Recombinant CD99 proteins induced the upregulation of IL-6 and TNF- expression, but not IFN-, in anti-CD3 monoclonal antibody activated T cells. subject. Statistical analysis was carried out by unpaired t test. ns = not statistically significant.(TIF) pone.0217393.s001.tif (902K) GUID:?B2D2B12B-6BE9-4C7C-B8DF-ED2E99D2EF65 S2 Fig: Flow cytometric gating strategy for analysis of intracellular cytokine expression. Size (ahead scatter; FSC) and granularity (part scatter; SSC) of peripheral blood mononuclear cells (PBMCs) were plotted and utilized for cell gating as indicated. (A) The gated cells were plotted against part scatter (SSC) and CD14. Monocytes were discriminated from lymphocytes based on CD14 manifestation and then CD14+ monocytes were further plotted against cytokine manifestation and CD14. (B) The gated cells were plotted against CD3 and CD19 and then CD3-CD19+ B cells were further plotted against cytokine manifestation and CD19. (C) The gated cells were plotted against CD3 and CD56 and then CD3-CD56+ NK cells were further plotted against cytokine manifestation and CD56. The cytokine manifestation in term of level of manifestation and rate of recurrence Ubiquitin Isopeptidase Inhibitor I, G5 in each human population were investigated.(TIF) pone.0217393.s002.tif (5.5M) Ubiquitin Isopeptidase Inhibitor I, G5 GUID:?6EB40A33-0E1E-455B-ACD1-DC219503C0EE S3 Fig: Flow Ubiquitin Isopeptidase Inhibitor I, G5 cytometric gating Ubiquitin Isopeptidase Inhibitor I, G5 strategy for analysis of CD99 ligand expression. Size (ahead scatter; FSC) and granularity (part scatter; SSC) of peripheral blood mononuclear cells (PBMCs) were plotted and utilized for cell gating as indicated. (A) The gated cells were plotted against CD3 and CD56. The CD3+CD56- T cell and CD3-CD56+ NK cells were further gated. (B) The gated cells were plotted against CD14 and CD19. CD14+ monocytes and CD19+ B cells were further gated. (C) Dendritic cells were recognized by CD3-CD14-CD16-CD19-CD56- and HLA-DR+ cells. The gated cells were plotted against CD56 and CD3, CD14, CD16, CD19 for lineage bad cell gating. The lineage bad gated cells were plotted against SSC and HLA-DR and dendritic cells were further gated. In each gated human population (i.e. NK cells, T cells, Monocytes, B cells and dendritic cells), the percentage of phycoerythrin (PE) positive cells were investigated.(TIF) pone.0217393.s003.tif (6.1M) GUID:?BA7C2A75-2FCE-478A-B969-CF941C0F8FEA Data Availability StatementAll relevant data are within the manuscript. Abstract CD99 has been reported to be involved in T cell rules. CD99 ligand involvement in the rules of T cell activation has been postulated. In this study, recombinant CD99 proteins were produced and used as a tool for determining the part of CD99 and its ligand interaction. Recombinant CD99 proteins induced the upregulation of IL-6 and TNF- manifestation, but not IFN-, in anti-CD3 monoclonal antibody triggered T cells. The cytokine alteration was not observed in unstimulated T cells indicating the cytokine upregulation required the signal from T cell activation. The upregulation of IL-6 and TNF- was, in addition, observed in CD3- mononuclear cell human population including monocytes and NK cells. The recombinant CD99 proteins, however, did not impact either CD25, CD69 or MHC class II manifestation or T cell proliferation, upon T cell activation. The CD99 ligands were demonstrated to be indicated on monocytes, NK cells and dendritic cells, but not on B and T cells. Our results indicated the presence of CD99 ligands on leukocyte surface. Interaction between CD99 and its ligands entails the rules of cytokine production. Introduction Over the last several decades, ligands of several leukocyte surface molecules including T cell rules have been recognized [1C3]. Uncovering these ligands is essential for understanding the precise immunoregulation mechanism [4]. In the achievement of this, SCDO3 the discovery of various leukocyte surface molecules and its ligands connection will lead to the development of fresh methods for Ubiquitin Isopeptidase Inhibitor I, G5 treatment of various diseases, including inflammatory diseases and cancers. The PD-1/PD-L1 immune checkpoint blockage in malignancy therapy [5C7], the interfering CD28 and CD80/CD86 binding with CTLA-4-Ig in the treatment of rheumatoid arthritis [8, 9] and using anti-CTLA-4 monoclonal antibody (mAb) for malignancy treatment [5, 6, 10] are the best examples. CD99.
VIP Receptors