Immunogenicity and protective efficacy of the recombinant Pasteurella lipoprotein E and outer membrane protein H from Pasteurella multocida A:3 in mice

Abstract

Pasteurella multocida serotype A:3 is a Gram-negative bacterial pathogen, one of the causative agents of shipping fever of cattle. In this study, outer membrane protein H (ompH) and Pasteurella lipoprotein E (plpE) genes were cloned and plpEC-ompH fusion was constructed and expressed in Escherichia coli. Recombinant PlpE, OmpH and PlpEC-OmpH fusion proteins were purified and formulated with oil-based and oil-based CpG ODN adjuvants. Antibody responses in mice vaccinated with recombinant PlpE and PlpEC-OmpH proteins formulated with both adjuvants were significantly (p<0.05) increased. However, a significant (p<0.05) increment in serum IFN-γ level was only observed upon immunization with oil-based CpG formulations. Protectivity of the vaccines were evaluated via intraperitoneal challenge of mice with 10 LD50 of P. multocida A:3. The recombinant proteins PlpE and PlpEC-OmpH fusion conferred 100% protection when formulated with oil-based CpG ODN while the protectivity was found to be 80% and 60%, respectively when only oil-based adjuvant was used in respective formulations. These findings indicated that the recombinant PlpE or PlpEC-OmpH fusion proteins formulated with oil-based CpG ODN adjuvant are possible acellular vaccine candidates against shipping fever.

Pasteurella multocida: from Zoonosis to Cellular Microbiology

“…However, a plpE knockout mutant strain retained full virulence (416). Conjugated vaccines comprised of multiple antigens, such as OmpH plus PlpE peptides, have also shown promise (445,565).…”

Evaluation of immunogenicity and protective efficacy of recombinant outer membrane proteins of Haemophilus parasuis serovar 5 in a murine model

“…Both of LppC and HAPS_0926 are outer membrane lipoproteins. Lipoproteins are widely distributed in Gram-negative bacteria, which are involved in diverse mechanisms of physiology/pathogenesis and are considered potential target antigens for vaccine development against many infectious diseases [30–31]. Our previous study demonstrated that the VacJ lipoprotein was a good vaccine candidate against Glässer's disease [18].…”

Immune responses elicited by the recombinant Erp, HspR, LppX, MmaA4, and OmpA proteins from Mycobacterium tuberculosis in mice

“…Purification of the recombinant proteins was performed according to Okay et al [15]. The recombinant E. coli BL21(DE3) cells with pET-28a(+) vectors carrying erp, hspR, lppX, mmaA4, and ompA genes were grown in Luria Broth (Merck, Germany) containing kanamycin (30 μg/ml) at 37°C until logarithmic growth phase.…”

“…The secondary antibody, alkaline phosphatase-conjugated anti-mouse IgG (Sigma, Germany), was used at a dilution of 1:10,000. The AP Conjugate Substrate Kit (Bio-Rad, CA, USA) was used for visualization of the bands [15].…”

“…The humoral immune response elicited by the vaccination groups was detected measuring the serum IgG titers via the enzyme-linked immunosorbent assay (ELISA) [15]. The 96-well ELISA plates were coated with 1 μg/well of purified recombinant proteins.…”