The Analysis Of Lymphocyte Subsets And IgA₁ Levels In Tonsil Tissues Of IgA Nephropathy Patients

?Objective?To observe and compare IgA nephropathy patients who had tonsillectomy with chronic tonsillitis who have no kidney disease.The histopathological differences of tonsils were analyzed,Tonsil tissues were divided into two parts:the germinal center and interfollicular area to be observed.The expression of lymphocyte subsets CD4,CD19,CD20,CD21 and IgA₁ in different areas of tonsils were analyzed.to confirm the relationships between the disease and the distribution of T?B lymphocytes in different areas in tonsils.The research is to find the effect of mucosal infection in immunological factors that cause the disease,it provides a theoretical basis to further explore the pathogenesis of IgA nephropathy.?Methods?A total of 33 patients underwent tonsillectomy with renal biopsy from August 2014 to August 2016 in Shenzhen Second People's Hospital.Cases of chronic tonsillitis who had tonsillectomy as the control group.HE staining was used to observethehistopathologicalchangesoftonsilsineachgroup.CD4,CD19,CD20,CD21 and IgA₁ staining were performed by MaxVision immunohistochemical method.Image-J analysis software was used to analyze the expression of labeled cells in the germinal center and interfollicular area in tonsil tissues.According to the history,IgA nephropathy patients were divided into two groups,20 patients had no tonsillitis induced macroscopic hematuria?non-HU-IgAN group?,13 patients had macroscopic hematuria due to tonsillitis?HU-IgAN group?,using the above indicators for subgroup analysis.Finally,correlation analysis was made between tonsil lymphocyte subsets/IgA₁ level and renal function,to know whether they have relationships.?Results?1.Baseline data:there has no differences between the two groups of patients'age and weight included in the study,the blood pressure of all patients were within the normal range,but the systolic blood pressure in the IgA nephropathy group was higher than that in the control group?P<0.05?.There was no significant difference in diastolic blood pressure between the two groups?P=0.342?.The level of serum creatinine in IgA nephropathy group was significantly higher than that in control group,and the level of eGFR was significantly decreased?P<0.01?.There was no significant difference in age and body weight between non-HU-IgAN group and HU-IgAN group in subgroup analysis.The systolic and diastolic blood pressure of non-HU-IgAN group was higher than that of HU-IgAN group?P<0.01?.There was no significant difference in urinary protein and serum IgA betweennon-HU-IgAN group and HU-IgAN group?P=0.076,P=0.438?.The serum creatinine was higher in non-HU-IgAN group than that in HU-IgAN group,but there was no difference in eGFR level.?P=0.132?.2.Tonsil pathology changes:tonsil tissue structure have varying degrees of proliferation,germinal center area expanded.Compared with the control group,tonsillar germinal center boundary is unclear and the expansion is not obvious in IgA nephropathy,but interfollicular area is enlarged.Subgroup analysis of non-HU-IgAN and HU-IgAN group was not obvious.3.Tonsil lymphocyte subsets changes:?CD4 staining in germinal center was significantly weaker in IgA nephropathy group than in control group?P<0.001?.CD4 staining was also weakly expressed in interfollicular area(P_MOD=0.0116,PAPSAP</sub>=0.0081).In the subgroup analysis,the mean optical density and average positive staining area percentage of non-HU-IgAN group were not statistically different from HU-IgAN group in germinal center or interfollicular area.(GC:PMOD</sub>=0.3118,PAPSAP</sub>=0.2569,P>0.05;IFA:PMOD</sub>=0.0826,PAPSAP</sub>=0.1234,P>0.05)?The average optical density and average positive staining area percentageof CD19 staining in interfollicular area were significantly higher in IgA nephropathy patients than those in control group?P<0.001?,but in germinal center the IgA nephropathy group was weaker than control group?P<0.001?.Insubgroup analysis,the mean optical density and average positive staining area percentage of germinal center and interfollicular area were not significantly different in two groups.(GC:PMOD</sub>=0.2856,PAPSAP</sub>=0.3230,P>0.05;IFA:PMOD</sub>=0.9692,PAPSAP</sub>=0.8839,P>0.05)?The average optical density and average positive staining area percentage ofCD20 staining in interfollicular area were significantly higher in IgA nephropathy patients than those in control group?P<0.001?,there were no differences in germinal center.?P=0.1179,P=0.5164,P>0.05?Insubgroup analysis,the mean optical density and average positive staining area percentage of germinal center and interfollicular area were not significantly different in two groups.(GC:PMOD</sub>=0.1179,PAPSAP</sub>=0.5164,P>0.05;IFA:PMOD</sub>=0.1392,PAPSAP</sub>=0.4452,P>0.05)?The average optical density and average positive staining area percentageof CD21 staining in interfollicular area were significantly higher in IgA nephropathy patients than those in control group?P<0.05?,but there was no significant difference in germinal center between the two groups.?P=0.160,P=0.0622,P>0.05?In subgroup analysis,the average optical density and average positive staining area percentage of CD21 staining in HU-IgAN group were significantly lower than those in non-HU-IgAN group?P<0.05?,but there was no statistical Differences in interfollicular area.(IFA:PMOD</sub>=0.385,PAPSAP</sub>=0.293,P>0.05)4.Tonsil IgA1 staining:IgA1 was strongly stained in patients with IgA nephropathy.The staining was significantly stronger in interfollicular area and germinal center than in control group?P<0.001?.IgA1 positive cells accounted for very low proportion in control group.In the subgroup analysis,the mean optical density and average positive staining area percentage of IgA1 staining in HU-IgAN group were not significantly different from those in non-HU-IgAN group.(GC:PMOD</sub>=0.2442,PAPSAP</sub>=0.536,P>0.05;IFA:PMOD</sub>=0.691,PAPSAP</sub>=0.8447,P>0.05)?5.Correlation analysis of lymphocyte subsets distribution in tonsils:the intensity of CD4 and CD19 staining in germinal center in tonsil tissues were positively correlated with eGFR,and the staining intensity of CD19 and CD20 in interfollicular area were negatively correlated with eGFR.The intensity of IgA1 staining was negatively correlated with eGFR in all parts of tonsils.?Conclusion??1?In this study,the number of CD4⁺T cells in germinal center and interfollicular area of tonsil in IgA nephropathy patients was significantly reduced compare with chronic tonsillitis patients without nephropathy.The number of CD19⁺B cells,CD20⁺B cells and CD21⁺cells were significantly increased in interfollicular area,while the number of CD19⁺B cells in the germinal center was significantly reduced.?2?In the tonsil tissues of IgA nephropathy patients who were induced by prodromal infection?tonsillitis?,the number of CD21⁺cells in germinal center was significantly lower than those who have no macroscopic hematuria.?3?Compared with chronic tonsillitis patients without nephropathy,the number of IgA1⁺cells in the tonsil tissue germinal center and interfollicular area of IgA nephropathy was significantly higher,IgA1⁺cells accounted for very low proportion in non-nephropathy patients.?4?There was no significant differences in the number of IgA1⁺cells in tonsillar germinal center or interfollicular area between patients with IgA nephropathy induced by prodromal infection?tonsillitis?or not.?5?The number of CD4⁺T cells and CD19⁺B cells in germinal center were positively correlated with eGFR,but the number of CD19⁺B cells and CD20⁺B cells were negatively correlated with eGFR.The number of IgA1⁺cells in various parts of tonsils were negatively correlated with eGFR...</sub>