title: Evaluation of Immunogenicity of Cocktail DNA Vaccine Containing Plasmids Encoding Complete GRA5, SAG1, and ROP2 Antigens of Toxoplasma gondii in BALB/C Mice.
abstract: BACKGROUND
  Severe and fatal complications of toxoplasmosis urge development of effective vaccines against the disease. The current study was performed to evaluate cocktail DNA vaccine containing plasmids encoding GRA5, SAG1, and ROP2 genes of Toxoplasma gondii in BALB/c mice in Tarbiat Modares University in 2012.
  
  
  METHODS
  The plasmids containing complete GRA5, SAG1, and ROP2 genes were mass extracted and then the recombinant plasmids were administered via intramuscular injections according to immunized mice three times with three-week intervals. Then splenocytes were cultured, and proliferation as well as cytokine assays were carried out. The other mice in each group were inoculated by the parasite and mortality of the mice was evaluated on a daily basis.
  
  
  RESULTS
  The results of cytokine assay for INF-γ were higher in the mice that received the cocktail DNA containing recombinant plasmids. Evaluation of proliferation of splenocytes using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay indicated induction of cellular response. Measurement of total IgG and the isotypes of IgG1 and IgG2a showed that the cocktail DNA stimulated IgG and IgG2a production in comparison with the control groups (P<0.05). Furthermore, the survival rate of mice in the groups that received the cocktail DNA was significantly higher than that in the control groups (P<0.05).
  
  
  CONCLUSIONS
  Administration of the cocktail DNA vaccine led to production of higher levels of IFN-γ, confirmed by secretion of IgG2a, and the immune response was shifted toward Th1. Thus, the cocktail DNA containing the recombinant plasmids can be an appropriate candidate for immunization against toxoplasmosis.
authors.affiliation: Department of Medical Parasitology, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran.
authors.forename: Razi
authors.initials: R
authors.lastname: Naserifar
authors.affiliation: Department of Parasitology and Entomology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Ghaffarifar
authors.affiliation: Department of Parasitology and Entomology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Abdolhosein
authors.initials: A
authors.lastname: Dalimi
authors.affiliation: Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
authors.forename: Zohreh
authors.initials: Z
authors.lastname: Sharifi
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, Jahrom University of Medical Sciences, Jahrom, Iran.
authors.forename: Kavous
authors.initials: K
authors.lastname: Solhjoo
authors.affiliation: Department of Parasitology and Entomology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Kami
authors.initials: K
authors.lastname: Hosseinian Khosroshahi
keywords: Cocktail DNA Vaccine; Toxoplasma gondii; pcGRA5; pcROP2; pcSAG1
references.citation: World J Gastroenterol. 2006 Apr 21;12(15):2363-8
references.pmid: 16688826
references.citation: Infect Immun. 1996 Sep;64(9):3858-62
references.pmid: 8751939
references.citation: Parasitol Today. 1994 Sep;10(9):358-60
references.pmid: 15275418
references.citation: Vaccine. 2011 Jan 17;29(4):778-83
references.pmid: 21095254
references.citation: Res Immunol. 1993 Jan;144(1):41-4
references.pmid: 8451517
references.citation: Mol Biochem Parasitol. 1988 Jun;29(2-3):203-11
references.pmid: 3045541
references.citation: Parasitol Today. 1994 Aug;10(8):293-6
references.pmid: 15275422
references.citation: Exp Parasitol. 2010 Mar;124(3):272-8
references.pmid: 19874823
references.citation: Parasite Immunol. 1999 May;21(5):261-72
references.pmid: 10320624
references.citation: Exp Parasitol. 2008 Mar;118(3):386-92
references.pmid: 18154953
references.citation: Genet Mol Res. 2008 Apr 08;7(2):305-13
references.pmid: 18551396
references.citation: Ann Inst Pasteur Immunol. 1988 Jan-Feb;139(1):109-17
references.pmid: 2451923
references.citation: Infect Immun. 1999 Dec;67(12):6358-63
references.pmid: 10569750
references.citation: Infect Immun. 2000 Feb;68(2):969-72
references.pmid: 10639474
references.citation: Parasitology. 1988 Aug;97 ( Pt 1):1-10
references.pmid: 3174230