Designing and evaluation of an antibody-targeted chimeric recombinant vaccine encoding Shigella flexneri outer membrane antigens

Kazi, Ada; Ismail, Che Muhammad Khairul Hisyam; Anthony, Amy Amilda; Chuah, Candy; Leow, Chiuan Herng; Lim, Boon Huat; Singh, Kirnpal Kaur Banga

doi:10.1016/j.meegid.2020.104176

Cited by 10 publications

(5 citation statements)

References 63 publications

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“…Furthermore, numerous studies have reported that the outer membrane channel protein (TolC) is involved in multidrug resistance in gram-negative bacteria including S. flexneri . TolC is also involved in the binding of bacteria to host cells, secretion of bacterial toxins, and excretion of antibiotics from bacteria 8 , 68 .…”

Section: Discussionmentioning

confidence: 99%

“…This protein can create a water-filled channel across the inner and outer membranes, leading molecules to secret from the bacteria without entering the periplasmic space. Therefore, the TolC plays a critical role in increasing multidrug resistance and is important for the survival of pathogenic bacteria during infection 8 , 9 . Another virulence factor contains Invasive plasmid antigens (Ipa) that enter the host cell through the Type III Secretion System (T3SS) 10 .…”

Section: Introductionmentioning

confidence: 99%

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An in silico Design, Expression and Purification of a Chimeric Protein as an Immunogen Candidate Consisting of IpaD, StxB, and TolC Proteins from Shigella spp

Fathi¹,

Nazarian²,

Kordbacheh³

et al. 2022

AJMB

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Background: Shigella spp. is the cause of dysentery and is widespread worldwide. On the other hand, antibiotic resistance is increasing in this bacterium. Bioinformatics is a new approach to vaccine and drug design involving the selection of appropriate antigens. This study aimed to design a chimeric protein consisting of IpaD, StxB, and TolC proteins from Shigella through a bioinformatics approach as an immunogen candidate. Methods: The sequences of ipaD, stxB, and tolC genes were obtained. Additionally, the immunogenic regions of the associated protein, physicochemical characteristics, protein structures, B and T cells epitopes, and molecular docking were determined using in silico servers. Besides, the chimeric gene was synthesized following sequence optimization by utilizing the codon usage of Escherichia coli (E. coli). The expression of the recombinant protein was confirmed via SDS-PAGE and Western blot technique. Results: The residues 41-160 of IpaD, 21-89 of StxB, and 40-335 of TolC were selected. According to half-life, instability, and buried indices, IpaD-StxB-TolC was selected as the best arrangement. The Ramachandran plot showed that 97.077% of the amino acids were in the favored area. Linear and conformational epitopes were also present throughout the chimeric protein sequence. Moreover, the C-ImmSim server indicated that IgG and IgM titers could reach desirable values by the third injection. Furthermore, the stability of the mRNA-optimized gene was enhanced, increasing the Codon Adaptive Index (CAI) to 0.9. Finally, the chimeric gene was transferred to E. coli BL21, and the expression of the 60.6 kDa recombinant protein was confirmed. Conclusion: The results indicated that the recombinant protein could act as a proper immunogen candidate against Shigella spp.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

An in silico Design, Expression and Purification of a Chimeric Protein as an Immunogen Candidate Consisting of IpaD, StxB, and TolC Proteins from Shigella spp

Fathi¹,

Nazarian²,

Kordbacheh³

et al. 2022

AJMB

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“…Quiroz et al, as well as Mendiola et al, reported the inhibition of shikimate dehydrogenase as a potential drug target to inhibit E. coli and MRSA, whereas other studies have reported it inhibiting the pantothenate synthetase against MTB to minimize the prolonged TB treatment [44]. However, Jebastin et al and Kazi et al, described the role of outer membrane proteins such as Emrs, OMPs, and Mdts as a potent vaccine candidate for chimeric vaccine designing against Shigella flexneri [56] as well as Salmonella Typhi [57]. Similarly, the identified Mdtc protein can be considered for future vaccine design against S. Typhi.…”

Section: Discussionmentioning

confidence: 99%

Identification of a Novel Therapeutic Target against XDR Salmonella Typhi H58 Using Genomics Driven Approach Followed Up by Natural Products Virtual Screening

et al. 2021

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Typhoid fever is caused by a pathogenic, rod-shaped, flagellated, and Gram-negative bacterium known as Salmonella Typhi. It features a polysaccharide capsule that acts as a virulence factor and deceives the host immune system by protecting phagocytosis. Typhoid fever remains a major health concern in low and middle-income countries, with an estimated death rate of ~200,000 per annum. However, the situation is exacerbated by the emergence of the extensively drug-resistant (XDR) strain designated as H58 of S. Typhi. The emergence of the XDR strain is alarming, and it poses serious threats to public health due to the failure of the current therapeutic regimen. A relatively newer computational method called subtractive genomics analyses has been widely applied to discover novel and new drug targets against pathogens, particularly drug-resistant ones. The method involves the gradual reduction of the complete proteome of the pathogen, leading to few potential and novel drug targets. Thus, in the current study, a subtractive genomics approach was applied against the Salmonella XDR strain to identify potential drug targets. The current study predicted four prioritized proteins (i.e., Colanic acid biosynthesis acetyltransferase wcaB, Shikimate dehydrogenase aroE, multidrug efflux RND transporter permease subunit MdtC, and pantothenate synthetase panC) as potential drug targets. Though few of the prioritized proteins are treated in the literature as the established drug targets against other pathogenic bacteria, these drug targets are identified here for the first time against S. Typhi (i.e., S. Typhi XDR). The current study aimed at drawing attention to new drug targets against S. Typhi that remain largely unexplored. One of the prioritized drug targets, i.e., Colanic acid biosynthesis acetyltransferase, was predicted as a unique, new drug target against S. Typhi XDR. Therefore, the Colanic acid was further explored using structure-based techniques. Additionally, ~1000 natural compounds were docked with Colanic acid biosynthesis acetyltransferase, resulting in the prediction of seven compounds as potential lead candidates against the S. Typhi XDR strain. The ADMET properties and binding energies via the docking program of these seven compounds characterized them as novel drug candidates. They may potentially be used for the development of future drugs in the treatment of Typhoid fever.

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“…Recently, using proteomics, iron-regulating bacteria molecules were found to exhibit antigenic and immunogenic characteristics (Ansari et al 2019 ). Five Outer membrane proteins ( OMPs ) with significant potential for vaccine development against S. flexneri infection were discovered due to the reverse vaccination strategy used to identify Shigella flexneri outer membrane antigens as cell-dependent vaccine and antibody candidates (Kazi et al 2020 ). The Brucella lumazine synthase (BLS) is a viable surface-exposed alternative because of its practical expression, simple subsequent purification, and excellent molecular mass for distribution (Badmasti et al 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Oral vaccination with novel Lactococcus lactis mucosal live vector-secreting Brucella lumazine synthase (BLS) protein induces humoral and cellular immune protection against Brucella abortus

2023

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This work aimed to provide recombinant Lactococcus lactis as a potential live vector for the manufacture of recombinant Brucella abortus (rBLS-Usp45). The sequences of the genes were collected from the GenBank database. Using Vaxijen and ccSOL, the proteins' immunogenicity and solubility were evaluated. Mice were given oral vaccinations with recombinant L. lactis . Anti-BLS-specific IgG antibodies were measured by ELISA assay. Cytokine reactions were examined using real-time PCR and the ELISA technique. The BLS protein was chosen for immunogenicity based on the vaccinology screening findings since it had maximum solubility and antigenic values of 99% and 0.75, respectively. The BLS gene, digested at 477 bp, was electrophoretically isolated to demonstrate that the recombinant plasmid was successfully produced. Protein-level antigen expression showed that the target group produced the 18 kDa-sized BLS protein, whereas the control group did not express any proteins. In the sera of mice given the L. lactis -pNZ8148-BLS-Usp45 vaccine 14 days after priming, there was a significant level of BLS-specific IgG1, IgG2a ( P < 0.001) compared to the PBS control group. Vaccinated mice showed higher levels of IFN-γ, TNFα, IL-4, and IL-10 in samples obtained on days 14 and 28, after receiving the L. lactis -pNZ8148-BLS-Usp45 and IRBA vaccines ( P < 0.001). The inflammatory reaction caused less severe spleen injuries, alveolar edema, lymphocyte infiltration, and morphological damage in the target group's spleen sections. Based on our findings, an oral or subunit-based vaccine against brucellosis might be developed using L. lactis -pNZ8148-BLS-Usp45 as a novel, promising, and safe alternative to the live attenuated vaccines now available. Supplementary Information The online version contains supplementary material available at 10.1007/s00203-023-03471-6.

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Designing and evaluation of an antibody-targeted chimeric recombinant vaccine encoding Shigella flexneri outer membrane antigens

Cited by 10 publications

References 63 publications

An in silico Design, Expression and Purification of a Chimeric Protein as an Immunogen Candidate Consisting of IpaD, StxB, and TolC Proteins from Shigella spp

An in silico Design, Expression and Purification of a Chimeric Protein as an Immunogen Candidate Consisting of IpaD, StxB, and TolC Proteins from Shigella spp

Identification of a Novel Therapeutic Target against XDR Salmonella Typhi H58 Using Genomics Driven Approach Followed Up by Natural Products Virtual Screening

Oral vaccination with novel Lactococcus lactis mucosal live vector-secreting Brucella lumazine synthase (BLS) protein induces humoral and cellular immune protection against Brucella abortus

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