Proteomic Analysis of Kveim Reagent Identifies Targets of Cellular Immunity in Sarcoidosis.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:28114394", "kind": "science-record", "revision": 7, "content_hash": "sha256:ff64e1db943166d583fe87830307c5fbac5350c66db75816bf3212a554fa8c7f", "created_at": "2026-11-01T15:40:22Z", "updated_at": "2026-11-01T21:59:15Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "Proteomic Analysis of Kveim Reagent Identifies Targets of Cellular Immunity in Sarcoidosis.", "doi": "10.1371/journal.pone.0170285", "pmid": "28114394", "authors": ["Christian Eberhardt", "Muhunthan Thillai", "Robert Parker", "Nazneen Siddiqui", "Lee Potiphar", "Rob Goldin", "John F Timms", "Athol U Wells", "Onn M Kon", "Melissa Wickremasinghe", "Donald Mitchell", "Mark E Weeks", "Ajit Lalvani"], "journal": "PloS one", "year": 2017, "abstract": "BACKGROUND: Kveim-reagent (Kv) skin testing was a historical method of diagnosing sarcoidosis. Intradermal injection of treated sarcoidosis spleen tissue resulted in a granuloma response at injection site by 4-6 weeks. Previous work indicates proteins as the possible trigger of this reaction. We aimed to identify Kv-specific proteins and characterise the ex vivo response of Peripheral Blood Mononuclear Cells (PBMCs) from sarcoidosis, tuberculosis and healthy control patients when stimulated with both Kv and selected Kv-specific proteins.\nMETHODS: Kv extracts were separated by 1D-SDS-PAGE and 2D-DIGE and then underwent mass spectrometric analysis for protein identification. Sarcoidosis and control PBMCs were first stimulated with Kv and then with three selected recombinant protein candidates which were identified from the proteomic analysis. PBMC secreted cytokines were subsequently measured by Multiplex Cytokine Assay.\nRESULTS: We observed significantly increased IFN-γ and TNF-α secretion from Kv-stimulated PBMCs of sarcoidosis patients vs. PBMCs from healthy volunteers (IFN-γ: 207.2 pg/mL vs. 3.86 pg/mL, p = 0.0018; TNF-α: 2375 pg/mL vs. 42.82 pg/mL, p = 0.0003). Through proteomic approaches we then identified 74 sarcoidosis tissue-specific proteins. Of these, 3 proteins (vimentin, tubulin and alpha-actinin-4) were identified using both 1D-SDS-PAGE and 2D-DIGE. Data are available via ProteomeXchange with identifier PXD005150. Increased cytokine secretion was subsequently observed with vimentin stimulation of sarcoidosis PBMCs vs. tuberculosis PBMCs (IFN-γ: 396.6 pg/mL vs 0.1 pg/mL, p = 0.0009; TNF-α: 1139 pg/mL vs 0.1 pg/mL, p<0.0001). This finding was also observed in vimentin stimulation of sarcoidosis PBMCs compared to PBMCs from healthy controls (IFN-γ: 396.6 pg/mL vs. 0.1 pg/mL, p = 0.014; TNF-α: 1139 pg/mL vs 42.29 pg/mL, p = 0.027). No difference was found in cytokine secretion between sarcoidosis and control PBMCs when stimulated with either tubulin or alpha-actinin-4.\nCONCLUSIONS: Stimulation with both Kveim reagent and vimentin induces a specific pro-inflammatory cytokine secretion from sarcoidosis PBMCs. Further investigation of cellular immune responses to Kveim-specific proteins may identify novel biomarkers to assist the diagnosis of sarcoidosis.", "claims": [{"id": "claim-method", "statement": "Statistical significance to look for differences between groups was determined by Mann-Whitney test.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:28114394", "locator": "sections[7] \"Multiplex cytokine analysis\" paragraph 2", "quote": "Statistical significance to look for differences between groups was determined by Mann-Whitney test."}], "state": "complete"}, {"id": "claim-data-access", "statement": "All relevant data are within the paper and its Supporting Information file.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:28114394", "locator": "sections[21] \"Data Availability\" paragraph 2", "quote": "All relevant data are within the paper and its Supporting Information file."}], "state": "complete"}, {"id": "claim-target-result", "statement": "The paper used Mann–Whitney tests for between-group comparisons and made its mass-spectrometry source data available through ProteomeXchange’s PRIDE repository under identifier PXD005150 (10.6019/PXD005150); the headline finding was that both Kveim reagent and vimentin induced a specific pro-inflammatory cytokine response in sarcoidosis PBMCs.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:28114394", "locator": "sections[0] \"Abstract\" paragraph 3", "quote": "We observed significantly increased IFN-γ and TNF-α secretion from Kv-stimulated PBMCs of sarcoidosis patients vs. PBMCs from healthy volunteers (IFN-γ: 207.2 pg/mL vs. 3.86 pg/mL, p = 0.0018; TNF-α: 2375 pg/mL vs. 42.82 pg/mL, p = 0.0003)."}, {"asset": "source:pmid:28114394", "locator": "sections[0] \"Abstract\" paragraph 3", "quote": "Data are available via ProteomeXchange with identifier PXD005150."}, {"asset": "source:pmid:28114394", "locator": "sections[0] \"Abstract\" paragraph 4", "quote": "Stimulation with both Kveim reagent and vimentin induces a specific pro-inflammatory cytokine secretion from sarcoidosis PBMCs."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "partial", "objective": "The paper used Mann–Whitney tests for between-group comparisons and made its mass-spectrometry source data available through ProteomeXchange’s PRIDE repository under identifier PXD005150 (10.6019/PXD005150); the headline finding was that both Kveim reagent and vimentin induced a specific pro-inflammatory cytokine response in sarcoidosis PBMCs.", "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:28114394"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. 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