Development of recombinant vaccines composed of PlpE and OmpH from pasteurella multocida A:3

Okay, Sezer
Pasteurella multocida serotype A:3 is a gram-negative bacterial pathogen which is one of the causative agents of shipping fever in cattle. In this study, ompH and two fragments of plpE gene (plpEN and plpEC) were cloned from the genomic DNA of P. multocida P-1062 (ATCC 15743, serotype A:3) and plpEN-ompH and plpEC-ompH fusions were constructed. In vitro expression of the genes was shown in HEK-293 cells. Later, full-length plpE gene was cloned and the recombinant proteins were expressed in E. coli and purified. Three DNA vaccine formulations, namely pCMV-ompH, pCMV-plpEN-ompH and pCMV-plpEC-ompH and five recombinant protein based vaccines, PlpEN-OmpH, PlpEC-OmpH, OmpH, PlpEC and PlpE were generated. Recombinant proteins were formulated with at least one of the adjuvants: alum, CpG, alum-CpG, oil based and oil based-CpG. BALB/c mice were immunized with these vaccine formulations and their sera were used for the evaluation of antibody and serum IFN-γ titers. Protective capacities of the vaccines were also evaluated via challenge of mice with 10 LD50 of P. multocida A:3. DNA vaccines induced immune responses, but did not provide protection. All protein vaccine formulations increased antibody levels and CpG containing formulations enhanced serum IFN-γ titers. 100 µg of PlpEC-OmpH protein adsorbed on alum adjuvant conferred 40% protection while no protection was obtained with PlpEN-OmpH. Next, the effects of CpG, or its alum and oil based combinations as adjuvants were investigated on PlpEC-OmpH mediated protection. The vaccine formulation composed of PlpEC-OmpH and oil based-CpG adjuvant conferred 100% protection. Finally, the mice were vaccinated with recombinant OmpH, PlpEC and PlpE formulated with oil based-CpG adjuvant. OmpH, PlpEC and PlpE formulations provided 50%, 60% and 100% protection, respectively. These findings implicated that recombinant PlpE and PlpEC-OmpH fusion proteins when formulated with oil based-CpG adjuvant are potent acellular vaccine formulation candidates against shipping fever.


Immunogenicity and protective efficacy of the recombinant Pasteurella lipoprotein E and outer membrane protein H from Pasteurella multocida A:3 in mice
Okay, Sezer; Ozcengiz, Erkan; GÜRSEL, İHSAN; Özcengiz, Gülay (2012-12-01)
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 ...
Development of recombinant vaccine candidates composed of ltka from mannheimia haemolytica A1 and PlpEC AND OmpH from Pasteurella Multocida A:3 against bovine respiratory disease
Çırçır, Ayça; Özcengiz, Gülay; Department of Biology (2014)
Mannheimia haemolytica A1 and Pasteurella multocida serotype A:3 are gramnegative bacterial pathogens which are considered the causative agents of bovine respiratory disease (BRD) or shipping fever in cattle. The foremost of several virulence factors of M. haemolytica A1 which allows lung colonization and establishment of infection is leukotoxin (Lkt). The others include adhesin, capsule, outer membrane proteins, and various proteases used for attachment, spreading and evading innate and adaptive host immun...
Investigation of the Ligand Binding Characteristics of Staphylococcus aureus NorA Efflux Pump
Işık , Esra Büşra; Serçinoğlu, Onur (Orta Doğu Teknik Üniversitesi Enformatik Enstitüsü; 2022-10)
Staphylococcus aureus is a gram-positive bacterial pathogen which is highly adaptive to environmental conditions and causes various disorders. Excessive usage of antibiotics may result in development of antibiotic resistance in S. aureus. One of the resistance mechanisms is increase in the activity of transmembrane multi-drug efflux pumps. NorA is the most studied efflux pump in S. aureus, which belongs to Major Facilitator Superfamily (MFS). NorA has been shown to contribute to resistance against a variety...
Development of an experimental recombinant vaccine formulation composed of LKTA from Mannheimia Haemolytica A1 and P31 and LPPB from Histophilus Somni 8025 against Hovine Respiratory Disease
Türkmen, Nazlı Hilal; Özcengiz, Gülay; Department of Molecular Biology and Genetics (2020-9)
Two major bacterial pathogens of Bovine Respiratory Disease (BRD), Mannheimia haemolytica and Histophilus somni are Gram-negative, opportunistic bacteria that live in upper respiratory tracts of ruminants commensally. The one of the most important virulence factor of M. haemolytica, leukotoxin, is responsible for lung colonization and establishment of infection. On the other hand, H. somni OMPs have a significant contribution to the pathogenicity of the organism. As the disease results in a destroyed animal...
Differential immune activation following encapsulation of immunostimulatory CpG oligodeoxynucleotide in nanoliposomes.
Erikçi, E; Gürsel, Mayda; Gürsel, I (2011-02-01)
The immunogenicity of a vaccine formulation is closely related to the effective internalization by the innate immune cells that provide prolonged and simultaneous delivery of antigen and adjuvant to relevant antigen presenting cells. Endosome associated TLR9 recognizes microbial unmethylated CpG DNA. Clinical applications of TLR9 ligands are significantly hampered due to their pre-mature in vivo digestion and rapid clearance. Liposome encapsulation is a powerful tool to increase in vivo stability as well as...
Citation Formats
S. Okay, “Development of recombinant vaccines composed of PlpE and OmpH from pasteurella multocida A:3,” Ph.D. - Doctoral Program, Middle East Technical University, 2011.