THE POTENTIALS OF PASTEURELLA MULTOCIDA OMPA PROTEIN AS THE CANDIDATE FOR SUB-UNIT VACCINE AND FOR THE DEVELOPMENT OF AN ELISA KIT TO EVALUATE THE VACCINE RESPONSE IN THE ANIMALS
Main Article Content
Keywords
Animal vaccines, Immune responses, Farm animals, Respiratory infections, Pasteurellosis.
Abstract
Pasteurella multocida is the primary causative agent of Hemorrhagic Septicemia (HS) in cattle and Buffalo. It is responsible for causing huge economic loses every year. The P. multocida vaccines are used for livestock, but because sensitive and specific serological tests are not available, sero- conversion in these animals is unknown. As a result, the vaccine and it’s the immune response cannot be evaluated. So, the present study was designed to evaluate immunogenic potential of OmpA gene of P. multocida. PCR primers were designed to have restriction sites to cut amplicon and ligate product into expression vector. Purified PCR product was subjected to cloning PCR and the product was sequenced. The product was excised from cloning vector and ligated into expression vector (pET 40b (+)). Expression vector was transformed to chemically competent Escherichia coli strain DH5-α by heat shock method. Plasmid was extracted and ligation was confirmed by restriction digestion. IPTG was used to trigger expression. Total cell protein and medium were SDS-PAGE was used to analyze the expression. Recombinant proteins were injected into mice to test their immunogenicity. OmpA was proved to be highly immunogenic in nature, making it a more suitable candidate for sub-unit vaccine preparation and development of ELISA kit for the detection of immune responses in animals.
References
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31. Sarah S, Zamri-Saad M, Zunita Z, and Raha AJ. 2006. Molecular cloning and sequence analysis of gdhA gene of Pasteurella multocida B: 2. J Anim Vet Adv 5:1146-1149.
32. Shivachandra S, Viswas K, and Kumar A. 2011. A review of hemorrhagic septicemia in cattle and buffalo. Animal Health Research Reviews 12:67-82.
33. Simpson RJ. 2006. SDS-PAGE of proteins. Cold Spring Harbor Protocols 2006:pdb. prot4313- pdb. prot4313.
34. Snyder E, and Credille B. 2020. Mannheimia haemolytica and Pasteurella multocida in bovine respiratory disease: how are they changing in response to efforts to control them? Veterinary Clinics: Food Animal Practice 36:253-268.
35. Su A, Tong J, Fu Y, Müller S, Weldearegay YB, Becher P, Valentin-Weigand P, Meens J, and Herrler G. 2020. In- fection of bovine well-differentiated airway epithelial cells by Pasteurella multocida: actions and counteractions in the bacteria–host interactions. Veterinary research 51:1- 11.
36. Tomer P, Chaturvedi G, Malik P, and Monga D. 2002. Comparative analysis of the outer membrane protein pro- files of isolates of the Pasteurella multocida (B: 2) associated with haemorrhagic septicaemia. Veterinary research communications 26:513-522.
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38. Viale AM, and Evans BA. 2020. Microevolution in the major outer membrane protein OmpA of Acinetobacter baumannii. Microbial genomics 6.
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Clinical microbiology reviews 26:631-655.
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41. Xing J, Ma L, Cheng X, Ma J, Wang R, Xu K, Mymryk JS, and Zhang Z. 2021. Expression and functional analysis of the argonaute protein of thermus thermophilus (TtAgo) in E. coli BL21 (DE3). Biomolecules 11:524.
42. Yaman T, Büyükbayram H, Özyıldız Z, Terzi F, Uyar A, Keles ÖF, Özsoy ŞY, and Yener Z. 2018. Detection of bovine respiratory syncytial virus, Pasteurella multocida, and Mannheimia haemolytica by immunohistochemical method in naturally-infected cattle. Journal of veterinary Research 62:439.
2. lecular biology tools for rapid identification and characterization of Pasteurella spp. Veterinary world 11:1006.
3. Ahmad TA, Rammah SS, Sheweita SA, Haroun M, and El-Sayed LH. 2014. Development of immunization trials against Pasteurella multocida. Vaccine 32:909-917.
4. Ahmed N, Abusalah MAHA, Farzand A, Absar M, Yusof NY, Rabaan AA, AlSaihati H, Alshengeti A, Alwarthan S, and Alsuwailem HS. 2022. Updates on Epstein–Barr Virus (EBV)- Associated Nasopharyngeal Carcinoma: Emphasis on the Latent Gene Products of EBV. Medicina 59:2.
5. Ahmed N, Rabaan AA, Alwashmi AS, Albayat H, Mashraqi MM, Alshehri AA, Garout M, Abduljabbar WA, Yusof NY, and Yean CY. 2023. Immunoinformatic Execution and Design of an Anti-Epstein–Barr Virus Vaccine with Multiple Epitopes Triggering Innate and Adaptive Immune Responses. Microorganisms 11:2448.
6. Al-Hasani K, Boyce J, McCarl VP, Bottomley S, Wilkie I, and Adler B. 2007. Identification of novel immunogens in Pasteurella multocida. Microbial Cell Factories 6:3.
7. Ayalew S, Shrestha B, Montelongo M, Wilson AE, and Confer AW. 2011. Immunogenicity of Mannheimia haemolytica recombinant outer membrane proteins serotype 1-specific antigen, OmpA, OmpP2, and OmpD15. Clinical Vaccine Immunology 18:2067-2074.
8. Borah P. 2011. Primer designing for PCR. Science Vision 11:134-136.
9. Chang AY, Chau V, Landas JA, and Pang Y. 2017. Preparation of calcium competent Escherichia coli and heat-shock transformation. JEMI methods 1:22-25.
10. Dabo SM, Confer A, Montelongo M, York P, and Wyckoff III JH. 2008. Vaccination with Pasteurella multocida recombinant OmpA induces strong but non-protective and deleterious Th2- type immune response in mice. Vaccine 26:4345-4351.
11. De Alwis M. 1992. Haemorrhagic septicaemia—a general review. British Veterinary Journal
148:99-112.
12. Farooq U, Hussain M, Irshad H, Badar N, Munir R, and Ali Q. 2007. Status of haemorrhagic septicaemia based on epidemiology in Pakistan. Pakistan Veterinary Journal 27:67.
13. Fischer MD, Mgboji E, and Liu Z. 2018. Pyrite cloning: a single tube and programmed reaction cloning with re-striction enzymes. Plant methods 14:1-8.
14. Gao P, Dong X, Wang Y, and Wei G-H. 2021. Optimized CRISPR/Cas9-mediated single nucleotide mutation in adherent cancer cell lines. STAR protocols 2:100419.
15. Harper M, Boyce JD, and Adler B. 2006. Pasteurella multocida pathogenesis: 125 years after Pasteur. FEMS micro-biology letters 265:1-10.
16. Hatfaludi T, Al-Hasani K, Boyce JD, and Adler B. 2010. Outer membrane proteins of Pasteurella multocida. Vet-erinary microbiology 144:1-17.
17. Kharb S. 2015. Development of ELISA techniques for haemorrhagic septicaemia. International Journal of Bioassays 4:4574-4577.
18. Liu Y, Pang A, Bao G, Wei Q, Xiao C, and Ji Q. September 3 - 6, 2012. Development of an ELISA using recombi-nant Ompa protein for the detection of Pasteurella multocida infections in rabbits. In: Sheikh SE-, editor. 10th World Rabbit Congress. Egypt: World Rabbit Science Association. p 1219-1223.
19. Martin T, Abdelmalek J, Yee B, Lavergne S, and Ritter M. 2018. Pasteurella multocida line infection: a case report and review of literature. BMC infectious diseases 18:1-4.
20. Mayahi V, Esmaelizad M, and Harzandi N. 2018. Designing a novel recombinant HN protein with multi neutral- izing antigenic sites and auto tag removal ability based on NDV-VIIj for diagnosis and vaccination application. Indian journal of microbiology 58:326-331.
21. Mohamed R, and Abdelsalam E. 2008. A review on pneumonic pasteurellosis (respiratory mannheimiosis) with emphasis on pathogenesis, virulence mechanisms and predisposing factors. Bulgarian Journal of Veterinary Medicine 11:139-160.
22. Mostaan S, Ghasemzadeh A, Sardari S, Shokrgozar MA, Brujeni GN, Abolhassani M, Ehsani P, and Karam MRA. 2020. Pasteurella multocida vaccine candidates: A systematic review. Avicenna journal of medical biotechnology 12:140.
23. Mushtaque A, Sheikh AA, Ghafoor A, Shehzad W, and Khan MR. 2022. Optimization of In- House Indirect ELISA for Pasteurella multocida B: 2 Based on Recombinant Outer Membrane Protein H to Detect Post Vaccination Immune Response in Bovine in Pakistan. Pakistan Veterinary Journal 42:281-284.
24. Naveed M, Ali U, Karobari MI, Ahmed N, Mohamed RN, Abullais SS, Kader MA, Marya A, Messina P, and Scardina GA. 2022a. A vaccine construction against COVID-19-associated mucormycosis contrived with immunoinformatics-based scavenging of potential Mucoralean Epitopes. Vaccines 10:664.
25. Naveed M, Hassan J-u, Ahmad M, Naeem N, Mughal MS, Rabaan AA, Aljeldah M, Shammari BRA, Alissa M, and Sabour AA. 2022b. Designing mRNA-and Peptide-Based Vaccine Construct against Emerging Multidrug-Resistant Citrobacter freundii: A Computational-Based Subtractive Proteomics Approach. Medicina 58:1356.
26. Naveed M, Jabeen K, Naz R, Mughal MS, Rabaan AA, Bakhrebah MA, Alhoshani FM, Aljeldah M, Shammari BRA, and Alissa M. 2022c. Regulation of Host Immune Response against Enterobacter cloacae Proteins via Computational mRNA Vaccine Design through Transcriptional Modification. Microorganisms 10:1621.
27. Naveed M, Yaseen AR, Khalid H, Ali U, Rabaan AA, Garout M, Halwani MA, Al Mutair A, Alhumaid S, and Al Alawi Z. 2022d. Execution and Design of an Anti HPIV-1 Vaccine with Multiple Epitopes Triggering Innate and Adaptive Immune Responses: An Immunoinformatic Approach. Vaccines 10:869.
28. Novagen I. 2002. pET system manual. Novagen Madison, WI.
29. Peng Z, Wang X, Zhou R, Chen H, Wilson BA, and Wu B. 2019. Pasteurella multocida: genotypes and genomics. Microbiology and Molecular Biology Reviews 83:e00014-00019.
30. Ruffolo CG, and Adler B. 1996. Cloning, sequencing, expression, and protective capacity of the oma87 gene encoding the Pasteurella multocida 87-kilodalton outer membrane antigen. Infection and immunity 64:3161-3167.
31. Sarah S, Zamri-Saad M, Zunita Z, and Raha AJ. 2006. Molecular cloning and sequence analysis of gdhA gene of Pasteurella multocida B: 2. J Anim Vet Adv 5:1146-1149.
32. Shivachandra S, Viswas K, and Kumar A. 2011. A review of hemorrhagic septicemia in cattle and buffalo. Animal Health Research Reviews 12:67-82.
33. Simpson RJ. 2006. SDS-PAGE of proteins. Cold Spring Harbor Protocols 2006:pdb. prot4313- pdb. prot4313.
34. Snyder E, and Credille B. 2020. Mannheimia haemolytica and Pasteurella multocida in bovine respiratory disease: how are they changing in response to efforts to control them? Veterinary Clinics: Food Animal Practice 36:253-268.
35. Su A, Tong J, Fu Y, Müller S, Weldearegay YB, Becher P, Valentin-Weigand P, Meens J, and Herrler G. 2020. In- fection of bovine well-differentiated airway epithelial cells by Pasteurella multocida: actions and counteractions in the bacteria–host interactions. Veterinary research 51:1- 11.
36. Tomer P, Chaturvedi G, Malik P, and Monga D. 2002. Comparative analysis of the outer membrane protein pro- files of isolates of the Pasteurella multocida (B: 2) associated with haemorrhagic septicaemia. Veterinary research communications 26:513-522.
37. Townsend KM, O'Boyle D, Phan TT, Hanh TX, Wijewardana TG, Wilkie I, Trung NT, and Frost AJ. 1998. Acute septicaemic pasteurellosis in Vietnamese pigs. Veterinary microbiology 63:205- 215.
38. Viale AM, and Evans BA. 2020. Microevolution in the major outer membrane protein OmpA of Acinetobacter baumannii. Microbial genomics 6.
39. Wilson BA, and Ho M. 2013. Pasteurella multocida: from zoonosis to cellular microbiology.
Clinical microbiology reviews 26:631-655.
40. Xie X, Muruato A, Lokugamage KG, Narayanan K, Zhang X, Zou J, Liu J, Schindewolf C, Bopp NE, and Aguilar PV. 2020. An infectious cDNA clone of SARS-CoV-2. Cell host microbe 27:841- 848. e843.
41. Xing J, Ma L, Cheng X, Ma J, Wang R, Xu K, Mymryk JS, and Zhang Z. 2021. Expression and functional analysis of the argonaute protein of thermus thermophilus (TtAgo) in E. coli BL21 (DE3). Biomolecules 11:524.
42. Yaman T, Büyükbayram H, Özyıldız Z, Terzi F, Uyar A, Keles ÖF, Özsoy ŞY, and Yener Z. 2018. Detection of bovine respiratory syncytial virus, Pasteurella multocida, and Mannheimia haemolytica by immunohistochemical method in naturally-infected cattle. Journal of veterinary Research 62:439.
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Nov 4, 2023
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Muhammad Ahsan Nawaz, Ramsha Shoaib, Azka Rizvi, & Aamir Ghafoor. (2023). THE POTENTIALS OF PASTEURELLA MULTOCIDA OMPA PROTEIN AS THE CANDIDATE FOR SUB-UNIT VACCINE AND FOR THE DEVELOPMENT OF AN ELISA KIT TO EVALUATE THE VACCINE RESPONSE IN THE ANIMALS. Journal of Population Therapeutics and Clinical Pharmacology, 30(18), 1517–1526. https://doi.org/10.53555/jptcp.v30i18.3305
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Author Biographies
Muhammad Ahsan Nawaz
University Diagnostic Laboratory, University of Veterinary and Animal Sciences, Lahore, Pakistan
Ramsha Shoaib
University Diagnostic Laboratory, University of Veterinary and Animal Sciences, Lahore, Pakistan
Azka Rizvi
University Diagnostic Laboratory, University of Veterinary and Animal Sciences, Lahore, Pakistan
Aamir Ghafoor
University Diagnostic Laboratory, University of Veterinary and Animal Sciences, Lahore, Pakistan