Coppi A;Natarajan R;Pradel G;Bennett BL;James ER;Roggero MA;Corradin G;Persson C;Tewari R;Sinnis P
通讯作者：
Sinnis P
影响因子：
8
作者：
Epstein, Judith E.;Paolino, Kristopher M.;Hoffman, Stephen L.
通讯作者：
Hoffman, Stephen L.