The role of TLR9 and CD40 signaling axis in regulation of IL10+ CD3- subsets from PBMC of children ITP patients
ZHENG Mengmeng1, SHI Jiandong2, ZHOU Haixia2
1.Flow Cytometry Core Facility, Scientific Research Center, Wenzhou Medical University, Wenzhou 325035, China; 2.Department of Hematology, the Second Affiliated Hospital & Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou 325027, China
ZHENG Mengmeng,SHI Jiandong,ZHOU Haixia. The role of TLR9 and CD40 signaling axis in regulation of IL10+ CD3- subsets from PBMC of children ITP patients[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2020, 50(7): 568-572.
Abstract：Objective: To study the effects of TLR9 combined with CD40 signal on the CD3- population and IL-10 production in the PBMCs from children with ITP. Methods: The plasma IL-10 level in 8 ITP children and healthy controls were determined by using flow cytometry based LEGENplex panel technology. PBMCs from 8 ITP children and 7 healthy controls were isolated by using human lymphocytes separation buffer, and subjected to CpG plus CD40L stimulation invitro for 5 hours. Then cells were analyzed by flow cytometry after staining with the indicated markers. Results: Compared with controls (5.677±0.808), elevated plasma IL-10 level (8.063±0.892) was observed in ITP children. The percentage of CD3- cells in PBMCs was significantly increased in both CpG plus CD40L stimulation and unstimulated group. However, percentage of CD3+ cells was decreased after stimulation. Moreover, the IL-10 production was robustly enhanced in stimulated PBMCs from ITP children. In addition, compared with unstimulated control samples, the unstimulated PBMCs samples from ITP children showed reduction in cell viability. Conclusion: The TLR9 signal together with CD40 signal is crucial for triggering both IL-10 production and cell-death in CD3- cells from children with ITP.
