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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

Immunization with membrane protein fractions derived from biofilm and mid-log and stationary planktonic cultures. Naive adult female BALB/c mice were subcutaneously immunized with 1 µg membrane proteins. Negative controls consisted of adjuvant only (Alum) and PBS, while an acellular vaccine (aP) containing 0.5 µg Ptx, 0.5 µg FHA and 0.16 µg Prn was used as a positive control. Subsequently, mice were infected intranasally with 2×107 CFU of B. pertussis strain B1917. The bacterial load in the lungs and nose was quantified three (A and C) and seven (B and D) days after challenge. Each symbol represents one mouse. Horizontal lines represent the mean. Dashed lines indicate the lower limit of detection. **P<0.005 relative to PBS mice; two-tailed Mann–Whitney U test. FHA, filamentous hemagglutinin.
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fig1: Immunization with membrane protein fractions derived from biofilm and mid-log and stationary planktonic cultures. Naive adult female BALB/c mice were subcutaneously immunized with 1 µg membrane proteins. Negative controls consisted of adjuvant only (Alum) and PBS, while an acellular vaccine (aP) containing 0.5 µg Ptx, 0.5 µg FHA and 0.16 µg Prn was used as a positive control. Subsequently, mice were infected intranasally with 2×107 CFU of B. pertussis strain B1917. The bacterial load in the lungs and nose was quantified three (A and C) and seven (B and D) days after challenge. Each symbol represents one mouse. Horizontal lines represent the mean. Dashed lines indicate the lower limit of detection. **P<0.005 relative to PBS mice; two-tailed Mann–Whitney U test. FHA, filamentous hemagglutinin.

Mentions: To evaluate the potential of biofilm-grown bacteria to confer protection against respiratory tract infection in mice, we cultured a B. pertussis clinical isolate (B1917, isolated in 2000) under biofilm-forming and planktonic conditions. This particular strain belongs to the P3 lineage which emerged in the 1980s and currently predominates in many vaccinated populations.33 For planktonic conditions, bacteria were grown to mid-log and stationary phase. Following growth, bacteria were harvested and cytosolic and membrane (-associated) proteins were extracted.34 The membrane fractions were adjuvanted with Alhydrogel and the equivalent of 1 µg of protein was used to vaccinate female BALB/c mice 2× by subcutaneous injection, with 2-week intervals. In parallel, groups of mice were vaccinated with a currently licensed three-component aP (Infanrix) or mock-vaccinated with PBS or adjuvant only. Immunized mice were then challenged by intranasal infection with strain B1917, after which the lung and nose bacterial load was determined three and seven days later. Mice vaccinated with aP showed a significant reduction (>220-fold) in lung CFUs at day 3 and day 7 (Figures 1A and 1B). At three days following infection, significant protection was observed in the lungs of mice immunized with proteins derived from planktonic as well as biofilm-grown bacteria (Figure 1A). After seven days, protection was more pronounced, with 29-fold, 43-fold and 170-fold reduction in bacterial load in mice immunized with biofilm, mid-log and stationary proteins, respectively (Figure 1B). The differences in protection induced by the three membrane fractions were not significant. Of note, we observed that none of the vaccinated mice, including those vaccinated with aP, were protected against infection with B. pertussis in the upper respiratory tract (Figures 1C and 1D). Taken together, these data suggest that immunization with biofilms can induce protection against infection.


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)

Immunization with membrane protein fractions derived from biofilm and mid-log and stationary planktonic cultures. Naive adult female BALB/c mice were subcutaneously immunized with 1 µg membrane proteins. Negative controls consisted of adjuvant only (Alum) and PBS, while an acellular vaccine (aP) containing 0.5 µg Ptx, 0.5 µg FHA and 0.16 µg Prn was used as a positive control. Subsequently, mice were infected intranasally with 2×107 CFU of B. pertussis strain B1917. The bacterial load in the lungs and nose was quantified three (A and C) and seven (B and D) days after challenge. Each symbol represents one mouse. Horizontal lines represent the mean. Dashed lines indicate the lower limit of detection. **P<0.005 relative to PBS mice; two-tailed Mann–Whitney U test. FHA, filamentous hemagglutinin.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig1: Immunization with membrane protein fractions derived from biofilm and mid-log and stationary planktonic cultures. Naive adult female BALB/c mice were subcutaneously immunized with 1 µg membrane proteins. Negative controls consisted of adjuvant only (Alum) and PBS, while an acellular vaccine (aP) containing 0.5 µg Ptx, 0.5 µg FHA and 0.16 µg Prn was used as a positive control. Subsequently, mice were infected intranasally with 2×107 CFU of B. pertussis strain B1917. The bacterial load in the lungs and nose was quantified three (A and C) and seven (B and D) days after challenge. Each symbol represents one mouse. Horizontal lines represent the mean. Dashed lines indicate the lower limit of detection. **P<0.005 relative to PBS mice; two-tailed Mann–Whitney U test. FHA, filamentous hemagglutinin.
Mentions: To evaluate the potential of biofilm-grown bacteria to confer protection against respiratory tract infection in mice, we cultured a B. pertussis clinical isolate (B1917, isolated in 2000) under biofilm-forming and planktonic conditions. This particular strain belongs to the P3 lineage which emerged in the 1980s and currently predominates in many vaccinated populations.33 For planktonic conditions, bacteria were grown to mid-log and stationary phase. Following growth, bacteria were harvested and cytosolic and membrane (-associated) proteins were extracted.34 The membrane fractions were adjuvanted with Alhydrogel and the equivalent of 1 µg of protein was used to vaccinate female BALB/c mice 2× by subcutaneous injection, with 2-week intervals. In parallel, groups of mice were vaccinated with a currently licensed three-component aP (Infanrix) or mock-vaccinated with PBS or adjuvant only. Immunized mice were then challenged by intranasal infection with strain B1917, after which the lung and nose bacterial load was determined three and seven days later. Mice vaccinated with aP showed a significant reduction (>220-fold) in lung CFUs at day 3 and day 7 (Figures 1A and 1B). At three days following infection, significant protection was observed in the lungs of mice immunized with proteins derived from planktonic as well as biofilm-grown bacteria (Figure 1A). After seven days, protection was more pronounced, with 29-fold, 43-fold and 170-fold reduction in bacterial load in mice immunized with biofilm, mid-log and stationary proteins, respectively (Figure 1B). The differences in protection induced by the three membrane fractions were not significant. Of note, we observed that none of the vaccinated mice, including those vaccinated with aP, were protected against infection with B. pertussis in the upper respiratory tract (Figures 1C and 1D). Taken together, these data suggest that immunization with biofilms can induce protection against infection.

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