Immunogenic and Invasive Properties of Brucella melitensis 16M Outer Membrane Protein Vaccine Candidates Identified via a Reverse Vaccinology Approach

Abstract

Brucella is the etiologic agent of brucellosis, one of the most common and widely distributed zoonotic diseases. Its highly infectious nature, the insidious, systemic, chronic, debilitating aspects of the disease and the lack of an approved vaccine for human use in the United States are features that make Brucella a viable threat to public health. One of the main impediments to vaccine development is identification of suitable antigens. In order to identify antigens that could potentially be used in a vaccine formulation, we describe a multi-step antigen selection approach. We initially used an algorithm (Vaxign) to predict ORF encoding outer membrane proteins with antigenic determinants. Differential gene expression during acute infection and published evidence for a role in virulence were used as criteria for down-selection of the candidate antigens that resulted from in silico prediction. This approach resulted in the identification of nine Brucella melitensis outer membrane proteins, 5 of which were recombinantly expressed and used for validation. Omp22 and Hia had the highest in silico scores for adhesin probability and also conferred invasive capacity to E. coli overexpressing recombinant proteins. With the exception of FlgK in the goat, all proteins reacted to pooled sera from exposed goats, mice, and humans. BtuB, Hia and FlgK stimulated a mixed Th1–Th2 response in splenocytes from immunized mice while BtuB and Hia elicited NO release from splenocytes of S19 immunized mice. The results support the applicability of the current approach to the identification of antigens with immunogenic and invasive properties. Studies to assess immunogenicity and protective efficacy of individual proteins in the mouse are currently underway.

Pathogenesis and Immunobiology of Brucellosis

“…6,8,13,16,18,32,53e60 This broad spectrum of data sets was analyzed to identify candidate genes and biomarkers of Brucella and hosts, 61 to analyze 62 and predict Brucella antigenic proteins, to identify subunit vaccines, 63 to understand gene regulatory networks, 54 and to characterize the stringent Brucella stress response 64 and modulation of host responses. 53 Time-series studies of in vitro or in vivo experimental infection data were performed with a systems-based perspective of complex biological organizations to categorize the interactions of individual proteins within Brucellaehost proteine protein networks.…”

New Scenarios for Brucella suis and Brucella melitensis

“…Reverse vaccinology is based on in silico genome analysis and the identification of Brucella antigens with characteristics such as immunogenicity or structure, requirements that are necessary if they are to be proposed as vaccines [63].…”

Small Ruminants and Zoonotic Infections: Live or Dead—Direct or Indirect

“…For human consumption, unpasteurized milk and milk products should be avoided. No human vaccine exists, however recent results are promising in developing a recombinant vaccine against B. melitensis (Gomez et al 2013). …”