Journal of Pure and Applied MicrobiologyVol. 9 No. 3

Immunization of Balb/C mice by Uropathogenic E.coli Recombinant PapG.AcmA Protein

Fatemeh Ashrafi1*, Mohammad Reza Masoumian2, Ramin Fallahzadeh3 and Mohammad D. Ghafari4

1Department of Microbiology, North Tehran branch, Islamic Azad University, Tehran, Iran. 2Department of Chemical Engineering - Biotechnology, Faculty of Biosciences and Biotechnology, Malek-ashtar University of Technology, Tehran, Iran. 3Department of Genetic Engineering, Faculty of Biosciences and Biotechnology, Malek-ashtar University of Technology, Tehran, Iran. 4Researchers and Elites Club, North Tehran Branch, Islamic Azad University, Tehran, Iran.

Received on 06 June 2015 and accepted on 17 August 2015

 

ABSTRACT

Urinary tract infection (UTI) caused by Escherichia coli (UPEC) is one of the most common infections in the world today, which has become resistant to most antibiotics. Despite many efforts, there is no vaccine to protect humans against the infection. PapG.AcmA recombinant protein can be a particular vaccine to treat this infection. Because it helps prevent bacteria from sticking to the surface of the host cell. In this study, after the purification of the protein with Ni-NTA column, the immunity of protein was assessed in Balb/C mice. Experimental mice were divided into three groups and were immunized with Freund adjuvanted PapG.AcmA protein or alum adjuvanted protein or PBS (as control group). Then, lymphocyte proliferation assay was conducted via Brdu/ELISA based methods, IL-4 and IFN-g cytokines secretion was quantified with commercial ELISA kit; in addition, total antibody, IgG1 and IgG2a subtyping were assayed using ELISA method. The results show that recombinant PapG.AcmA protein was resulted to induction of cellular and humoral immune responses in the mice. This effect of nano-vaccination strategy (with TLR5 agonists) also varies on different parameters of the immune system; in this regard, the cellular immune responses had a higher stimulation level compared with the humoral immune responses.

Keywords : Uropathogenic E.coli, UTI, PapG.AcmA, expression, immune responses.