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http://hdl.handle.net/1942/34352
Title: | Biological Characterization of Commercial Recombinantly Expressed Immunomodulating Proteins Contaminated with Bacterial Products in the Year 2020: The SAA3 Case | Authors: | Salama, SA DE BONDT, Mirre Berghmans, N GOUWY, Mieke de Oliveira, VLS Oliveira, SC Amaral, FA Proost, P. Van Damme, J Struyf, S De Buck, M |
Issue Date: | 2020 | Publisher: | HINDAWI LTD | Source: | MEDIATORS OF INFLAMMATION, (Art N° 6087109) | Abstract: | The serum amyloid A (SAA) gene family is highly conserved and encodes acute phase proteins that are upregulated in response to inflammatory triggers. Over the years, a considerable amount of literature has been published attributing a wide range of biological effects to SAAs such as leukocyte recruitment, cytokine and chemokine expression and induction of matrix metalloproteinases. Furthermore, SAAs have also been linked to protumorigenic, proatherogenic and anti-inflammatory effects. Here, we investigated the biological effects conveyed by murine SAA3 (mu rSAA3) recombinantly expressed inEscherichia coli. We observed the upregulation of a number of chemokines including CCL2, CCL3, CXCL1, CXCL2, CXCL6 or CXCL8 following stimulation of monocytic, fibroblastoid and peritoneal cells with mu rSAA3. Furthermore, this SAA variant displayed potentin vivorecruitment of neutrophils through the activation of TLR4. However, a major problem associated with proteins derived from recombinant expression in bacteria is potential contamination with various bacterial products, such as lipopolysaccharide, lipoproteins and formylated peptides. This is of particular relevance in the case of SAA as there currently exists a discrepancy in biological activity between SAA derived from recombinant expression and that of an endogenous source, i.e. inflammatory plasma. Therefore, we subjected commercial recombinant mu rSAA3 to purification to homogeneity via reversed-phase high-performance liquid chromatography (RP-HPLC) and re-assessed its biological potential. RP-HPLC-purified mu rSAA3 did not induce chemokines and lackedin vivoneutrophil chemotactic activity, but retained the capacity to synergize with CXCL8 in the activation of neutrophils. In conclusion, experimental results obtained when using proteins recombinantly expressed in bacteria should always be interpreted with care. | Document URI: | http://hdl.handle.net/1942/34352 | ISSN: | 0962-9351 | e-ISSN: | 1466-1861 | DOI: | 10.1155/2020/6087109 | ISI #: | WOS:000553448500001 | Rights: | Copyright © 2020 Sara Abouelasrar Salama et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. | Category: | A1 | Type: | Journal Contribution |
Appears in Collections: | Research publications |
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File | Description | Size | Format | |
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6087109.pdf | Published version | 1.84 MB | Adobe PDF | View/Open |
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