Bioinformatics design of recombinant chimeric protein containing SipD and LptD immunogens and evaluation of its immunogenicity against Salmonella Typhimurium

Microb Pathog. 2023 Feb:175:105959. doi: 10.1016/j.micpath.2022.105959. Epub 2022 Dec 26.

Abstract

The growing emergence of resistant bacteria is the current global concern for the humans and animals. Vaccination could be the desirable method to preventing such infectious diseases. Safe and effective vaccines are urgently needed to manage and prevent Salmonella contamination. Subunit vaccines are safe approaches for the protection against Salmonella spp. The bioinformatics methods were performed to determine the gene structure. Gene cassette (rLPSI) was ordered in pET28a (+), and cloned into E.coli BL21 (DE3), and the recombinant protein was expressed using IPTG (1 mM). Mice were immunized by subcutaneous administration of recombinant protein. Then, the mice were challenged by oral administration of 100LD50 of live S. Typhimurium. The Codon adaptation index of the chimeric gene was multiplied by 0.92. Validation results showed that >90% of residues lie in the desired or extra allowed area of the Ramachandran plot. The recombinant protein (65.9 kDa) was expressed in E.coli. Antibody titers in vaccinated mice were significantly different from those in the control groups. Recombinant protein immunization of the mice provided 90% and 70% protection against 10LD50 and 100LD50 of S. Typhimurium, respectively. In general, the results showed the high efficiency of rLPSI chimeric protein as a protective antigen against S. Typhimurium infection. The rLPSI chimeric protein could be an effective recombinant vaccine candidate against S. Typhimurium infection, but more research is needed.

Keywords: In silico design; LpsD-SipD chimeric antigen; Protective immunity; Recombinant protein; Salmonella Typhimurium; Vaccine candidate.

MeSH terms

  • Animals
  • Antibodies, Bacterial / immunology
  • Bacterial Outer Membrane Proteins / immunology
  • Escherichia coli / genetics
  • Escherichia coli Proteins*
  • Immunization
  • Mice
  • Mice, Inbred BALB C
  • Recombinant Fusion Proteins / genetics
  • Recombinant Proteins / genetics
  • Salmonella Vaccines*
  • Salmonella typhimurium* / genetics
  • Salmonella typhimurium* / immunology
  • Vaccines, Attenuated
  • Vaccines, Synthetic

Substances

  • Antibodies, Bacterial
  • Bacterial Outer Membrane Proteins
  • Escherichia coli Proteins
  • LptD protein, E coli
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Salmonella Vaccines
  • Vaccines, Attenuated
  • Vaccines, Synthetic
  • SspD protein, Salmonella typhimurium