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Anthrax is primarily a disease of herbivorous animals including cattle, buffalo, sheep, and goat. Anthrax causes per acute
death in the animals, hence treatment is neither possible nor pragmatic approach to control the disease. So vaccination is
considered as a key strategy for prevention and control of anthrax in ruminants. In the present study, bicistronic DNA vaccine
construct for anthrax using protective antigen (pag) and catalytically inactivated mutated lethal factor (mlef) along with
monocistronic construct encoding pag and mlef were developed. The lethal factor was catalytically inactivated by mutating the
GAA codon to TGC codon (E687C) by primer based site directed mutagenesis. Simultaneously both pag and mlef were cloned
and expressed in prokaryotic expression system and recombinant protein was used for booster immunization. Prepared DNA
vaccine constructs were tested for their functional expression by transfection of MDBK cell line followed by indirect immune-
fluorescence analysis. The immune response study was done in mice with these DNA vaccines boosted with DNA vaccine or
recombinant protein or anthrax spore vaccine on 14
DPV and monitoring of immune response was done by ELISA.
Antibodies were detectable from 7
day onwards and peaked on 42
day. Cell mediated immune response was assessed by
quantitating level of IFN
, IL-2 and IL-4. The level of these three cytokines were high in the (pMlef-Pag+ rPag+ rMlef) and
(pPag +rPag) groups. The immunized mice were challenged with 1000 LD
of Bacillus anthracis IVRI strain on 42
the various vaccine groups, pMlef-Pag+rPag+rMlef and pPag+rPag groups were found to provide maximum protection (66.66%)
while, pMlef-Pag + pMlef-Pag, pPag+pPag and AVA induced only 50%, 33.33%, and 50% protection respectively. Considering
the protective efficacy, immune responses and stimulation of cytokines pMlef- pPag+rPag+rMlef vaccine seems to be better than
other vaccines. Among the DNA vaccine group pMlef-Pag had shown better immune response and protective efficacy than
pPag indicating synergestic effect of pMlef on immune response and protective efficacy.
Bincy Joseph has completed graduation in veterinary sciences (BVSc & AH) from Kerala Agricultural University with KAU merit scholarship. She
has completed post graduation in Veterinary Bacteriology from Indian Veterinary Research Institute with ICAR Junior Research Fellowship and
completed doctorate in veterinary bacteriology from Indian veterinary research institute with Senior Research Fellowship. She has published 8
research articles in various reputed journals and participated in more than 10 national and international conferences and presented research papers.
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