A detailed protocol for the measurement of TGF-β1 in human blood samples

Reinhold, Dirk; Bank, Ute; Bühling, Frank; Junker, U.; Kekow, Jörn; Schleicher, Erwin; Ansorge, Siegfried

doi:10.1016/s0022-1759(97)00160-9

Cited by 38 publications

(19 citation statements)

References 4 publications

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“…The samples were collected into sterile citrate tubes, which were immediately immersed in ice, and processed within 30 min after collection. Cells were sedimented by centrifugation at 700 g for 10 min at 4°C; then supernatant plasma was collected and respun for another 20 min at 1300 g to sediment platelets (23). Cell-free plasma was aliquoted into 0.5 mL samples and stored at Ϫ80°C until measurements.…”

Section: Methodsmentioning

confidence: 99%

Immune Mediators in Idiopathic Nephrotic Syndrome: Evidence for a Relation Between Interleukin 8 and Proteinuria

et al. 2008

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ABSTRACT:The pathogenesis of idiopathic nephrotic syndrome (INS) remains unknown. Several findings suggest a role for the immune system. This study aimed to evaluate immune mediators in INS by measuring plasma and urinary levels of transforming growth factor ␤ 1 (TGF-␤ 1 ), monocyte chemoattractant protein-1 (MCP-1/ CCL2), regulated on activation normal T-cell expressed and secreted (RANTES/CCL5) and IL-8 (IL-8/CXCL8) in pediatric patients with INS and in age-matched healthy controls. Patients were divided according to their response to corticosteroids: steroid-sensitive (SS, n ϭ 8), or steroid-resistant (SR, n ϭ 24). Immune mediators were also compared in regard with disease activity (relapse and remission). Immune mediators were measured by ELISA. Plasma TGF-␤ 1 levels in SR patients were approximately 2.8-fold higher than control values (p Ͻ 0.05). Urinary IL-8/CXCL8 was 2.9-fold higher in INS patients in relapse (proteinuria Ͼ100 mg/m 2 /24 h) when compared with patients in remission (p Ͻ 0.05), and levels had a positive correlation with individual proteinuria values (p Ͻ 0.05). Urinary IL-8/CXCL8 was significantly higher in relapsed SR than in SS patients in remission. No changes in MCP-1/CCL2 and RANTES/CCL5 levels were detected. Our findings suggest that IL-8/CXCL8 and TGF-␤ 1 are involved in the pathogenesis of INS: IL-8/CXCL8 associated with local changes in glomerular permeability and TGF-␤ 1 could be related to worse response to corticosteroids. (Pediatr Res 64: 637-642, 2008)

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Section: Methodsmentioning

confidence: 99%

Immune Mediators in Idiopathic Nephrotic Syndrome: Evidence for a Relation Between Interleukin 8 and Proteinuria

et al. 2008

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“…Intraassay and interassay coefficients of variations of these measurements were less than 10% in each determination. The total concentrations of latent transforming growth factor (TGF) b1 were measured as described previously [5].…”

Section: Measurement Of Cytokine Concentrationsmentioning

confidence: 99%

Effect of 2 anesthetic techniques on the postoperative proinflammatory and anti-inflammatory cytokine response and cellular immune function to minor surgery

Schneemilch

Ittenson

Ansorge

et al. 2005

Journal of Clinical Anesthesia

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“…Plasma was obtained from deeply anesthetized mice by drawing 0.5 ml cardiac blood into a syringe containing 50 l citrate as anticoagulant (ACD solution A; Becton Dickinson, Franklin Lakes, NJ). For the ELISA, a mouse monoclonal anti-TGF-␤1, -␤2, -␤3 Ab (Genzyme) and a chicken anti-TGF-␤1 Ab (R&D Systems) were used (40). To release latent TGF-␤1, samples were tested before and after transient acidification (40).…”

Section: Cytokine Measurementsmentioning

confidence: 99%

“…For the ELISA, a mouse monoclonal anti-TGF-␤1, -␤2, -␤3 Ab (Genzyme) and a chicken anti-TGF-␤1 Ab (R&D Systems) were used (40). To release latent TGF-␤1, samples were tested before and after transient acidification (40). TGF-␤1 concentrations in CNS and plasma were compared by a one-way ANOVA and the Tukey test for multiple pairwise comparisons (41).…”

Section: Cytokine Measurementsmentioning

confidence: 99%

Targeting Dipeptidyl Peptidase IV (CD26) Suppresses Autoimmune Encephalomyelitis and Up-Regulates TGF-β1 Secretion In Vivo

Steinbrecher

Reinhold

Quigley

et al. 2001

The Journal of Immunology

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139

115

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CD26 or dipeptidyl peptidase IV (DP IV) is expressed on various cell types, including T cells. Although T cells can receive activating signals via CD26, the physiological role of CD26/DP IV is largely unknown. We used the reversible DP IV inhibitor Lys[Z(NO2)]-pyrrolidide (I40) to dissect the role of DP IV in experimental autoimmune encephalomyelitis (EAE) and to explore the therapeutic potential of DP IV inhibition for autoimmunity. I40 administration in vivo decreased and delayed clinical and neuropathological signs of adoptive transfer EAE. I40 blocked DP IV activity in vivo and increased the secretion of the immunosuppressive cytokine TGF-β1 in spinal cord tissue and plasma during acute EAE. In vitro, while suppressing autoreactive T cell proliferation and TNF-α production, I40 consistently up-regulated TGF-β1 secretion. A neutralizing anti-TGF-β1 Ab blocked the inhibitory effect of I40 on T cell proliferation to myelin Ag. DP IV inhibition in vivo was not generally immunosuppressive, neither eliminating encephalitogenic T cells nor inhibiting T cell priming. These data suggest that DP IV inhibition represents a novel and specific therapeutic approach protecting from autoimmune disease by a mechanism that includes an active TGF-β1-mediated antiinflammatory effect at the site of pathology.

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A detailed protocol for the measurement of TGF-β1 in human blood samples

Cited by 38 publications

References 4 publications

Immune Mediators in Idiopathic Nephrotic Syndrome: Evidence for a Relation Between Interleukin 8 and Proteinuria

Immune Mediators in Idiopathic Nephrotic Syndrome: Evidence for a Relation Between Interleukin 8 and Proteinuria

Effect of 2 anesthetic techniques on the postoperative proinflammatory and anti-inflammatory cytokine response and cellular immune function to minor surgery

Targeting Dipeptidyl Peptidase IV (CD26) Suppresses Autoimmune Encephalomyelitis and Up-Regulates TGF-β1 Secretion In Vivo

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