title: Immunogenicity and protective efficacy of recombinant major envelope protein (rH3L) of buffalopox virus in animal models.
abstract: Buffalopox virus, a zoonotic Indian vaccinia-like virus, is responsible for contagious disease affecting mainly buffaloes, cattle and humans. H3L gene, encoding for an immunodominant major envelope protein of intracellular mature virion of orthopoxviruses, is highly conserved and found to elicit neutralizing antibodies. Therefore in the present study, the immunogenicity and protective efficacy of the recombinant H3L protein of buffalopox virus in laboratory animal models has been evaluated. A partial H3L gene encoding for the C-terminal truncated ectodomain of H3L protein (1M to I280) of BPXV-Vij/96 strain was cloned, over-expressed and purified as histidine-tagged fusion protein (50 kDa) from Escherichia coli using Ni-NTA affinity chromatography. The purified rH3L protein was further used for active immunization of guinea pig (250 μg/dose) and adult mice (10 μg and 50 μg/dose) with or without adjuvants (alum, Freund's Complete Adjuvant and CpG). Subsequently, a gradual increase in antigen specific serum IgG as well as neutralizing antibody titres measured by using indirect-ELISA and serum neutralization test respectively, was noted in both guinea pigs and mouse models. Suckling mice immunized passively with anti-H3L serum showed 80% pre-exposure prophylaxis upon challenge with virulent buffalopox virus strain. An indirect-ELISA based on rH3L protein showed no cross-reactivity with hyperimmune sera against sheeppox virus (SPPV), goatpox virus (GTPV), orf virus (ORFV), foot- and- mouth disease virus (FMDV), peste des petits ruminants virus (PPRV) and bluetongue virus (BTV) during the course of study. The study highlights the potential utility of rH3L protein as a safer prophylactic and diagnostic reagent for buffalopox.
authors.affiliation: Pox Virus Laboratory, Division of Virology, ICAR-Indian Veterinary Research Institute (IVRI), Regional Campus, Mukteswar, Nainital (District) 263138, Uttarakhand (UK), India.
authors.forename: Amit
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Pox Virus Laboratory, Division of Virology, ICAR-Indian Veterinary Research Institute (IVRI), Regional Campus, Mukteswar, Nainital (District) 263138, Uttarakhand (UK), India.
authors.forename: Revanaiah
authors.initials: R
authors.lastname: Yogisharadhya
authors.affiliation: Pox Virus Laboratory, Division of Virology, ICAR-Indian Veterinary Research Institute (IVRI), Regional Campus, Mukteswar, Nainital (District) 263138, Uttarakhand (UK), India.
authors.forename: Gnanavel
authors.initials: G
authors.lastname: Venkatesan
authors.affiliation: Pox Virus Laboratory, Division of Virology, ICAR-Indian Veterinary Research Institute (IVRI), Regional Campus, Mukteswar, Nainital (District) 263138, Uttarakhand (UK), India. Electronic address: bhanu6467@gmail.com.
authors.forename: Veerakyathappa
authors.initials: V
authors.lastname: Bhanuprakash
authors.affiliation: Pox Virus Laboratory, Division of Virology, ICAR-Indian Veterinary Research Institute (IVRI), Regional Campus, Mukteswar, Nainital (District) 263138, Uttarakhand (UK), India.
authors.forename: Sathish Bhadravati
authors.initials: SB
authors.lastname: Shivachandra
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D000917:Antibody Formation / Q000276:immunology*; D001483:Base Sequence; D001820:Bluetongue virus / Q000235:genetics / Q000276:immunology*; D018151:Capripoxvirus / Q000276:immunology; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D005260:Female; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology; D006168:Guinea Pigs; D007074:Immunoglobulin G / Q000097:blood; D008297:Male; D051379:Mice; D023421:Models, Animal; D009923:Orf virus / Q000276:immunology; D018107:Peste-des-petits-ruminants virus / Q000276:immunology; D011213:Poxviridae Infections / Q000175:diagnosis / Q000517:prevention & control; D065129:Pre-Exposure Prophylaxis; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D014611:Vaccination; D014615:Vaccinia / Q000276:immunology / Q000517:prevention & control / Q000821:virology*; D014616:Vaccinia virus / Q000235:genetics / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D002352:Carrier Proteins; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C406065:envelope H3L protein, Vaccinia virus
keywords: Animal models; Buffalopox virus; Immunogenicity; Indirect ELISA; Protective efficacy; Recombinant H3L protein