A novel chimeric flagellum fused with the multi-epitope vaccine CTB-UE prevents Helicobacter pylori-induced gastric cancer in a BALB/c mouse model

Abstract

Helicobacter pylori (H. pylori) infection causes peptic ulcers, gastric adenocarcinoma, and mucosa-associated lymphoid tissue (MALT) lymphoma. The eradication of H. pylori might be an effective means of preventing gastric cancer. A dual-antigen epitope and dual-adjuvant vaccine called CTB-UE-CF (CCF) was constructed by combining a multi-epitope vaccine CTB-UE with a novel chimeric flagellum (CF) to simultaneously activate Toll-like receptor (TLR) 5-agonist activity and preserve the immunogenicity of H. pylori flagellum FlaA. The evaluation of efficacy to reduce H. pylori colonization was performed using BALB/c mice by oral immunization with a triple dose of this vaccine strain. Two weeks after the last immunization, mice were sacrificed to determine specific antibody levels and proinflammatory cytokine production. To determine the presence of H. pylori, we detected the number of H. pylori by real-time quantitative PCR (qPCR) and measured the urease activity in the gastric tissue. The results showed that the immunogenicity and mucosal immune responses of CCF performed significantly better than those of CTB-UE. This dual-antigen epitope and dual-adjuvant system might greatly contribute to the development of a safe and efficient therapeutic vaccine for humans against H. pylori infection.

Inflammation, immunity, and vaccines for Helicobacter pylori infection

“…This particular vaccine induced CD4(+) T cells, serum‐ and mucosal‐specific antibodies against native H. pylori urease, decreased H. pylori colonization, and relieved gastritis by regulation of IFN‐γ and IL‐17 in BALB/c mice. Similar effects on reduction of H. pylori colonization by improving the efficacy of CTB‐UE in this mouse strain were obtained by Song et al . using a dual‐antigen epitope and dual‐adjuvant vaccine called CTB‐UE‐CF that was constructed by combining the multi‐epitope vaccine CTB‐UE with a novel chimeric flagellum (CF).…”

Toxic adjuvants alter the function and phenotype of dendritic cells to initiate adaptive immune responses induced by oral Helicobacter pylori vaccines

“…25 Briefly, after an 8-h induction with 1 mmol/L IPTG, cells were harvested and purified using nickel affinity chromatography with a His-Trap excel column (GE Healthcare Bioscience, Piscataway, NJ) and anion-exchange chromatography with DEAE Sepharose FF (Amersham Pharmacia Biotech AB, Sweden). 25 Briefly, after an 8-h induction with 1 mmol/L IPTG, cells were harvested and purified using nickel affinity chromatography with a His-Trap excel column (GE Healthcare Bioscience, Piscataway, NJ) and anion-exchange chromatography with DEAE Sepharose FF (Amersham Pharmacia Biotech AB, Sweden).…”

“…Heat-shock protein (Hsp), neutrophil-activating protein (Nap), lipoprotein 20 (Lpp20), and chimeric flagellum (CF) were expressed in Escherichia coli Rosetta cells and purified as described previously. 25 Briefly, after an 8-h induction with 1 mmol/L IPTG, cells were harvested and purified using nickel affinity chromatography with a His-Trap excel column (GE Healthcare Bioscience, Piscataway, NJ) and anion-exchange chromatography with DEAE Sepharose FF (Amersham Pharmacia Biotech AB, Sweden). Purified proteins were dialyzed against phosphate-buffered saline (PBS) and stored at −80°C.…”

Vaccine‐induced gastric CD4+ tissue‐resident memory T cells proliferate in situ to amplify immune response against Helicobacter pylori insult

“…The CCF (CTB‐UE‐CF) protein that constructed by cholera toxin B, multi‐epitopes from H pylori urease, and self‐adjuvant regions from S typhimurium phase I flagellin FliC was expressed by Escherichia coli Rosetta (DE3) cells with pET‐28a‐CCF. The purification of CCF was as described previously . Briefly, the protein was first purified by nickel affinity chromatography (GE Healthcare), followed by anion‐exchange chromatography with DEAE Sepharose FF (Amersham Pharmacia Biotech AB).…”