Plasmodium berghei sporozoite specific genes- PbS10 and PbS23/SSP3 are required for the development of exo-erythrocytic forms

Togiri, Jyothi; Segireddy, Rameswara R; Mastan, Babu S.; Singh, Dipti; Kolli, Surendra Kumar; Ghosh, Ankit; Al-Nihmi, Faisal Mohammed Abdul; Maruthi, Mulaka; Choudhary, Hadi Hasan; Dey, Sandeep Kumar; Mishra, Satish; Kumar, Kota Arun

doi:10.1016/j.molbiopara.2019.111198

Cited by 7 publications

(15 citation statements)

References 43 publications

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“…Of the markers in cluster 2, PBANKA_1002500 (Sporozoite-specific protein S10), was the most prominent. Previously shown to be upregulated in oocysts 38 , we detected the transcript in 77% (836/1,086) of sporozoites in cluster 2, compared to only 13% of sporozoites in cluster 1, in addition to being differentially expressed (average logFC 3.49; adjusted P value = 1.34 × 10 -215 ) ( Figs Integration of mixed sporozoite scRNA-seq data with Malaria Cell Atlas allows for fine-tuning of clusters. Cell annotation in single-cell datasets remains a challenging task, especially for species with limited or incomplete gene models such as Plasmodium species.…”

Section: Strategy To Capture Sporozoites and Measure Gene Expression mentioning

confidence: 76%

Single-cell RNA sequencing reveals developmental heterogeneity among Plasmodium berghei sporozoites

Ruberto

Bourke

Mérienne

et al. 2021

Sci Rep

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In the malaria-causing parasite’s life cycle, Plasmodium sporozoites must travel from the midgut of a mosquito to the salivary glands before they can infect a mammalian host. However, only a fraction of sporozoites complete the journey. Since salivary gland invasion is required for transmission of sporozoites, insights at the molecular level can contribute to strategies for malaria prevention. Recent advances in single-cell RNA sequencing provide an opportunity to assess sporozoite heterogeneity at a resolution unattainable by bulk RNA sequencing methods. In this study, we use a droplet-based single-cell RNA sequencing workflow to analyze the transcriptomes of over 8000 Plasmodium berghei sporozoites derived from the midguts and salivary glands of Anopheles stephensi mosquitoes. The detection of known marker genes confirms the successful capture and sequencing of samples composed of a mixed population of sporozoites. Using data integration, clustering, and trajectory analyses, we reveal differences in gene expression profiles of individual sporozoites, and identify both annotated and unannotated markers associated with sporozoite development. Our work highlights the utility of a high-throughput workflow for the transcriptomic profiling of Plasmodium sporozoites, and provides new insights into gene usage during the parasite’s development in the mosquito.

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Section: Strategy To Capture Sporozoites and Measure Gene Expression mentioning

confidence: 76%

Single-cell RNA sequencing reveals developmental heterogeneity among Plasmodium berghei sporozoites

Ruberto

Bourke

Mérienne

et al. 2021

Sci Rep

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“…To analyze the morphology of mutant sporozoites and liver stages, IFA was performed as described earlier (Jillapalli et al, 2019; Togiri et al, 2019). The zygotes and ookinetes were stained with anti‐p28 antibody (mouse) and immunoreactivity was revealed using goat anti‐mouse Alexa 594.…”

Section: Methodsmentioning

confidence: 99%

A conserved Plasmodium structural integrity maintenance protein (SIMP) is associated with sporozoite membrane and is essential for maintaining shape and infectivity

Singh

Patri

Narahari

et al. 2022

Molecular Microbiology

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Plasmodium sporozoites are extracellular forms introduced during mosquito bite that selectively invade mammalian hepatocytes. Sporozoites are delimited by a cell membrane that is linked to the underlying acto‐myosin molecular motor. While membrane proteins with roles in motility and invasion have been well studied, very little is known about proteins that maintain the sporozoite shape. We demonstrate that in Plasmodium berghei (Pb) a conserved hypothetical gene, PBANKA_1422900 specifies sporozoite structural integrity maintenance protein (SIMP) required for maintaining the sporozoite shape and motility. Sporozoites lacking SIMP exhibited loss of regular shape, extensive membrane blebbing at multiple foci, and membrane detachment. The mutant sporozoites failed to infect hepatocytes, though the altered shape did not affect the organization of cytoskeleton or inner membrane complex (IMC). Interestingly, the components of IMC failed to extend under the membrane blebs likely suggesting that SIMP may assist in anchoring the membrane to IMC. Endogenous C‐terminal HA tagging localized SIMP to membrane and revealed the C‐terminus of the protein to be extracellular. Since SIMP is highly conserved among Plasmodium species, these findings have important implications for utilizing it as a novel sporozoite‐specific vaccine candidate.

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“…For six proteins, SPELD, GEST, SSP3, SPATR, HT, MAEBL, SIAP1 and SIAP2, evidence has been presented for a sporozoite surface location [27][28][29][30][31][32][33][34][35][36][37]. Absence of SPELD, a sporozoiteand liver-specific protein, leads to abrogation of liver stage maturation [27].…”

Section: Selection Of Fourteen P Falciparum Pre-erythrocytic Vaccine Candidate Antigensmentioning

confidence: 99%

“…GEST plays a role in egress of gametes from the infected RBC and in sporozoite traversal [34,35]. SSP3, a sporozoite-and liver-stage specific protein, plays a role in sporozoite gliding motility and liver stage development [30,36]. SPATR is expressed both in blood stages and sporozoites.…”

Section: Selection Of Fourteen P Falciparum Pre-erythrocytic Vaccine Candidate Antigensmentioning

confidence: 99%

“…Early transcribed membrane protein 10.3 PF3D7_1016900 PBANKA_0501200 No Yes PV [45] yes GEST Gamete egress and sporozoite traversal protein PF3D7_1449000 PBANKA_1312700 Yes No Spz surface [34,35] no SSP3 Sporozoite surface protein 3 PF3D7_0812300 PBANKA_1425200 Yes Yes Spz surface [30,34,36] no SIAP1 Sporozoite invasion-associated protein 1 PF3D7_0408600 PBANKA_1006200 Yes No Spz surface [29,34] no MAEBL Membrane associated erythrocyte binding-like protein PF3D7_1147800 PBANKA_0901300 Yes Yes Spz surface [32,37] no P36 6-cysteine protein P36 PF3D7_0404400 PBANKA_1002100 No Yes Micronemes, PV [43] no P52 6-cysteine protein P52 PF3D7_0404500 PBANKA_1002200 No Yes Micronemes, PV no…”

Section: Selection Of Fourteen P Falciparum Pre-erythrocytic Vaccine Candidate Antigensmentioning

confidence: 99%

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Screening of viral-vectored P. falciparum pre-erythrocytic candidate vaccine antigens using chimeric rodent parasites

et al. 2021

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To screen for additional vaccine candidate antigens of Plasmodium pre-erythrocytic stages, fourteen P. falciparum proteins were selected based on expression in sporozoites or their role in establishment of hepatocyte infection. For preclinical evaluation of immunogenicity of these proteins in mice, chimeric P. berghei sporozoites were created that express the P. falciparum proteins in sporozoites as an additional copy gene under control of the uis4 gene promoter. All fourteen chimeric parasites produced sporozoites but sporozoites of eight lines failed to establish a liver infection, indicating a negative impact of these P. falciparum proteins on sporozoite infectivity. Immunogenicity of the other six proteins (SPELD, ETRAMP10.3, SIAP2, SPATR, HT, RPL3) was analyzed by immunization of inbred BALB/c and outbred CD-1 mice with viral-vectored (ChAd63 or ChAdOx1, MVA) vaccines, followed by challenge with chimeric sporozoites. Protective immunogenicity was determined by analyzing parasite liver load and prepatent period of blood stage infection after challenge. Of the six proteins only SPELD immunized mice showed partial protection. We discuss both the low protective immunogenicity of these proteins in the chimeric rodent malaria challenge model and the negative effect on P. berghei sporozoite infectivity of several P. falciparum proteins expressed in the chimeric sporozoites.

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Plasmodium berghei sporozoite specific genes- PbS10 and PbS23/SSP3 are required for the development of exo-erythrocytic forms

Cited by 7 publications

References 43 publications

Single-cell RNA sequencing reveals developmental heterogeneity among Plasmodium berghei sporozoites

Single-cell RNA sequencing reveals developmental heterogeneity among Plasmodium berghei sporozoites

A conserved Plasmodium structural integrity maintenance protein (SIMP) is associated with sporozoite membrane and is essential for maintaining shape and infectivity

Screening of viral-vectored P. falciparum pre-erythrocytic candidate vaccine antigens using chimeric rodent parasites

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