Extraction and purification of hepatitis B virus-like M particles from a recombinant Saccharomyces cerevisiae strain using alumina powder

Hadiji-Abbes, Nadia; Martin, Marta; Benzina, Wafa; Karray-Hakim, Héla; Gergely, Csilla; Gargouri, Ali; Mokdad-Gargouri, Raja

doi:10.1016/j.jviromet.2012.09.023

Cited by 18 publications

(9 citation statements)

References 31 publications

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“…Despite debate on the use of hydroxychloroquine in sepsis, especially in acute respiratory distress syndrome from Coronavirus (COVID-19) infection [86,87], there might be some sepsis conditions that are beneficial from TLR-9 inhibition and bacterial DNA neutralization [88]. Several extracorporeal blood purification procedures reduce the free DNA particles in either sepsis [29] or non-sepsis (viral infection) [89][90][91], which could be adapted for the removal of bacteria-free DNA in sepsis.…”

Section: Discussionmentioning

confidence: 99%

Blood Bacteria-Free DNA in Septic Mice Enhances LPS-Induced Inflammation in Mice through Macrophage Response

Kaewduangduen

Visitchanakun

Saisorn

et al. 2022

IJMS

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Although bacteria-free DNA in blood during systemic infection is mainly derived from bacterial death, translocation of the DNA from the gut into the blood circulation (gut translocation) is also possible. Hence, several mouse models with experiments on macrophages were conducted to explore the sources, influences, and impacts of bacteria-free DNA in sepsis. First, bacteria-free DNA and bacteriome in blood were demonstrated in cecal ligation and puncture (CLP) sepsis mice. Second, administration of bacterial lysate (a source of bacterial DNA) in dextran sulfate solution (DSS)-induced mucositis mice elevated blood bacteria-free DNA without bacteremia supported gut translocation of free DNA. The absence of blood bacteria-free DNA in DSS mice without bacterial lysate implies an impact of the abundance of bacterial DNA in intestinal contents on the translocation of free DNA. Third, higher serum cytokines in mice after injection of combined bacterial DNA with lipopolysaccharide (LPS), when compared to LPS injection alone, supported an influence of blood bacteria-free DNA on systemic inflammation. The synergistic effects of free DNA and LPS on macrophage pro-inflammatory responses, as indicated by supernatant cytokines (TNF-α, IL-6, and IL-10), pro-inflammatory genes (NFκB, iNOS, and IL-1β), and profound energy alteration (enhanced glycolysis with reduced mitochondrial functions), which was neutralized by TLR-9 inhibition (chloroquine), were demonstrated. In conclusion, the presence of bacteria-free DNA in sepsis mice is partly due to gut translocation of bacteria-free DNA into the systemic circulation, which would enhance sepsis severity. Inhibition of the responses against bacterial DNA by TLR-9 inhibition could attenuate LPS-DNA synergy in macrophages and might help improve sepsis hyper-inflammation in some situations.

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Section: Discussionmentioning

confidence: 99%

Blood Bacteria-Free DNA in Septic Mice Enhances LPS-Induced Inflammation in Mice through Macrophage Response

Kaewduangduen

Visitchanakun

Saisorn

et al. 2022

IJMS

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“…The study also showed that the HBsAg did not assemble into VLPs within the yeast cells, and the particles are most likely formed during the downstream purification process. Purification of HBsAg VLPs produced by Saccharomyces cerevisiae involved laborious steps, which include the lysis of cells with detergent and alumina, followed by lyophilisation of the crude lysate, and separation of the particles using sepharyl-S column chromatography and size-exclusion chromatography [23]. Since the 1990s, expression of HBsAg VLPs has been performed in plants, particularly the tobacco [24].…”

Section: Discussionmentioning

confidence: 99%

Immunological Analysis of the Hepatitis B Virus “a” Determinant Displayed on Chimeric Virus-Like Particles of Macrobrachium rosenbergii Nodavirus Capsid Protein Produced in Sf9 Cells

Ninyio

Ong

et al. 2020

Vaccines

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Chimeric virus-like particles (VLPs) have been widely exploited for various purposes including their use as vaccine candidates, particularly due to their ability to induce stronger immune responses than VLPs consisting of single viral proteins. In the present study, VLPs of the Macrobrachium rosenbergii nodavirus (MrNV) capsid protein (Nc) displaying the hepatitis B virus “a” determinant (aD) were produced in Spodoptera frugiperda (Sf9) insect cells. BALB/c mice immunised with the purified chimeric Nc-aD VLPs elicited a sustained titre of anti-aD antibody, which was significantly higher than that elicited by a commercially available hepatitis B vaccine and Escherichia coli-produced Nc-aD VLPs. Immunophenotyping showed that the Sf9-produced Nc-aD VLPs induced proliferation of cytotoxic T-lymphocytes and NK1.1 natural killer cells. Furthermore, enzyme-linked immunospot (ELISPOT)analysis showed the presence of antibody-secreting memory B cells in the mice splenocytes stimulated with the synthetic aD peptide. The significant humoral, natural killer cell and memory B cell immune responses induced by the Sf9-produced Nc-aD VLPs suggest that they present good prospects for use as a hepatitis B vaccine candidate.

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“…of HBsAg-recombinant vaccinia viruses exhibited high yields of HBsAg production. Primary hepatocytes secreted HBsAg efficiently to the cell culture supernatant with high concentrations of 8 μg/ml (= 8 mg/l), which exceeded, for example, the yield of the total protein extraction of Hepatitis B virus-like M particles from the Saccharomyces cerevisiae strain using alumina powder 8 times [14]. Thus, expression of HBsAg particles by primary hepatocytes following vaccinia viral infection could be an alternative source of S-antigen for further research or diagnostic purposes.…”

Section: Discussionmentioning

confidence: 99%

“…The limited supply of plasma from chronically infected humans and safety concerns demanded an alternative to plasma-derived particles serving as vaccines [7]. Since then, the S-gene has been expressed in many different systems, such as prokaryotic cells [8], yeast [9–14], stably transfected mammalian cells including mouse fibroblasts [15,16] and chinese hamster ovary (CHO) cells [17], mammalian cells infected with recombinant vaccinia viruses [18–22], insect cells infected with recombinant baculoviruses [23–25], and plants [26], in order to produce a sufficient amount of safe and effective HBsAg-based recombinant vaccine. Meanwhile, HBV surface antigen (HBsAg) lipoprotein particles are the basic components in almost all experimental and commercially used HBV candidate vaccine preparations.…”

Section: Introductionmentioning

confidence: 99%

Comparative purification and characterization of hepatitis B virus-like particles produced by recombinant vaccinia viruses in human hepatoma cells and human primary hepatocytes

et al. 2019

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This study describes the comparative expression and purification of hepatitis B surface antigen (HBsAg) particles produced upon infection of human primary hepatocytes and human hepatoma cell lines (HuH-7 and HepG2) with recombinant vaccinia viruses. The highest levels of HBsAg expression were found in HuH-7 hepatoma cells following infection with recombinant vaccinia viruses, which contain the S gene under control of a 7.5 k-promoter. Four different methods for purification of the HBsAg particles were examined: isopycnic ultracentrifugation, sucrose cushion sedimentation, isocratic column gel filtration, and binding to anti-HBs-coated microparticles. The highest degree of purity of HBsAg particles was reached by the method based on anti-HBs-coated microparticles. The resulting product was >98% pure. Biochemical analysis and characterization of purified HBsAg particles were performed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), western blotting, and electron microscopy. The HBsAg, purified from human hepatoma cell lines and from human primary hepatocytes, consisted of both the non-glycosylated (p25) and the glycosylated (gp27) form and assembled into typical 22-nm particles, and thus may be of great interest and importance for research, diagnostics, and medical treatments.

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Extraction and purification of hepatitis B virus-like M particles from a recombinant Saccharomyces cerevisiae strain using alumina powder

Cited by 18 publications

References 31 publications

Blood Bacteria-Free DNA in Septic Mice Enhances LPS-Induced Inflammation in Mice through Macrophage Response

Blood Bacteria-Free DNA in Septic Mice Enhances LPS-Induced Inflammation in Mice through Macrophage Response

Immunological Analysis of the Hepatitis B Virus “a” Determinant Displayed on Chimeric Virus-Like Particles of Macrobrachium rosenbergii Nodavirus Capsid Protein Produced in Sf9 Cells

Comparative purification and characterization of hepatitis B virus-like particles produced by recombinant vaccinia viruses in human hepatoma cells and human primary hepatocytes

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