Identification of H. pylori epitopes responsible for host immuno-response modulation through ORF-filtered phage display libraries and Interactome-sequencing
Altro Prodotto di Ricerca
Data di Pubblicazione:
2015
Citazione:
Identification of H. pylori epitopes responsible for host immuno-response modulation through ORF-filtered phage display libraries and Interactome-sequencing / S. Puccio, M.F. Soluri, E. Touati, E. Pinatel, C. Della Bella, M.M. D’Elios, D. Sblattero, C. Peano. ((Intervento presentato al 6. convegno Congress of European Microbiologists tenutosi a Maastricht nel 2015.
Abstract:
Background
To elucidate the molecular mechanisms involved in persistency/latency of the H. pylori infection or in its progression towards serious diseases, it is necessary to analyze the host pathogen interaction in vivo. The circulating antibody repertoire represents an important source of diagnostic information, serving as biomarker to provide a “disease signature”.
Objectives
The aim of this work is the identification of H. pylori epitopes responsible for host immunoresponse modulation though a discoverydriven approach that couples “phage display” and deep sequencing.
Methods
We used an approach for identifying novel antigens by screening gDNA libraries created from the pathogen genome, directly with sera from infected patients. Three phage display libraries from three H. pylori strains (HP26695, HPB128, HPSS1) have been constructed by using ßlactamase ORF selection vectors (Di Niro et al., 2010). Genomic DNA was sonicated, fragments cloned into the filtering vector, after transformation libraries of 1x106 clones were obtained and sequenced by 454 technology.
Conclusions
More than 93% of HP CDSs were represented in the phage genomic library therefore being representative of the whole H. pylori antigenic ORFeome. Putative antigens were selected from libraries using sera from patients affected by H. pylori presenting increasing degrees of infection: i) autoimmune gastritis and pernicious anemia; ii) gastric adenocarcinoma; iii) MALT lymphoma. The results show that the diversity of the libraries obtained after selection is significantly reduced. Furthermore, individual ranks, for each infection condition, have been compared highlighting the pattern of putative antigens, shared by all the conditions, and some that can distinguish the different stages of infection.
Tipologia IRIS:
14 - Intervento a convegno non pubblicato
Elenco autori:
S. Puccio, M.F. Soluri, E. Touati, E. Pinatel, C. Della Bella, M.M. D’Elios, D. Sblattero, C. Peano
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