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. Molecular & Cellular

The Immunomodulatory Imbalance in Patients with Ketamine Cystitis

“…The differentiation of CD4 + cells generates the T H 1 and T H 2 phenotypes with the same receptor specificity, indicating the competition for antigenic stimulation, mediated by antigen-presenting cells (APCs), combined with cytokine-mediated cross-suppression between phenotypes to yield a response that is eventually dominated by T helper cells that are uniform in both receptor specificity and cytokine secretion phenotype. We found a different T helper cell phenotype distribution that the T H 1, T H 2, and T H 17 receptors were all expressed in PBMCs ( Figure 2 ), consistent with recent data [ 17 ]. We subsequently analyzed the serum cytokine secreted from the T helper cells in KC patients.…”

Human Regulatory T Cells From Umbilical Cord Blood Display Increased Repertoire Diversity and Lineage Stability Relative to Adult Peripheral Blood

“…For C06 in particular, the top five differentially expressed (DE) genes (ranked on p-value) were CCL5, GZMK, CXCR3, LYAR, and NKG7 ( Supplementary Table S2), as shown in the UMAP plots colored by relative expression ( Figure 2C). CXCR3, CCL5, and NKG7 have all been associated previously with T H 1 migratory capacity (55,56) and phenotype (57), while the expression of GZMK and LYAR likely indicate a cytotoxic and activated population (58,59). Figure 2C also depicts the canonical Treg markers, FOXP3 and IKZF2, which were highly expressed in the majority of APB and CB Treg clusters but only lowly expressed in C06.…”

Proteomics and Lipidomics in Inflammatory Bowel Disease Research: From Mechanistic Insights to Biomarker Identification

“…Studies have implicated Th17 T-cells in IBD pathogenesis, particularly in CD-associated dysregulation of immune responses [ 55 , 56 ]. Riaz et al compared the proteomes of human Th1 and Th1/Th17 clones derived from intestinal biopsies of CD patients [ 57 ]. In total, 7401 proteins have been quantified; 334 were differentially expressed between Th1 and Th1/Th17 clones.…”

“…IBD biomarker products. References [ 51 , 54 , 57 , 58 , 59 , 60 , 61 , 62 , 67 , 68 , 69 , 70 , 73 , 74 , 75 , 77 , 78 ] are cited in the supplementary materials.…”