{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Txt":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/checksum.txt"],"Xml":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137228.xml","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137224.xml","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137237.xml","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137242.xml","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137233.xml","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137238.xml","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137230.xml","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137235.xml","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137240.xml","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137236.xml"],"Raw":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/00212169saj_270CAstV.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/00212171saj_277K.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/00212166saj_264CAstV.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/00212173saj_279K.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/00212167saj_265CAstV.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/00212170saj_276K.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/00212165saj_263CAstV.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/00212172saj_278K.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/00212168saj_268CAstV.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/00212174saj_280K.raw"],"Mgf":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137233.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137238.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137228.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137242.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137224.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137237.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137240.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137236.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137230.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD074601/F137235.mgf"]},"type":"primary"},"statusCodeValue":200,"statusCode":"OK"}],"scores":null,"additional":{"labhead_mail":["joanna.sajewicz@piwet.pulawy.pl"],"submitter":["Agata Malinowska"],"technology_type":["Data-dependent acquisition","Mass Spectrometry"],"software":[""],"submitter_keywords":["Chicken astrovirus","Spleen","Label-free proteomics"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD074601"],"tissue":["Spleen"],"sample_protocol":["Approximately 50–100 mg of spleen tissue per sample was homogenized in lysis buffer (1% SDS, 30 mM Tris-HCl pH 8.0) using an MP FastPrep homogenizer. Lysates were cleared by centrifugation (14,000 × g, 20 min, 4 °C), and protein concentrations were determined via BCA assay (Thermo Fisher Scientific). Mass spectrometry experiments were carried out at the Mass Spectrometry Laboratory, Institute of Biochemistry and Biophysics PAS. Protein concentrations were determined using the Pierce BCA Protein Assay Kit (Thermo Scientific). For each sample, 100 ug of protein was diluted to a final volume of 55 ul in urea buffer (8 M urea in 100 mM ammonium bicarbonate). Protein digestion was performed according to the FASP protocol with minor modifications (Wiśniewski et al. 2009). Reduction of disulfide bonds was achieved by incubating samples for 1 hour with 50 mM tris(2-carboxyethyl)phosphine (TCEP) at 37 oC, followed by transfer onto Vivacon 30 kDa molecular weight cut-off filters (Sartorius). Samples were centrifuged at 14 000 g for 30 min and washed twice with 200 µl of urea buffer. Cysteine residues were blocked by a 15 min incubation at room temperature with 100 mM s-methylmethanethiosulfonate. Filters were subsequently washed three times with urea buffer and three times with 100 mM ammonium bicarbonate to remove residual denaturant. After each buffer addition, centrifugation was performed for 15 to 40 min until the filter was dry. Proteolytic digestion was carried out overnight using 4 ug of trypsin/LysC mix (Promega) at 37 oC with vortexing. Peptides were eluted from the FASP filters by consecutive centrifugation steps using 100 mM ammonium bicarbonate and 0.5 M sodium chloride. Peptide concentrations were measured with the Pierce Quantitative Colorimetric Peptide Assay (Thermo Scientific).  2 ug of peptides were analysed using an LC-MS system consisting of a UPLC chromatograph (nanoAcquity, Waters) directly coupled to a Q Exactive mass spectrometer (Thermo). Samples were loaded onto an RP-18 pre-column (Waters, Milford, MA) using water containing 0.1% FA as the mobile phase and subsequently separated on a nano-HPLC RP-18 analytical column (75 µm x 250 mm, 1.7 µm, Waters) with an ACN gradient (0–35% ACN over 160 min) in the presence of 0.1% FA at a flow rate of 250 nl/min. Data acquisition was performed in a data-dependent mode with the top 12 precursors selected for MS2 analysis following CID fragmentation with an NCE of 27. Full MS scans were acquired over the mass range of 300–1650 m/z at a resolution of 70,000, with a maximum injection time of 60 ms and an AGC target value of 1e6. MS2 scans were acquired with a maximum injection time of 60 ms, an AGC target value of 5e5, and an isolation window of 3.0 m/z. Dynamic exclusion was set to 30 s. Blank injections and QC samples were used to confirm instrument stability and minimal carryover."],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["Raw MS data were searched in Mascot Server v2.8 against the Gallus gallus UniProt reference proteome (release 2019_10, 27,542 sequences; 17,434,812 residues). Trypsin specificity was enforced with one missed cleavage permitted. Carbamidomethylation of cysteine was set as a fixed modification, while methionine oxidationwas included as variable modification. Data were re-calibrated offline with resulting peptide mass tolerance of 5 ppm and fragment mass tolerance of 0.01. False discovery rate was estimated with target/decoy strategy. Protein identification required a false discovery rate (FDR)≤1% at peptide  levels and the presence of at least two unique peptides. Using these criteria, 2,529 proteins were identified across alle spleen samples.  The mass calibration and data filtering described above were carried out with MScan software, developed in-house (http://proteom.ibb.waw.pl/mscan/). For quantitative analysis, proteins detected in at least four of the five biological replicates per group were retained, resulting in a final set of 1,118 quantified proteins. The list of identified peptides was overlayed onto 2-D heatmaps generated from LC-MS datasets to obtain LC peak volumes fro monoisotopic peaks later used for quantification.Statistical analysis was performed with Diffprot algorithm with LOWESS normalization of quantitative values. Benjamini-Hochberg correction for multiple-testing was performed. Proteins with q≤0.05 were classified as differentially expressed (DEPs)."],"omics_type":["Proteomics"],"labhead":["Joanna Sajewicz-Krukowska"],"instrument_platform":[""],"submission_type":["COMPLETE"],"labhead_affiliation":["Department of Virology and Viral Animal Diseases, National Veterinary Research Institute - State Research Institute, Pulawy, Poland"],"species":["Gallus Gallus (chicken)"],"publication":["Not available"],"submitter_mail":["esme@ibb.waw.pl"],"submitter_affiliation":["IBB PAS"],"submitter_country":["Poland"],"doi":["10.6019/PXD074601"],"additional_accession":[]},"is_claimable":false,"name":"Proteomic remodeling of the chicken spleen during chicken astrovirus infection reveals coordinated interferon activation and stromal restructuring","description":"Chicken astrovirus (CAstV) is an enteric pathogen associated with white chick syndrome (WCS), a condition characterized by reduced hatchability, hypopigmentation, developmental delay, and increased early mortality. While transcriptomic studies have provided insight into host responses to CAstV infection, protein-level alterations underlying systemic pathology remain poorly defined. Here, we performed label-free quantitative proteomic profiling of the chicken spleen, a central lymphoid organ coordinating systemic immune regulation. Twenty-one-day-old SPF White Leghorn chickens were orally infected with CAstV strain PL/G059/2014 or inoculated with PBS. Spleens collected at 4 days post-infection were analyzed by nanoLC–MS/MS, and relative protein abundance was compared between infected (CA) and control (K) groups. In total, 2,529 proteins were identified (≥2 unique peptides, FDR≤1%), of which 1,118 met criteria for quantitative comparison. Among these, 33 proteins (~3%) exhibited significant differential abundance (q≤0.05), indicating a selective proteomic response to infection. Proteins with increased abundance formed a coherent type I interferon–associated antiviral module, including IFI27L1 (ISG12), OASL, STAT1, MOV10, and IFITM1. In contrast, proteins with reduced abundance were enriched for extracellular matrix components and cytoskeletal regulators, including the small leucine-rich proteoglycans decorin, lumican, and mimecan, as well as actin-associated structural proteins, indicating decreased cytoskeletal tension and stromal remodeling. Increased abundance of HSP90, together with discordant transcript–protein regulation of selected chaperones and metabolic enzymes, suggested selective modulation of proteostasis and mitochondrial metabolism. Functional enrichment and protein interaction analyses revealed two distinct response modules: a transcriptionally driven interferon-mediated antiviral program and a post-transcriptionally regulated structural and metabolic remodeling program. Integration with previously generated RNA-seq data confirmed concordant transcript–protein induction of interferon-stimulated genes, alongside transcript–protein decoupling for extracellular matrix, cytoskeletal, and mitochondrial proteins. Together, these findings demonstrate that CAstV infection induces a systemic antiviral state accompanied by stromal and metabolic reprogramming, providing a mechanistic framework linking splenic proteome remodeling to the pathogenesis of white chick syndrome.","dates":{"publication":"2026-06-08","submission":"2026-02-18"},"accession":"PXD074601","cross_references":{"TAXONOMY":["NEWT:108018","NEWT:377960","NEWT:1131","NEWT:2042546","NEWT:295546","NEWT:258594","NEWT:1126","NEWT:1129","NEWT:309807","NEWT:89184","NEWT:309800","NEWT:1323429","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:10034","NEWT:661410","NEWT:224326","NEWT:376619","NEWT:3765","NEWT:3767","NEWT:433727","NEWT:135622","NEWT:672127","NEWT:656061","NEWT:37965","NEWT:656064","NEWT:3760","NEWT:36630","NEWT:381198","NEWT:418106","NEWT:10029","NEWT:913645","NEWT:2492962","NEWT:641809","NEWT:44250","NEWT:317447","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:12637","NEWT:2164133","NEWT:2767846","NEWT:314344","NEWT:108061","NEWT:224308","NEWT:49928","NEWT:160621","NEWT:110368","NEWT:1916073","NEWT:1310161","NEWT:1230383","NEWT:77133","NEWT:1310165","NEWT:114727","NEWT:5516","NEWT:498211","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:1310173","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:419481","NEWT:5507","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:1143","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:709964","NEWT:3712","NEWT:3711","NEWT:3714","NEWT:3715","NEWT:2065263","NEWT:1549675","NEWT:177437","NEWT:9014","NEWT:10891","NEWT:226126","NEWT:1616117","NEWT:34865","NEWT:28174","NEWT:550537","NEWT:284812","NEWT:3701","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:1246791","NEWT:9005","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:1883426","NEWT:657321","NEWT:10886","NEWT:3708","NEWT:536231","NEWT:219813","NEWT:215451","NEWT:460519","NEWT:272844","NEWT:1348799","NEWT:55784","NEWT:485","NEWT:2499525","NEWT:142786","NEWT:2853422","NEWT:487","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:46835","NEWT:216778","NEWT:376686","NEWT:237610","EFO:0001352","NEWT:219809","NEWT:219808","NEWT:119562","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:141469","NEWT:987059","NEWT:491","NEWT:460523","NEWT:85694","NEWT:85692","NEWT:3750","NEWT:291146","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:278161","NEWT:415978","NEWT:410289","NEWT:191701","NEWT:9054","NEWT:472","NEWT:29459","NEWT:1380096","NEWT:28128","NEWT:470","NEWT:41122","NEWT:1328300","NEWT:748693","NCBITaxon:50557","NEWT:29491","NEWT:911022","NEWT:446","NEWT:153481","NEWT:445","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:215402","NEWT:3726","NEWT:30455","NEWT:28141","NEWT:9031","NEWT:3728","NEWT:108931","NEWT:219799","NEWT:219798","NEWT:548","NEWT:150475","NEWT:256737","NEWT:51350","NEWT:8694","NEWT:8692","NEWT:542","NEWT:394503","NEWT:13076","NEWT:219785","NEWT:451515","NEWT:8673","NEWT:550","NEWT:10804","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:42410","NEWT:1225786","NEWT:771870","NEWT:51338","NEWT:9995","NEWT:28112","NEWT:49012","NEWT:28110","NEWT:10832","NEWT:507601","NEWT:520","NEWT:64657","NEWT:186441","NEWT:2585209","NEWT:643680","NEWT:214092","NEWT:40697","NEWT:13095","NEWT:162425","NEWT:242124","NEWT:9986","NEWT:8654","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:80833","NEWT:479096","NEWT:28108","NEWT:28104","NEWT:1114957","NEWT:568703","NEWT:568708","NEWT:52641","NEWT:353152","NEWT:353153","NEWT:227377","NEWT:1774284","NEWT:27689","NEWT:207769","NEWT:40674","NEWT:51329","NCBITaxon:1763","NEWT:443906","NEWT:13037","NEWT:519","NEWT:7394","NEWT:6063","NEWT:404330","NEWT:1328388","NEWT:300641","NEWT:39655","NEWT:38323","NEWT:206411","NEWT:80869","NEWT:214053","NEWT:80863","NEWT:665890","NEWT:52639","NEWT:52638","NEWT:8697","NEWT:861109","NEWT:8695","NEWT:1196866","NEWT:510516","NEWT:1045775","NEWT:1123869","NCBITaxon:1335626","NEWT:7370","NEWT:83906","NEWT:99488","NEWT:1134506","NEWT:255470","NEWT:27697","NEWT:86185","NEWT:9935","NEWT:287889","NEWT:1028307","NEWT:52603","NEWT:92867","NEWT:92866","NEWT:1344414","NEWT:202950","NEWT:741158","NEWT:1293091","NEWT:202956","NEWT:315750","NEWT:632957","NEWT:9925","NCBITaxon:9606","NEWT:1436183","NEWT:4232","NEWT:9922","NEWT:416348","NEWT:11298","NEWT:4236","NEWT:398031","NEWT:196627","NEWT:523840","NEWT:1076878","NEWT:200308","NEWT:200302","NEWT:9913","NEWT:9915","NEWT:105841","NEWT:2666255","NEWT:999810","NEWT:28995","NEWT:1392998","NEWT:1150621","NEWT:1085791","NEWT:745531","NEWT:226900","NEWT:231490","NEWT:590651","NEWT:449239","NEWT:244704","NEWT:9909","NEWT:9902","NEWT:430615","NEWT:563041","NEWT:36599","NEWT:41949","NEWT:326423","NEWT:326424","NEWT:13013","NEWT:749906","NEWT:418985","NEWT:749907","NEWT:431947","NEWT:142809","NEWT:1095183","NEWT:302409","NEWT:9979","NEWT:130821","NEWT:2320211","NEWT:27606","NEWT:290610","NEWT:1519788","NEWT:59202","NEWT:9975","NEWT:8643","NEWT:1801631","NEWT:1159899","NEWT:502780","NEWT:860688","NEWT:35231","NEWT:1774208","NEWT:8644","NEWT:1125630","NEWT:28930","NEWT:202914","NEWT:222523","NEWT:27602","NEWT:50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