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Effect of serum from patients with mesangial proliferative glomerulonephritis on cultured rat mesangial cells
1Department of Pediatrics, Hirosaki University School of Medicine, Japan.
Abstract:
We studied the effect of serum from patients with renal diseases characterized by mesangial proliferation on the proliferation of rat mesangial cells (MC) in culture. Indirect immunofluorescence technique with human anti-proliferating cell nuclear antigen (PCNA) antibody was utilized to detect proliferating MC in S-phase on microscope slides. MC from serum-starved culture medium containing 0.5% fetal calf serum (FCS) showed 6 +/- 2% of PCNA-positive MC, whereas those from complete medium containing 20% FCS showed 45 +/- 7%. This was confirmed by flow cytometry for MC from both conditions, in which 10.9% and 55.2% of S-phase MC were detected, respectively. Serum from 17 patients with IgA nephropathy as a prototype of mesangial proliferative glomerulonephritis produced more PCNA-positive MC than serum from 18 healthy controls (mean +/- S.D. = 21.5 +/- 9.9% vs. 4.4 +/- 5.9%, p < 0.001). In IgA nephropathy, this effect correlated tentatively with active histologic lesion in the glomeruli, but not with severity of urinary abnormalities. Serum from membrano-proliferative glomerulonephritis (MPGN) also produced significant PCNA-positive MC (mean +/- S.D. = 15.6 +/- 9.3%), compared to controls (p < 0.01), independent of the severity of urinary abnormalities. These findings suggest that serum effect on the proliferation of cultured rat MC may reflect an active histologic lesion in the glomeruli, and that anti-PCNA antibody to detect proliferating MC is, if not quantitative, relevant methodology to enumerate proliferating MC in culture.
Insights
Patient serum from IgA nephropathy and MPGN induced significant rat mesangial cell proliferation, detected by anti-proliferating cell nuclear antigen (PCNA) antibody. This proliferation may reflect active kidney disease lesions.
Area of Science:
- Nephrology
- Cell Biology
- Immunology
Background:
- Mesangial cell proliferation is a hallmark of several glomerulonephritides.
- Understanding factors driving mesangial cell proliferation is crucial for disease management.
Purpose of the Study:
- To investigate the effect of serum from patients with mesangial proliferative renal diseases on rat mesangial cell (MC) proliferation.
- To assess the correlation between serum-induced MC proliferation and histologic activity in IgA nephropathy.
Main Methods:
- Cultured rat MC proliferation was assessed using indirect immunofluorescence for anti-proliferating cell nuclear antigen (PCNA) antibody.
- Flow cytometry was used to quantify S-phase MC.
- Serum from patients with IgA nephropathy and membrano-proliferative glomerulonephritis (MPGN) was tested against serum from healthy controls.
Main Results:
- Serum from patients with IgA nephropathy significantly increased PCNA-positive MC compared to controls (21.5% vs 4.4%, p < 0.001).
- Serum from MPGN patients also significantly increased PCNA-positive MC compared to controls (15.6% vs 4.4%, p < 0.01).
- In IgA nephropathy, serum-induced proliferation correlated with active glomerular histology but not urinary abnormalities.
Conclusions:
- Serum from patients with mesangial proliferative glomerulonephritis stimulates rat MC proliferation in vitro.
- This in vitro proliferation may serve as a biomarker for active renal histologic lesions.
- Anti-PCNA antibody is a relevant method for enumerating proliferating MC in culture.