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

doi:10.1016/S0022-1759(97)00160-9

Journal of Immunological Methods

Volume 209, Issue 2, 1 December 1997, Pages 203-206

https://doi.org/10.1016/S0022-1759(97)00160-9 Get rights and content

Abstract

Quantification of the multifunctional cytokine Transforming Growth Factor-β1 (TGF-β1) in blood samples has aroused increasing interest in recent years, since an abnormal regulation of this cytokine appears to play a key role in the pathogenesis of different diseases, such as autoimmundiseases or malignant tumors. The measurement of TGF-β1 is complicated by a lot of problems concerning the collection, preparation and handling of blood samples, the platelet contamination, and the TGF-β1 activation procedure. Here, we recommend detailed instructions for measurement of TGF-β1 in blood plasma samples which should be followed to exclude the determination of false positive or negative results.

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Acknowledgements

This work was supported by the Deutsche Forschungsgemeinschaft, SFB 387, grant No. A4. The authors thank Mrs. Ines Meinert and Ms. Kathleen Wichmann for the skilled technical assistance.

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Chronic heart failure (CHF) leads to complex effects distant from the heart. As these changes may be reflected in the balance of systemic inflammatory and fibrotic immunomodulators we measured these potential biomarkers in ambulatory CHF patients. Using the New York Heart Association (NYHA; levels II–IV) functional classification, 30 CHF patients were compared with 21 age and gender matched controls. Peripheral blood levels of regulatory cytokines (TNF-α, TGF-β, KGF, IL-8, IL-10 and IL-12) and markers of cellular activation (CD11b, CD16, CD18, CD34, HLADR, CXCR1 and CCR5) were analysed by ELISA and flow cytometry, respectively. NYHA classification, which reflected increasing pulmonary microvascular pressure (E:E′) but not ejection fraction, was positively associated with TGF-β and IL-10 (p ⩽ 0.03). Similarly, monocytes, as well as cell surface expression of the neutrophil adhesion molecule CD11b, and the macrophage complement receptor complex (CD11b/CD18), were increased in CHF patients (p ⩽ 0.03), while the chemokine receptor CXCR1 was decreased on cells of CHF patients. Twenty month follow-up of CHF subjects identified monocyte number as a powerful prognostic factor for cardio-pulmonary adverse events (p = 0.001); however, no concurrent relationship with cellular activation marker expression was found. In subjects with CHF, monocytes, TGF-β, IL-10, CD11b/CD18 and CXCR1 expression in peripheral blood may act as novel biomarkers of immune activation and remodelling. Given the importance of dyspnea and the relationship of pulmonary microvascular pressure to the NYHA classification, we suggest these findings may reflect a contribution by the lung.
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The aim of this study was to investigate the influence of 2 established anesthetic techniques: total intravenous anesthesia and balanced inhalation anesthesia (BAL) on the perioperative-induced changes of peripheral blood mononuclear cells (PBMCs), changes in lymphocyte subsets, and the balance of proinflammatory and anti-inflammatory cytokines.
This is a prospective, randomized, clinical comparison study.
This study was set at a university hospital.
This study involved 50 patients with American Society of Anesthesiologists physical status I who were scheduled for elective minimal invasive partial diskectomy.
There was no intervention involved in this study.
Changes in differential counts, lymphocyte subsets, and proliferation rates were determined before surgery and in the early postoperative period. Plasma concentrations of proinflammatory cytokines (IL-2, IL-6, IL-12, interferon γ) and anti-inflammatory cytokines (IL-10, IL-1RA, transforming growth factor β), and plasma concentrations of cortisol, epinephrine, and norepinephrine were measured before, during, and after surgery.
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Letter to the EditorsA detailed protocol for the measurement of TGF-β1 in human blood samples

Abstract

Section snippets

Acknowledgements

J. Immunol. Methods

Cytokine

Heparin-induced leukocyte lysis in vitro

J. Pharmacobiodyn.

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