Molecular Cloning and Immunogenicity Evaluation of PpiC, GelE, and VS87_01105 Proteins of Enterococcus faecalis as Vaccine Candidates

Abstract

Background:

Among the enterococci strains, Enterococcus faecalis is considered as one of the important nosocomial pathogens affecting immunocompromised patients. In this study, the immunogenicity of PpiC, GelE, and VS87_01105 proteins against enterococcal infection was investigated in a mice model.

Methods:

The genes encoding these proteins were cloned into pET21a expression vector, and the recombinant proteins were produced. Mice and rabbits were immunized with the purified recombinant proteins, and subsequently, mice were challenged with E. faecalis for the evaluation of their survival and bacterial clearances. The antibody responses to recombinant proteins were determined by ELISA assay, and opsonophagocytic activities of the antibodies were also measured. Passive immunization was performed using purified antibodies. Mice were challenged, and their survival and bacterial clearance were determined.

Results:

Immunized mice with PpiC, GelE, and VS87_01105 recombinant proteins showed 80%, 70%, and 40% survival rate, respectively. The survival rates among passively immunized mice that received 500 µg of IgG fraction in 100 µl PBS buffer of each of anti-PpiC, anti-GelE, and anti-VS87_01105 were 60%, 50%, and 20%, respectively. The rates of opsonization with anti-PpiC, anti-GelE, and anti-VS87_01105 antibodies at 1/10 dilution were 77%, 64%, and 23%, respectively.

Conclusion:

Based on our findings, PpiC, and GelE proteins can protect the mice against E. faecalis ATCC 29212 and effectively induce a protective antibody response. Thus, these proteins could be used as an additional therapeutic tool against enterococcal infections. Further studies to determine the role of PpiC in ligand binding and demonstration of epitope mapping may establish a credible target for vaccination.

• Authors: Hamid Kazemian, Mohammad Reza Pourmand, Seyed Davar Siadat, Mehdi Mahdavi, Mohammad Hossein Yazdi, Peyman Avakh Majelan, Davoud Afshar, Mehdi Yaseri, Mehdi Davari, Muhammad Ibrahim Getso
• Journal: Iranian biomedical journal (Iran Biomed J) Vol. 23 Issue 5 Pg. 344-53 (09 2019) ISSN: 2008-823X [Electronic] Iran
• PMID: 31103023 (Publication Type: Journal Article)

Chemical References

• Antibodies, Bacterial
• Antigens, Bacterial
• Bacterial Proteins
• Bacterial Vaccines
• Cytokines
• Opsonin Proteins
• Recombinant Proteins

Topics

• Animals
• Antibodies, Bacterial (immunology)
• Antigens, Bacterial (immunology)
• Bacterial Proteins (immunology)
• Bacterial Vaccines (immunology)
• Cloning, Molecular
• Cytokines (metabolism)
• Enterococcus faecalis (immunology)
• Gram-Positive Bacterial Infections (immunology, microbiology)
• Immunity
• Immunization
• Kaplan-Meier Estimate
• Male
• Mice, Inbred BALB C
• Opsonin Proteins (metabolism)
• Phagocytosis
• Rabbits
• Recombinant Proteins (isolation & purification)
• Survival Analysis