Dataset Information

The AMA1-RON complex drives Plasmodium sporozoite invasion in the mosquito and mammalian hosts

ABSTRACT: Sporozoites and merozoites share a number of proteins that are expressed by both stages, including the Apical Membrane Antigen 1 (AMA1) and the Rhoptry Neck Proteins (RONs). Although AMA1 and RONs are essential for merozoite invasion of erythrocytes during asexual blood stage replication of the malaria parasite, their precise function in sporozoites is unclear. Here we performed RFP-trap immunoprecipitation experiments using lysates from transgenic sporozoites expressing RON4 fused to mCherry. RON4, RON2, RON5 and AMA1 were the main proteins identified by mass spectrometry among co-precipitated proteins, showing that AMA1-RON interactions are conserved in salivary gland sporozoites.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Plasmodium Berghei Anka

DISEASE(S): Malaria

SUBMITTER: Soumia HAMADA

LAB HEAD: Solenne Chardonnet

PROVIDER: PXD031463 | Pride | 2022-08-12

REPOSITORIES: Pride

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Publications

The AMA1-RON complex drives Plasmodium sporozoite invasion in the mosquito and mammalian hosts.

Fernandes Priyanka P Loubens Manon M Le Borgne Rémi R Marinach Carine C Ardin Béatrice B Briquet Sylvie S Vincensini Laetitia L Hamada Soumia S Hoareau-Coudert Bénédicte B Verbavatz Jean-Marc JM Weiner Allon A Silvie Olivier O

PLoS pathogens 20220622 6

Plasmodium sporozoites that are transmitted by blood-feeding female Anopheles mosquitoes invade hepatocytes for an initial round of intracellular replication, leading to the release of merozoites that invade and multiply within red blood cells. Sporozoites and merozoites share a number of proteins that are expressed by both stages, including the Apical Membrane Antigen 1 (AMA1) and the Rhoptry Neck Proteins (RONs). Although AMA1 and RONs are essential for merozoite invasion of erythrocytes durin ...[more]

PMID: 35731833

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Project description:Tuberculosis remains a major cause of death from an infectious disease worldwide, yet only 10% of people infected with Mycobacterium tuberculosis develop disease. Defining both necessary and sufficient immunologic determinants of protection remains a great scientific challenge. Analysis of peripheral blood gene expression profiles of active tuberculosis patients has identified correlates of risk for disease or pathogenesis. We sought to identify human potential candidate markers of host defense by studying gene expression profiles of macrophages, cells which, upon infection by M. tuberculosis, can mount an antimicrobial response. Weighted gene co-expression network analysis revealed an association between the cytokine, IL-32, and the vitamin D antimicrobial pathway in a network of IFN-γ and IL-15 induced ‘defense response’ genes. IL-32 was sufficient for induction of the vitamin D-dependent antimicrobial peptides, cathelicidin and DEFB4, and generation of antimicrobial activity in vitro, dependent on the presence of adequate 25-hydroxyvitamin D. The IL-15 induced ‘defense response’ macrophage gene network was integrated with ranked pairwise comparisons of gene expression from five different clinical data sets of latent vs. active tuberculosis or healthy controls, and a co-expression network derived from gene expression in patients with tuberculosis undergoing chemotherapy. Together, these analyses identified eight common genes, including IL-32, as molecular markers of latent tuberculosis and the IL-15 induced gene network. Inferring that maintaining M. tuberculosis in a latent state and preventing transition to active disease represents host resistance, we believe these results identify IL-32 as one functional marker and potential correlate of protection against active tuberculosis. Adherent peripheral blood monuclear cells were derived by Ficoll-Hypaque from the whole blood of four healthy donors. Cells adhered to tissue culture-treated plates for 2 h in 1% Fetal Bovine Serum (FBS) in RPMI. Cells were washed then stimulated with IL-10 (10ng/ml), IL-15 (10ng/ml) (R&D Systems), or IL-4 (1U/ml) in RPMI 1640 supplemented with 10% FBS at 37°C, 5% CO2. Cells were harvested at 6 h and 24 h after stimulation and monocytes purified by CD14 microbeads (Miltenyi Biotec) for a confirmed monocyte purity of at least 90%. RNA from purified monocytes extracted by Trizol and purified by Qiagen RNeasy Kit. RNA probe and microarray performed by UCLA Clinical Microarray Core using Ambion labeling kit and Affymetrix Human U133 Plus 2.0 array.

Dataset Information

The AMA1-RON complex drives Plasmodium sporozoite invasion in the mosquito and mammalian hosts

Publications

The AMA1-RON complex drives Plasmodium sporozoite invasion in the mosquito and mammalian hosts.

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