Use of immunogenic moiety of Pseudomonas aeruginosa exotoxin A as a DNA vaccine in experimentally contaminated mice

Document Type : Immunology

Authors

1 Department of Microbiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

2 Department of Immunology, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran.

3 Department of Horticulture, Faculty of Agriculture, Urmia University, Urmia, Iran.

Abstract

Background: DNA immunization is an appropriate method to produce an immunological response. Pseudomonas aeruginosa produces exotoxin A which is highly cytotoxic for eukaryotic cells. Since domains II (translocation domain) and 1b of the toxin have antigenic qualities, so they could be  useful candidates to protect against pseudomonas infections. Objectives: To evaluate if recombinant plasmid containing immunogenic domain of exotoxin A might be protective against Pseudomonas aeruginosa infections. Methods: To study the biologic and immunological effects of antigenic domains of exotoxin A, plasmid expression vector (pET28a) containing domain II and 1b of exotoxin was constructed. To evaluate the effects of intracellular recombinant gene expression, BALB/C mice were immunized with the recombinant plasmid and then subjected to third degree thermal injury and the humoral immunity responses were assayed. Results: Immunization with the recombinant plasmid containing translocation and 1b domains of exotoxin A resulted in  increasing antibodies production (IgA and IgG) against Pseudomonas aeruginosa. DNA immunization significantly decreased the bacterial count liver, spleen, blood and inoculated burns after challenging with P. aeruginosa and dramatically improved the survival rate of burn-injured mice. Conclusions: Finally, immunization by gene encoding antigenic products may be a good technique for protection against P. aeruginosa infections.
 

Keywords


 
Baxter, D. (2007) Active and passive immunity, vaccine types, excipients and licensing. Occup Med (Lond). 57: 552-56.
Bayat, E., Kamali, M., Zareei  Mahmoodabadi,  A., Mortazavi, Y., Ebrahim Habibi, A., Amini,  B., Javadi, H.R., Farhadi, N., Faghihi, H. (2010) Isolation, determination and cloning of translocation domain of  exotoxin A  from  Pseudomonas aeruginosa. Kowsar Medical Journal. 15: 149-54.
Chen, T.Y., Lin, C.P., Loa, C.C., Chen, T.L., Shang, H.F., Hwang, J., Hui, C.F. (1999) A nontoxic Pseudomonas aeruginosa exotoxin A induces active Immunity and passive protective antibody against Pseudomonas exotoxin A intoxication. J Biomed Sci. 6: 357-63.
Denis-Mize, K.S., Price, B.M., Baker, N.R., Galloway, D. (1999) Analysis of Immunization  with DNA encoding Pseudomonas aeruginosa exotoxin A. FEMS Immunol Med Microbiol.  27: 147- 54.
Fujimura, T., Suzuki, T., Mitsuyama, M, Saito,  H.,  Nomoto, K. (1989) Antibody independent   Protection against  Pseudomonas aeruginosa infection  in mice after  treatment  with a  homologus strain vaccine. J Med Microbial. 28: 101-8.
Harkness,  J.E. (1993) A practitioners guide to Domestic Rodents. American Animal Hospital Association. Lakewood, USA.
Hertle, R., Mrsny, R., Fitzgerald, D.J. (2001) Dual–function vaccine for Pseudomonas aeruginosa: characterization of chimeric exotoxin A– Pilin Protection. Infect Immun. 69: 6962-69.
Hosseini  Jazani,  N., Parsania,  S., Sohrabpour, M., Mazloomi,  E., Karimzad, M., Shahabi,  SH. (2010) Naloxone and alum synergistically augment adjuvant activities of each other in a mouse vaccine  model of Sallmonella typhimurium infection. Immunobiology. 6: 744-51.
Hung, C.F., Cheng, W.F., Hsu, K.F., Chai, C.Y., He, L., Ling, M., WU, T.C. (2001) Cancer immunotherapy using a DNA vaccine encoding the  translocation  domain of a bacterial toxin linked to a tumor antigen. Cancer Res. 61: 3698-3703.
Khan, A., Cerniglia, C. (1994) Detection of Pseudomonas aeruginosa from clinical environmental samples by amplification of the exotoxin A gene using PCR. Appl Environ Microbiol. 60: 3739-45.
Kinoshita, M., Shinomiya, N., Ono,  S., Tsujmoto, H., Kawabata, T., Matsumoto, A., Hiraide, H., Seki, S. (2006) Restoration of natural IgM production from liver B cells by Exogenous Il-18 improves the survival of burn injured mice infected with  Pseudomonas aeruginosa. J Immunol. 177: 4627-35.
Lang, A.B., Horn,  M.P., Imboden,  M.A., Zuercher,  A.W. (2004) Prophylaxis and therapy of Pseudomonas aeruginosa infection  in cystic fibrosis and unocompromised patients. Vaccine. 22: S44-S48.
Manafi, A., Kohanteb, J., Mehrabani, D., Japoni, A., Amini, M., Naghmachi, M., Zaghi,  A.H., Khalili, N. (2009) Active immunization using exotoxin  A confers protection against Pseudomonas aeruginosa infection in mouse burn model. BMC Microbiol. 9: 23-27.
Mcvay, C.S., Velasquez, M., Fralik, J.A. (2007) Phage therapy of Pseudomonas aeruginosa  infection in a mouse burn wound model. Antimicrob Agents Chemother. 51: 1934-38.
Nouri Gharajelar, S., Ahmadi, M., Hosseini, B. (2013) Cloning and expression of the immunogenic moiety of Pseudomonas aeruginosa exotoxin A. Biological Journal of Microorganism. 1: 7-14.
Pavlovskis, O.R., Edman, D.C., Leppla, SH., Wretlind, B., Lewis, L.R., Martin, K.E. (1981)Protection against toxoids, experimental Pseudomonas aeruginosa infection in mice by active immunization with Exotoxin A. Infect Immun. 32: 681-89.
Pollack, M. (1983) The role of exotoxin A in Pseudomonas disease And  Immunity. Rev Infect Dis. 5: S979-84.
Rumbaugh, K.P., Griswold, J.A., Iglewski, B.H., Hamood, A.N. (1999) Contribution of quorum sensing to the virulence of  Pseudomonas aeruginosa in burn wound infections. Infect Immun. 67: 5854-62.
Staczek, J., Gilleland, L.B., Vanderhyde, H.C., Gilleland, J.R. (2003) DNA vaccines against chronic lung infections by Pseudomonas aeruginosa. FEMS Immunol Med Microbiol. 37: 147-53.
Wahren, B., Liu, M.A. (2014) DNA vaccines: Recent developments and the future. Vaccines. 2: 785-796.
Whalen, R.G. (1996) DNA vaccines for emerging infectious diseases. Emerg Infect Dis. 2: 168-175.