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
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 life, its control is crucial for cattle industries. Inactive bacterin and attenuated bacteria are widely used today against BRD. However, while inactive bacterins are insufficient for protection, attenuated ones can cause outbreak in ruminants. Therefore, there is a need for development of safe and potent recombinant vaccines. The most important epitope region of LktA protein of M. haemolytica, and two critical immunogens of H. somni, p31 and LppB OMPs, were chosen for the present experimental vaccine. lktA-p31, and lktA-lppB fusions were constructed, and expressed in E. coli BL21 cells. Three different experimental vaccines; LktA-p31, LktA-LppB, and LktA-p31 + LktA-LppB were formulated with an oil-based adjuvant, and tested on mice to assess the type of immune responses induced. As a result, all three formulations led to a significant increase in the level of total antigen specific IgG antibodies. Also, our combined LktA-p31 + LktA-LppB formulation induced significant levels of IgG2a antibodies indicating the induction of cellular immunity. Additionally, combined vaccine showed 52% bactericidal activity against H. somni as compared to the control group, allowing an assessment of the target specificity and functional activity of bactericidal antibodies. The bactericidal killing assay and the antigen-specific IgG2a response proved that our combined vaccine possesses dual protectivity against infection with M. haemolytica and H. somni.


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...
Vaccination of dairy cows with recombinant Streptococcus uberis adhesion molecule induces antibodies that reduce adherence to and internalization of S. uberis into bovine mammary epithelial cells
Prado, M. E.; Almeida, R. A.; Özen, Can; Luther, D. A.; Lewis, M. J.; Headrick, S. J.; Oliver, S. P. (2011-06-15)
Streptococcus uberis is an important environmental mastitis pathogen that causes subclinical and clinical mastitis in lactating and nonlactating cows and heifers throughout the world. Previous work from our laboratory suggests that S. uberis adhesion molecule (SUAM) is involved in S. uberis pathogenesis and may be an excellent target for vaccine development. The objective of this study was to evaluate the antibody response of cattle vaccinated with recombinant SUAM (rSUAM). Uninfected primiparous dairy cows...
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 vaccines composed of PlpE and OmpH from pasteurella multocida A:3
Okay, Sezer; Özcengiz, Gülay; Department of Biology (2011)
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 purif...
Citation Formats
N. H. Türkmen, “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,” M.S. - Master of Science, Middle East Technical University, 2020.