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Comparison of in vitro tests for antioxidant and immunomodulatory capacities of compounds.
Phytomedicine 2014; 21(2):164-71P

Abstract

Oxidative stress is considered to be critically involved in the normal aging process but also in the development and progression of various human pathologies like cardiovascular and neurodegenerative diseases, as well as of infections and malignant tumors. These pathological conditions involve an overwhelming production of reactive oxygen species (ROS), which are released as part of an anti-proliferative strategy during pro-inflammatory immune responses. Moreover, ROS themselves are autocrine forward regulators of the immune response. Most of the beneficial effects of antioxidants are considered to derive from their influence on the immune system. Due to their antioxidant and/or radical scavenging nature, phytochemicals, botanicals and herbal preparations can be of great importance to prevent oxidation processes and to counteract the activation of redox-regulated signaling pathways. Antioxidants can antagonize the activation of T-cells and macrophages during the immune response and this anti-inflammatory activity could be of utmost importance for the treatment of above-mentioned disorders and for the development of immunotolerance. Herein, we provide an overview of in vitro assays for the measurement of antioxidant and anti-inflammatory activities of plant-derived substances and extracts, by discussing possibilities and limitations of these methods. To determine the capacity of antioxidants, the oxygen radical absorbance capacity (ORAC) assay and the cell-based antioxidant activity (CAA) assay are widely applied. To examine the influence of compounds on the human immune response more closely, the model of mitogen stimulated human peripheral blood mononuclear (PBMC) cells can be applied, and the production of the inflammatory marker neopterin as well as the breakdown of the amino acid tryptophan in culture supernatants can be used as readout to indicate an immunomodulatory potential of the tested compound. These two biomarkers of immune system activation are robust and correlate with the course of cardiovascular, neurodegenerative and malignant tumor diseases, but also with the normal aging process, and they are strongly predictive. Thus, while the simpler ORAC and CAA assays provide insight into one peculiar chemical aspect, namely the neutralization of peroxyl radicals, the more complex PBMC assay is closer to the in vivo conditions as the assay comprehensively enlights several properties of immunomodulatory test compounds.

Authors+Show Affiliations

Division of Medical Biochemistry, Biocenter, Medical University Innsbruck, Innsbruck, Austria.Division of Biological Chemistry, Medical University Innsbruck, Innsbruck, Austria.Division of Medical Biochemistry, Biocenter, Medical University Innsbruck, Innsbruck, Austria.Division of Biological Chemistry, Medical University Innsbruck, Innsbruck, Austria.Central Institute of Blood Transfusion and Immunology, University Hospital Innsbruck, Innsbruck, Austria.Division of Medical Biochemistry, Biocenter, Medical University Innsbruck, Innsbruck, Austria.Division of Biological Chemistry, Medical University Innsbruck, Innsbruck, Austria. Electronic address: dietmar.fuchs@i-med.ac.at.

Pub Type(s)

Comparative Study
Journal Article
Review

Language

eng

PubMed ID

24041614

Citation

Becker, Kathrin, et al. "Comparison of in Vitro Tests for Antioxidant and Immunomodulatory Capacities of Compounds." Phytomedicine : International Journal of Phytotherapy and Phytopharmacology, vol. 21, no. 2, 2014, pp. 164-71.
Becker K, Schroecksnadel S, Gostner J, et al. Comparison of in vitro tests for antioxidant and immunomodulatory capacities of compounds. Phytomedicine. 2014;21(2):164-71.
Becker, K., Schroecksnadel, S., Gostner, J., Zaknun, C., Schennach, H., Uberall, F., & Fuchs, D. (2014). Comparison of in vitro tests for antioxidant and immunomodulatory capacities of compounds. Phytomedicine : International Journal of Phytotherapy and Phytopharmacology, 21(2), pp. 164-71. doi:10.1016/j.phymed.2013.08.008.
Becker K, et al. Comparison of in Vitro Tests for Antioxidant and Immunomodulatory Capacities of Compounds. Phytomedicine. 2014 Jan 15;21(2):164-71. PubMed PMID: 24041614.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Comparison of in vitro tests for antioxidant and immunomodulatory capacities of compounds. AU - Becker,Kathrin, AU - Schroecksnadel,Sebastian, AU - Gostner,Johanna, AU - Zaknun,Cathrine, AU - Schennach,Harald, AU - Uberall,Florian, AU - Fuchs,Dietmar, Y1 - 2013/09/14/ PY - 2013/02/18/received PY - 2013/07/09/revised PY - 2013/08/09/accepted PY - 2013/9/18/entrez PY - 2013/9/18/pubmed PY - 2014/9/10/medline KW - ARE KW - CAA KW - Cell-based antioxidant activity (CAA) KW - GTP-CH1 KW - GTP-cyclohydrolase I KW - IDO KW - IFN-γ KW - IL-2 KW - Immunomodulation KW - In vitro test systems KW - Inflammation KW - Kyn/Trp KW - NO KW - Nf-κB KW - ORAC KW - Oxidative stress KW - Oxygen radical absorbance capacity (ORAC) assay KW - PBMC KW - Peripheral blood mononuclear cells KW - ROS KW - Reactive oxygen species KW - TNF-α KW - antioxidant response element KW - cell-based antioxidant activity KW - iNOS KW - indoleamine 2,3-dioxygenase KW - inducible nitric oxide synthase KW - interferon-gamma KW - interleukine-2 KW - kynurenine to tryptophan ratio KW - nitric oxide KW - nuclear factor-κB KW - oxygen radical absorbance capacity KW - peripheral blood mononuclear cells KW - reactive oxygen species KW - tumor necrosis factor-alpha SP - 164 EP - 71 JF - Phytomedicine : international journal of phytotherapy and phytopharmacology JO - Phytomedicine VL - 21 IS - 2 N2 - Oxidative stress is considered to be critically involved in the normal aging process but also in the development and progression of various human pathologies like cardiovascular and neurodegenerative diseases, as well as of infections and malignant tumors. These pathological conditions involve an overwhelming production of reactive oxygen species (ROS), which are released as part of an anti-proliferative strategy during pro-inflammatory immune responses. Moreover, ROS themselves are autocrine forward regulators of the immune response. Most of the beneficial effects of antioxidants are considered to derive from their influence on the immune system. Due to their antioxidant and/or radical scavenging nature, phytochemicals, botanicals and herbal preparations can be of great importance to prevent oxidation processes and to counteract the activation of redox-regulated signaling pathways. Antioxidants can antagonize the activation of T-cells and macrophages during the immune response and this anti-inflammatory activity could be of utmost importance for the treatment of above-mentioned disorders and for the development of immunotolerance. Herein, we provide an overview of in vitro assays for the measurement of antioxidant and anti-inflammatory activities of plant-derived substances and extracts, by discussing possibilities and limitations of these methods. To determine the capacity of antioxidants, the oxygen radical absorbance capacity (ORAC) assay and the cell-based antioxidant activity (CAA) assay are widely applied. To examine the influence of compounds on the human immune response more closely, the model of mitogen stimulated human peripheral blood mononuclear (PBMC) cells can be applied, and the production of the inflammatory marker neopterin as well as the breakdown of the amino acid tryptophan in culture supernatants can be used as readout to indicate an immunomodulatory potential of the tested compound. These two biomarkers of immune system activation are robust and correlate with the course of cardiovascular, neurodegenerative and malignant tumor diseases, but also with the normal aging process, and they are strongly predictive. Thus, while the simpler ORAC and CAA assays provide insight into one peculiar chemical aspect, namely the neutralization of peroxyl radicals, the more complex PBMC assay is closer to the in vivo conditions as the assay comprehensively enlights several properties of immunomodulatory test compounds. SN - 1618-095X UR - https://www.unboundmedicine.com/medline/citation/24041614/Comparison_of_in_vitro_tests_for_antioxidant_and_immunomodulatory_capacities_of_compounds_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0944-7113(13)00311-5 DB - PRIME DP - Unbound Medicine ER -