Molecular Signatures of the Evolving Immune Response in Mice following a Bordetella pertussis Infection

Abstract

Worldwide resurgence of pertussis necessitates the need for improvement of pertussis vaccines and vaccination strategies. Since natural infections induce a longer-lasting immunity than vaccinations, detailed knowledge of the immune responses following natural infection can provide important clues for such improvement. The purpose was to elucidate the kinetics of the protective immune response evolving after experimental Bordetella pertussis (B. pertussis) infection in mice. Data were collected from (i) individual analyses, i.e. microarray, flow cytometry, multiplex immunoassays, and bacterial clearance; (ii) twelve time points during the infection; and (iii) different tissues involved in the immune responses, i.e. lungs, spleen and blood. Combined data revealed detailed insight in molecular and cellular sequence of events connecting different phases (innate, bridging and adaptive) of the immune response following the infection. We detected a prolonged acute phase response, broad pathogen recognition, and early gene signatures of subsequent T-cell recruitment in the lungs. Activation of particular transcription factors and specific cell markers provided insight into the time course of the transition from innate towards adaptive immune responses, which resulted in a broad spectrum of systemic antibody subclasses and splenic Th1/Th17 memory cells against B. pertussis. In addition, signatures preceding the local generation of Th1 and Th17 cells as well as IgA in the lungs, considered key elements in protection against B. pertussis, were established. In conclusion, molecular and cellular immunological processes in response to live B. pertussis infection were unraveled, which may provide guidance in selecting new vaccine candidates that should evoke local and prolonged protective immune responses.

Bordetella pertussis outer membrane vesicle vaccine confers equal efficacy in mice with milder inflammatory responses compared to a whole-cell vaccine

“…RNA isolation from lung and spleen tissue and a RNA quality check (concentration and integrity) were performed as described before24. Microarray analysis was performed on RNA concentrates from spleen tissue of each omvPV-mice and wPV-mice (n = 3) sacrificed at 16 time points: D0, D28, D28(4 h), D29, D30, D31, D32, D35, D42, D49, D56, D56(4 h), D58, D63, D70 and D77 (Fig.…”

“…RNA samples extracted from lung and spleen tissue of N.I.-mice were pooled (n = 3) for according time points described before. Amplification, labeling and hybridization of RNA samples for either lung or spleen tissue to microarray chips (NimbleGen 126135 k Mus musculus, Roche, Germany) was carried out at the Microarray Department of the University of Amsterdam, The Netherlands, as described previously24.…”

“…Quality control and normalization of raw microarray data was performed as described before24. To identify differentially expressed genes (DEGs) between experimental groups (naive and various time points post immunization or challenge) an ANOVA was applied.…”

“…Moreover, systems biology can serve to compare molecular signatures induced by distinct vaccines2122 and gain insight into vaccine safety23. Previously we used a systems approach in mice to study B. pertussis infection-induced responses24.…”

Pertussis: Microbiology, Disease, Treatment, and Prevention

“…Vaccines with improved adverse event profiles and that provide higher efficacy will provide protection to vulnerable populations, including neonates, very young infants, and older adults (575). In these populations, research to understand B. pertussis virulence factors associated with vaccine-associated ad-verse events may take advantage of immunosignature analysis (576). Immunosignatures in different age groups will also aid the understanding of factors driving humoral and cell-mediated immunity among infants, children, and adults with naturally acquired pertussis and following pertussis vaccination (577).…”