TNF Inhibitory Activity in the Urine of Chronic Renal Failure Patients with Glomerulonephritis

Tomizawa, Shuichi; Suzuki, Jun; Arai, Hideyuki; Seki, Yuhsuke; Shimabukuro, N.; Maruyama, Kazuyasu; Kuroume, Takayoshi

doi:10.1159/000187155

Cited by 3 publications

(6 citation statements)

References 6 publications

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“…Condensed and dialyzed urine was loaded on to 4 liters of DEAE-Sepharose CL-6B column (10x50 cm), eluted with stepwise gradient of NaCI (100 mM, 1.5 M) at a flow rate of 400 ml/h. Protein concentration and TN F inhibitory activity [14] of each fraction was measured.…”

Section: Resultsmentioning

confidence: 99%

“…TNF inhibitory activity was measured as previously described [14], One unit of TNF inhibitory activity was defined as the reciprocal of dilution that blocked 30% of TNF-a-induced cytotoxicity.…”

Section: Bioassay O F Tnf Inhibitorsmentioning

confidence: 99%

“…All the urine of the patient, who showed the highest TNF inhibitory activity among the chronic GN patients [14], was collected and stored at -20 °C.…”

Section: Collection O F the Urinementioning

confidence: 99%

“…Each 5 ml of eluted fraction was lyophilized, then recon stituted with PBS. TNF inhibition rate [14] and ODjso of all the fractions were measured. Active fractions (peaks I and II) were pooled from 20 separations.…”

Section: Fig 2 Purification Of Tnf Inhibitors (Second Step)mentioning

confidence: 99%

“…We have previously reported that the urine from chronic renal failure (CRF) patients with glomerulonephritis (GN) having showed mesangial proliferation and chronic inflam matory changes demonstrated higher titer of TNF inhibi tory activity than CRF patients with congenital renal dis eases [14], In the present communication we report on the purification of two types of TNF inhibitors from the urine of the CRF patient with GN in order to clarify whether TNF inhibitory activity in the urine of these patients depends on the existence of TNF inhibitors/soluble TNF receptors or not. …”

Section: Introductionmentioning

confidence: 99%

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Purification of Two Types of TNF Inhibitors in the Urine of the Patient with Chronic Glomerulonephritis

Suzuki

Tomizawa²,

Arai³

et al. 1994

Nephron

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In order to clarify whether TNF inhibitory activity in the urine of glomerulonephritis (GN) patients depends on the existence of TNF inhibitors/soluble TNF receptors, we purified two types of TNF inhibitors from the urine of chronic renal failure (CRF) with GN patient. Using four steps of chromatography (anion exchange chromatography, preparative reverse phase chromatography, TNF affinity chromatography, analytical reverse phase chromatography), two types of 99% pure TNF inhibitors were purified. Amino terminal amino acid sequence revealed that one inhibitor was identical to soluble TNF receptor, p55, however, the other was homologous but not identical to soluble TNF receptor, p75. Our experiments demonstrated that TNF inhibitory activity in the urine of CRF patients depended partly on the existence of soluble TNF receptors in the urine.

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Section: Resultsmentioning

confidence: 99%

Section: Bioassay O F Tnf Inhibitorsmentioning

confidence: 99%

“…All the urine of the patient, who showed the highest TNF inhibitory activity among the chronic GN patients [14], was collected and stored at -20 °C.…”

Section: Collection O F the Urinementioning

confidence: 99%

Section: Fig 2 Purification Of Tnf Inhibitors (Second Step)mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Purification of Two Types of TNF Inhibitors in the Urine of the Patient with Chronic Glomerulonephritis

Suzuki

Tomizawa²,

Arai³

et al. 1994

Nephron

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Increased Excretion of Tumor Necrosis Factor Alpha and Interleukin 1 β in Urine from Patients with IgA Nephropathy and Schönlein-Henoch Purpura

Wu¹,

Wu²,

Huang³

et al. 1996

Nephron

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Urinary proteins (5 mg/ml) collected from a group of 16 patients including 13 with IgA nephropathy and 3 with Schönlein-Henoch purpura (SHP) and from a control group consisting of 6 patients with diabetic nephropathy, 5 patients with hypertensive nephrosclerosis, and 5 healthy hospital staff members were studied for the contents of interleukins (IL) 1β, 2, 4, 6, and 12 and tumor necrosis factor alpha (TNF-α). Eleven patients with IgA nephropathy or SHP (11/16) but only 1 of the controls (1/16) had TNF-α activity in urinary proteins (p < 0.01). The IL-lβ activity exhibited a similar tendency but to a lesser extent (10 of 16 patients with IgA nephropathy or SHP vs. 2 of 16 with other conditions, p < 0.05). Conversely, the detection rates of IL-2, IL-4, and IL-6 in both groups were not significantly different. IL-12 was not found in any of the samples from both groups. Sera and nonpurified urine samples from the same individuals were also measured for cytokines. IL-lβ, IL-2, IL-4, and IL-12 were absent in all these samples, but TNF-α was found in four of the serum samples from patients with IgA nephropathy. Urinary proteins (2 mg/ml) were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, whereby peptides of 52, 49, 45, 34, 30, and 11 kD could be demonstrated in the patients with IgA nephropathy or SHP. Urinary proteins (200 μg/ml) from patients with IgA nephropathy or SHP exerted a mitogen-like effect on the normal human mononuclear cells, as demonstrated by ³H-thymidine incorporation. In addition, these urinary proteins (400 μg/ml) enhanced the prolifera-tive activity of the cultured rat glomerular mesangial cells. The exaggerated proliferation of rat glomerular mesangial cells exerted by urine proteins from 2 patients with active disease was markedly suppressed after treatment with glu-cocorticoids/cyclophosphamide. These results suggest that patients with IgA nephropathy or SHP can excrete excessive amounts of TNF-α and IL-1 β in the urine. The inconsistent presence of these two cytokines in urine and serum may indicate that they can be produced locally and that they are implicated in the development of mesangial inflammation and glomerular damage.

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Late Expression of Tumor Necrosis Factor-α IS Markedly Depressed in Patients with IGA Nephropathy

Bull

Julian

Jackson

1998

Immunological Investigations

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In this study we demonstrate that peripheral blood mononuclear cells (PBMC) of patients with IgA nephropathy (IgAN) express mRNA for tumor necrosis factor alpha (TNF-alpha) at a significantly lower frequency after stimulation for 24 hours with pokeweed mitogen than do PBMC of a control population. These differences were not seen in patient samples stimulated for only 3 hours. To identify the cells responsible for TNF-alpha expression after overnight mitogen stimulation, we performed reverse transcriptase polymerase chain reaction analysis after preparative flow cytometry of PBMC labelled with fluorescent monoclonal antibodies. After stimulation for 24 hours, PBMC of all patients tested displayed no detectable TNF-alpha mRNA; in the control PBMC, TNF-alpha mRNA was present in CD4+ and CD8+ T cells, as well as in CD19+ B cells in some samples. These results suggest that diminished expression of TNF-alpha by T cells may play a role in the pathogenesis of IgAN.

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TNF Inhibitory Activity in the Urine of Chronic Renal Failure Patients with Glomerulonephritis

Cited by 3 publications

References 6 publications

Purification of Two Types of TNF Inhibitors in the Urine of the Patient with Chronic Glomerulonephritis

Purification of Two Types of TNF Inhibitors in the Urine of the Patient with Chronic Glomerulonephritis

Increased Excretion of Tumor Necrosis Factor Alpha and Interleukin 1 β in Urine from Patients with IgA Nephropathy and Schönlein-Henoch Purpura

Late Expression of Tumor Necrosis Factor-α IS Markedly Depressed in Patients with IGA Nephropathy

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TNF Inhibitory Activity in the Urine of Chronic Renal Failure Patients with Glomerulonephritis

Cited by 3 publications

References 6 publications

Purification of Two Types of TNF Inhibitors in the Urine of the Patient with Chronic Glomerulonephritis

Purification of Two Types of TNF Inhibitors in the Urine of the Patient with Chronic Glomerulonephritis

Increased Excretion of Tumor Necrosis Factor Alpha and Interleukin 1 &beta; in Urine from Patients with IgA Nephropathy and Sch&ouml;nlein-Henoch Purpura

Late Expression of Tumor Necrosis Factor-α IS Markedly Depressed in Patients with IGA Nephropathy

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Increased Excretion of Tumor Necrosis Factor Alpha and Interleukin 1 β in Urine from Patients with IgA Nephropathy and Schönlein-Henoch Purpura