An anti-amoebic vaccine: generation of the recombinant antigen LC3 from Entamoeba histolytica linked to mutated exotoxin A (PEΔIII) via the Pichia pastoris system

Abstract

We report for the first time the design and expression of the recombinant E. histolytica LC3 protein fused to PEΔIII and KDEL3, with potential application as an immunogen.

Fed-batch cultivation and operational conditions for the production of a recombinant anti-amoebic vaccine in Pichia pastoris system.

“…The specific production rate (q P ) increased 35% when it was used 3% of methanol in comparison with the use of 0.5 and 1%, methanol, which reached similar q P around 0.1 mg/g DCW h in both methanol concentrations. In comparison, production of VAEh in the shake flask cultures was improved with those previous reported results of 3.8 µg/mL (Martínez-Hernández et al, 2017). The higher methanol concentration supplied in shake flask cultures is in the range for the Mut + strain at which it can be assimilated adequately without causing cell intoxication or lysis (Cos et al, 2006).…”

“…A recombinant strain P. pastoris GS115 containing the LC3 gene from E. histolytica was used. The characteristics of chimeric gene for the vaccine (VAEh) is described in previous work (Martínez-Hernández et al, 2017).…”

“…However, these drugs display adverse side effects and are expensive and not easily available in certain countries and areas; thus, the application of new immunogenic schemes as vaccines are of high interest to solve such issues (Bansal et al, 2006). Recently it has been developed a new recombinant P. pastoris Mut + strain to produce a recombinant Entamoeba histolytica (E. histolytica) LC3 protein with a potential application as an immunogen (Martínez-Hernández et al, 2017). The candidate for a vaccine against E. histolytica (VAEh) produced, is a chimeric construction between E. histolytica LC3 fragment; the retrograde delivery domains (I-II) from Pseudomonas aeruginosa exotoxin A and the carboxy-terminal signal KDEL3 (Martínez-Hernández et al, 2017).…”

Entamoeba histolytica: Membrane and Non-Membrane Protein Structure, Function, Immune Response Interaction, and Vaccine Development

“…In a study conducted by Martinez-Hernandez et al (2017; 2021) [34,35], the newly developed recombinant chimeric vaccine (PE∆III-LC3-KDEL3) produced from the LC3 fragment of E. histolytica, domains I and II of Pseudomonas aeruginosa, and the carboxy-terminal signal KDEL3 in Pichia pastoris has potential as an immunogen and showed the inhibition of the adherence of trophozoites to the HepG2 cell monolayer in a hamster model. Furthermore, the vaccine protects against liver tissue damage and uncontrolled inflammation.…”