title: Self Replicating Gene Vaccine Carrying P1-2A Gene of FMDV Serotype O and its Effects on the Immune Responses of Cattle.
abstract: DNA vaccines are considered as alternatives to live attenuated ones for those diseases like foot-and-mouth disease (FMD) where the production and application of live vaccines have been found unsuccessful. However, stability of DNA and the quantity of antigen expressed are the major limitation with naked DNA vaccines. To address these issues self replicating gene vaccine construct was made for foot-and-mouth disease virus (FMDV) type 'O' and studied. The vector for vaccine construct, designated as pSinCMVVac carried CMV promoter and Poly(A) signal sequences at 5' and 3' end of Sindbis replicase gene respectively. Gene for structural protein precursor (P1-2A) of FMDV serotype 'O' was inserted into pSinCMVVac under subgenomic promoter. 5'UTR (untranslated region) of FMDV was introduced upstream of P1-2A to enhance the level of expression of cloned gene. Functionality of the vaccine construct was confirmed in vitro and in vivo. The self-replicating gene vaccine construct was tested in cattle in comparison with naked DNA vaccine carrying P1-2A and 3CD (pUP3CD). Humoral immune response by ELISA and SNT and cellular response by lymphoproliferation assay using MTT were studied. The default approach of using self replicating gene vaccine in high dose and multiple injection in cattle as followed in our studies might result in immunosuppression as this was observed in our subsequent experiments in guinea pigs. Hence based on dose response studies, vaccine strategy needs to be decided. However, the approach of using Sindbis polymerase gene and UTR in FMDV vaccine is the first report and shows future scope of developing such vaccines.
authors: National Research Center on Camel, Post Bag No 7, Jorbeer, Bikaner, 334001 Rajasthan India.
authors: G
authors: G
authors: Nagarajan
authors: P
authors: P
authors: Ravikumar
authors: C
authors: C
authors: Ashok Kumar
authors: G R
authors: GR
authors: Reddy
authors: H J
authors: HJ
authors: Dechamma
authors: V V S
authors: VV
authors: Suryanarayana
keywords: Cattle; FMD; Immune response; Self replicating RNA vaccine; Sindbis replicase gene
references: J Virol Methods. 1999 Apr;79(1):75-84
references: 10328537
references: Science. 1989 Mar 3;243(4895):1188-91
references: 2922607
references: Vaccine. 1992;10(14):1022-6
references: 1335197
references: J Virol. 1997 Apr;71(4):3138-45
references: 9060677
references: Antiviral Res. 1995 Oct;28(2):147-57
references: 8585768
references: Nat Med. 1999 Jul;5(7):823-7
references: 10395329
references: Virology. 1989 Nov;173(1):35-45
references: 2554577
references: J Immunol. 1991 Jan 15;146(2):749-55
references: 1702816
references: J Gen Virol. 2001 May;82(Pt 5):1013-1025
references: 11297676
references: Virology. 1989 Jul;171(1):189-204
references: 2525837
references: Intervirology. 2000;43(4-6):247-57
references: 11251380
references: Virology. 1999 Jun 20;259(1):129-34
references: 10364496
references: J Gen Virol. 1988 Sep;69 ( Pt 9):2313-25
references: 2842438
references: Comp Immunol Microbiol Infect Dis. 1999 Oct;22(4):231-46
references: 10465327
references: Biochem Int. 1992 May;26(6):1003-16
references: 1378734
references: Nature. 1970 Aug 15;227(5259):680-5
references: 5432063
references: Dev Biol Stand. 1999;101:39-43
references: 10566773
references: Ann N Y Acad Sci. 1995 Nov 27;772:261-4
references: 8546403
references: Nucleic Acids Res. 1984 Aug 24;12(16):6587-601
references: 6089122
references: J Virol. 1996 Jan;70(1):508-19
references: 8523564
references: J Virol. 1998 Feb;72(2):950-8
references: 9444987
references: Cell. 1994 Nov 4;79(3):497-506
references: 7954815
references: Vaccine. 1994 Dec;12(16):1510-4
references: 7879415
references: Nat Biotechnol. 1998 Jun;16(6):562-5
references: 9624688
references: Biotechnology (N Y). 1991 Dec;9(12):1356-61
references: 1370252
references: J Virol. 1992 Apr;66(4):1835-40
references: 1312607
references: Trends Biotechnol. 1996 Apr;14(4):130-4
references: 8936434
references: Prog Biophys Mol Biol. 1993;60(3):241-60
references: 8396787
references: Vaccine. 1999 Dec 10;18(9-10):765-77
references: 10580187
references: J Virol. 1991 Dec;65(12):6572-80
references: 1658362
references: J Immunol. 1995 Jul 1;155(1):259-65
references: 7602102