The pre-induction temperature affects recombinant HuGM-CSF aggregation in thermoinducible Escherichia coli

Restrepo-Pineda, Sara; Sánchez-Puig, Nuria; Pérez, Nestor O.; García‐Hernández, Enrique; Valdez‐Cruz, Norma A.; Trujillo‐Roldán, Mauricio A.

doi:10.1007/s00253-022-11908-z

Cited by 7 publications

(5 citation statements)

References 127 publications

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“…Macrophages are the first encounters with invading bacteria and initiate immune responses to pathogens delivered through circulation (1)(2)(3). The function of macrophages is finely tuned by multiple environmental factors, such as temperature, pH, and oxygen concentration (4)(5)(6). Recently, several studies have shown that multiple types of mechanical stresses, including hydrostatic pressure, shear stress, and tensile force, are closely associated with inflammatory responses (7)(8)(9)(10).…”

Section: Introductionmentioning

confidence: 99%

Ion channel Piezo1 activation promotes aerobic glycolysis in macrophages

et al. 2022

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Altered microenvironmental stiffness is a hallmark of inflammation. It is sensed by the mechanically activated cation channel Piezo1 in macrophages to induce subsequent immune responses. However, the mechanism by which the mechanosensitive signals shape the metabolic status of macrophages and tune immune responses remains unclear. We revealed that Piezo1-deficient macrophages exhibit reduced aerobic glycolysis in resting or liposaccharide (LPS)-stimulated macrophages with impaired LPS-induced secretion of inflammatory cytokines in vitro. Additionally, pretreatment with the Piezo1 agonist, Yoda1, or cyclical hydrostatic pressure (CHP) upregulated glycolytic activity and enhanced LPS-induced secretion of inflammatory cytokines. Piezo1-deficient mice were less susceptible to dextran sulfate sodium (DSS)-induced colitis, whereas Yoda1 treatment aggravated colitis. Mechanistically, we found that Piezo1 activation promotes aerobic glycolysis through the Ca2+-induced CaMKII-HIF1α axis. Therefore, our study revealed that Piezo1-mediated mechanosensitive signals Piezo1 can enhance aerobic glycolysis and promote the LPS-induced immune response in macrophages.

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Section: Introductionmentioning

confidence: 99%

Ion channel Piezo1 activation promotes aerobic glycolysis in macrophages

et al. 2022

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“…The recombinant proteins produced in the bioreactor and isolated from IBs were solubilized, separated by gel chromatography, and recovered in MQ water by electro-elution [ 54 ]. Around 50% of the recombinant proteins were recovered.…”

Section: Resultsmentioning

confidence: 99%

“…Recombinant RBD-P and RBD expression was induced by adding isopropyl-β- d -1-thiogalactopyranoside (IPTG, 0.1 mM, Merck-Sigma-Aldrich, USA) after 6 h of culture. The composition of the mineral media used was described by Restrepo-Pineda et al [ 54 ]. The growth of E. coli cultures was followed by optical density and converted to dry cell weight (DCW) by a linear-correlation standard curve, where 1 A.U.…”

Section: Methodsmentioning

confidence: 99%

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Oral administration of a recombinant modified RBD antigen of SARS-CoV-2 as a possible immunostimulant for the care of COVID-19

Valdez‑Cruz,

Rosiles-Becerril,

Martínez-Olivares

et al. 2024

Microb Cell Fact

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Background Developing effective vaccines against SARS-CoV-2 that consider manufacturing limitations, equitable access, and acceptance is necessary for developing platforms to produce antigens that can be efficiently presented for generating neutralizing antibodies and as a model for new vaccines. Results This work presents the development of an applicable technology through the oral administration of the SARS-CoV-2 RBD antigen fused with a peptide to improve its antigenic presentation. We focused on the development and production of the recombinant receptor binding domain (RBD) produced in E. coli modified with the addition of amino acids extension designed to improve antigen presentation. The production was carried out in shake flask and bioreactor cultures, obtaining around 200 mg/L of the antigen. The peptide-fused RBD and peptide-free RBD proteins were characterized and compared using SDS-PAGE gel, high-performance chromatography, and circular dichroism. The peptide-fused RBD was formulated in an oil-in-water emulsion for oral mice immunization. The peptide-fused RBD, compared to RBD, induced robust IgG production in mice, capable of recognizing the recombinant RBD in Enzyme-linked immunosorbent assays. In addition, the peptide-fused RBD generated neutralizing antibodies in the sera of the dosed mice. The formulation showed no reactive episodes and no changes in temperature or vomiting. Conclusions Our study demonstrated the effectiveness of the designed peptide added to the RBD to improve antigen immunostimulation by oral administration. Graphical Abstract

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“…In this way, cI857 permits transcription by the RNA polymerase at elevated temperatures. Due to this property, the λpL/pR‐cI857 system was developed as a useful thermo‐regulated expression system for heterologous protein production in Escherichia coli , phages and viruses (Ahmad et al ., 2018 ; Menart et al ., 2003 ; Restrepo‐Pineda et al ., 2022 ; Rosano and Ceccarelli, 2014 ; Valdez‐Cruz et al ., 2010 ). Here, we have explored the possibility of engineering the λpL/pR‐cI857 expression system to function as thermo‐regulator of transgene expression in plastids.…”

Section: Introductionmentioning

confidence: 99%

A heat‐inducible expression system for external control of gene expression in plastids

Xu,

Li,

Bock

et al. 2023

Plant Biotechnology Journal

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SummaryInducible expression systems can overcome the trade‐off between high‐level transgene expression and its pleiotropic effects on plant growth. In addition, they can facilitate the expression of biochemical pathways that produce toxic metabolites. Although a few inducible expression systems for the control of transgene expression in plastids have been developed, they all depend on chemical inducers and/or nuclear transgenes. Here we report a temperature‐inducible expression system for plastids that is based on the bacteriophage λ leftward and rightward promoters (pL/pR) and the temperature‐sensitive repressor cI857. We show that the expression of green fluorescent protein (GFP) in plastids can be efficiently repressed by cI857 under normal growth conditions, and becomes induced over time upon exposure to elevated temperatures in a light‐dependent process. We further demonstrate that by introducing into plastids an expression system based on the bacteriophage T7 RNA polymerase, the temperature‐dependent accumulation of GFP increased further and was ~24 times higher than expression driven by the pL/pR promoter alone, reaching ~0.48% of the total soluble protein. In conclusion, our heat‐inducible expression system provides a new tool for the external control of plastid (trans) gene expression that is cost‐effective and does not depend on chemical inducers.

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The pre-induction temperature affects recombinant HuGM-CSF aggregation in thermoinducible Escherichia coli

Cited by 7 publications

References 127 publications

Ion channel Piezo1 activation promotes aerobic glycolysis in macrophages

Ion channel Piezo1 activation promotes aerobic glycolysis in macrophages

Oral administration of a recombinant modified RBD antigen of SARS-CoV-2 as a possible immunostimulant for the care of COVID-19

A heat‐inducible expression system for external control of gene expression in plastids

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