Gold Nanoparticles Conjugating Recombinant Nontoxic Pseudomonas Exotoxin A as a Vaccine Candidate for Pseudomonas aeruginosa Infections.

Autor: Abbasi, M., Kafilzadeh, F., Tanomand, A., Zolghadri, S., Hosainzadegan, H.
Zdroj: Molecular Genetics, Microbiology & Virology; Dec2021 Supplement 1, Vol. 36 Issue 1, pS7-S12, 6p
Abstrakt: Background:Pseudomonas aeruginosa, is an opportunistic pathogen with high resistance to many antibiotics which can cause health-care associated infections in debilitated patients. The rapid development and spread of resistant strains in hospitals has created many problems in the treatment of infections caused by this bacterium in high-risk patients. In this study, the immunogenicity of recombinant non-toxic Pseudomonas exotoxin A conjugate with nanogold as a new nanovaccine candidate have been investigated in animal models. Methods: The sequence for ExoA domain I-II gene was amplified from P. aeruginosa PAO1, ligated to pET22b experssion vector and transformed into the E. coli BL21. Recombinant truncated ExoA expression was assayed by SDS-PAGE. The Ni-NTA affinity chromatography was applied for recombinant protein purification and confirmed by western blot. The gold nanoparticles were prepared using Turkevich method and conjugated to recombinant domain I-II by the diluted NaBH4 and electrostatic forces. Nanoparticles size was determined before and after conjugation by electron microscopy. Conjugation was also confirmed by Fourier transform infrared spectrometry (FTIR) and then antigens injected to mice groups to evaluate its immunogenicity by assessing the antibody response and animal protection by challenge against 2× LD50Pseudomonas infection. Results: The results indicated that synthesized Gold nanoparticles were conjugated to purified recombinant Exo A domain I-II. Immunogenicity studies showed that antibody titers produced against domain I-II and its conjugate with nanogold are significant in comparison with controls (P < 0/005). The animal challenges showed significant protection against 2× LD50 P. aeruginosa infection in animals immunized with domain I-II alone and nanogold-domain I-II conjugates in comparison with controls (P = 0/00). Conclusion: Results of this study indicate that recombinant ExoA domain I-II alone or in conjugation with nanogold could act as a safe and immunogenic vaccine against Pseudomonas infections. Although, further studies are needed to make a final judgment. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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