Cloning and expression of g-gene of chandipura virus and evaluation of immunogenicity in mice

Abstract

Chandipura virus (CHPV) belongs to family Rhabdovoridae and has emerged as an important encephalitis causing pathogen with high mortality (55-75%) among pediatric population (aged between 2months to 15 years) from three Indian states. Epidemic causing potentiality and rapid onset of disease (death between 24 hours to 48 of hospitalization) are some of the important attributes of CHPV. Till date there is no vaccine available and the treatment is only symptomatic. So far, the disease is prevalent in restricted areas of the country and vaccination of children may prove beneficial in the control / elimination of this life-threatening and rapidly progressing disease killing large proportion of encephalitis cases. Considering the well-documented immunological efficacy of Glycoprotein in other closely related viruses i.e. Rabies, Vescular stomatitis virus etc. the recombinant Glycoprotein (rGp) was the protein of choice in our study. The entire G-gene of CHPV (CIN0327R strain from Andhrapradesh epidemic, 2003) was cloned and expressed by Baculovirus expression system. The expression of rGp was confirmed by immunoblot analysis. It was pleasing to note that the protein was predominantly present in the supernatant of the SF9 cells. The rGp was purified by HPLC and used for mice immunization, 3 doses, and 4 weeks apart. The sero-conversion was immunogen (rGp) concentration dependent.