Using Plasmodium knowlesi as a model for screening Plasmodium vivax blood-stage malaria vaccine targets reveals new candidates.
Using Plasmodium knowlesi as a model for screening Plasmodium vivax blood-stage malaria vaccine targets reveals new candidates.
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DOI：
10.1371/journal.ppat.1008864
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发表时间：
2021-07
期刊：
影响因子：
6.7
通讯作者：
Rayner JC
中科院分区：
文献类型：
--
作者：
Ndegwa DN;Kundu P;Hostetler JB;Marin-Menendez A;Sanderson T;Mwikali K;Verzier LH;Coyle R;Adjalley S;Rayner JC
间日疟原虫是非洲以外大多数疟疾病例的罪魁祸首。与恶性疟原虫不同，间日疟原虫的生命周期包括一个休眠的肝脏阶段，即催眠期，它可以在没有蚊子传播的情况下引起感染。针对间日疟原虫血分期的有效疫苗将限制此类复发性感染的症状和病理，因此可能在控制该物种方面发挥关键作用。然而，间日疟原虫的疫苗开发远远落后于恶性疟原虫，后者有许多已确定的目标，其中一些已过渡到第二阶段测试。相比之下，只有一种基于Duffy结合蛋白（PvDBP）的间日疟原虫血期候选疫苗达到了i期，这在很大程度上是因为缺乏连续的间日疟原虫体外培养系统限制了对新候选疫苗的系统筛选。我们利用间日疟原虫和诺氏疟原虫之间密切的系统发育关系，在人红细胞中建立了体外培养系统，以测试系统反向疫苗学的可扩展性，以识别和优先考虑间日疟原虫的血期靶点。在哺乳动物表达系统中，以全长重组外结构域的形式表达了一组预测在红细胞侵袭中起作用的间日疟原虫蛋白。其中8种抗原用于生成多克隆抗体，并对其识别诺氏疟原虫同源蛋白的能力进行了筛选。然后测试这些抗体对野生型和嵌合型诺氏疟原虫的生长和侵袭的抑制作用，这些嵌合型诺氏疟原虫使用CRISPR/Cas9修饰，与间日疟原虫同源物交换诺氏疟原虫基因。鉴定出了诱导抗体抑制侵袭的候选抗体，其水平与PvDBP相似，证实了诺氏疟原虫作为间日疟原虫疫苗开发模型的效用，并确定了进一步随访的优先抗原。疟疾寄生虫在入侵人类红细胞后引起疾病，这意味着阻断这一过程的疫苗可能在疟疾控制中发挥重要作用。多种疟原虫可引起人类疟疾，非洲以外的大多数疟疾是由间日疟原虫引起的。目前还没有针对任何疟疾寄生虫血液阶段的有效疫苗，间日疟原虫疫苗开发的进展尤其受到阻碍，因为这种寄生虫不能在实验室长时间培养。我们探索了一种可以在体外人红细胞中繁殖的诺氏疟原虫是否可以用于筛选间日疟原虫疫苗的潜在靶点。我们培养了针对间日疟原虫蛋白的抗体，并测试了它们识别和阻止诺氏疟原虫入侵人类红细胞的能力，从而确定了多种新的候选疫苗。
Plasmodium vivax is responsible for the majority of malaria cases outside Africa. Unlike P. falciparum, the P. vivax life-cycle includes a dormant liver stage, the hypnozoite, which can cause infection in the absence of mosquito transmission. An effective vaccine against P. vivax blood stages would limit symptoms and pathology from such recurrent infections, and therefore could play a critical role in the control of this species. Vaccine development in P. vivax, however, lags considerably behind P. falciparum, which has many identified targets with several having transitioned to Phase II testing. By contrast only one P. vivax blood-stage vaccine candidate based on the Duffy Binding Protein (PvDBP), has reached Phase Ia, in large part because the lack of a continuous in vitro culture system for P. vivax limits systematic screening of new candidates. We used the close phylogenetic relationship between P. vivax and P. knowlesi, for which an in vitro culture system in human erythrocytes exists, to test the scalability of systematic reverse vaccinology to identify and prioritise P. vivax blood-stage targets. A panel of P. vivax proteins predicted to function in erythrocyte invasion were expressed as full-length recombinant ectodomains in a mammalian expression system. Eight of these antigens were used to generate polyclonal antibodies, which were screened for their ability to recognize orthologous proteins in P. knowlesi. These antibodies were then tested for inhibition of growth and invasion of both wild type P. knowlesi and chimeric P. knowlesi lines modified using CRISPR/Cas9 to exchange P. knowlesi genes with their P. vivax orthologues. Candidates that induced antibodies that inhibited invasion to a similar level as PvDBP were identified, confirming the utility of P. knowlesi as a model for P. vivax vaccine development and prioritizing antigens for further follow up. Malaria parasites cause disease after invading human red blood cells, implying that a vaccine that interrupts this process could play a significant role in malaria control. Multiple Plasmodium parasite species can cause malaria in humans, and most malaria outside Africa is caused by Plasmodium vivax. There is currently no effective vaccine against the blood stage of any malaria parasite, and progress in P. vivax vaccine development has been particularly hampered because this parasite species cannot be cultured for prolonged periods of time in the lab. We explored whether a related species, P. knowlesi, which can be propagated in human red blood cells in vitro, can be used to screen for potential P. vivax vaccine targets. We raised antibodies against selected P. vivax proteins and tested their ability to recognize and prevent P. knowlesi parasites from invading human red blood cells, thereby identifying multiple novel vaccine candidates.
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DOI：
10.1073/pnas.1702944114
发表时间：
2017-11-07
影响因子：
11.1
作者：
Bustamante LY;Powell GT;Lin YC;Macklin MD;Cross N;Kemp A;Cawkill P;Sanderson T;Crosnier C;Muller-Sienerth N;Doumbo OK;Traore B;Crompton PD;Cicuta P;Tran TM;Wright GJ;Rayner JC
通讯作者：
Rayner JC
影响因子：
3.7
作者：
Healer J;Thompson JK;Riglar DT;Wilson DW;Chiu YH;Miura K;Chen L;Hodder AN;Long CA;Hansen DS;Baum J;Cowman AF
通讯作者：
Cowman AF
影响因子：
64.8
作者：
Carlton, Jane M.;Adams, John H.;Silva, Joana C.;Bidwell, Shelby L.;Lorenzi, Hernan;Caler, Elisabet;Crabtree, Jonathan;Angiuoli, Samuel V.;Merino, Emilio F.;Amedeo, Paolo;Cheng, Qin;Coulson, Richard M. R.;Crabb, Brendan S.;del Portillo, Hernando A.;Essien, Kobby;Feldblyum, Tamara V.;Fernandez-Becerra, Carmen;Gilson, Paul R.;Gueye, Amy H.;Guo, Xiang;Kang'a, Simon;Kooij, Taco W. A.;Korsinczky, Michael;Meyer, Esmeralda V. -S.;Nene, Vish;Paulsen, Ian;White, Owen;Ralph, Stuart A.;Ren, Qinghu;Sargeant, Tobias J.;Salzberg, Steven L.;Stoeckert, Christian J.;Sullivan, Steven A.;Yamamoto, Marcio M.;Hoffman, Stephen L.;Wortman, Jennifer R.;Gardner, Malcolm J.;Galinski, Mary R.;Barnwell, John W.;Fraser-Liggett, Claire M.
通讯作者：
Fraser-Liggett, Claire M.
影响因子：
4.6
作者：
Cheng Y;Lu F;Wang B;Li J;Han JH;Ito D;Kong DH;Jiang L;Wu J;Ha KS;Takashima E;Sattabongkot J;Cao J;Nyunt MH;Kyaw MP;Desai SA;Miller LH;Tsuboi T;Han ET
通讯作者：
Han ET
影响因子：
3.8
作者：
França CT;Hostetler JB;Sharma S;White MT;Lin E;Kiniboro B;Waltmann A;Darcy AW;Li Wai Suen CS;Siba P;King CL;Rayner JC;Fairhurst RM;Mueller I
通讯作者：
Mueller I