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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

bipA expression during infection. (A) Naive adult female BALB/c mice were infected intranasally with B. pertussis strain B1917. After seven days, bacteria were collected through BAL and used for in vivo transcriptional analysis using bipA-specific primers as described in the text. Expression levels were compared to in vitro Bvg+, Bvgi and Bvg− conditions. The transcription data are expressed as 40−ΔCt value, which is a measure of expression relative to the recA gene (ΔCt=Cttarget–CtrecA). *P<0.05, **P<0.005, ***P<0.0005 relative to the Bvg− condition; Student's t-test with Welch's correction. (B) Human serum, from 10 confirmed B. pertussis-infected individuals (Bp+) and four non-infected individuals (Bp−) was diluted and incubated on ELISA plates coated with recombinant BipA or Ptx and the titers were determined. Horizontal lines represent the geometrical mean. **P<0.005, ***P<0.0005 relative to Bp- sera; one-tailed Mann–Whitney U test. BAL, bronchoalveolar lavage.
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fig3: bipA expression during infection. (A) Naive adult female BALB/c mice were infected intranasally with B. pertussis strain B1917. After seven days, bacteria were collected through BAL and used for in vivo transcriptional analysis using bipA-specific primers as described in the text. Expression levels were compared to in vitro Bvg+, Bvgi and Bvg− conditions. The transcription data are expressed as 40−ΔCt value, which is a measure of expression relative to the recA gene (ΔCt=Cttarget–CtrecA). *P<0.05, **P<0.005, ***P<0.0005 relative to the Bvg− condition; Student's t-test with Welch's correction. (B) Human serum, from 10 confirmed B. pertussis-infected individuals (Bp+) and four non-infected individuals (Bp−) was diluted and incubated on ELISA plates coated with recombinant BipA or Ptx and the titers were determined. Horizontal lines represent the geometrical mean. **P<0.005, ***P<0.0005 relative to Bp- sera; one-tailed Mann–Whitney U test. BAL, bronchoalveolar lavage.

Mentions: To determine whether BipA is expressed during natural infection, naive adult BALB/c mice were infected intranasally with B. pertussis strain B1917. Seven days after infection, bacteria were isolated from the lungs and gene expression of bipA was analyzed by quantitative polymerase chain reaction and compared to Bvg+, Bvgi and Bvg−in vitro conditions. As a negative control, the kpsT gene encoding a Bvg− protein involved in capsule biosynthesis was included.40 Transcriptional analysis showed that bipA is expressed in the lungs of mice, at a level also observed under virulent in vitro Bvg+ and Bvgi conditions (Figure 3A). Thus, these data indicate that BipA is expressed during respiratory infection in mice.


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)

bipA expression during infection. (A) Naive adult female BALB/c mice were infected intranasally with B. pertussis strain B1917. After seven days, bacteria were collected through BAL and used for in vivo transcriptional analysis using bipA-specific primers as described in the text. Expression levels were compared to in vitro Bvg+, Bvgi and Bvg− conditions. The transcription data are expressed as 40−ΔCt value, which is a measure of expression relative to the recA gene (ΔCt=Cttarget–CtrecA). *P<0.05, **P<0.005, ***P<0.0005 relative to the Bvg− condition; Student's t-test with Welch's correction. (B) Human serum, from 10 confirmed B. pertussis-infected individuals (Bp+) and four non-infected individuals (Bp−) was diluted and incubated on ELISA plates coated with recombinant BipA or Ptx and the titers were determined. Horizontal lines represent the geometrical mean. **P<0.005, ***P<0.0005 relative to Bp- sera; one-tailed Mann–Whitney U test. BAL, bronchoalveolar lavage.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig3: bipA expression during infection. (A) Naive adult female BALB/c mice were infected intranasally with B. pertussis strain B1917. After seven days, bacteria were collected through BAL and used for in vivo transcriptional analysis using bipA-specific primers as described in the text. Expression levels were compared to in vitro Bvg+, Bvgi and Bvg− conditions. The transcription data are expressed as 40−ΔCt value, which is a measure of expression relative to the recA gene (ΔCt=Cttarget–CtrecA). *P<0.05, **P<0.005, ***P<0.0005 relative to the Bvg− condition; Student's t-test with Welch's correction. (B) Human serum, from 10 confirmed B. pertussis-infected individuals (Bp+) and four non-infected individuals (Bp−) was diluted and incubated on ELISA plates coated with recombinant BipA or Ptx and the titers were determined. Horizontal lines represent the geometrical mean. **P<0.005, ***P<0.0005 relative to Bp- sera; one-tailed Mann–Whitney U test. BAL, bronchoalveolar lavage.
Mentions: To determine whether BipA is expressed during natural infection, naive adult BALB/c mice were infected intranasally with B. pertussis strain B1917. Seven days after infection, bacteria were isolated from the lungs and gene expression of bipA was analyzed by quantitative polymerase chain reaction and compared to Bvg+, Bvgi and Bvg−in vitro conditions. As a negative control, the kpsT gene encoding a Bvg− protein involved in capsule biosynthesis was included.40 Transcriptional analysis showed that bipA is expressed in the lungs of mice, at a level also observed under virulent in vitro Bvg+ and Bvgi conditions (Figure 3A). Thus, these data indicate that BipA is expressed during respiratory infection in mice.

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