The effect of homologous and heterologous Bacterial DNA on the development of immune responses against Pasteurella multocida in BALB / c mice
Subject Areas : Molecular MicrobiologyYahya Tahamtan 1 , Maryam Homayoon 2 , Mohammad Kargar 3
1 - Associate Professor, Department of Microbiology, Shiraz Branch, Razi Vaccine and Serum Research Institute, Agriculture Research, Education and Extension Organization (AREEO), Shiraz, Iran.
2 - Ph.D., Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
3 - Professor, Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran.
Keywords: Pasteurella multocida, Adjuvant, Vaccine, DTH, Ferric chloride, Bacterial DNA,
Abstract :
Background & Objectives: Pasteurella multocida is a principal pathogen of domestic animals causing agents of pneumonia and hemorrhagic septicemia in cattle, sheep, and goats, fowl cholera in chickens, and progressive atrophic rhinitis in swine and as well as an opportunistic pathogen of humans. In this study, we investigated the humoral and cellular immune responses and protective immunity conferred by an iron-inactivated vaccine with two different bacterial DNA as an adjuvant. Materials & Methods: P. multocida was grown in BHI broth, inactivated with FeCl3, adjuvanted with alum and P. multocida A (homologous DNA) and P. multocida B (heterologous DNA) bacterial DNAs. BALB/c mice were immunized with two whole-cell inactivated vaccine doses at 2 weeks apart. The animals were challenged 4 weeks after booster immunization. The serum antibodies titer was tested by ELISA. At 28 days post-immunization, cell mediates immunity responses were measured by assay of DTH and IL-6 and IL-12 in the serum samples. Results: Our results showed the levels of antibodies in bDNA adjuvant groups were higher than the alum adjuvant vaccine group. Peak antibody titers of 0.372 were obtained in the IIA+AbDNA group. The protection rate of the AbDNA adjuvant vaccine was better than of other adjuvant vaccines and they protected 100% of mice challenge groups. Peak serum IL-6 and IL-12 titers were achieved in the IIA+AbDNA groups. Conclusion: These studies indicate that bDNA is effective as immune adjuvants and because of its stimulating properties it can be used as an inducer of humoral and cellular immune responses for vaccination applications. The findings also showed a better ability of homologous bDNA to induce immune responses.
multocida. Sci World J. 2014; 9(2): 1-7.
2. Shivachandra SB, Kumar A, Yogisharadhya R, Viswas KN. Immunogenicity of highly
conserved recombinant VacJ outer membrane lipoprotein of Pasteurella multocida. Vaccine.
2014; 32(2): 290-296.
3. Wilson BA, Ho M. Pasteurella multocida: from zoonosis to cellular microbiology. Clin
Microbiol Rev. 2013; 26(3): 631-55.
4. Jarvinen LZ, Hogenesch H, Suckow MA, Bowersock TL. Induction of protective immunity in
rabbits by coadministration of inactivated Pasteurella multocida toxin and potassium
thiocyanate extract. Infect Immun. 1998; 66(8): 3788-3795.
5. Lee KE, Jeoung HY, Lee JY, Lee MH, Choi HW, Chang KS. Phenotypic characterization and
random amplified polymorphic DNA (RAPD) analysis of Pasteurella multocida isolated from
Korean pigs. J Vet Med Sci. 2012; 74(5): 567-573.
6. Xie Z, Li H, Chen J, Zhang H-b, Wang Y-Y, Chen Q, Zhao ZZ, Cheng C, Zhang H, Yang
Y, Wang HN, Gao R. Shuffling of pig interleukin-2 gene and its enhancing of immunity in
mice to Pasteurella multocida vaccine. Vaccine. 2007; 25(48): 8163-8171.
7. Dagleish MP, Christopher Hodgson J, Ataei S, Finucane A, Finlayson J, Sales J. Safety and
protective efficacy of intramuscular vaccination with a live aroA derivative of Pasteurella
multocida B:2 against experimental hemorrhagic septicemia in calves. Infect Immu. 2007; 75
(12): 5837-5844.
8. Trevani AS, Chorny A, Salamone G, Vermeulen M, Gamberale R, Schettini J, Raiden S,
Geffner J. Bacterial DNA activates human neutrophils by a CpG-independent pathway. Eur J
Immunol. 2003; 33(11): 3164-3174.
9. Mapletoft JW, Oumouna M, Kovacs-Nolan J, Latimer L, Mutwiri G, Babiuk LA, van Drunen
Littel-van den Hurk S. Intranasal immunization of mice with a formalin-inactivated bovine
respiratory syncytial virus vaccine co-formulated with CpG oligodeoxynucleotides and
polyphosphazenes results in enhanced protection. J Gen Virol. 2008; 89 (Pt 1): 250-260.
10. Tang ML, Lahtinen SJ, Boyle RJ. Probiotics and prebiotics: clinical effects in allergic disease.
Curr Opinion Pediatr. 2010; 22(5): 626-634.
11. Bode C, Zhao G, Steinhagen F, Kinjo T, Klinman DM. CpG DNA as a vaccine adjuvant.
Expert Rev Vaccines. 2011; 10(4): 499-511.
12. Kumar D, Singh A. Salmonella typhimurium grown in iron-rich media, inactivated with ferric
chloride and adjuvanted with homologous bacterial DNA is potent and efficacious vaccine in
mice. Vaccine. 2005; 23(48-49): 5590-5598.
13. Homayoon M, Tahamtan Y, Kargar M, Hosseini SMH, Akhavan SA. Adjuvant activity of
Pasteurella maltocida A strain, Pasteurella maltocida B strain and salmonella typhimurium
bacterial and on cellular responses against Pasteurella maltocida specific strain infectionsin
Balb/C mice. Trop Med Asian Pac J. 2018; 11(5): 336-341.
14. Cheng HR, Jiang N. Extremely rapid extraction of DNA from bacteria and yeasts. Biotechnol
Lett. 2006; 28(1): 55-59.
15. Chaiyotwittayakun A, Burton L, Weber PSD, Kizilkaya K, Cardoso FF, Erskine RJ.
Hyperimmunization of steers with J5 Escherichia coli bacterin: effects on isotype-specific
serum antibody responses and cross reactivity with heterogeneous Gram-negative bacteria. J
Dairy Sci. 2004; 87(10): 3375-3385.
16. Ahmad TA, Rammah SS, Sheweita SA, Haroun M, El-Sayed LH. Development of
immunization trials against Pasteurella multocida. Vaccine. 2014; 32(8): 909-917.
17. Radhi SN. Optimization of heavy metals chlorides resistance by Staphylococcus aureus and its
ability to remove them. Iraqi J Sci. 2012; 53(4): 778-785.
18. Herath C, Kumar P, Singh M, Kumar D, Ramakrishnan S, Goswami TK, Singh A, Ram GC.
Experimental iron-inactivated Pasteurella multocida A: 1 vaccine adjuvanted with bacterial
DNA is safe and protects chickens from fowl cholera. Vaccine. 2010; 28(11): 2284-2289.
19. Homayoon M, Tahamtan Y, Kargar M, Hosseini SMH, Akhavan S.A. Pasteurella multocida
inactivated with ferric chloride and adjuvanted with bacterial DNA is a potent and efficacious
vaccine in Balb/c mice. J Med Microbiol. 2018; 67(9): 1383-1390.
20. Freitas E, Marinho AC, Albuquerque D, Teles L, Sindeaux M, Salles MT, Sousa DC, Lima
MM, Silva MG, Fernandes D. Adjuvant activity of peanut, cottonseed and rice oils on cellular
and humoral response. Vacci Monitor. 2013; 22(1): 4-9.
21. Sayed HE, Ashry E, Ahmad TA. The use of propolis as vaccine’s adjuvant. Vaccine. 2012; 31
(1): 31-39.
22. Wu C, Qin X, Li P, Pan T, Ren W, Li N, Peng Y. Transcriptomic analysis on responses of
murine lungs to Pasteurella multocida infection. Front Cell Infect Microbiol. 2017; 7: 251.
23. Nakae S, Komiyama Y, Nambu A, Sudo K, Iwase M, Homma I, Sekikawa K, Asano M,
Iwakura Y. Antigen-specific T cell sensitization is impaired in IL-17-deficient mice, causing
suppression of allergic cellular and humoral responses. Immunity. 2002; 17(3): 375-387.
24. Al-Samarrae EAA, Al-Shweey SHA. The synergistic effect of BCG and Pasteurella multocida
vaccines in local rabbits. Al-Anbar J Vet Sci. 2012; 5(1): 1-6.
25. Hanagata N. CpG oligodeoxynucleotide nanomedicines for the prophylaxis or treatment of
cancers, infectious diseases, and allergies. Int J Nanomedicine. 2017; 12: 515-531.
26. Hirahara K, Poholek A, Vahedi G, Laurence A, Kanno Y, Milner JD, O'Shea JJ. Mechanisms
underlying helper T-cell plasticity: implications for immune-mediated disease. J Allergy Clin
Immunol. 2013; 131(5): 1276-1287.
27. Okay S, Özcengiz E, Gürsel I, Özcengiz G. Immunogenicity and protective efficacy of the
recombinant Pasteurella lipoprotein E and outer membrane protein H from Pasteurella
multocida A:3 in mice. Res Vet Sci. 2012; 93(3): 1261-1265.
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multocida. Sci World J. 2014; 9(2): 1-7.
2. Shivachandra SB, Kumar A, Yogisharadhya R, Viswas KN. Immunogenicity of highly
conserved recombinant VacJ outer membrane lipoprotein of Pasteurella multocida. Vaccine.
2014; 32(2): 290-296.
3. Wilson BA, Ho M. Pasteurella multocida: from zoonosis to cellular microbiology. Clin
Microbiol Rev. 2013; 26(3): 631-55.
4. Jarvinen LZ, Hogenesch H, Suckow MA, Bowersock TL. Induction of protective immunity in
rabbits by coadministration of inactivated Pasteurella multocida toxin and potassium
thiocyanate extract. Infect Immun. 1998; 66(8): 3788-3795.
5. Lee KE, Jeoung HY, Lee JY, Lee MH, Choi HW, Chang KS. Phenotypic characterization and
random amplified polymorphic DNA (RAPD) analysis of Pasteurella multocida isolated from
Korean pigs. J Vet Med Sci. 2012; 74(5): 567-573.
6. Xie Z, Li H, Chen J, Zhang H-b, Wang Y-Y, Chen Q, Zhao ZZ, Cheng C, Zhang H, Yang
Y, Wang HN, Gao R. Shuffling of pig interleukin-2 gene and its enhancing of immunity in
mice to Pasteurella multocida vaccine. Vaccine. 2007; 25(48): 8163-8171.
7. Dagleish MP, Christopher Hodgson J, Ataei S, Finucane A, Finlayson J, Sales J. Safety and
protective efficacy of intramuscular vaccination with a live aroA derivative of Pasteurella
multocida B:2 against experimental hemorrhagic septicemia in calves. Infect Immu. 2007; 75
(12): 5837-5844.
8. Trevani AS, Chorny A, Salamone G, Vermeulen M, Gamberale R, Schettini J, Raiden S,
Geffner J. Bacterial DNA activates human neutrophils by a CpG-independent pathway. Eur J
Immunol. 2003; 33(11): 3164-3174.
9. Mapletoft JW, Oumouna M, Kovacs-Nolan J, Latimer L, Mutwiri G, Babiuk LA, van Drunen
Littel-van den Hurk S. Intranasal immunization of mice with a formalin-inactivated bovine
respiratory syncytial virus vaccine co-formulated with CpG oligodeoxynucleotides and
polyphosphazenes results in enhanced protection. J Gen Virol. 2008; 89 (Pt 1): 250-260.
10. Tang ML, Lahtinen SJ, Boyle RJ. Probiotics and prebiotics: clinical effects in allergic disease.
Curr Opinion Pediatr. 2010; 22(5): 626-634.
11. Bode C, Zhao G, Steinhagen F, Kinjo T, Klinman DM. CpG DNA as a vaccine adjuvant.
Expert Rev Vaccines. 2011; 10(4): 499-511.
12. Kumar D, Singh A. Salmonella typhimurium grown in iron-rich media, inactivated with ferric
chloride and adjuvanted with homologous bacterial DNA is potent and efficacious vaccine in
mice. Vaccine. 2005; 23(48-49): 5590-5598.
13. Homayoon M, Tahamtan Y, Kargar M, Hosseini SMH, Akhavan SA. Adjuvant activity of
Pasteurella maltocida A strain, Pasteurella maltocida B strain and salmonella typhimurium
bacterial and on cellular responses against Pasteurella maltocida specific strain infectionsin
Balb/C mice. Trop Med Asian Pac J. 2018; 11(5): 336-341.
14. Cheng HR, Jiang N. Extremely rapid extraction of DNA from bacteria and yeasts. Biotechnol
Lett. 2006; 28(1): 55-59.
15. Chaiyotwittayakun A, Burton L, Weber PSD, Kizilkaya K, Cardoso FF, Erskine RJ.
Hyperimmunization of steers with J5 Escherichia coli bacterin: effects on isotype-specific
serum antibody responses and cross reactivity with heterogeneous Gram-negative bacteria. J
Dairy Sci. 2004; 87(10): 3375-3385.
16. Ahmad TA, Rammah SS, Sheweita SA, Haroun M, El-Sayed LH. Development of
immunization trials against Pasteurella multocida. Vaccine. 2014; 32(8): 909-917.
17. Radhi SN. Optimization of heavy metals chlorides resistance by Staphylococcus aureus and its
ability to remove them. Iraqi J Sci. 2012; 53(4): 778-785.
18. Herath C, Kumar P, Singh M, Kumar D, Ramakrishnan S, Goswami TK, Singh A, Ram GC.
Experimental iron-inactivated Pasteurella multocida A: 1 vaccine adjuvanted with bacterial
DNA is safe and protects chickens from fowl cholera. Vaccine. 2010; 28(11): 2284-2289.
19. Homayoon M, Tahamtan Y, Kargar M, Hosseini SMH, Akhavan S.A. Pasteurella multocida
inactivated with ferric chloride and adjuvanted with bacterial DNA is a potent and efficacious
vaccine in Balb/c mice. J Med Microbiol. 2018; 67(9): 1383-1390.
20. Freitas E, Marinho AC, Albuquerque D, Teles L, Sindeaux M, Salles MT, Sousa DC, Lima
MM, Silva MG, Fernandes D. Adjuvant activity of peanut, cottonseed and rice oils on cellular
and humoral response. Vacci Monitor. 2013; 22(1): 4-9.
21. Sayed HE, Ashry E, Ahmad TA. The use of propolis as vaccine’s adjuvant. Vaccine. 2012; 31
(1): 31-39.
22. Wu C, Qin X, Li P, Pan T, Ren W, Li N, Peng Y. Transcriptomic analysis on responses of
murine lungs to Pasteurella multocida infection. Front Cell Infect Microbiol. 2017; 7: 251.
23. Nakae S, Komiyama Y, Nambu A, Sudo K, Iwase M, Homma I, Sekikawa K, Asano M,
Iwakura Y. Antigen-specific T cell sensitization is impaired in IL-17-deficient mice, causing
suppression of allergic cellular and humoral responses. Immunity. 2002; 17(3): 375-387.
24. Al-Samarrae EAA, Al-Shweey SHA. The synergistic effect of BCG and Pasteurella multocida
vaccines in local rabbits. Al-Anbar J Vet Sci. 2012; 5(1): 1-6.
25. Hanagata N. CpG oligodeoxynucleotide nanomedicines for the prophylaxis or treatment of
cancers, infectious diseases, and allergies. Int J Nanomedicine. 2017; 12: 515-531.
26. Hirahara K, Poholek A, Vahedi G, Laurence A, Kanno Y, Milner JD, O'Shea JJ. Mechanisms
underlying helper T-cell plasticity: implications for immune-mediated disease. J Allergy Clin
Immunol. 2013; 131(5): 1276-1287.
27. Okay S, Özcengiz E, Gürsel I, Özcengiz G. Immunogenicity and protective efficacy of the
recombinant Pasteurella lipoprotein E and outer membrane protein H from Pasteurella
multocida A:3 in mice. Res Vet Sci. 2012; 93(3): 1261-1265.