Protectome Analysis: A New Selective Bioinformatics Tool for Bacterial Vaccine Candidate Discovery

New generation vaccines are in demand to include only the key antigens sufficient to confer protective immunity among the plethora of pathogen molecules. In the last decade, large-scale genomics-based technologies have emerged. Among them, the Reverse Vaccinology approach was successfully applied to the development of an innovative vaccine against Neisseria meningitidis serogroup B, now available on the market with the commercial name BEXSERO® (Novartis Vaccines). The limiting step of such approaches is the number of antigens to be tested in in vivo models. Several laboratories have been trying to refine the original approach in order to get to the identification of the relevant antigens straight from the genome. Here we report a new bioinformatics tool that moves a first step in this direction. The tool has been developed by identifying structural/functional features recurring in known bacterial protective antigens, the so called “Protectome space,” and using such “protective signatures” for protective antigen discovery. In particular, we applied this new approach to Staphylococcus aureus and Group B Streptococcus and we show that not only already known protective antigens were re-discovered, but also two new protective antigens were identified.

[1]  P. Dull,et al.  Meningococcal vaccine development--from glycoconjugates against MenACWY to proteins against MenB--potential for broad protection against meningococcal disease. , 2012, Vaccine.

[2]  Scott A Lesley,et al.  The Polymerase Incomplete Primer Extension (PIPE) method applied to high-throughput cloning and site-directed mutagenesis. , 2009, Methods in molecular biology.

[3]  G. Grandi,et al.  Staphylococcus aureus FhuD2 is involved in the early phase of staphylococcal dissemination and generates protective immunity in mice. , 2012, The Journal of infectious diseases.

[4]  J. Venter,et al.  Identification of vaccine candidates against serogroup B meningococcus by whole-genome sequencing. , 2000, Science.

[5]  H. Tettelin,et al.  Identification of a Universal Group B Streptococcus Vaccine by Multiple Genome Screen , 2005, Science.

[6]  J. Musser,et al.  Multi High-Throughput Approach for Highly Selective Identification of Vaccine Candidates: the Group A Streptococcus Case , 2012, Molecular & Cellular Proteomics.

[7]  I. Margarit,et al.  Surfome Analysis as a Fast Track to Vaccine Discovery , 2009, Molecular & Cellular Proteomics.

[8]  M. Schirle,et al.  Mining the bacterial unknown proteome: identification and characterization of a novel family of highly conserved protective antigens in Staphylococcus aureus. , 2013, The Biochemical journal.

[9]  Fang Chen,et al.  VIOLIN: vaccine investigation and online information network , 2007, Nucleic Acids Res..

[10]  Irini A. Doytchinova,et al.  BMC Bioinformatics BioMed Central Methodology article VaxiJen: a server for prediction of protective antigens, tumour , 2007 .

[11]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[12]  I. Margarit,et al.  Capturing host‐pathogen interactions by protein microarrays: identification of novel streptococcal proteins binding to human fibronectin, fibrinogen, and C4BP , 2009, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[13]  G. Bensi,et al.  Characterization and identification of vaccine candidate proteins through analysis of the group A Streptococcus surface proteome , 2006, Nature Biotechnology.

[14]  M. Hanner,et al.  Antigenome technology: a novel approach for the selection of bacterial vaccine candidate antigens. , 2005, Vaccine.

[15]  B. Gonik,et al.  Preventing the broad spectrum of perinatal morbidity and mortality through group B streptococcal vaccination. , 2013, Vaccine.

[16]  J. Musser,et al.  Protein Array Profiling of Tic Patient Sera Reveals a Broad Range and Enhanced Immune Response against Group A Streptococcus Antigens , 2009, PloS one.

[17]  J. V. van Dijl,et al.  The cell surface proteome of Staphylococcus aureus , 2011, Proteomics.

[18]  G. Grandi,et al.  Approach to discover T- and B-cell antigens of intracellular pathogens applied to the design of Chlamydia trachomatis vaccines , 2011, Proceedings of the National Academy of Sciences.

[19]  Mary Ann Robinson,et al.  Identification of a Novel Staphylococcus aureus Two-Component Leukotoxin Using Cell Surface Proteomics , 2010, PloS one.

[20]  R. Fleischmann,et al.  Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. , 1995, Science.

[21]  R. Mott,et al.  Accurate formula for P-values of gapped local sequence and profile alignments. , 2000, Journal of molecular biology.

[22]  R. Rappuoli,et al.  The new multicomponent vaccine against meningococcal serogroup B, 4CMenB: immunological, functional and structural characterization of the antigens. , 2012, Vaccine.

[23]  María Martín,et al.  Activities at the Universal Protein Resource (UniProt) , 2013, Nucleic Acids Res..

[24]  G. Grandi,et al.  Proteomics Characterization of Outer Membrane Vesicles from the Extraintestinal Pathogenic Escherichia coli ΔtolR IHE3034 Mutant*S , 2008, Molecular & Cellular Proteomics.

[25]  Peer Bork,et al.  SMART 7: recent updates to the protein domain annotation resource , 2011, Nucleic Acids Res..

[26]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[27]  G. Grandi Bacterial vaccine discovery: From “brute force” to high selectivity , 2010, Human vaccines.

[28]  M. Hecker,et al.  Profiling the surfacome of Staphylococcus aureus , 2010, Proteomics.

[29]  M. Larsen,et al.  Improved accuracy of cell surface shaving proteomics in Staphylococcus aureus using a false‐positive control , 2010, Proteomics.

[30]  E. Birney,et al.  Pfam: the protein families database , 2013, Nucleic Acids Res..

[31]  R. Rappuoli,et al.  A universal vaccine for serogroup B meningococcus. , 2006, Proceedings of the National Academy of Sciences of the United States of America.