Immunoproteomic Analysis of Human Serological Antibody Responses to Vaccination with Whole-Cell Pertussis Vaccine (WCV)

Abstract

BackgroundPertussis (whooping cough) caused by Bordetella pertussis (B.p), continues to be a serious public health threat. Vaccination is the most economical and effective strategy for preventing and controlling pertussis. However, few systematic investigations of actual human immune responses to pertussis vaccines have been performed. Therefore, we utilized a combination of two-dimensional electrophoresis (2-DE), immunoblotting, and mass spectrometry to reveal the entire antigenic proteome of whole-cell pertussis vaccine (WCV) targeted by the human immune system as a first step toward evaluating the repertoire of human humoral immune responses against WCV.Methodology/Principal FindingsImmunoproteomic profiling of total membrane enriched proteins and extracellular proteins of Chinese WCV strain 58003 identified a total of 30 immunoreactive proteins. Seven are known pertussis antigens including Pertactin, Serum resistance protein, chaperonin GroEL and two OMP porins. Sixteen have been documented to be immunogenic in other pathogens but not in B.p, and the immunogenicity of the last seven proteins was found for the first time. Furthermore, by comparison of the human and murine immunoproteomes of B.p, with the exception of four human immunoreactive proteins that were also reactive with mouse immune sera, a unique group of antigens including more than 20 novel immunoreactive proteins that uniquely reacted with human immune serum was confirmed.Conclusions/SignificanceThis study is the first time that the repertoire of human serum antibody responses against WCV was comprehensively investigated, and a small number of previously unidentified antigens of WCV were also found by means of the classic immunoproteomic strategy. Further research on these newly identified predominant antigens of B.p exclusively against humans will not only remarkably accelerate the development of diagnostic biomarkers and subunit vaccines but also provide detailed insight into human immunity mechanisms against WCV. In particular, this work highlights the heterogeneity of the B.p immunoreactivity patterns of the mouse model and the human host.

A rapid method for capture and identification of immunogenic proteins in Bordetella pertussis enriched membranes fractions: A fast-track strategy applicable to other microorganisms

“…Several major Bp Ag (s) were identified including pertactin, filamentous hemagglutinin, and adenylate cyclase toxin. Previously, researchers have used 2D-GE, immunoblotting, and MS to identify immunoproteins of Bp [3][4][5][6][7]. Using our strategy, we identified many of the same proteins further validating and demonstrating the usefulness of this approach (see proteins with superscript in Table 1).…”

“…Their potential as candidate diagnostics, biomedical therapeutics, pathogenic markers and vaccines is an on-going subject of investigation within academia and pharmaceutical companies. Traditional strategies of identifying these important antigens (Ag) (s) usually involve a series of sodium dodecyl sulfate polyacrylamide 1-or 2-dimensional gel electrophoresis (SDS-PAGE 1D/2D-GE), immunoblotting, followed by mass spectrometry (MS) [1][2][3][4][5][6][7]. These procedures are time-consuming, laborious, and results are challenging to reproduce.…”

Characterization of co-purified acellular pertussis vaccines

“…[37][38][39] This protein is also found to be recognized by the human immune system and identified as human immunoreactive proteins of B. pertussis. 40 Since this antigen could not be sourced, direct assessment of antibody production and protective effect of the antigen were not possible. However, although the precise role of sphB1 in pertussis immune response and protection deserve further investigation, it cannot be ruled out that it may serve as one of the possible explanations for the observed differences in protection between S2 and S3.…”

Outer membrane protein OmpQ of Bordetella bronchiseptica is required for mature biofilm formation

“…On the one hand, Finn et al (1995) showed that the absence of OmpQ does not affect the capacity of B. pertussis to colonize mouse lung tissue, discarding the possibility that this protein may play a role in the survival of the bacterium inside the host. On the other hand, recent proteomic studies proposed OmpQ as a novel protective antigen candidate (de Gouw et al, 2014;Packard et al, 2004;Tefon et al, 2011;Williamson et al, 2012;Zhu et al, 2010) and even suggested that OmpQ could play a role in the pathogenesis of Bordetella (Tefon et al, 2011).…”

“…In the first studies of OmpQ, Finn et al (1995) reported that the absence of this protein did not affect the ability of B. pertussis 18323 to survive in the mouse respiratory tract. However, more recently, several reports have recognized OmpQ as an immunodominant antigen and, moreover, as a potential novel protective antigen (de Gouw Packard et al, 2004;Tefon et al, 2011;Williamson et al, 2012;Zhu et al, 2010). In addition, it has been proposed that OmpQ should play a role in the pathogenesis of Bordetella, perhaps during the first steps of infection or in the establishment of a carrier state (Tefon et al, 2011).…”