Immunoinformatics and analysis of antigen distribution of Ureaplasma diversum strains isolated from different Brazilian states

Manoel Neres Santos Junior,Izadora Souza Rezende,Bruna Carolina De Brito Guimarães,Lucas Miranda Marques,Lucas Santana Coelho Silva,Bruno Lopes Bastos,Caline Novaes Teixeira Oliveira,Maysa Santos Barbosa,Janaina Marinho Fernandes,Guilherme Barreto Campos,Wanderson Souza Neves,Jorge Timenetsky,Ronaldo Silva Santos,Camila Pacheco Gomes,Nayara Silva De Macêdo Neres

doi:10.1186/s12917-020-02602-1

Abstract

BackgroundUreaplasma diversum has numerous virulence factors that contribute to pathogenesis in cattle, including Lipid-associated membrane proteins (LAMPs). Therefore, the objectives of this study were to evaluate in silico important characteristics for immunobiological applications and for heterologous expression of 36 LAMPs of U. diversum (UdLAMPs) and, also, to verify by conventional PCR the distribution of these antigens in strains of Brazilian states (Bahia, Minas Gerais, São Paulo, and Mato Grosso do Sul). The Manatee database was used to obtain the gene and peptide sequences of the antigens. Similarity and identity studies were performed using BLASTp and direct antigenicity was evaluated by the VaxiJen v2.0 server. Epitope prediction for B lymphocytes was performed on the BepiPred v2.0 and CBTOPE v1.0 servers. NetBoLApan v1.0 was used to predict CD8+ T lymphocyte epitopes. Subcellular location and presence of transmembrane regions were verified by the software PSORTb v3.0.2 and TMHMM v2.2 respectively. SignalP v5.0, SecretomeP v2.0, and DOLOP servers were used to predict the extracellular excretion signal. Physico-chemical properties were evaluated by the web-software ProtParam, Solpro, and Protein-sol.ResultsIn silico analysis revealed that many UdLAMPs have desirable properties for immunobiological applications and heterologous expression. The proteins gudiv_61, gudiv_103, gudiv_517, and gudiv_681 were most promising. Strains from the 4 states were PCR positive for antigens predicted with immunogenic and/or with good characteristics for expression in a heterologous system.ConclusionThese works contribute to a better understanding of the immunobiological properties of the UdLAMPs and provide a profile of the distribution of these antigens in different Brazilian states.

Highlights

Ureaplasma diversum has numerous virulence factors that contribute to pathogenesis in cattle, including Lipid-associated membrane proteins (LAMPs)
U. diversum antigens have low similarity with bovine proteome proteins BLASTp analyses of the 36 UdLAMPs with bovine proteomes revealed that the maximum similarity occurred between the lipoprotein gudiv_159 and the Tinken-1 protein from Bos taurus taurus (29%)
In silico analysis showed that U. diversum antigens have epitopes for B and T lymphocytes Conformational and linear B cell epitopes were evaluated for the number of regions and the total percentage of amino acids in epitope regions

Summary

Introduction

Ureaplasma diversum has numerous virulence factors that contribute to pathogenesis in cattle, including Lipid-associated membrane proteins (LAMPs). Epitope prediction for B lymphocytes was performed on the BepiPred v2.0 and CBTOPE v1.0 servers. U. diversum infection is not conditioned by the presence of clinical symptoms, it can colonize the respiratory and genital/reproductive systems of cattle, generating severe inflammatory conditions often culminating in abortion [3]. It is considered an opportunistic pathogen found in the mucosa and secretions of the vulva, vagina, and udder of cows and secretion of the respiratory tract of calves [1]. Infection of semen for artificial insemination and in vitro fertilization results in serious obstacles to modern bovine reproduction techniques [6]

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