Immunization with the MAEBL M2 Domain Protects against Lethal Plasmodium yoelii Infection

Leite, Juliana Almeida; Bargieri, Daniel Youssef; Carvalho, Bruna O.; Albrecht, Letusa; Lopes, Stefanie Costa Pinto; Kayano, Ana Carolina Andrade Vitor; Farias, Alessandro S.; Chia, Wan Ni; Claser, Carla; Malleret, Benoît; Russell, Bruce; Castiñeiras, Catarina; Santos, Leonilda Maria Barbosa dos; Brocchi, Marcelo; Wunderlich, Gerd; Soares, Irene da Silva; Rodrigues, Maurício M.; Rénia, Laurent; Costa, Fábio Trindade Maranhão

doi:10.1128/iai.00262-15

Cited by 13 publications

(18 citation statements)

References 46 publications

(50 reference statements)

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“…To investigate the functional implications of the AMA4-RON2 L1 complex in T. gondii, we first established that TgAMA4 is part of a highly divergent AMA clade that includes the functionally important malaria vaccine candidate Plasmodium merozoite apical erythrocyte-binding ligand (MAEBL) (18)(19)(20) and that TgRON2 L1 displays a similar divergence consistent with coevolution of receptor and ligand. We then show that TgAMA4 and TgRON2 L1 form a high-affinity binary complex and probe its overall architecture and underlying mechanism of assembly using crystal structures of TgAMA4 in the apo and TgRON2 L1 D3 bound forms.…”

Section: Significancementioning

confidence: 99%

Dissecting the interface between apicomplexan parasite and host cell: Insights from a divergent AMA–RON2 pair

Parker

Penarete-Vargas

Hamilton

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Plasmodium falciparum and Toxoplasma gondii are widely studied parasites in phylum Apicomplexa and the etiological agents of severe human malaria and toxoplasmosis, respectively. These intracellular pathogens have evolved a sophisticated invasion strategy that relies on delivery of proteins into the host cell, where parasite-derived rhoptry neck protein 2 (RON2) family members localize to the host outer membrane and serve as ligands for apical membrane antigen (AMA) family surface proteins displayed on the parasite. Recently, we showed that T. gondii harbors a novel AMA designated as TgAMA4 that shows extreme sequence divergence from all characterized AMA family members. Here we show that sporozoite-expressed TgAMA4 clusters in a distinct phylogenetic clade with Plasmodium merozoite apical erythrocyte-binding ligand (MAEBL) proteins and forms a high-affinity, functional complex with its coevolved partner, TgRON2 L1 . High-resolution crystal structures of TgAMA4 in the apo and TgRON2 L1 -bound forms complemented with alanine scanning mutagenesis data reveal an unexpected architecture and assembly mechanism relative to previously characterized AMA-RON2 complexes. Principally, TgAMA4 lacks both a deep surface groove and a key surface loop that have been established to govern RON2 ligand binding selectivity in other AMAs. Our study reveals a previously underappreciated level of molecular diversity at the parasite-host-cell interface and offers intriguing insight into the adaptation strategies underlying sporozoite invasion. Moreover, our data offer the potential for improved design of neutralizing therapeutics targeting a broad range of AMA-RON2 pairs and apicomplexan invasive stages.Apicomplexa | Toxoplasma gondii | invasion | moving junction | X-ray crystallography P hylum Apicomplexa comprises >5,000 parasitic protozoan species, many of which cause devastating diseases on a global scale. Two of the most prevalent species are Toxoplasma gondii and Plasmodium falciparum, the causative agents of toxoplasmosis and severe human malaria, respectively (1, 2). The obligate intracellular apicomplexan parasites lead complex and diverse lifestyles that require invasion of many different cell types. Despite this diversity of target host cells, most apicomplexans maintain a generally conserved mechanism for active invasion (3). The parasite initially glides over the surface of a host cell and then reorients to place its apical end in close contact to the hostcell membrane. After this initial attachment, a circumferential ring of adhesion (termed the moving or tight junction) is formed, through which the parasite actively propels itself while concurrently depressing the host-cell membrane to create a nascent protective vacuole (4).Formation of the moving junction relies on a pair of highly conserved parasite proteins: rhoptry neck protein 2 (RON2) and apical membrane antigen 1 (AMA1). Initially, parasites discharge RON2 into the host cell membrane where an extracellular portion (domain 3; D3) serves as a ligand for AMA1 displa...

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Section: Significancementioning

confidence: 99%

Dissecting the interface between apicomplexan parasite and host cell: Insights from a divergent AMA–RON2 pair

Parker

Penarete-Vargas

Hamilton

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…It was demonstrated in P. berghei that the disruption or deletion of some regions of the CS protein affects the formation and maturation of sporozoites, escape from the oocyst, and subsequent progression of the Plasmodium life cycle [ 38 , 39 ]. Likewise, several other gene deletions have been described that affect P. berghei oocyst formation and consequent sporozoite escape: an oocyst-specific papain-like cysteine protease, known as the egress cysteine protease (ECP1), oocyst capsule protein (PbCAP380), fertilization gene (Pb GEX), lectin adhesive proteins (PbLAPs), protein kinases (PbCDLK), and nuclear forming-like protein (PbMISFIT) [ 40 – 48 ]. The results showed that P. berghei sporozoites are liberated from the oocyst by decortication and subsequent dissolution of the oocyst wall, which is consistent with a mechanism involving a proteolytic activity as has been proposed for P. berghei [ 42 ].…”

Section: Discussionmentioning

confidence: 99%

Species-specific escape of Plasmodium sporozoites from oocysts of avian, rodent, and human malarial parasites

Orfanó

Nacif-Pimenta

Duarte

et al. 2016

Malar J

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BackgroundMalaria is transmitted when an infected mosquito delivers Plasmodium sporozoites into a vertebrate host. There are many species of Plasmodium and, in general, the infection is host-specific. For example, Plasmodium gallinaceum is an avian parasite, while Plasmodium berghei infects mice. These two parasites have been extensively used as experimental models of malaria transmission. Plasmodium falciparum and Plasmodium vivax are the most important agents of human malaria, a life-threatening disease of global importance. To complete their life cycle, Plasmodium parasites must traverse the mosquito midgut and form an oocyst that will divide continuously. Mature oocysts release thousands of sporozoites into the mosquito haemolymph that must reach the salivary gland to infect a new vertebrate host. The current understanding of the biology of oocyst formation and sporozoite release is mostly based on experimental infections with P.berghei, and the conclusions are generalized to other Plasmodium species that infect humans without further morphological analyses.ResultsHere, it is described the microanatomy of sporozoite escape from oocysts of four Plasmodium species: the two laboratory models, P. gallinaceum and P. berghei, and the two main species that cause malaria in humans, P.vivax and P. falciparum. It was found that sporozoites have species-specific mechanisms of escape from the oocyst. The two model species of Plasmodium had a common mechanism, in which the oocyst wall breaks down before sporozoites emerge. In contrast, P. vivax and P. falciparum sporozoites show a dynamic escape mechanism from the oocyst via polarized propulsion.ConclusionsThis study demonstrated that Plasmodium species do not share a common mechanism of sporozoite escape, as previously thought, but show complex and species-specific mechanisms. In addition, the knowledge of this phenomenon in human Plasmodium can facilitate transmission-blocking studies and not those ones only based on the murine and avian models.

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“…C57BL/6 mice with 5–7 weeks-old were injected subcutaneously four times at 3 weeks intervals with 5 μg of rPyM2-MAEBL emulsified in 1:1 (vol/vol) complete Freund’s adjuvant (CFA) for the first dose or incomplete Freund’s adjuvant (IFA) in the subsequent doses [ 4 ].…”

Section: Methodsmentioning

confidence: 99%

“…Well diameters were established with the aid of a Dako-Pen (Dako), and blocking was performed by 30-min incubation at 37 °C with PBS containing 3% BSA (USB). C57BL/6 mice with 5–7 weeks-old were immunized as described elsewhere [ 4 ] and pooled sera from the different immunization groups were diluted 1:50 in PBS supplemented with 3% BSA and applied to the slides for 1 h at 37 °C. Slides were washed 3X in PBS and incubated with Alexa-568 goat anti-mouse IgG (Invitrogen) for 1 h at 37 °C in the dark, then washed 3X in PBS and incubated with DAPI (4′,6-diamidino-2-phenylindole, dihydrochloride-(Invitrogen) diluted in ultrapure (Millipore) water for 10 min at room temperature.…”

Section: Methodsmentioning

confidence: 99%

“…Recently, the immunogenicity of M2 MAEBL (Merozoite Adhesive Erythrocytic Binding Protein) domain of Plasmodium yoelii has been demonstrated. This vaccine candidate conferred 90% protection in immunized mice after lethal challenge and corresponding antisera inhibited significantly erythrocyte invasion by P. yoelii [ 4 ]. MAEBL is a membrane protein that belongs to the erythrocyte binding protein ( ebl ) family [ 5 – 8 ] and exhibits homologous regions to the DBL-EBP (Duffy binding like–Erythrocyte binding protein) and to the apical membrane antigen 1 (AMA1) [ 7 – 9 ].…”

Section: Introductionmentioning

confidence: 99%

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In silico epitope mapping and experimental evaluation of the Merozoite Adhesive Erythrocytic Binding Protein (MAEBL) as a malaria vaccine candidate

Cravo

Machado

Leite

et al. 2018

Malar J

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BackgroundTechnical limitations for culturing the human malaria parasite Plasmodium vivax have impaired the discovery of vaccine candidates, challenging the malaria eradication agenda. The immunogenicity of the M2 domain of the Merozoite Adhesive Erythrocytic Binding Protein (MAEBL) antigen cloned from the Plasmodium yoelii murine parasite, has been previously demonstrated.ResultsDetailed epitope mapping of MAEBL through immunoinformatics identified several MHCI, MHCII and B cell epitopes throughout the peptide, with several of these lying in the M2 domain and being conserved between P. vivax, P. yoelii and Plasmodium falciparum, hinting that the M2-MAEBL is pan-reactive. This hypothesis was tested through functional assays, showing that P. yoelii M2-MAEBL antisera are able to recognize and inhibit erythrocyte invasion from both P. falciparum and P. vivax parasites isolated from Thai patients, in ex vivo assays. Moreover, the sequence of the M2-MAEBL is shown to be highly conserved between P. vivax isolates from the Amazon and Thailand, indicating that the MAEBL antigen may constitute a vaccine candidate outwitting strain-specific immunity.ConclusionsThe MAEBL antigen is promising candidate towards the development of a malaria vaccine.Electronic supplementary materialThe online version of this article (10.1186/s12936-017-2144-x) contains supplementary material, which is available to authorized users.

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Immunization with the MAEBL M2 Domain Protects against Lethal Plasmodium yoelii Infection

Cited by 13 publications

References 46 publications

Dissecting the interface between apicomplexan parasite and host cell: Insights from a divergent AMA–RON2 pair

Dissecting the interface between apicomplexan parasite and host cell: Insights from a divergent AMA–RON2 pair

Species-specific escape of Plasmodium sporozoites from oocysts of avian, rodent, and human malarial parasites

In silico epitope mapping and experimental evaluation of the Merozoite Adhesive Erythrocytic Binding Protein (MAEBL) as a malaria vaccine candidate

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