title: Quantitative Proteomics of Gut-Derived Th1 and Th1/Th17 Clones Reveal the Presence of CD28+ NKG2D- Th1 Cytotoxic CD4+ T cells.
abstract: T-helper cells are differentiated from CD4+ T cells and are traditionally characterized by inflammatory or immunosuppressive responses in contrast to cytotoxic CD8+ T cells. Mass-spectrometry studies on T-helper cells are rare. In this study, we aimed to identify the proteomes of human Th1 and Th1/Th17 clones derived from intestinal biopsies of Crohn's disease patients and to identify differentially expressed proteins between the two phenotypes. Crohn's disease is an inflammatory bowel disease, with predominantly Th1- and Th17-mediated response where cells of the "mixed" phenotype Th1/Th17 have also been commonly found. High-resolution mass spectrometry was used for protein identification and quantitation. In total, we identified 7401 proteins from Th1 and Th1/Th17 clones, where 334 proteins were differentially expressed. Major differences were observed in cytotoxic proteins that were overrepresented in the Th1 clones. The findings were validated by flow cytometry analyses using staining with anti-granzyme B and anti-perforin and by a degranulation assay, confirming higher cytotoxic features of Th1 compared with Th1/Th17 clones. By testing a larger panel of T-helper cell clones from seven different Crohn's disease patients, we concluded that only a subgroup of the Th1 cell clones had cytotoxic features, and these expressed the surface markers T-cell-specific surface glycoprotein CD28 and were negative for expression of natural killer group 2 member D.
authors.affiliation: From the Centre for Immune Regulation and Department of Immunology, University of Oslo and Oslo University Hospital-Rikshospitalet, 0372 Oslo, Norway; Tahira.riaz@medisin.uio.no.
authors.forename: Tahira
authors.initials: T
authors.lastname: Riaz
authors.affiliation: From the Centre for Immune Regulation and Department of Immunology, University of Oslo and Oslo University Hospital-Rikshospitalet, 0372 Oslo, Norway;
authors.forename: Ludvig Magne
authors.initials: LM
authors.lastname: Sollid
authors.affiliation: From the Centre for Immune Regulation and Department of Immunology, University of Oslo and Oslo University Hospital-Rikshospitalet, 0372 Oslo, Norway; Section for Immunology, Norwegian Veterinary Institute, Ullevaalsveien 68, 0454 Oslo, Norway.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Olsen
authors.affiliation: From the Centre for Immune Regulation and Department of Immunology, University of Oslo and Oslo University Hospital-Rikshospitalet, 0372 Oslo, Norway;
authors.forename: Gustavo Antonio
authors.initials: GA
authors.lastname: de Souza
mesh_terms: D018106:CD28 Antigens / Q000378:metabolism*; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D003424:Crohn Disease / Q000378:metabolism* / Q000473:pathology; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D013058:Mass Spectrometry; D055655:NK Cell Lectin-Like Receptor Subfamily K / Q000172:deficiency*; D020543:Proteome / Q000276:immunology / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D013602:T-Lymphocytes, Cytotoxic / Q000378:metabolism; D018417:Th1 Cells / Q000378:metabolism*; D058504:Th17 Cells / Q000378:metabolism*
chemical_list: D018106:CD28 Antigens; C527085:KLRK1 protein, human; D055655:NK Cell Lectin-Like Receptor Subfamily K; D020543:Proteome