Limits...
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 rBipA protects against B. pertussis infection. Naive adult female BALB/c mice were subcutaneously immunized with rBipA or controls as described in Figure 1 and 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 difference between rBipA and aP and between vaccinated and PBS control mice; two-tailed Mann–Whitney U test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4150286&req=5

fig4: Immunization with rBipA protects against B. pertussis infection. Naive adult female BALB/c mice were subcutaneously immunized with rBipA or controls as described in Figure 1 and 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 difference between rBipA and aP and between vaccinated and PBS control mice; two-tailed Mann–Whitney U test.

Mentions: To determine the protective effects of BipA, we constructed a recombinant His-tagged fusion protein of BipA (rBipA). After purification and refolding, the purity of the produced protein was confirmed by Coomassie staining and immunoblotting (Supplementary Figure S1). rBipA was then adjuvanted with Alhydrogel and used as stand-alone antigen (5 µg) to vaccinate female adult BALB/c mice. Mice were immunized 2× in 2-week intervals, after which they were challenged by intranasal infection with B1917. Vaccination with rBipA resulted in a small but significant reduction of bacterial load in the lungs as compared to the PBS-vaccinated mice (Figures 4A and 4B, 5.4 and 3.4-fold at days 3 and 7, respectively) which was not observed in the nasopharynx (Figures 4C and 4D). Thus, these data show that rBipA is able to reduce bacterial load.


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 rBipA protects against B. pertussis infection. Naive adult female BALB/c mice were subcutaneously immunized with rBipA or controls as described in Figure 1 and 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 difference between rBipA and aP and between vaccinated and PBS control mice; two-tailed Mann–Whitney U test.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig4: Immunization with rBipA protects against B. pertussis infection. Naive adult female BALB/c mice were subcutaneously immunized with rBipA or controls as described in Figure 1 and 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 difference between rBipA and aP and between vaccinated and PBS control mice; two-tailed Mann–Whitney U test.
Mentions: To determine the protective effects of BipA, we constructed a recombinant His-tagged fusion protein of BipA (rBipA). After purification and refolding, the purity of the produced protein was confirmed by Coomassie staining and immunoblotting (Supplementary Figure S1). rBipA was then adjuvanted with Alhydrogel and used as stand-alone antigen (5 µg) to vaccinate female adult BALB/c mice. Mice were immunized 2× in 2-week intervals, after which they were challenged by intranasal infection with B1917. Vaccination with rBipA resulted in a small but significant reduction of bacterial load in the lungs as compared to the PBS-vaccinated mice (Figures 4A and 4B, 5.4 and 3.4-fold at days 3 and 7, respectively) which was not observed in the nasopharynx (Figures 4C and 4D). Thus, these data show that rBipA is able to reduce bacterial load.

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