| Minimal change disease(MCD)is one of the common causes in children and adult-onset nephrotic syndrome,accounting for 70-90%of cases in children under 10 years old and 10%-15%of cases primary nephrotic syndrome in adults.MCD is clinically manifested as massive proteinuria and pathologically manifested as podocyte injure and no immune complex deposition.Compared with MCD in children,MCD in adults is a more challenging and serious disease and is more likely to steroid-dependent or frequent relapse and has higher risks of acute kidney injury.The long-time prognosis of adult-onset MCD is poor and a considerable number of MCD patients in adults progressed to end-stage renal disease(ESRD)or death.The mechanism underling MCD remains unknown.Our study aims to explore the pathogenesis of adult-onset MCD.We designed the study in two parts:Part One:The significance of cytokines expression of Th1/Th2/Th17 in patients with adult-onset minimal change diseaseObjectiveIt is well known that minimal change disease is a T cell-related immune disease.Although it is considered that T cell disorder and circulating cytokines play a key role in MCD,it is still unknown a cell compartment or a particular cytokine by which causes proteinuria in MCD.We have evaluated the changes in lymphocyte subsets composition and the cytokines expression of Th1/Th2/Th17 to explore its significance in MCD.MethodologyA total of 28 subjects were included in this study(15 males and 13 females;average age:34.1 years,age range:18-56 years),including 10 patients with MCD in relapse,nine patients with MCD in remission and nine healthy controls.Lymphocyte subsets,including CD3+T cells,CD3+CD4+T cells,CD3+CD8+T cells,CD3-CD19+ and CD3-CD16/CD56+ NK cells,were evaluated using flow cytometry analysis.The cytokine levels of MCD patients in relapse(n=6),MCD patients in remission(n=3)and healthy controls(n=3)were detected by cytokine antibody array,including 34 human Th1-,Th2-and Th17-related cytokines.ResultsNo differences in CD3+and CD3+CD4+cells proportions were found between MCD patients and healthy controls(P=0.445 and P=0.802).MCD patients in relapse had higher proportions of CD3+CD8+cells compared to MCD in remission and healthy controls(42.7±2.29%vs 27.42± 1.51%,P=0.008 and 42.7±2.29%vs 27.97±2.34%,P<0.001).Furthermore,the CD4+/CD8+ratio was lower in MCD in relapse compared with MCD in remission(0.84±0.09 vs 1.45±0.14,P=0.014)and healthy controls(0.84±0.09 vs 1.4±0.12,P=0.005).There were no differences in CD3-CD 16/CD56+NK cell and CD3-CD19+B cell populations between MCD patients and healthy controls(P=0.199 and p=0.445).Our results indicated that the levels of GM-CSF and TRNACE(TNFSF11)in patients with MCD in relapse increased 1.5 times higher than in patients with MCD in remission.GM-CSF,il-10,il-22 and TNF beta levels were increased significantly in the patients with MCD in relapse compared to healthy controls.ConclusionThe cytokines GM-CSF and TRNACE are increased in adult-onset MCD patient in relapse,indicating that Th cell dysfunction may trigger the releasing of GM-CSF and TRNACE which may make urinary proteinuria production in the in adult-onset MCD patient in relapse.Part Two:The value of urinary CD80 excretion in patients with adult-onset minimal change diseaseObjectivePodocyte is an important element of the glomerular filtration membrane and it is involved in charge barrier and mechanical barrier of glomerular filtration membrane.Minimal change disease(MCD)has been postulated to be a podocytopathy while CD80 plays an important role in podocyte injury.But the mechanism of CD80 in adult-onset MCD is incompletely understood.The aim of this study is to explore the value of CD80 in patients with adult-onset minimal change disease.MethodologyA total of 28 subjects were included in this study(15 males and 13 females;average age:34.1 years,age range:18-56 years),including 10 patients with MCD in relapse,nine patients with MCD in remission and nine healthy controls.CD80 and CTLA-4 in serum and urine were analyzed using Enzyme linked immunosorbent assay method.The expression of CD80,WT-1 and synaptopodin in glomeruli was detected by immunofluorescence technique.ResultsAn evident increase in the excretion of urinary CD80 was found in MCD patients during relapse when compared with MCD patients during remission(598.4± 115.8 ng/g vs 81.78±7.04 ng/g creatinine,P<0.001)and controls(598.4±115.8 ng/g vs.67.44±8.94 ng/g creatinine,P<0.001).The excretion of urinary CD80 showed no statistical difference between MCD patients in remission and healthy control subjects(81.78±7.04 vs 67.44±8.94 ng/g creatinine,P=0.269).No differences were found in serum CD80 concentrations among patients with MCD during relapse,patients with MCD during remission,and healthy control subjects(379.9±32.3,287.6± 19.48,and 342.4±28.43,pg/ml,P=0.081,respectively).There was no correlation between urinary CD80 and proteinuria in MCD patients in relapse(r=-0.32,P=0.366).Urinary CTLA-4 levels were not significantly increased in patients in relapse when compared with MCD patients in remission(173.7±8.73 vs 155.0±8.54 ng/g creatinine,P=0.098)and healthy control subjects(173.7±8.73 vs.160.0± 10.85 ng/g creatinine,P=0.253).Serum CTLA-4 concentrations were not different among patients with MCD in relapse,patients in remission and controls(149±9.9,143±11.7 and 148± 12.3 pg/ml;P=0.949).Compared with MCD patient during remission,the urinary CD80 to CTLA-4 ratio was elevated in MCD during relapse(3.48±0.71 vs 0.53±0.03,p=0.004).There were no differences in the serum CD80/CTLA-4 ratio among MCD patients in relapse,MCD in remission and controls(2.61±0.24,2.10±0.16,and 2.52±0.25,mean±SEM,P=0.278,respectively).The double immunostaining for CD80 and WT-1 in the glomerulus of MCD patient during relapse showed colocalization.CD80 and synaptopodin co-localized in the glomerulus of MCD patient during relapse.ConclusionThe urinary CD80 is elevated in adult-onset MCD patient in relapse and it is expressed on podocyte,implicating it may be relevant to relapse and diagnosis and prognosis. |
PDF Full Text Request