Prideapplication/xmlftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/checksum.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/Pv_ID.csvftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/FL1029_IPP114_PB_CB_vivax_2_odd.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/FL1029_IPP114_PB_CB_vivax_8_odd.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/FL1029_IPP114_PB_CB_vivax_1_even.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/FL1029_IPP114_PB_CB_vivax_7_odd.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/FL1029_IPP114_PB_CB_vivax_3_odd.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/FL1029_IPP114_PB_CB_vivax_4_even.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/FL1029_IPP114_PB_CB_vivax_2_even.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/FL1029_IPP114_PB_CB_vivax_8_even.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/FL1029_IPP114_PB_CB_vivax_1_odd.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/FL1029_IPP114_PB_CB_vivax_7_even.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/FL1029_IPP114_PB_CB_vivax_6_even.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/FL1029_IPP114_PB_CB_vivax_6_odd.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/FL1029_IPP114_PB_CB_vivax_4_odd.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD077321/FL1029_IPP114_PB_CB_vivax_3_even.rawprimaryOK200nicola.ternette@ndm.ox.ac.ukNicola TernetteImmunopeptidomicsData-dependent acquisitionMass SpectrometryPlasmodium Vivax MalariaNot availableImmunopeptidomicsPlasmodium vivaxHlahttps://www.ebi.ac.uk/pride/archive/projects/PXD077321ReticulocyteBlood100mL of blood was collected from infected individuals and controls. First, mononuclear cells were separated from peripheral blood (PBMC). For this, the blood was diluted in a 1: 1 ratio, was gently added to a tube containing 15mL of Ficoll (GE Healthcare) and the protocol previously described was followed58. The red blood cell pellet resulting from PBMC purification was resuspended in RPMI culture medium in a 1:4 ratio. Diluted blood was added carefully into a 50mL tube containing Percoll 45% (GE Healthcare, USA) at 5x the volume of the red blood cell pellet. Samples were centrifuged for 15 minutes at 2000 rpm. After centrifugation, the reticulocyte interface was collected. Reticulocyte purified samples had 99% purity, identified by Giemsa-stained thin blood smear. Pv-infected reticulocytes were lysed in 500µl lysis buffer (1% Igepal, 300mM NaCl, 100mM Tris, pH 8.0) supplemented with cOmplete™ Inhibitor Cocktail (Roche). HLA complexes were immunoprecipitated using 1mg monoclonal antibody W6/32 against pan-HLA class-I complexes (Biolegend) cross-linked to Protein A Sepharose beads using dimethylpimelimidate (DMP, Sigma). Cell lysates were incubated overnight with W6/32 beads. Beads were subsequently washed with 10 column volumes of 2 × 150mM NaCl in 50mM Tris, 1 × 450mM NaCl in 50mM Tris and 50mM Tris buffer without salt. Peptides bound to the HLA groove were released with 5mL 10% acetic acid. To separate peptides from the α-chain and β2-microglobulin, samples were further purified by HPLC (Ultimate 3000, Thermo Scientific) on a ProSwift RP-1S 4.6 × 50mm column (Thermo Scientific) by applying a linear gradient of 2–35% (v/v) acetonitrile in 0.1% (v/v) formic acid in water over 10 min. Samples were dried in speed vacuum and suspended in 20μl buffer A (1% acetonitrile, 0.1% TFA in water). Eluted peptide fractions were analyzed by nUPLC-MS/MS. An Ultimate 3000 RSLCnano System coupled with an Orbitrap Fusion Lumos Tribrid mass spectrometer (Thermo Scientific) were used to perform mass spectrometry.PrideNot availableSequence interpretation of MS/MS spectra were performed using a database containing all annotated human Uniprot entries (Uniprot- UP000005640 -09-2019) and P. vivax Uniprot entries (Uniprot- UP000008333- Version 08-2019) combined using PEAKS 7 (Bioinformatics Solutions Inc).ProteomicsNicola TernetteUniversity of Oxford University of DundeePARTIALHomo Sapiens (human)nicola.ternette@ndm.ox.ac.uk42386975 Barbosa CRR, de Lacerda LB, Bettencourt PJG, Morrow D, Pereira DB, Aleshnick M, Mitchell JL, Poulton NC, Gomes C, Cordeiro LPB, Doumbia K, Ntalla C, Arama C, Zhao Z, Maia GC, Almeida GG, Schrimpf MR, Hart TF, Haumpy D, Medeiros-Rodrigues BC, Costa CM, Nicastri A, Gilbride RM, Schell JB, Kirtley P, Antonelli LRV, Gaiha GD, Hansen SG, Lieberman J, Gazzinelli RT, Niangaly M, Woodford J, Goldberg J, Früh K, Portugal S, Duffy PE, Ternette N, Wilder B, Hill AVS, Junqueira C. Identification of cross-stage, cross-species malaria CD8<sup>+</sup> T cell antigens. Nature. 2026 655(8123):747-757 10.1038/s41586-026-10730-1Human proteome projectBiology/disease-driven human proteome project (b/d-hpp)Human immuno-peptidome project (hupo-hipp) (b/d-hpp)University of OxfordUnited KingdomA major limitation on the development of a malaria vaccine is the lack of validated T cell epitope targets. Plasmodium falciparum is the most prevalent malaria parasite affecting humans in Africa, whereas Plasmodium vivax is more widespread and is the main species that causes malaria in the Americas and Asia¹. P. vivax exclusively infects peripheral-blood reticulocytes, which retain RNA and the capacity for host protein synthesis². We previously reported that reticulocytes infected with P. vivax express human leukocyte antigen class I (HLA-I), which enables recognition and killing of the parasite by CD8⁺ T cells³. Here we use immunopeptidomics to identify Plasmodium-antigen-derived peptides presented by HLA-I on infected reticulocytes. We identified 453 unique peptides, mapping to 166 proteins. Seventy-five antigens were housekeeping proteins that are constitutively expressed at multiple stages of the parasite's life cycle and are highly conserved between Plasmodium species. Identical peptides were presented in different individuals by the same or distinct HLA-A, HLA-B and HLA-C alleles, as well as by the non-classical HLA-E allele. The antigenicity of the newly identified epitopes was validated in samples from both P. vivax-infected and P. falciparum-infected individuals. Furthermore, T cell responses to several of these antigens were observed in the blood and liver of non-human primates after infection with Plasmodium or immunization with attenuated parasites. Two antigens also induced protective CD8⁺ T cell-mediated immunity in rodents. Thus, these antigens have the potential for use in a cross-stage and cross-species malaria vaccine.Identification of cross-stage, cross-species malaria CD8<sup>+</sup> T cell antigens.Barbosa Camila R R CRR, de Lacerda Luna B LB, Bettencourt Paulo J G PJG, Morrow David D, Pereira Dhelio B DB, Aleshnick Maya M, Mitchell Julie L JL, Poulton Nicholas C NC, Gomes Cristopher C, Cordeiro Lídia P B LPB, Doumbia Kadia K, Ntalla Christina C, Arama Charles C, Zhao Zezhou Z, Maia Guilherme C GC, Almeida Gregório G GG, Schrimpf Marie Rose MR, Hart Thalia F TF, Haumpy Derek D, Medeiros-Rodrigues Beatriz C BC, Costa Camila M CM, Nicastri Annalisa A, Gilbride Roxanne M RM, Schell John B JB, Kirtley Payton P, Antonelli Lis R V LRV, Gaiha Gaurav D GD, Hansen Scott G SG, Lieberman Judy J, Gazzinelli Ricardo T RT, Niangaly Moussa M, Woodford John J, Goldberg Joel J, Früh Klaus K, Portugal Silvia S, Duffy Patrick E PE, Ternette Nicola N, Wilder Brandon B, Hill Adrian V S AVS, Junqueira Caroline CfalseImmmunopeptidomics data from reticulocytes of Plasmodium vivax infected human donorsPeripheral blood samples from 7 acute malaria patients with random HLA typing were analysed by immunopeptidomics. The donors were recruited in Porto Velho, Rondônia, a malaria endemic region in Brazil, were confirmed to be infected with Pv, and not Pf, with parasitemia ranging from 2-3 crosses (501-100,000 parasites/mm3). Highly purified Retics contained 4.3x107 to 7x108 cells, accounting for 98.11.2% of iRetics, 1.31.1% of uninfected RBCs, and 0.010.008% of nucleated cell 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T:2186NEWT:8724NEWT:8723NEWT:283166NEWT:37334NCBITaxon:10357NCBITaxon:10114NCBITaxon:10359NEWT:1203258NEWT:242619NEWT:49317NEWT:160791NEWT:34073NEWT:696811NEWT:373995NEWT:5689NEWT:8715NEWT:544404NEWT:92519NEWT:37349NEWT:4113NEWT:837NEWT:36015NEWT:81866NEWT:5691NEWT:48479NEWT:61435NEWT:627025NEWT:1097677NEWT:375939NEWT:61674NEWT:1117957NEWT:6526NEWT:8709NEWT:4100NEWT:1159556NEWT:5671NEWT:1076NEWT:4102NEWT:4101NEWT:6763NEWT:8700NEWT:803NEWT:29722NEWT:1692259NEWT:1486917NEWT:419947NEWT:468911NEWT:817NEWT:7604NEWT:818NEWT:135588NEWT:1843183NEWT:5661NEWT:51866NEWT:29719NEWT:58002NEWT:5665NEWT:6754NEWT:6997NEWT:452589NEWT:509068NEWT:5666NEWT:4577NEWT:813NEWT:5664NEWT:2157NEWT:146479NEWT:1042986NEWT:1911079NEWT:69373NEWT:1843192NEWT:206189NEWT:264203NEWT:1480154NEWT:1274414NEWT:9612NEWT:38626NEWT:38865NEWT:3077NEWT:9615NEWT:10299NEWT:6100NEWT:713604NEWT:112273NEWT:1147787NEWT:1440772NEWT:1355477NEWT:8759NEWT:9605NEWT:9606NEWT:90371NEWT:641501NEWT:4153NEWT:9601NEWT:278839NEWT:35128NEWT:7668NEWT:8756NEWT:915099NEWT:74940NEWT:13551NEWT:10041NEWT:9600NEWT:1450511NEWT:1143193NEWT:1122233NEWT:890382NEWT:174934NEWT:179392NEWT:2823525NEWT:9838NEWT:9839NEWT:546414NEWT:227086NEWT:3055NEWT:11167NEWT:9835NEWT:4146NEWT:5478NEWT:6326NEWT:5476NEWT:1450520NEWT:1042311NEWT:1882945NEWT:299767NEWT:235909NEWT:28835NEWT:47311NEWT:1274432NEWT:2053667NEWT:1274426NEWT:9825NEWT:1274423NEWT:393765NEWT:39946NEWT:9823NEWT:95602NEWT:8732NEWT:314539NEWT:39947NEWT:718274NEWT:1274420NEWT:578458NEWT:60520423869750000-0002-9283-0743