Plataforma de Produção e Infecção de Vetores da Malária (PIVEM)/Laboratório de Entomologia, Fiocruz Rondônia, Porto Velho, Rondônia, Brazil.
Programa de Pós-Graduação em Saúde Publica, Faculdade de Saúde Pública, Universidade de São Paulo, São Paulo, Brazil.
Sci Rep. 2023 Oct 24;13(1):18207. doi: 10.1038/s41598-023-44556-y.
Obtaining Plasmodium vivax sporozoites is essential for in vitro culture of liver stage parasites, not only to understand fundamental aspects of parasite biology, but also for drug and vaccine development. A major impediment to establish high-throughput in vitro P. vivax liver stage assays for drug development is obtaining sufficient numbers of sporozoites. To do so, female anopheline mosquitoes have to be fed on blood from P. vivax-infected patients through an artificial membrane-feeding system, which in turns requires a well-established Anopheles colony. In this study we established conditions to provide a robust supply of P. vivax sporozoites. Adding a combination of serum replacement and antibiotics to the membrane-feeding protocol was found to best improve sporozoite production. A simple centrifugation method appears to be a possible tool for rapidly obtaining purified sporozoites with a minimal loss of yield. However, this method needs to be better defined since sporozoite viability and hepatocyte infection were not evaluated.
获取间日疟原虫（Plasmodium vivax）的孢子虫是体外培养肝期寄生虫的必要条件，不仅可以帮助我们理解寄生虫生物学的基本方面，还有助于药物和疫苗的开发。然而，建立高通量的间日疟原虫肝期药物开发体外检测方法的主要障碍是获得足够数量的孢子虫。为此，需要通过人工膜饲系统让雌性按蚊吸食来自间日疟原虫感染患者的血液，这反过来又需要一个成熟的按蚊群体。在本研究中，我们建立了提供大量间日疟原虫孢子虫的条件。在膜饲方案中添加血清替代物和抗生素的组合被发现是提高孢子虫产量的最佳方法。一种简单的离心方法似乎是一种快速获得高纯度孢子虫的可行工具，且对产量的损失最小。然而，由于尚未评估该方法对孢子虫活力和肝细胞感染的影响，因此需要进一步优化。
