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Peripheral Blood Mononuclear Cells versus Myelomonocytic Cell Line THP-1 to Test for Immumodulatory Properties of Chemicals

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In the last decade there has been a strong increase in the use of engineered nanoparticles, but despite their growing presence and their exposure to humans only little is known about the potential risks for human health. Immune system regulatory circuits play a crucial role in the pathogenesis of a variety of human diseases including autoimmune syndromes and cancer. Interferences of nanoparticles with the immune system may begin at lower dose than toxicity of vital organ function becomes detectable. Moreover, when nanoparticles interfere with immunosurveillance, this will enhance their disease-propagating activities, e.g., when genotoxic effects may promote tumorigenesis. This review article discusses two cell-based methods which could be easily applied for the detection of immunomodulatory properties of nanoparticles. Th1-type cytokine interferon-γ (IFN-γ) is a central regulator within the pro-inflammatory immune response. Among other biochemical events, IFN-γ induces the enzymes GTP-cyclohydrolase I, which gives rise to neopterin production by activated human macrophages, and indoleamine 2,3-dioxygenase (IDO), which converts tryptophan to kynurenine. Accordingly, enhanced tryptophan degradation and neopterin formation are detected in the blood during several disease states involving a Th1-type immune activation such as virus infections, cancer and cardiovascular diseases and are of strong predictive value. To study the immunomodulatory potential of chemicals, unstimulated and stimulated peripheral blood mononuclear cells (PBMC) and myelomonocytic THP-1-Blue cells are applied in vitro. Neopterin formation (by ELISA) and tryptophan degradation (by HPLC) are monitored in the supernatants. In addition, expression of the central signal-transduction element nuclear factor kappa B (NF-κB) was recorded in THP-1-Blue cells, which are transfected with a corresponding NF-κB gene reporter system. Stimulation of PBMC with mitogen phytohaemagglutinin-L (PHA) strongly induces neopterin formation and tryptophan degradation in vitro. Similar effects are observed in THP-1-Blue cells stimulated with lipopolysaccharide and the close correlation with activation of NF-κB becomes evident. Cannabis compound cannabidiol was used to demonstrate the dose-dependent suppression of all the biochemical read-outs in both in vitro cell-based assays. Analysis of neopterin production and tryptophan degradation in freshly isolated human PBMCs is a useful way to measure the influence of compounds or chemicals on the interaction between T-cells and macrophages. The myelomonocytic cell line THP-1-Blue can be used as a convenient alternative, and both assays can be easily applied for the testing of chemicals but the effects of compounds on the T-cell/macrophage interaction will be missed when THP-1-Blue cells are used.
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Document Type: Research Article

Publication date: December 1, 2012

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  • Bionanoscience attempts to harness various functions of biological macromolecules and integrate them with engineering for technological applications. It is based on a bottom-up approach and encompasses structural biology, biomacromolecular engineering, material science, and engineering, extending the horizon of material science. The journal aims at publication of (i) Letters (ii) Reviews (3) Concepts (4) Rapid communications (5) Research papers (6) Book reviews (7) Conference announcements in the interface between chemistry, physics, biology, material science, and technology. The use of biological macromolecules as sensors, biomaterials, information storage devices, biomolecular arrays, molecular machines is significantly increasing. The traditional disciplines of chemistry, physics, and biology are overlapping and coalescing with nanoscale science and technology. Currently research in this area is scattered in different journals and this journal seeks to bring them under a single umbrella to ensure highest quality peer-reviewed research for rapid dissemination in areas that are in the forefront of science and technology which is witnessing phenomenal and accelerated growth.
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