Assessment of immune response of recombinant glycoprotein D (gD) of infectious laryngotracheitis (ILT) virus expressed in Pichia pastoris
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Date
2020-02
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Abstract
Infectious laryngotracheitis (ILT), caused by Gallid herpesvirus1, is one of the most important diseases
of poultry that affects chickens of all age groups. The present study was aimed at elucidation of immune
response of recombinant glycoprotein D (gD) of ILT virus through in vitro and in vivo studies. Hence, the
truncated ectodomain of gD was cloned in pPICZαA shuttle vector and was integrated into Pichia pastoris
strain GS115 yeast chromosomal DNA. The recombinant protein (eu-rgD) was expressed in 72 hours of
methanol metabolism and was analyzed by SDS-PAGE followed by western blotting. Further, the protein
was confirmed by MALDI-TOF-MS/MS analysis. The immunogenicity of eu-rgD was assessed through
immunization trial in specific pathogen free (SPF) chicken model. The antigen induced cellular proliferation of
peripheral blood mononuclear cells (PBMCs) was appreciated by lymphocyte proliferation assay. Assessment
of the antigen specific immune response by IFN-γ cytokine profiling in PBMCs through competitive ELISA
had revealed a significantly higher level of IFN-γ response in PBMCs primed with orally administered
antigen to those primed parenterally. On assessment of seroconversion by indirect ELISA developed in the
study, it was observed that there was no significant difference across the parenterally inoculated groups (high
dose, medium dose and low dose groups), which indicated a lower dose would be sufficient for an efficient
seroconversion. However, there was poor seroconversion found in the orally administered group. In the
present study, the seroconversion to the yeast expressed recombinant glycoprotein antigen was assessed to
forecast its immunogenicity. It was found optimum in producing ideal seroconversion in parenteral route
administration strategies with adjuvant.
Description
TNV_20thMVC_PP_Feb-2020_OP21
Keywords
Veterinary Science