Biological Characterization of Commercial Recombinantly Expressed Immunomodulating Proteins Contaminated with Bacterial Products in the Year 2020: The SAA3 Case
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 recrui...
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2020-01-01
|
| Series: | Mediators of Inflammation |
| Online Access: | http://dx.doi.org/10.1155/2020/6087109 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849399514244317184 |
|---|---|
| author | Sara Abouelasrar Salama Mirre De Bondt Nele Berghmans Mieke Gouwy Vivian Louise Soares de Oliveira Sergio C. Oliveira Flavio A. Amaral Paul Proost Jo Van Damme Sofie Struyf Mieke De Buck |
| author_facet | Sara Abouelasrar Salama Mirre De Bondt Nele Berghmans Mieke Gouwy Vivian Louise Soares de Oliveira Sergio C. Oliveira Flavio A. Amaral Paul Proost Jo Van Damme Sofie Struyf Mieke De Buck |
| author_sort | Sara Abouelasrar Salama |
| collection | DOAJ |
| description | 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 in Escherichia 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 potent in vivo recruitment 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 lacked in vivo neutrophil 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. |
| format | Article |
| id | doaj-art-9d98eb1390c849428a2061beef83c39f |
| institution | Kabale University |
| issn | 0962-9351 1466-1861 |
| language | English |
| publishDate | 2020-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Mediators of Inflammation |
| spelling | doaj-art-9d98eb1390c849428a2061beef83c39f2025-08-20T03:38:19ZengWileyMediators of Inflammation0962-93511466-18612020-01-01202010.1155/2020/60871096087109Biological Characterization of Commercial Recombinantly Expressed Immunomodulating Proteins Contaminated with Bacterial Products in the Year 2020: The SAA3 CaseSara Abouelasrar Salama0Mirre De Bondt1Nele Berghmans2Mieke Gouwy3Vivian Louise Soares de Oliveira4Sergio C. Oliveira5Flavio A. Amaral6Paul Proost7Jo Van Damme8Sofie Struyf9Mieke De Buck10KU Leuven, Department of Microbiology and Immunology, Rega Institute, Laboratory of Molecular Immunology, Herestraat 49, Box 1042, 3000 Leuven, BelgiumKU Leuven, Department of Microbiology and Immunology, Rega Institute, Laboratory of Molecular Immunology, Herestraat 49, Box 1042, 3000 Leuven, BelgiumKU Leuven, Department of Microbiology and Immunology, Rega Institute, Laboratory of Molecular Immunology, Herestraat 49, Box 1042, 3000 Leuven, BelgiumKU Leuven, Department of Microbiology and Immunology, Rega Institute, Laboratory of Molecular Immunology, Herestraat 49, Box 1042, 3000 Leuven, BelgiumImunofarmacologia, Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, Pampulha, Belo Horizonte, 31270-901 Minas Gerais, BrazilDepartamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, Pampulha, Belo Horizonte, 31270-901 Minas Gerais, BrazilImunofarmacologia, Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, Pampulha, Belo Horizonte, 31270-901 Minas Gerais, BrazilKU Leuven, Department of Microbiology and Immunology, Rega Institute, Laboratory of Molecular Immunology, Herestraat 49, Box 1042, 3000 Leuven, BelgiumKU Leuven, Department of Microbiology and Immunology, Rega Institute, Laboratory of Molecular Immunology, Herestraat 49, Box 1042, 3000 Leuven, BelgiumKU Leuven, Department of Microbiology and Immunology, Rega Institute, Laboratory of Molecular Immunology, Herestraat 49, Box 1042, 3000 Leuven, BelgiumKU Leuven, Department of Microbiology and Immunology, Rega Institute, Laboratory of Molecular Immunology, Herestraat 49, Box 1042, 3000 Leuven, BelgiumThe 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 in Escherichia 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 potent in vivo recruitment 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 lacked in vivo neutrophil 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.http://dx.doi.org/10.1155/2020/6087109 |
| spellingShingle | Sara Abouelasrar Salama Mirre De Bondt Nele Berghmans Mieke Gouwy Vivian Louise Soares de Oliveira Sergio C. Oliveira Flavio A. Amaral Paul Proost Jo Van Damme Sofie Struyf Mieke De Buck Biological Characterization of Commercial Recombinantly Expressed Immunomodulating Proteins Contaminated with Bacterial Products in the Year 2020: The SAA3 Case Mediators of Inflammation |
| title | Biological Characterization of Commercial Recombinantly Expressed Immunomodulating Proteins Contaminated with Bacterial Products in the Year 2020: The SAA3 Case |
| title_full | Biological Characterization of Commercial Recombinantly Expressed Immunomodulating Proteins Contaminated with Bacterial Products in the Year 2020: The SAA3 Case |
| title_fullStr | Biological Characterization of Commercial Recombinantly Expressed Immunomodulating Proteins Contaminated with Bacterial Products in the Year 2020: The SAA3 Case |
| title_full_unstemmed | Biological Characterization of Commercial Recombinantly Expressed Immunomodulating Proteins Contaminated with Bacterial Products in the Year 2020: The SAA3 Case |
| title_short | Biological Characterization of Commercial Recombinantly Expressed Immunomodulating Proteins Contaminated with Bacterial Products in the Year 2020: The SAA3 Case |
| title_sort | biological characterization of commercial recombinantly expressed immunomodulating proteins contaminated with bacterial products in the year 2020 the saa3 case |
| url | http://dx.doi.org/10.1155/2020/6087109 |
| work_keys_str_mv | AT saraabouelasrarsalama biologicalcharacterizationofcommercialrecombinantlyexpressedimmunomodulatingproteinscontaminatedwithbacterialproductsintheyear2020thesaa3case AT mirredebondt biologicalcharacterizationofcommercialrecombinantlyexpressedimmunomodulatingproteinscontaminatedwithbacterialproductsintheyear2020thesaa3case AT neleberghmans biologicalcharacterizationofcommercialrecombinantlyexpressedimmunomodulatingproteinscontaminatedwithbacterialproductsintheyear2020thesaa3case AT miekegouwy biologicalcharacterizationofcommercialrecombinantlyexpressedimmunomodulatingproteinscontaminatedwithbacterialproductsintheyear2020thesaa3case AT vivianlouisesoaresdeoliveira biologicalcharacterizationofcommercialrecombinantlyexpressedimmunomodulatingproteinscontaminatedwithbacterialproductsintheyear2020thesaa3case AT sergiocoliveira biologicalcharacterizationofcommercialrecombinantlyexpressedimmunomodulatingproteinscontaminatedwithbacterialproductsintheyear2020thesaa3case AT flavioaamaral biologicalcharacterizationofcommercialrecombinantlyexpressedimmunomodulatingproteinscontaminatedwithbacterialproductsintheyear2020thesaa3case AT paulproost biologicalcharacterizationofcommercialrecombinantlyexpressedimmunomodulatingproteinscontaminatedwithbacterialproductsintheyear2020thesaa3case AT jovandamme biologicalcharacterizationofcommercialrecombinantlyexpressedimmunomodulatingproteinscontaminatedwithbacterialproductsintheyear2020thesaa3case AT sofiestruyf biologicalcharacterizationofcommercialrecombinantlyexpressedimmunomodulatingproteinscontaminatedwithbacterialproductsintheyear2020thesaa3case AT miekedebuck biologicalcharacterizationofcommercialrecombinantlyexpressedimmunomodulatingproteinscontaminatedwithbacterialproductsintheyear2020thesaa3case |