ABSTRACT
IL-6 and IL-23 are both pleiotropic cytokines involved in the regulation of the immune response, inflammation, and hematopoeisis. They also could mediate effector cells and tolerance mediated by cells with regulatory function. Inflammatory bowel disease (IBD) is associated with a reduced ratio of Treg cells ato Th17 effector cells in peripheral blood and is characterised by a pro-inflammatory cytokine microenvironment which supports the continued generation of Th17 cells. It is well described in adults but little is known in a pediatric population. This study was aimed to investigate the role of IL-6, IL-23 and its association with Treg and Th17 subsets in pediatric IBD patients. Peripheral blood mononuclear cells from patients and controls were stimulated with PMA, ionomycin, and brefeldin A. The frequencies of CD4+Foxp3+ cells, and CD4+IL17a+ cells were analyzed by flow cytometry. The serum level of IL-6 and IL-23 was determined by Elisa kit. The mRNA expression of Foxp3, IL-17a, IL-6 and IL-23 was detected by real-time quantitative PCR. The ratio of Treg/Th17 decreased in pediatric IBD patients, and it strongly correlated with IL-6 and IL-23. The present study provides a quantitative analysis regarding the Th17/Treg cell balance in peripheral blood of children with IBD and its association with serum IL-6 and IL-23 level.
Subject(s)
Inflammatory Bowel Diseases/physiopathology , Interleukin-23/blood , Interleukin-6/blood , T-Lymphocytes, Regulatory/immunology , Adolescent , Case-Control Studies , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Humans , Inflammatory Bowel Diseases/blood , Inflammatory Bowel Diseases/immunology , Leukocytes, Mononuclear/metabolism , Real-Time Polymerase Chain Reaction , Th17 Cells/immunologyABSTRACT
Regulatory B cells (Breg) are a distinct B cell subset, which contribute to the pathogenesis of autoimmune disorders. Interleukin-10 (IL-10) plays a pivotal function to Breg. It is well described in adults but little is known in a pediatric population. This study was to investigate the role of IL-10-producing B cell (B10) and its association with Treg and Th17 subsets in the children with inflammatory bowel diseases (IBD). Peripheral blood mononuclear cells from IBD children patients and controls were stimulated with PMA, ionomycin, and brefeldin A. The frequencies of CD19+IL-10+ B cells, CD3+CD4+IL-17+Th17 cells, and CD4+ CD25(hi)Foxp3+ Treg cells were analyzed by flow cytometry. The mRNA expression of Foxp3, IL-17a and RORγt was detected by real-time quantitative PCR. The number of B10 cells was elevated in IBD children patients. There was a positive correlation between B10 cells and Tregs in IBD. The ratio of Treg/Th17 decreased in IBD, and it strongly correlated with B10 cells. The frequency of B10 cells is elevated in IBD and it correlates with both the Tregs counts and the Treg/Th17 ratio. B10 cells to regulate functional T cell subsets might be impaired in paediatric patients with IBD.
