The Mechanisms Of Chronic Tonsillitis Affect Immune Function

PartⅠThe changes of periheral blood mononuclear cells in chronic tonsillitispatientsObjective: To explore the immune status and function of peripheral blood mononuclearcells (PBMC) in chronic tonsillitis patients.Methods: The peripheral blood mononuclear cells were isolated from patients with chronictonsillitis and healthy volunteers. Proliferation of PBMCs were determined by the MTT test,cell cycle and apoptosis of PBMCs were accessed by Flow cytometry test, the capacity ofcytokine secretion of PBMCs was accessed by enzyme-linked-immunosorbent serologicassay (ELISA), and cell activity and phagocytosis ability of PBMCs were test at the sametime.Results: There was no significant difference in proliferation and cell cycle of the PBMCsbetween patients with chronic tonsillitis and healthy volunteers. Comparing with healthycontrol group, the cell activity and phagocytosis ability of PBMCs were significant lowerin chronic tonsillitis patients (P＜0.05). However, the level of cytokine produtions and apoptosis of PBMCs were significantly higher in chonic tonsillitis patients than that ofhealthy controls (P＜0.05).Conclusions: Chronic tonsillitis has effect on the immune status and function of theperipheral blood mononuclear cells. It can increase PBMCs apoptosis, upregulate thecytokine secretion of PBMCs, while decrease the activity and phagocytosis ability ofPBMCs.PartⅡPeripheral T cell subsets in Chronic Tonsillitis patientsObjective: To investigate the changing characteristics of T lymphocytic subsets in chronictonsillitis patients in order to evaluating their clinical implication.Method: Fresh peripheral blood samples were obtained from 54 chronic tonsillitis patientsand 52 healthy counterparts. CD4~+ and CD8~+ T lymphocyte subsets including the naive(CD4~+CD45RA~+ and CD8~+CD45RA~+), memory (CD4~+CD45RO~+ and CD8~+CD45RO~+),functional (CD4~+CD28~+ and CD8~+CD28~+), activated (CD4~+CD25~+, CD8~+CD38~+,CD4~+HLA-DR~+ and CD8~+HLA-DR~+) and apoptosis (CD4~+CD95~+ and CD8~+CD95~+) Tlymphocytes, were analyzed by flow cytometry, respectively. The clinical data such asserum Cystatin C concentration, ASO and ESR were simultaneously recorded from eachchronic tonsillitis patient.Result: The CD4~+T cells, rate of CD4~+/CD8~+ and CD4~+CD45RA~+/CD4~+CD45RO~+, CD4~+CD45RA~+ T cells and CD4~+ CD25~+ T cells in chronic tonsillitis were significantly lowerthan those of the control group(P＜0.05) while an obviously increasing percentage ofmemory (CD45RO~+) and apoptosis (CD95~+) T lymphocytes in chronic tonsillitis patients,and there were significant differences between the patients with chronic tonsillitis and the healthy volunteers (P＜0.05). Furthermore, In the chronic tonsillitis patients, Serum CystatinC level was negatively correlated with CD4~+CD45RA~+ T (P＜0.05), and not significantlycorrelated with other T lymphocyte subtypes (P＞0.05).Conclusion: Chronic tonsillitis causes immune disorder in the peripheral blood of patients.Our data may provide valuable information for evaluation of disease progression of chronictonsillitis patients.PartⅢSerum chemokines profile of patients with chronic tonsillitisObjective: To investigate the expression of chemokines in peripheral blood of patients withchronic tonsillitis and their relationship with clinical indicators.Methods: Serum interleukin (IL)-8, growth-related gene product (GRO)-a, granulocytechemotactic protein (GCP)-2 and Monocyte chemoattractant protein (MCP)-1 weremeasured by ELISA assays in 60 chronic tonsillitis patients and 50 healthy volunteers.Serum chemokine levels of patients and heathy volunteers were compared, and thechemokine levels were correlated with anti-streptolysin O (ASO), erythrocytesedimentation rate (ESR) and Cystatin C. Levels of IL-8、GRO-α、GCP-2 and MCP-1were analysis with ROC curve, in order to explore a new diagnosis indicator.Results: Serum levels of IL-8、GRO-αand MCP-1 were significantly higher in patientswith chronic tonsillitis compared with healthy group (P＜0.05). There was no significantdifference in serum GCP-2 level between patients with chronic tonsillitis and healthy group(P＞0.05). Serum IL-8 was correlated positively with ESR of chronic tonsillitis patients.And serum MCP-1 was correlated positively with serum Cystatin C. One month afterTonsillectomy, serum IL-8, GRO-α, GCP-2, MCP-1 and Cystatin C were alldownregulated (P＜0.05). The cutoff values of 516 pg/ml for MCP-1, had 100% sensitivity and 82.93% specificity for renal dysfunction.Conclusions: Chronic tonsillitis can upregulate Serum levels of chemokines, whiletonsillectomy can downregulate them. MCP-1 in patients with chronic tonsillitis canpredict renal dysfunction and can correlate strongly with renal dysfunction severity. MCP-1may play a key role in the pathophysiology of renal impairment associated with chronictonsillitis and are potentially new targets for therapeutic approaches.PartⅣChronic tonsillitis elevate MCP-1 secretion of peripheral bloodmononuclear cells by active NF-κB pathwayObjective: To study the mechanism of chronic tonsillitis increasing MCP-1 secretionfrom peripheral blood mononuclear cells.Methods: The peripheral blood mononuclear cells were separated from patients withchronic tonsillitis and healthy volunteers. MCP-1 in peripheral blood mononuclear cellsculture supernatant was detected by ELISA assay. MCP-1 mRNA expression of peripheralblood mononuclear cells was accessed by RT-PCR. IκB、p-IκB and NF-κB in peripheralblood mononuclear cells cytoplasm and nuclear were analysis by Western Blot test. At thesame time, TNF-a or PDTC was added in culture solution to observe and compare thechanges that have taken place.Results: The level of MCP-1 secreted by peripheral blood mononuclear cells was higher inpatients with chronic tonsillitis than that in healthy volunteers (P＜0.05). In chronictonsillitis group, IκB in cytoplasm was dowregulated while p-IκB was upregulated, NF-κBwas decreased in cytoplasm while increased in nuclear, then MCP-1 mRNA expression waselevated through activation of NF-κB pathway (P＜0.05). TNF-a enhances the activation ofNF-κB pathway so that MCP-1 mRNA was increased (P＜0.05). On the contrary, PDTC inhibits the activation of NF-κB pathway so that MCP-1 mRNA was decreased (P＜0.05).Conclusions: NF-κB signal transduction pathway was actived by chronic tonsillitis toincrease the transcription and translation of MCP-1 in peripheral blood mononuclear cells.PartⅤCCR2 expression and its interaction with MCP-1 in peripheral bloodmononuclear cells of patients with chronic tonsillitisObjective: To investigate CCR2 expression and its interaction with MCP-1 in peripheral bloodmononuclear cells of patients with chronic tonsillitis.Methods: The expression of CCR2 mRNA in peripheral blood mononuclear cells was detectedby RT-PCR. And the expression of CCR2 protein in peripheral blood mononuclear cells wasaccessed by Western Blot test. Localization of the expression of CCR2 was detected byimmunohistochemical Staining. The interaction between MCP-1 and CCR2 was analysisby chemotaxis experiments.Results: The mRNA and protein of CCR2 were higher in the peripheral blood mononuclearcells of patients with chronic tonsillitis than that of healthy volunteers (P＜0.05). CCR2 expression waslocalizated on the membrane of peripheral blood mononuclear cells. A dose-dependent locomotion toMCP-1 with the typical bell-shaped curve was observed. However, blocking of CCR2 inhibitedsignificantly mononuclear cell migration in response to MCP-1.Conclusions: CCR2 was high expression on the membrane of peripheral blood mononuclearcells of patients with chronic tonsillitis. And the CCR2 positive peripheral blood mononuclear cells werechemotaxis by its autocrine or paracrine MCP-1.