Dissection-independent production of<i>Plasmodium</i>sporozoites from whole mosquitoes

Blight, Joshua; Sala, Katarzyna; Atcheson, Erwan; Kramer, Holger; El-Turabi, Aadil; Real, Eliana; Dahalan, Farah A.; Bettencourt, Paulo; Dickinson-Craig, Emma; Alves, Eduardo Rodrigues; Salman, Ahmed M.; Janse, Chris J.; Ashcroft, Frances M.; Hill, Adrian V. S.; Reyes-Sandoval, Arturo; Blagborough, Andrew M.; Baum, Jake

doi:10.26508/lsa.202101094

Cited by 2 publications

(1 citation statement)

References 67 publications

(109 reference statements)

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“…A single-shot selfboosting vaccine would have enormous (logistical) benefits for resource-poor settings over vaccines that need regular boosting. In addition, live sporozoite vaccine approaches are becoming more feasible for mass application in humans due to advances in GMP production technologies, production of sporozoites in insect cell lines 26 and the cryopreservation of sporozoites 27 . Although cross-species protection between P. cynomolgi and P. vivax remains to be ascertained and while the current approach cannot directly be translated to humans, given the initial success of the hypnoboost concept reported here, approaches using wild-type or engineered P. cynomolgi GAP parasites together with selected adjuvants may well be a viable option for the development of a highly efficacious P. vivax malaria vaccine.…”

Section: Discussionmentioning

confidence: 99%

Sterile protection against relapsing malaria with a single-shot vaccine

et al. 2022

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Vaccine development for Plasmodium vivax, an important human relapsing malaria, is lagging behind. In the case of the most deadly human malaria P. falciparum, unprecedented high levels of protection have been obtained by immunization with live sporozoites under accompanying chemoprophylaxis, which prevents the onset of blood-stage malaria. Such an approach has not been fully evaluated for relapsing malaria. Here, in the P. cynomolgi-rhesus macaque model for relapsing malaria, we employ the parasites’ natural relapsing phenotype to self-boost the immune response against liver-stage parasites, following a single-shot high-dose live sporozoite vaccination. This approach resulted in sterile protection against homologous sporozoite challenge in three out of four animals in the group that was also exposed for several days to blood stages during primary infection and relapses. One out of four animals in the group that received continuous chemoprophylaxis to abort blood-stage exposure was also protected from sporozoite challenge. Although obtained in a small number of animals as part of a Proof-of-Concept study, these results suggest that limited blood-stage parasite exposure may augment protection in this model. We anticipate our data are a starting point for further research into correlates of protection and extrapolation of the single-shot approach to develop efficacious malaria vaccines against relapsing human malaria.

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Section: Discussionmentioning

confidence: 99%

Sterile protection against relapsing malaria with a single-shot vaccine

et al. 2022

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An Optimized <em>P. berghei</em> Liver Stage–HepG2 Infection Model for Simultaneous Quantitative Bioimaging of Host and Parasite Nascent Proteomes

McLellan,

Garcia-Vilanova,

Hanson

2024

BIO-PROTOCOL

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The Plasmodium parasites that cause malaria undergo an obligate, asymptomatic developmental stage in the host liver before initiating the symptomatic blood-stage infection. The parasite liver stage is a key intervention point for antimalarial chemoprophylaxis: successful targeting of liver-stage parasites prevents disease development in individuals and can help to reduce parasite transmission in populations, as the gametocyte forms that transmit infection to mosquitos are exclusively found in the blood stage. Antimalarial drugs that can target multiple parasite stages are thus highly desirable, and one emerging cellular target for such multistage active compounds is the process of protein synthesis or translation. Quantitative study of liver stage translation, and thus mechanistic evaluation of translation inhibitors against liver stage parasites, is not amenable to the methods allowing quantification of asexual blood stage translation, such as radiolabeled amino acid incorporation or lysate-based translation of reporter transcripts. Here, we present a method using o-propargyl puromycin (OPP) labeling of host and parasite nascent proteomes in the P. berghei -HepG2 infection model, followed by automated confocal image acquisition and computational separation of P. berghei vs. H. sapiens nascent proteome signals to allow simultaneous readout of the effects of translation inhibitors on both host and parasite. This protocol details our HepG2 cell culture and infected monolayer handling optimized for microscopy, our OPP labeling workflow, and our approach to automated confocal imaging, image processing, and data analysis. Key features • Uses the o-propargyl puromycin labeling technique developed by Liu et al. to quantitatively analyze protein synthesis in Plasmodium berghei liver-stage parasites in actively translating hepatoma cells. • This quantitative approach should be adaptable for other puromycin-sensitive intracellular pathogens residing in actively translating host cells. • The P. berghei –infected HepG2 recovery and reseeding protocol presented here is of use in applications beyond nascent proteome labeling and quantification.

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Dissection-independent production ofPlasmodiumsporozoites from whole mosquitoes

Cited by 2 publications

References 67 publications

Sterile protection against relapsing malaria with a single-shot vaccine

Sterile protection against relapsing malaria with a single-shot vaccine

An Optimized <em>P. berghei</em> Liver Stage–HepG2 Infection Model for Simultaneous Quantitative Bioimaging of Host and Parasite Nascent Proteomes

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