دورية أكاديمية
Semi-stable Production of Bovine IL-4 and GM-CSF in The Mammalian Episomal Expression System.
العنوان: | Semi-stable Production of Bovine IL-4 and GM-CSF in The Mammalian Episomal Expression System. |
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المؤلفون: | Blanco FC; Institute of Agrobiotechnology and Molecular Biology, National Institute of Agricultural Technology-National Scientific and Technical Research Council, Buenos Aires, Argentina.; Institute of Biotechnology, Centre for Veterinary Scientific and Agronomics Research, National Institute of Agricultural Technology, Nicolás Repetto y De los Reseros s/n, Hurlingham 1686, Buenos Aires, Argentina., Vazquez CL; Institute of Agrobiotechnology and Molecular Biology, National Institute of Agricultural Technology-National Scientific and Technical Research Council, Buenos Aires, Argentina.; Institute of Biotechnology, Centre for Veterinary Scientific and Agronomics Research, National Institute of Agricultural Technology, Nicolás Repetto y De los Reseros s/n, Hurlingham 1686, Buenos Aires, Argentina., García JSY; Institute of Agrobiotechnology and Molecular Biology, National Institute of Agricultural Technology-National Scientific and Technical Research Council, Buenos Aires, Argentina.; Institute of Biotechnology, Centre for Veterinary Scientific and Agronomics Research, National Institute of Agricultural Technology, Nicolás Repetto y De los Reseros s/n, Hurlingham 1686, Buenos Aires, Argentina., Rocha RV; Institute of Agrobiotechnology and Molecular Biology, National Institute of Agricultural Technology-National Scientific and Technical Research Council, Buenos Aires, Argentina.; Institute of Biotechnology, Centre for Veterinary Scientific and Agronomics Research, National Institute of Agricultural Technology, Nicolás Repetto y De los Reseros s/n, Hurlingham 1686, Buenos Aires, Argentina., Gravisaco MJ; Institute of Agrobiotechnology and Molecular Biology, National Institute of Agricultural Technology-National Scientific and Technical Research Council, Buenos Aires, Argentina.; Institute of Biotechnology, Centre for Veterinary Scientific and Agronomics Research, National Institute of Agricultural Technology, Nicolás Repetto y De los Reseros s/n, Hurlingham 1686, Buenos Aires, Argentina., Forrellad MA; Institute of Agrobiotechnology and Molecular Biology, National Institute of Agricultural Technology-National Scientific and Technical Research Council, Buenos Aires, Argentina.; Institute of Biotechnology, Centre for Veterinary Scientific and Agronomics Research, National Institute of Agricultural Technology, Nicolás Repetto y De los Reseros s/n, Hurlingham 1686, Buenos Aires, Argentina., Magistrelli G; Light Chain Bioscience - Novimmune SA, 1228 Plan-Les-Ouates Geneva, Switzerland., Bigi F; Institute of Agrobiotechnology and Molecular Biology, National Institute of Agricultural Technology-National Scientific and Technical Research Council, Buenos Aires, Argentina.; Institute of Biotechnology, Centre for Veterinary Scientific and Agronomics Research, National Institute of Agricultural Technology, Nicolás Repetto y De los Reseros s/n, Hurlingham 1686, Buenos Aires, Argentina. |
المصدر: | Journal of veterinary research [J Vet Res] 2021 Aug 19; Vol. 65 (3), pp. 315-321. Date of Electronic Publication: 2021 Aug 19 (Print Publication: 2021). |
نوع المنشور: | Journal Article |
اللغة: | English |
بيانات الدورية: | Publisher: Sciendo Country of Publication: Poland NLM ID: 101696630 Publication Model: eCollection Cited Medium: Print ISSN: 2450-7393 (Print) Linking ISSN: 24507393 NLM ISO Abbreviation: J Vet Res Subsets: PubMed not MEDLINE |
أسماء مطبوعة: | Publication: [Warsaw, Poland] : Sciendo Original Publication: Pulawy : National Veterinary Research Institute, 2016- |
مستخلص: | Introduction: Granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) are cytokines widely used in ex vivo monocyte differentiation experiments, vaccine formulations and disease treatment. The aim of this study was to produce recombinant bovine GM-CSF and IL-4 in an episomal expression system that conserves the postransductional modification of the native proteins and to use the products to differentiate bovine monocytes into dendritic cells. Material and Methods: The recombinant proteins rGM-CSF and rIL-4 were expressed in PEAKrapid CRL-2828 human kidney cells, ATCC CRL-2828. The functional activity of the recombinant cytokines was monitored by registering morphological changes in bovine monocytes and assessing the expression of CD14 upon incubation with them. Results: Both recombinant proteins were detected in the cell culture supernatant of transfected cells. Culture supernatants of transfected cells induced in bovine monocytes morphological changes that resemble macrophages or dendritic cells. In addition, bovine cells treated with rGM-CSF and rIL-4 showed reduced expression of the macrophage surface marker CD14 compared with untreated cells. This effect indicates the expected differentiation. The expression of the cytokines was stable after many successive cell passages and a freeze/thaw cycle. Conclusions: The semi-stable mammalian episomal expression system used in this study allowed us to easily produce functional bovine rGM-CSF and rIL-4 without the need for protein purification steps. Competing Interests: Conflict of Interest Conflict of Interests Statement: The authors declare that there is no conflict of interests regarding the publication of this article. (© 2021 F.C. Blanco et al. published by Sciendo.) |
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فهرسة مساهمة: | Keywords: bovine GM-CSF; bovine IL-4; dendritic cells; macrophages; mammalian episomal expression |
تواريخ الأحداث: | Date Created: 20211217 Latest Revision: 20220429 |
رمز التحديث: | 20240628 |
مُعرف محوري في PubMed: | PMC8643090 |
DOI: | 10.2478/jvetres-2021-0043 |
PMID: | 34917844 |
قاعدة البيانات: | MEDLINE |
تدمد: | 2450-7393 |
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DOI: | 10.2478/jvetres-2021-0043 |