title: Evaluation of recombinant E2 protein-based and whole-virus inactivated candidate vaccines against chikungunya virus.
abstract: OBJECTIVE
  With the re-emergence of chikungunya virus (CHIKV) in an explosive form and in the absence of a commercially available vaccine, we aimed to develop candidate vaccines employing recombinant E2 protein or chemically inactivated whole virus.
  
  
  METHODS
  E2 gene of CHIKV isolate of ECSA genotype was cloned in pET15b vector, expressed and purified (rE2p). The virus was propagated in Vero cell line, purified and inactivated with formalin and BPL individually. Six to eight weeks old female BALB/c mice were immunized intramuscularly with two doses of 10μg, 20μg and 50μg of vaccine formulations with or without adjuvants, 2 weeks apart. The adjuvants evaluated were alum, Mw, CadB (rE2p), alum/Mw (formalin inactivated CHIKV) and alum (BPL-inactivated CHIKV). Humoral immunity was assessed by ELISA and in vitro neutralization test using homologous and heterologous (Asian genotype) strains of CHIKV. Two cohorts of vaccinated mice were challenged separately via intranasal route with homologous virus two and 20 weeks after the 2nd dose. Viral load (CHIKV RNA by real time PCR) was determined in the serum and tissues (muscle, brain, spleen) of the mice challenged with the homologous virus.
  
  
  RESULTS
  Anti-CHIK-antibody titres were dose dependent for all the immunogen formulations. BPL-inactivated vaccines led to the highest ELISA/neutralizing antibody (nAb) titres while alum was the most effective adjuvant. Asian genotype strain could be neutralized by the nAbs. In an adult mouse model, complete protection was offered by the alum-adjuvanted rE2p and both the inactivated vaccines as no virus was detected in the tissues and blood after challenge 2 weeks or 20 weeks-post-2nd dose. However, with rE2p-CadB, very low viremia was recorded on the 2nd day-post-challenge.
  
  
  CONCLUSIONS
  Both rE2p and BPL/formalin-inactivated virus are promising candidate vaccines deserving further evaluation.
authors.affiliation: National Institute of Virology, Microbial Containment Complex, Sus Road, Pashan, Pune 411021, India.
authors.forename: Manish
authors.initials: M
authors.lastname: Kumar
authors.forename: A B
authors.initials: AB
authors.lastname: Sudeep
authors.forename: Vidya A
authors.initials: VA
authors.lastname: Arankalle
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D018354:Alphavirus Infections / Q000517:prevention & control*; D000534:Alum Compounds / Q000008:administration & dosage; D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood; D065632:Chikungunya Fever; D002646:Chikungunya virus / Q000276:immunology*; D002522:Chlorocebus aethiops; D003429:Cross Reactions / Q000276:immunology; D016207:Cytokines / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D056724:Immunity, Humoral; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000276:immunology; D015164:Vaccines, Inactivated / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: D000276:Adjuvants, Immunologic; D000534:Alum Compounds; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D016207:Cytokines; D011994:Recombinant Proteins; D015164:Vaccines, Inactivated; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; C041524:aluminum sulfate