@article{Eberhardt:2017:10.1371/journal.pone.0170285,
author = {Eberhardt, C and Thillai, M and Parker, R and Siddiqui, N and Potiphar, L and Goldin, R and Timms, JF and Wells, AU and Kon, OM and Wickremasinghe, M and Mitchell, D and Weeks, ME and Lalvani, A},
doi = {10.1371/journal.pone.0170285},
journal = {PLOS ONE},
title = {Proteomic analysis of Kveim reagent identifies targets of cellular immunity in sarcoidosis},
url = {http://dx.doi.org/10.1371/journal.pone.0170285},
volume = {12},
year = {2017}
}

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TY  - JOUR
AB  - 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.Methods: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.Results: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 contr
AU  - Eberhardt,C
AU  - Thillai,M
AU  - Parker,R
AU  - Siddiqui,N
AU  - Potiphar,L
AU  - Goldin,R
AU  - Timms,JF
AU  - Wells,AU
AU  - Kon,OM
AU  - Wickremasinghe,M
AU  - Mitchell,D
AU  - Weeks,ME
AU  - Lalvani,A
DO  - 10.1371/journal.pone.0170285
PY  - 2017///
SN  - 1932-6203
TI  - Proteomic analysis of Kveim reagent identifies targets of cellular immunity in sarcoidosis
T2  - PLOS ONE
UR  - http://dx.doi.org/10.1371/journal.pone.0170285
UR  - http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000396129000033&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
UR  - http://hdl.handle.net/10044/1/45268
VL  - 12
ER  - 

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