Design of a predicted MHC restricted short peptide immunodiagnostic and vaccine candidate for Fowl adenovirus C in chicken infection.

Hugo Valdivia-Olarte,Francesca Falconi-Agapito,Manolo Fernández-Díaz,Iván Best,David Requena,Ray Izquierdo,Milagros Zavaleta,Mirko Zimic,Manuel Ramirez,Luis E Saravia

doi:10.6026/97320630011460

Hugo Valdivia-Olarte, Francesca Falconi-Agapito + Show 8 more

Open Access

https://doi.org/10.6026/97320630011460

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Abstract

Fowl adenoviruses (FAdVs) are the ethiologic agents of multiple pathologies in chicken. There are five different species of FAdVs grouped as FAdV-A, FAdV-B, FAdV-C, FAdV-D, and FAdV-E. It is of interest to develop immunodiagnostics and vaccine candidate for Peruvian FAdV-C in chicken infection using MHC restricted short peptide candidates. We sequenced the complete genome of one FAdV strain isolated from a chicken of a local farm. A total of 44 protein coding genes were identified in each genome. We sequenced twelve Cobb chicken MHC alleles from animals of different farms in the central coast of Peru, and subsequently determined three optimal human MHC-I and four optimal human MHC-II substitute alleles for MHC-peptide prediction. The potential MHC restricted short peptide epitope-like candidates were predicted using human specific (with determined suitable chicken substitutes) NetMHC MHC-peptide prediction model with web server features from all the FAdV genomes available. FAdV specific peptides with calculated binding values to known substituted chicken MHC-I and MHC-II were further filtered for diagnostics and potential vaccine epitopes. Promiscuity to the 3/4 optimal human MHC-I/II alleles and conservation among the available FAdV genomes was considered in this analysis. The localization on the surface of the protein was considered for class II predicted peptides. Thus, a set of class I and class II specific peptides from FAdV were reported in this study. Hence, a multiepitopic protein was built with these peptides, and subsequently tested to confirm the production of specific antibodies in chicken.

Highlights

Fowl adenoviruses (FAdVs) are widely distributed, and some species are associated with important poultry diseases, representing current threats of serious economic losses for the aviculture industry
Prediction of immunogenic MHC-I and MHC-II FAdV peptides Prediction of peptide affinity to MHC-I and MHC-II We explored the FAdV genomes looking for potential immunogenic peptides
The synthetic genes were cloned in pET28a-c Human MHC-I and MHC-II best chicken-substitute-alleles plasmid, using the NcoI and XhoI restriction sites, including a We found nine human MHC-I alleles highly similar to the six-His-Tag at the 3’ end for affinity chromatography chicken MHC-I

Summary

Introduction

FAdVs are widely distributed, and some species are associated with important poultry diseases, representing current threats of serious economic losses for the aviculture industry. Fowl adenovirus (FAdV) is classified into five different species Proteins are the essential immune-target structures, which in the MHC class I/II (MHC-I/II) pathway are processed to 811/15-18 mer peptides. In this way, peptides that bind to MHC molecules are presented and potentially recognized by cytotoxic T cells (CD8/CD4), which can lead to an immune response [9]. The most selective step in this antigen presentation is the peptide binding to MHC [10].

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