| IgAN is the most common form of progressive glomerulonephritis, characterized by mesangial IgA1deposits, although its mucosal immunizationpathogenesis is unclear.It is recognized as a tonsil-related disease since it often gets worse after and/or during acute tonsillitis and the disease progression is often arrested by tonsillectomy.On it's pathogenesis,Van Den Wall Bake,et al proposed the mucosa-bone marrow axis based on their series of elegant studies about20years ago. They also hypothesized an impaired IgA immune response in the'mucosa-bone-marrow axis'of IgAN patients. At that time, such proposals were based on experimental models of IgAN. However, the presence of such an axis has not yet been confirmed in human IgAN.Respectively,TLR9and memory B cell play a important role in mucosal(innate) and humoral immune response of mucosal and bone marrow,so that some experiments on them were carried out in human IgAN, which can provide some clues for the abnormal mucosa-bone marrow axis in the pathogenesis of IgA nephropathy, which can provide theoretical basis for tonsillectomy to cure IgAN patients. Chapter I The expression of TLR9in the tonsils and peripheral blood of IgA nephropathy patients and it's variation after tonsillectomyObjective To observe an abnormal expression of mucosal(innate) immune response which is induced by TLR9in the tonsils and peripheral blood of IgAN patients and it's variation after tonsillectomy.Methods55patients and10nomal human beings were enrolled into the study, they were diagnosed as IgAN (n=28),chronic tonsillitis(Non-IgAN)(n=27). The tonsils of chronic tonsillitis patients and peripheral blood of nomal human beings(n=10)were as control groups.Pathological changes of tonsillar tissues were observed by immunohistochemistry. The expression of TLR9of B cells in tonsil and peripheral blood was tested by flow cytometry(FCM),and the same method was used to test the variation of the expression of TLR9in peripheral blood of IgAN after tonsillectomy.Results Immunohistochemistry showed that the TLR9positive cells were mainly localized in the epithelium of tonsil,and minor part in germinal centres, follicular and interfollicular area of tonsil. Compared to non-IgAN group,TLR9positive cells of IgAN were significantly increased (P<0.05). Whether in the tonsils of IgAN vs Non-IgAN patients or the peripheral blood of IgAN patients vs Normal human being,TLR9mRNA have no significantly difference.But the expressions of TLR9of B cells by FCM,both in tonsil and peripheral blood of IgAN,were higher than control groups (P<0.05).After tonsillectomy,when remission of proteinuria and hematuria of IgAN patients, it showed that the expression of TLR9of B cells by FCM was significantly down-regulated,compared with that before tonsillectomy; Meanwhile,it had no significant effect to the expression of TLR9of B cells of normal human beings.Conclusion IgAN patients have an high expression of mucosal(innate) immune response which is induced by TLR9in the tonsils and peripheral blood. TLR9may induce humoral immune response in the pathogenesis of IgAN. After tonsillectomy,the abnormal response can be improved and reach to the level of normal human beings. Chapter Ⅱ The expression of memory B cells (pertaining to CD19+CD27+cells) in tonsil and PB of IgAN patients and it s variation after tonsillectomyObjective To observe an abnormal expression of humoral immune response which is induced by memory B cells (pertaining to CD19+CD27+cells)in the tonsils and peripheral blood of IgAN patients;To analysis the relationship of memory B cells and hematuria, proteinuria of IgAN patients; After tonsillectomy, to observation the variation of memory B cells (pertaining to CD19+CD27+cells) and try to know if there have a mucosa-bone marrow axis in IgAN.Methods55patients and10nomal human beings were enrolled into the study as the former chapter. The position of CD19+CD27+cells in tonsils were observed by confocal laser scanning microscope; The expression of memory B cells (pertaining to CD19CD27+cells) in tonsil and peripheral blood was tested by FCM(flow cytometry),and the same method was used to test the variation of the expression of memory B cells (pertaining to CD19+CD27+cells) in peripheral blood of IgAN after tonsillectomy;The relationship of memory B cells and hematuria, proteinuria and the variation of memory B cells in different groups of IgAN patients(the different groups can be seen in the second chapter).Results By confocal laser scanning microscope,it showed that the CD19+CD27+cells(include memory B cells and plasmablasts) were mainly localized in the Follicular germinal centers, Mantle zone of the follicle,follicular and interfollicular area of tonsil, memory B cells (pertaining to CD19+CD27+cells), compared to non-IgAN group, had a significantly increased in tonsil and peripheral blood of IgAN by FCM (P<0.05).Meanwhile,it have positive relationship in the expression of memory B cells in tonsil and peripheral blood and the urinary protein and microhematuria of IgAN;and with the more increasing number of the urinary protein and microhematuria of IgAN, the more increasing expression of memory B cells can be tested. After tonsillectomy,with the remission of proteinuria and hematuria of IgAN patients, it showed that the expression of memory B cells (pertaining to CD19+CD27+cells)by FCM was significantly down-regulated,compared with that before tonsillectomy。Conclusion IgAN patients have an high expression of humoral immune response which is induced by memory B cells and CD19+CD27+cells in the tonsils and peripheral blood. After tonsillectomy,the abnormal humoral immune response can be improved and reach to the level of normal human beings;It may have a abnomal mucosa-PB-bone marrow axis in the pathogenesis of IgAN;Memory B cells can be play a biomarker in the progression of proteinuria and hematuria of IgAN.There has a pathway between tonsil and PB for memory B cells in IgAN. Chapter Ⅲ The variation of memory B cells and TLR9in tonsil and PB of IgAN patients after HS's stimulation and IgAl's variation after tonsillectomyObjective To investigate the variation of memory B cells and TLR9in tonsil and PB of IgAN patients after HS's stimulation and the relationship of memory B cells and TLR9;In order to analyse the source of IgA1in PB, the concentraton of which was observed before and after tonsillectomy.Methods55patients and10nomal human beings were enrolled into the study as the former chapter. After12days' cultivation in vitro,we had a test to the apoptosis of lymphocytes by FCM;The same method was used to test the expression of TLR9and memory B cells and their relationship after7days'HS'stimulation. The concentraton of IgA1was observed before and after tonsillectomy by ELISA.Results The apoptosis of lymphocytes have no difference in12days' cultivation in vitro. Compared to control groups, TLR9and memory B cells of IgAN were significantly increased (P<0.05),and which have a positive relationship with each other in tonsil and PB. It has a decreased trend for the concentraton of IgAl in PB after tonsillectomyConclusion IgAN patients have an high expression of mucosal (innate) and humoral immune response which is induced by TLR9and memory B cells in the tonsils and peripheral blood after HS's stimulation.TLR9may induce the activation of memory B cells. IgA1in PB of IgAN come from tonsil;and may be from bone marrow. |
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