| Part ⅠPeripheral lymphocyte and subset alteration in chronic kidney diseaseObjective It is widely accepted that chronic renal failure is associated with severe alterations of immune system which is simultaneously presented with immune activation and immune deficiency. Most studies have focused on impaired immune system in dialysis patients. However few studies looked into the immune alteration in pre-dialysis patients. To characterize immune defect in these patients, we performed lymphocyte subsets analysis and explored its relationship with renal function and its related factors.Methods 458 CKD patients were enrolled in this study. Lymphocyte subsets (CD19+, CD3+, CD3+CD4+, CD3+CD8+, CD3-CD56+CD16+) were determined by flow cytometry. Clinical and laboratory data were collected. Patterns of immune cells in different stages of CKD were compared. Multivariate linear regression was used to evaluate the relationship between lymphocyte subset and renal function. Correlation analysis was used to assess the relationship between lymphocyte subset and other clinical and laboratory datas.Results1. The patients’ mean age was 51.1±16.2 years and the predominant group comprised male patients (60.9%). There were significant differences in every subset of lymphocyte compartment among 5 CKD stages (p<0.01). Linear correlation analysis showed that B cell count (r=101.258,p<0.001), CD3+T cell count (r=28.695,p<0.001), CD4+T cell count (r=41.983,p<0.001) and CD8+T cell count (r=45.419,p<0.001) were positively associated with eGFR while NK cell percentage (r=-1.460,p<0.001) was negatively associated with eGFR. Futher multivarious linear regression showed increased B cell (β=19.516,p=0.012), increased CD4+T cell (P=8.898,p=0.014) and decreased NK cell percentage (β=-12.295, p=0.010) were independently correlated with higher eGFR, respectively.2. B cell count was positively associated with Hb, TC, TG, HDL-C, LDL-C, CO2 and Ca and was negatively associated with age, history of hypertension and CVD, usage of CCB and level of SBP, Hey, BUN, sCr, Cystatin C, UA, P,02-M, hsCPR, NT-proBNP and iPTH (p<0.01). Futher multivarious linear regression showed age, β2-M, TC, HDL-C, NT-proBNP,25OH-vitD were independent influencing factors for B cell count (p<0.05).3. CD3+T cell count was positively associated with smoking habit, Hb, TC, TG, LDL-C, CO2 and Ca and was negatively associated with age, history of hypertension and CVD, usage of CCB, and level of SBP, Hcy, BUN, sCr, Cystatin C, UA, P,β2-M, hsCPR, NT-proBNP and iPTH (p<0.01). Futher multivarious linear regression showed age, gender, smoking habit, UA, eGFR, LDL-C, TG, Ca, Hb were independent influencing factors for CD3+T cell count (p<0.05).4. CD4+T cell count was positively associated with somking habit, Hb, TC, TG, LDL-C, CO2, Ca and was negatively associated with age, history of hypertension and CVD, usage of CCB, level of SBP, Hey, BUN, sCr, Cystatin C, UA, P, β2-M, hsCPR, NT-proBNP and iPTH (p<0.05). Futher multivarious linear regression showed age, smoking habit, LDL-C, TG, UA, Ca were independent influencing factors for CD4+T cell count (p<0.05).5. CD8+T cell count was positively associated with Hb, TC, TG, LDL-C, CO2, Ca and was negatively associated with age, history of hypertension, usage of CCB and level of SBP, Hcy, BUN, sCr, Cystatin C, UA, β2-M, NT-proBNP and iPTH (p<0.05). Futher multivarious linear regression showed age, gender, Hb, UA, eGFR were independent influencing factors for CD8+T cell (p<0.05).6. NK cell count was positively associated with age, prescription of anti-hypertension medicines including ACEI, ARB and P receptor blocker, serum level of Hb, Alb, Ca, TRF and 25OH-vitD and was negatively associated with DBP, P and NT-proBNP (p<0.05). Futher multivarious linear regression showed age, history of CVD, usage of β receptor blocker, Hb, Ca, eGFR, Hcy, iPTH were independent influencing factors for NK cell (p<0.05).7. NK cell percentage was positively associated with age, history of hypertension and CVD and anti-hypertension medicines including ACEI, ARB, CCB and p receptor blocker, serum level of Hb, Hey, Bun, sCr, UA, Cyctatin c, β2-M, TRF, NT-proBNP, iPTH and 25OH-vitD and was negatively associated with Hb, TC, TG, LDL-C and CO2 (p<0.05). Futher multivarious linear regression showed age, usage of p receptor blocker, Ca, eGFR, TG, iPTH, CO2 were independent influencing factors for NK cell percentage (p<0.05).Conclusions Lymphocyte subset alteration occoured long before dialysis and predominately in CKD3 stage. Decrease of B cell and CD4+T cell and increase of NK cell percentage were significantly correlated with deterioration of renal function. These immune cells fluctuation were closely linked to hypertension, dyslipidemia, hyperuricemia and disturbance of calcium and phosphorus metabolism.Part II The role of peripheral lymphocyte subset in glomerulopathies of primary glomerular diseaseObjective In China, primary glomerular disease remains the most common cause of End-stage renal disease. Our limited knowledge of underlying glomerular disease mechanisms is a major barrier to successful therapeutic intervention. In this study, we used lymphocyte subset condition as a marker of immune activation and explored its relationship to glomerulonephritis.Methods 251 patients hospitalized for renal biopsy during Oct 2012 to Jan 2014 were enrolled in this study. Lymphocyte subsets (CD19+, CD3+, CD3+CD4+,CD3+CD8+, CD3"CD56+CD16+) were determined by flow cytometry. Patterns of immune cells in different glomerulonephritis were compared. Multivariate logistic regression was used to evaluate the relationship between lymphocyte subsets and glomerulopathies.Results1. The patients’mean age was 44.0±16.0 years and male patients comprised 54.2% of the whole population. The glomerulopathies consisted of 42 FSGS (16.7%),97 IgAN (38.6%),52 MN (20.7%),31 MCD (12.4%),15 MsPGN (6.0%),9 SGN (3.6%) and 5 MPGN (2.0%).2. There were significant differences in CD3+T cell compartment among all these glomerulopathies groups (p<0.05). Further analysis showed CD3+T cell was signifantly lower in MN than MCD [(1.37±0.57)x109/L vs. (1.72±0.63)x109/L,p=0.025].3. There were significant differences in CD8+T cell compartment among all these glomerulopathies groups (p<0.01). Further analysis showed CD8+T cell was signifantly lower in MN than MCD [(0.45±0.28)x109/L vs. (0.59±0.22)x109/L,p=0.002].4. There were significant differences in NK cell compartment among all these glomerulopathies groups (p<0.01). Further analysis showed NK cell was signifantly lower in MCD than FSGS [(0.14±0.07)×109/L vs. (0.23±0.14)x109/L,p=0.004], IgAN [(0.14±0.07)x109/L vs. (0.20±0.10)x109/L,p=0.011] and MN [(0.14±0.07)x109/L vs. (0.22±0.12)x109/L, p=0.007]. Multivariate logistic regression analysis indicated that decreased NK cell was independently associated with specific glomerulopathy of MCD (p=0.044).5. There were significant differences in NK cell percentage among all these glomerulopathies groups (p<0.01). Further analysis showed NK cell was signifantly lower in MCD than FSGS (6.5±3.7% vs. 11.1±7.0%,p=0.001), IgAN (6.5±3.7% vs. 10.4±5.4%,p<0.001) and MN (6.5±3.7% vs.11.7±5.6%,p<0.001).Conclusions Lymphocyte subset is closely linked to primary glomerulopathies and decreased NK cell count was independently associated with specific glomerulopathy of MCD.Part Ⅲ The role of peripheral lymphocyte subset in proteinuria remission of primary glomerular diseaseObjective Proteinuria, as one of the most important presentation of glomerular disease, is also commonly used as an observation target for accessing treatment effect. Proteinurea remission always indicates effective treatment and favorable prognosis. Although it is widely accepted that immunoregulation is the key point concening the disease natural history and responsiveness to therapy, we still have little knowledge about the mechanism of underlying glomerular disease or immunosuppressive therapy. In this study, we used lymphocyte subset as a marker of immune condition and explored its relationship to proteinuria remission after 6 months of treatment.Methods Patients hospitalized for renal biopsy with pathological results supporting primary glomerulonephritis during Oct 2012 to Jun 2013 were enrolled in this study. Lymphocyte subsets (CD19+, CD3+, CD3+CD4+, CD3+CD8+, CD3CD56+CD16+) were determined by flow cytometry. Patients were followed for 6 months and 24-h urinary protein excretion and immunosuppressive therapy alteration were recorded. Multivariate logistic regression was used to evaluate the relationship between lymphocyte subsets and outcome of proteinuria.Results1. A total of 81 patients were enrolled in this study with mean age of 43.5±16.9 years and male patients comprised 58.0% of the whole population. The glomerulopathies consisted of 11 FSGS (13.6%),25 IgAN (30.9%),22 MN (27.2%), 16 MCD (19.8%),4 MsPGN (4.9%),1 SGN (1.2%) and 2 MPGN (2.5%).2. After 6 months of treatment, there are 43 patients receiving proteinuria remission. These patients were younger, had higher eGFR and less diabetes, less somking habbit and less usage of anti-hypertension medine including ACEI, ARB and CCB than those without proteinuria remission. There were less MN and more MCD in the goup of proteinuria remission. More patients took immunosuppressive agents in proteinuria non-remission group.3. B cell percentage was significantly higher in proteirnuria remission group than proteinuria non-remission group (16.7±5.7%vs.14.1±5.5%,p=0.045). NK cell count [(0.18±0.10)x109/L vs. (0.23±0.12)x109/L,p=0.027] and NK cell percentage (8.5±4.9% vs.11.1±5.4%, p=0.026)was significantly lower in proteirnuria remission group than proteinuria non-remission group. Further multivariate logistic regression analysis showed increased B cell perncentage (OR:10.779,95%CI [1.601-72.580], p=0.015) and decreased NK cell count (OR:13.581,95%CI [1.581～116.695],p=0.017) were indepentently associated with proteinuria remission after adjusting for age, gender, hypertension, diabetes, smoking habit, basic eGFR, glomerulopathies and treatmen of ACEI/ARB, prednisone and immunsupressive agents.Conclusions Lymphocyte subset is associated with proteinuria outcome in patients of primary glomerular disease. Higher B cell percentage and lower NK cell count were correlated to better proteinuria outcome and increased B cell percentage can independently predict proteinuria remission in these patients. |
PDF Full Text Request