Original Article

Rising Cellular Immune Response after Injection of pVax/iutA: A Genetic DNA Cassette as Candidate Vaccine against Urinary Tract Infection

Abstract

Background: Uropathogenic Escherichia coli (UPEC) are major bacterial agent of Urinary Tract Infection (UTI). This infection is more prevalent among women because approximately half of all women will experience a UTI in their life-time and near a quarter of them will have a recurrent infection within 6–12 months. IutA protein has a major role during UPEC pathogenesis and consequently infection. Therefore, the aim of current study was assessment of IutA protein roles as a potential candidate antigen based for vaccine designing.

Methods: This survey was conducted during 2014-2015 at the University of Tehran, Iran. Chromosomal DNA extracted from E. coli 35218 and iutA gene amplified by PCR. The amplicon cloned to pVax.1 eukaryotic expression vector and recombinant vector confirmed by sequencing. The iutA gene expression in genetic cassette of pVax/iutA was evaluated in COS7 cell line by RT-PCR. Then, injected to mouse model, which divided to three groups: injected with pVax vector, PBS and pVax/iutA cassette respectively in two stages (d 1 and 14). One week after the second injection, bleeding from immunized mouse was performed and IFN-gamma was measured.

Results: The mice immunized with pVax/iutA showed increased interferon-γ responses significantly higher than two non-immunized groups (P<0.05).

Conclusion: Cellular immune response has a main protective role against UTI. Raising this kind of immune response is important to preventing of recurrent infection. Moreover, the current DNA cassette will be valuable for more trying to prepare a new vaccine against UTI.

 

 

Johnson JR, Owens K, Gajewski A, Kuskowski MA (2005). Bacterial Characteristics in Relation to Clinical Source of Escherichia coli Isolates from Women with Acute Cystitis or Pyelonephritis and Uninfected Women. J Clin Microbiol, 43 (12): 6064-72.

Snyder JA, Lloyd AL, Lockatell CV, Johnson DE, Mobley HLT (2006). Role of Phase Variation of Type 1 Fimbriae in a Uropathogenic Escherichia coli Cystitis Isolate during Urinary Tract Infection. Infect Immun, 74 (2):1387-93.

Hooton TM. Recurrent urinary tract infection in women (2001). Int J Antimicrob Agents, 17 (4):259-68.

Dodson KW, Pinkner JS, Rose T, Magnusson G, Hultgren SJ, Waksman G (2001). Structural Basis of the Interaction of the Pyelonephritic E. coli Adhesin to Its Human Kidney Receptor. Cell, 105 (6):733-43.

Svanborg C, Godaly G (1997). Bacterial virulence in urinary tract infection. Infect Dis Clin North Am, 11 (3):513-29.

Yasmeen Kausar SKC, Shobha DN, LHHaleshand MRC handrasekhar (2009). Virulence factors, Serotypes and Antimicrobial Suspectibility Pattern of Escherichia coli in Urinary Tract Infections. Al Ameen J Med Sci, 2 (1):47-5.

Karr JF, Nowicki BJ, Truong LD, Hull RA, Moulds JJ, Hull SI (1990). pap-2-encoded fimbriaead here to the P blood group-related glycosphingolipids tage-specific embryonic antigen 4 in the human kidney. Infec Immun, 58 (12):4055-62.

Blanco M, Blanco JE, Alonso MP, Blanco J (1996). Virulence factors and O groups of Escherichia coli isolates from patients with acute pyelonephritis, cystitis and asymptomatic bacteriuria. Eur J Epidemiol, 12 (2):191-8.

Russo TA, McFadden CD, Carlino-Mac-Donald UB, Beanan JM, Barnard TJ (2002). IroN functions as a siderophore re¬ceptor and is a urovirulence factor in an extra intestinal pathogenic isolate of Esche-richia coli. Infect Immun, 70:7156-60.

Johnson JR, Jelacic S, Schoening LM, Clabots C, Shaikh N (2005). The IrgA homologue adhesin Iha is an Escherichia coli virulence factor in murine urinary tract infection. In¬fect Immun, 73:965–971.

Russo TA, Carlino UB, Johnson JR (2001). Identification of a new iron-regulated vir¬ulence gene, ireA, in an extra intestinal pathogenic isolate of Escherichia coli. Infect Immun, 69:6209–6216.

Hagan EC, Mobley HL (2009). Haem acqui¬sition is facilitated by a novel receptor Hma and required by uropathogenic Esch¬erichia coli for kidney infection. Mol Micro¬biol, 71 (1):79-91

Liu MA (2003) DNA vaccines: a review. J Internal Med, 253 (4): 402-410.

Alteri CJ, Hagan EC, Sivick KE, Smith SN, Mobley HLT (2009). Mucosal Immunization with Iron Receptor Antigens Protects against Urinary Tract Infection. PLoS Pathog, 5 (9):e1000586.

Cui ZH (2005). DNA vaccine. Adv Genet, 54: 257-289.

Martinez JJ, Mulvey MA, Schiling JD et al. (2000). Type l pilus mediated bacterial in¬vasion of bladder epithelial cells. EMBO J, 19: 2803-2812.

Sambrook J, Maccallum P, Russell D (2001). Molecular Cloning: A Laboratory Manual. 3rd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.

Kozak, M (2002). Pushing the limits of the scanning mechanism for initiation of translation. Gene, 299: 1-34.

Johnson JR (1991). Virulence factors in Esch¬erichia coli urinary tract infection. Clin Micro¬biol Rev, 4: 80.

Russo TA, McFadden CD, Carlino-Mac-Donald UB, Beanan JM, Olson R, Wilding GE (2003). The siderophore receptor IroN of extraintestinal pathogenic Esche¬richia coli is a potential vaccine candidate. In¬fect Immun, 71: 7164-7169.

Imani Fooladi AA, Bagherpour G, Khoramabadi N, Fallah Mehrabadi J, Mahdavi M, Halabian R (2014). Cellular immunity survey against urinary tract infection using pVax/fimH cassette with mammalian and wild type codon usage as a DNA vaccine. Clin Exp Vaccine Res, 3 (2):185-93.

Thumbikat P, Waltenbaugh C, Schaeffer AJ, Klumpp DJ (2006). Antigen-specific responses accelerate bacterial clearance in the bladder. J Immunol (Baltimore, Md : 1950), 176 (5):3080-6.

Jones-Carson J, Balish E, Uehling DT (1999). Susceptibility of immunodeficient gene-knockout mice to urinary tract infection. J Urol, 161 (1):338-41.

Wieser A, Romann E, Magistro G, Hoffmann C, Norenberg D, Weinert K, et al. (2010). A multiepitope subunit vaccine conveys protection against extraintestinal pathogenic Escherichia coli in mice. Infect Immun, 78 (8):3432-42.

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IssueVol 45 No 7 (2016) QRcode
SectionOriginal Article(s)
Keywords
Genetic vaccination Uropathogenic Escherichia coli IutA

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How to Cite
1.
BAKHTIARI R, AHMADIAN S, FALLAH MEHRABADI J. Rising Cellular Immune Response after Injection of pVax/iutA: A Genetic DNA Cassette as Candidate Vaccine against Urinary Tract Infection. Iran J Public Health. 2016;45(7):890-896.