Significance Of T-lymphocyte-associated Subsets In The Development Of Acute Pancreatitis

Objective: To study the changes of T lymphocyte related subsets in the pathogenesis of acute pancreatitis and its significance.Methods: Acute pancreatitis(AP)group and healthy control group were set up.In AP group,5 patients with acute pancreatitis were enrolled to collect patient information.Peripheral blood samples of 10 ml were collected from each patient at three time points: the first day,the third day and the seventh day after the onset of acute pancreatitis.After centrifugation,the whole blood cells were collected from the supernatant.The healthy control group included 5 health check-ups at the same time.10 ml peripheral blood samples were collected from 5 health check-ups and centrifuged to collect whole blood cells.A total of 20 blood samples were obtained.PBMC(peripheral blood mononuclear cells)was separated from blood samples by Ficoll density gradient centrifugation and then made into single cell suspension.43 immune-related sites or receptors were selected for intracellular and extracellular marker staining.Relevant data were collected on Cy TOF instrument(mass spectrometry flow cytometry),Xshift cluster analysis was used to identify the related subgroups.MST visualization and vi SNE scatter plot were used to analyze the expression of various subgroups and maker.The differences of T lymphocyte-based immune expression at different time points in patients with pancreatitis were deeply analyzed.The changes of all subgroups were analyzed by t-test to find out the meaningful subgroups and analyze the meaningful subgroups.Proportional changes,metabolic characteristics,effector factors and expression of co-suppressor molecules.Result:1.Compared with the healthy control group,the expression of AMS,CRP and neutrophil in serum of patients with acute pancreatitis increased significantly.2.T cell related subsets: There were significant changes in peripheral blood lymphocyte subsets in 10 groups of patients with acute pancreatitis and healthy control group:(1)The expression of C22,C17(neutrophil subsets),C26(FoxP3+,CD152-CTLA-4+RORr+,CD154-CD40L+eosinophil subsets)in the development of acute pancreatitis was significantly higher than that in the healthy control group.(2)The expression of C13(monocyte macrophage subsets),C14(CD38+,CD39+) conventional dendritic cell subsets,C27(eosinophil subsets),C28(CD8a+initial T cells),C31(CD8+effect memory T cells),C34(CD8+,h-m-Granzyme B+effect memory T cells)and C38(CD4+effect memory T cells)in the development of acute pancreatitis was significantly lower than that in the healthy control group.The difference was statistically significant(P < 0.05).3.Immune characteristics of 16 markers differentially expressed in healthy control group and acute pancreatitis group: CD27,CD14,Lag-3,CD39,PD-L1,CD59,Gata3-h/m,CD127,CTLA-4,Fox P3,CD137,RORy,CD154,h/m-Granzyme B,HLA-DR,CD8a(p < 0.05),which accords with the probability of the same distribution.It is generally believed that in one experiment,such sample group is really different).Among them,CD27,CD39,HLA-DR,CD8 a and CD127 were lower than those of healthy control group,while CD14,Lag-3,CD39,PD-L1,Gata3-h/m,h/m-Granzyme B,CD154,RORy,CD137,FoxP3 and CTLA-4 were higher than those of healthy control group.Co-inhibitors PD1,CD28 and Tim-3 were not expressed in the development of acute pancreatitis.Conclusion:1.Immunosuppression in acute pancreatitis may be related to the decrease of the percentage of CD38+,CD39+ dendritic cell subsets,CD8a+initial T cell subsets,CD8+effective memory T cells,CD8+,h-m-Granzyme B+effective memory T cell subsets and CD4+effective memory T cell subsets.2.The expression of CD27,CD39,HLA-DR,CD8 a and CD127 increased significantly in the development of acute pancreatitis,suggesting that CD27,CD39,HLA-DR,CD8 a and CD127 may be used as potential biomarkers of acute pancreatitis.CD14,Lag-3,CD39,PD-L1,Gata3-h/m,h/m-Granzyme B,CD154,RORy,CD137,FoxP3,CTLA-4 decreased significantly in the development of acute pancreatitis,indicating that they may be potential prognostic indicators or therapeutic targets of acute pancreatitis.3.The Co-stimulation factor PD1,CD28 and Tim-3 were not obvious expression in the development of acute pancreatitis.