Immunogenicity of Amino Acid Region 7601-8140 in Biofilm Associated Protein of Acinetobacter baumannii

Authors
M. Jalali 1
I. Rasooli 2
K. Ahmadi Zanoos 1
A. Jahangiri 1
M. Jalali Nadooshan 3
S. Darvish Alipour Astaneh 4
1 Department of Biology, Shahed University, Tehran, Iran
2 Department of Biology, Molecular Microbiology Research Center, Shahed University, Tehran, Iran
3 Department of Pathology, Faculty of Medicine, Molecular Microbiology Research Center, Shahed University, Tehran, Iran
4 Department of Microbiology, Shahed University, Tehran, Iran
Abstract
Objective: Acinetobacter baumannii (A. baumannii) is a major hospital pathogen with a high capacity to resist most common anti-microbial agents. A. baumannii is the etiologic agent for various illnesses including pneumonia, meningitis, and bloodstream infections. Biofilm associated proteins (Bap) are specific cell surface proteins essential for the formation of biofilm and play a main role in its pathogenicity. Previously, we have studied various regions of this protein. Considering different criteria, some regions were introduced as conserved and immunogenic. The immunogenicity of one of those regions pertaining to amino acids 706-1076 previously examined has shown that its expression triggers high antibody levels when injected to mice thereby protecting the animals against the bacterium. The present study examines region 4 of the Bap protein in order to validate the previous bioinformatics studies and its immunogenicity. Methods: In order to obtain immunity against this pathogen, a 1620 bp gene from Bap was amplified and cloned in pET32a. This region from Bap was cloned, expressed and verified by monoclonal antibodies. BALB/c mice were immunized by subcutaneous injection of the pure recombinant protein. Mice immune response was determined by ELISA. Results: High titer of raised antibodies implied that the recombinant protein was a strong antigen and immunogen. Conclusion: The results indicate that this protein can be a suitable choice for developing a new recombinant vaccine against A. baumannii.

Keywords

[1]     Beijerinck M. Pigmenten als oxydatieproducten gevormd door bacterien. Versl Koninklijke Akad Wetensch Amsterdam 1911; 19: 1092-103.
[2]     Towner K. The genus Acinetobacter. In: Dworkin M, Falkow S, Scheifer KH, Rosenberg E, Stackebrandt E. Prokaryotes, Volume 6, New York: Springer, 2006; p: 746-58.
[3]     Chaari A, Mnif B, Bahloul M, Mahjoubi F, Chtara K, Turki O, Gharbi N, Chelly H, Hammami A, Bouaziz M. Acinetobacter baumannii ventilator-associated pneumonia: epidemiology, clinical characteristics, and prognosis factors. Int J Infect Dis 2013; 17(12): e1225-8.
[4]     Peleg AY, Seifert H, Paterson DL. Acinetobacter baumannii: emergence of a successful pathogen. Clin Microbiol Rev 2008; 21(3): 538-82.
[5]     Towner KJ. Acinetobacter: an old friend, but a new enemy. J Hosp Infect 2009; 73(4): 355-63.
[6]     Chang HC, Wei YF, Dijkshoorn L, Vaneechoutte M, Tang CT, Chang TC. Species-level identification of isolates of the Acinetobacter calcoaceticus-Acinetobacterbaumannii complex by sequence analysis of the 16S-23S rRNA gene spacer region. J Clin Microbiol 2005; 43(4): 1632-9.
[7]     van den Broek PJ, Arends J, Bernards AT, De Brauwer E, Mascini EM, van der Reijden TJ, Spanjaard L, Thewessen EA, van der Zee A, van Zeijl JH, Dijkshoorn L. Epidemiology of multiple Acinetobacter outbreaks in The Netherlands during the period 1999-2001. Clin Microbiol Infect 2006; 12(9): 837-43.
[8]     Dijkshoorn L, Nemec A, Seifert H. An increasing threat in hospitals: multidrug-resistant Acinetobacter baumannii. Nat Rev Microbiol 2007; 5(12): 939-51.
[9]     Espinal P, Martí S, Vila J. Effect of biofilm formation on the survival of Acinetobacter baumannii on dry surfaces. J Hosp Infect 2012; 80(1): 56-60.
[10]  Jeong SH, Bae IK, Park KO, An YJ, Sohn SG, Jang SJ, Sung KH, Yang KS, Lee K, Young D, Lee SH. Outbreaks of imipenem-resistant Acinetobacter baumannii producing carbapenemases in Korea. J Microbiol 2006; 44(4): 423-31.
[11]  Gordon NC, Wareham DW. Multidrug-resistant Acinetobacter baumannii: mechanisms of virulence and resistance. Int J Antimicrob Agents 2010; 35(3): 219-26.
[12]  Cevahir N, Demir M, Kaleli I, Gurbuz M, Tikvesli S. Evaluation of biofilm production, gelatinase activity, and mannose-resistant hemagglutination in Acinetobacter baumannii strains. J Microbiol Immunol Infect 2008; 41(6): 513-8.
[13]  Lee HW, Koh YM, Kim J, Lee JC, Lee YC, Seol SY, Cho DT, Kim J. Capacity of multidrug-resistant clinical isolates of Acinetobacter baumannii to form biofilm and adhere to epithelial cell surfaces. Clin Microbiol Infect 2008; 14(1): 49-54.
[14]  Milletli Sezgin F, Coban AY, Gunaydin M. Investigation of biofilm formation in Acinetobacter baumannii isolates and their colistin susceptibilities in biofilm. Int J Antimicrob Agents 2013; 41(2): 199.
[15]  Tomaras AP, Dorsey CW, Edelmann RE, Actis LA. Attachment to and biofilm formation on abiotic surfaces by Acinetobacter baumannii: involvement of a novel chaperone-usher pili assembly system. Microbiology 2003; 149(Pt 12): 3473-84.
[16]  Loehfelm TW, Luke NR, Campagnari AA. Identification and characterization of an Acinetobacterbaumannii biofilm-associated protein. J Bacteriol 2008; 190(3): 1036-44.
[17]  Cucarella C, Solano C, Valle J, Amorena B, Lasa I, Penadés JR. Bap, a Staphylococcus aureus surface protein involved in biofilm formation. J Bacteriol 2001; 183(9): 2888-96.
[18]  Donlan RM, Costerton JW. Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev 2002; 15(2): 167-93.
[19]  Hall-Stoodley L, Costerton JW, Stoodley P. Bacterial biofilms: from the natural environment to infectious diseases. Nat Rev Microbiol 2004; 2(2): 95-108.
[20]  Luppens SB, Reij MW, van der Heijden RW, Rombouts FM, Abee T. Development of a standard test to assess the resistance of Staphylococcusaureus biofilm cells to disinfectants. Appl Environ Microbiol 2002; 68(9): 4194-200.
[21]  Stoodley P, Sauer K, Davies DG, Costerton JW. Biofilms as complex differentiated communities. Annu Rev Microbiol 2002; 56: 187-209.
[22]  Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: a common cause of persistent infections. Science 1999; 284(5418): 1318-22.
[23]  Blackwell HE. Bacterial crowd control with iron. Chem Biol 2005; 12(7): 721-3.
[24]  Lasa I, Penadés JR. Bap: a family of surface proteins involved in biofilm formation. Res Microbiol 2006; 157(2): 99-107.
[25]  Rahbar MR, Rasooli I, Mousavi Gargari SL, Amani J, Fattahian Y. In silico analysis of antibody triggering biofilm associated protein in Acinetobacter baumannii. J Theor Biol 2010; 266(2): 275-90.
[26]  Fattahian Y, Rasooli I, Mousavi Gargari SL, Rahbar MR, Darvish Alipour Astaneh S, Amani J. Protection against Acinetobacter baumannii infection via its functional deprivation of biofilm associated protein (Bap). Microb Pathog 2011; 51(6): 402-6.
Pathobiology Reserach
Volume 16, Issue 4
February 2014
Pages 15-26