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The vaccine potential of Bordetella pertussis biofilm-derived membrane proteins.

de Gouw D, Serra DO, de Jonge MI, Hermans PW, Wessels HJ, Zomer A, Yantorno OM, Diavatopoulos DA, Mooi FR - Emerg Microbes Infect (2014)

Bottom Line: As proof of concept, mice were vaccinated with recombinantly produced BipA.Immunization significantly reduced colonization of the lungs and antibodies to BipA were found to efficiently opsonize bacteria.Together, these data suggest that biofilm proteins and in particular BipA may be of interest for inclusion into future pertussis vaccines.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Pediatric Infectious Diseases, Department of Pediatrics, Radboud University Medical Center , Nijmegen 6500 HB, The Netherlands ; Laboratory of Medical Immunology, Department of Laboratory Medicine, Radboud University Medical Center , Nijmegen 6500 HB, The Netherlands.

ABSTRACT
Pertussis is an infectious respiratory disease of humans caused by the gram-negative pathogen Bordetella pertussis. The use of acellular pertussis vaccines (aPs) which induce immunity of relative short duration and the emergence of vaccine-adapted strains are thought to have contributed to the recent resurgence of pertussis in industrialized countries despite high vaccination coverage. Current pertussis vaccines consist of antigens derived from planktonic bacterial cultures. However, recent studies have shown that biofilm formation represents an important aspect of B. pertussis infection, and antigens expressed during this stage may therefore be potential targets for vaccination. Here we provide evidence that vaccination of mice with B. pertussis biofilm-derived membrane proteins protects against infection. Subsequent proteomic analysis of the protein content of biofilm and planktonic cultures yielded 11 proteins which were ≥three-fold more abundant in biofilms, of which Bordetella intermediate protein A (BipA) was the most abundant, surface-exposed protein. As proof of concept, mice were vaccinated with recombinantly produced BipA. Immunization significantly reduced colonization of the lungs and antibodies to BipA were found to efficiently opsonize bacteria. Finally, we confirmed that bipA is expressed during respiratory tract infection of mice, and that anti-BipA antibodies are present in the serum of convalescent whooping cough patients. Together, these data suggest that biofilm proteins and in particular BipA may be of interest for inclusion into future pertussis vaccines.

No MeSH data available.


Related in: MedlinePlus

Functional clustering of proteins identified with proteomics. 645 proteins were identified under all three growth conditions (biofilm, planktonic mid-log and planktonic stationary), of which 60 and 48 proteins were found to be upregulated or downregulated, respectively, in biofilms relative to planktonic conditions. The identified proteins were grouped by functional categories (A) and PSORTb-predicated subcellular localization (B). The relative frequencies of biofilm-up- or -downregulated proteins, compared to the total number of annotated proteins identified under all conditions for each functional class, are listed on the right-hand side. Asterisks indicate statistically significant enrichment of biofilm-regulated proteins in a certain class as determined by Fisher's exact test. **P<0.005.
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fig2: Functional clustering of proteins identified with proteomics. 645 proteins were identified under all three growth conditions (biofilm, planktonic mid-log and planktonic stationary), of which 60 and 48 proteins were found to be upregulated or downregulated, respectively, in biofilms relative to planktonic conditions. The identified proteins were grouped by functional categories (A) and PSORTb-predicated subcellular localization (B). The relative frequencies of biofilm-up- or -downregulated proteins, compared to the total number of annotated proteins identified under all conditions for each functional class, are listed on the right-hand side. Asterisks indicate statistically significant enrichment of biofilm-regulated proteins in a certain class as determined by Fisher's exact test. **P<0.005.

Mentions: As our initial results implied that biofilms contain protective antigens, we compared the proteome of cytosolic and membrane protein fractions of the mid-log, stationary and biofilm cultures by mass spectrometry, as described in Supplementary Information. This approach identified a total of 749, 729 and 825 proteins in the extracts from mid-log, stationary and biofilm cultures, respectively, with an overlap of 645 proteins (88%). This analysis covered 21%–24% of the predicted total of 3449 protein-coding open reading frames in the B. pertussis genome.35 IDEAL-Q36 was then used to identify proteins which were differentially regulated in biofilms (Supplementary Information). This method normalizes and quantifies comparable peptides between parallel runs of different samples.36 Proteins that were ≥three-fold more or less abundant under biofilm (either cytoplasmic or membrane fraction) conditions compared to either mid-log or stationary conditions, were defined as biofilm-upregulated and biofilm-downregulated proteins, respectively. This strategy yielded 60 biofilm-upregulated proteins. Aggregation based on function and predicted subcellular localization showed that these proteins were significantly enriched for (ribosomal) proteins involved in protein synthesis (N=17, 27.4%, P=7.7×10−4; Figure 2 and Supplementary Table S1). Additionally, 48 proteins were identified which were downregulated in biofilms. While this set of proteins was not significantly enriched for any specific functional classes, 17 of these proteins have previously been identified as Bvg-activated genes in strain B1917,10 including the virulence factors Vag8, FimX, and eight type III secretion system (T3SS) proteins (Supplementary Table S1).


The vaccine potential of Bordetella pertussis biofilm-derived membrane proteins.

de Gouw D, Serra DO, de Jonge MI, Hermans PW, Wessels HJ, Zomer A, Yantorno OM, Diavatopoulos DA, Mooi FR - Emerg Microbes Infect (2014)

Functional clustering of proteins identified with proteomics. 645 proteins were identified under all three growth conditions (biofilm, planktonic mid-log and planktonic stationary), of which 60 and 48 proteins were found to be upregulated or downregulated, respectively, in biofilms relative to planktonic conditions. The identified proteins were grouped by functional categories (A) and PSORTb-predicated subcellular localization (B). The relative frequencies of biofilm-up- or -downregulated proteins, compared to the total number of annotated proteins identified under all conditions for each functional class, are listed on the right-hand side. Asterisks indicate statistically significant enrichment of biofilm-regulated proteins in a certain class as determined by Fisher's exact test. **P<0.005.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150286&req=5

fig2: Functional clustering of proteins identified with proteomics. 645 proteins were identified under all three growth conditions (biofilm, planktonic mid-log and planktonic stationary), of which 60 and 48 proteins were found to be upregulated or downregulated, respectively, in biofilms relative to planktonic conditions. The identified proteins were grouped by functional categories (A) and PSORTb-predicated subcellular localization (B). The relative frequencies of biofilm-up- or -downregulated proteins, compared to the total number of annotated proteins identified under all conditions for each functional class, are listed on the right-hand side. Asterisks indicate statistically significant enrichment of biofilm-regulated proteins in a certain class as determined by Fisher's exact test. **P<0.005.
Mentions: As our initial results implied that biofilms contain protective antigens, we compared the proteome of cytosolic and membrane protein fractions of the mid-log, stationary and biofilm cultures by mass spectrometry, as described in Supplementary Information. This approach identified a total of 749, 729 and 825 proteins in the extracts from mid-log, stationary and biofilm cultures, respectively, with an overlap of 645 proteins (88%). This analysis covered 21%–24% of the predicted total of 3449 protein-coding open reading frames in the B. pertussis genome.35 IDEAL-Q36 was then used to identify proteins which were differentially regulated in biofilms (Supplementary Information). This method normalizes and quantifies comparable peptides between parallel runs of different samples.36 Proteins that were ≥three-fold more or less abundant under biofilm (either cytoplasmic or membrane fraction) conditions compared to either mid-log or stationary conditions, were defined as biofilm-upregulated and biofilm-downregulated proteins, respectively. This strategy yielded 60 biofilm-upregulated proteins. Aggregation based on function and predicted subcellular localization showed that these proteins were significantly enriched for (ribosomal) proteins involved in protein synthesis (N=17, 27.4%, P=7.7×10−4; Figure 2 and Supplementary Table S1). Additionally, 48 proteins were identified which were downregulated in biofilms. While this set of proteins was not significantly enriched for any specific functional classes, 17 of these proteins have previously been identified as Bvg-activated genes in strain B1917,10 including the virulence factors Vag8, FimX, and eight type III secretion system (T3SS) proteins (Supplementary Table S1).

Bottom Line: As proof of concept, mice were vaccinated with recombinantly produced BipA.Immunization significantly reduced colonization of the lungs and antibodies to BipA were found to efficiently opsonize bacteria.Together, these data suggest that biofilm proteins and in particular BipA may be of interest for inclusion into future pertussis vaccines.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Pediatric Infectious Diseases, Department of Pediatrics, Radboud University Medical Center , Nijmegen 6500 HB, The Netherlands ; Laboratory of Medical Immunology, Department of Laboratory Medicine, Radboud University Medical Center , Nijmegen 6500 HB, The Netherlands.

ABSTRACT
Pertussis is an infectious respiratory disease of humans caused by the gram-negative pathogen Bordetella pertussis. The use of acellular pertussis vaccines (aPs) which induce immunity of relative short duration and the emergence of vaccine-adapted strains are thought to have contributed to the recent resurgence of pertussis in industrialized countries despite high vaccination coverage. Current pertussis vaccines consist of antigens derived from planktonic bacterial cultures. However, recent studies have shown that biofilm formation represents an important aspect of B. pertussis infection, and antigens expressed during this stage may therefore be potential targets for vaccination. Here we provide evidence that vaccination of mice with B. pertussis biofilm-derived membrane proteins protects against infection. Subsequent proteomic analysis of the protein content of biofilm and planktonic cultures yielded 11 proteins which were ≥three-fold more abundant in biofilms, of which Bordetella intermediate protein A (BipA) was the most abundant, surface-exposed protein. As proof of concept, mice were vaccinated with recombinantly produced BipA. Immunization significantly reduced colonization of the lungs and antibodies to BipA were found to efficiently opsonize bacteria. Finally, we confirmed that bipA is expressed during respiratory tract infection of mice, and that anti-BipA antibodies are present in the serum of convalescent whooping cough patients. Together, these data suggest that biofilm proteins and in particular BipA may be of interest for inclusion into future pertussis vaccines.

No MeSH data available.


Related in: MedlinePlus