Assays for Determining Pertussis Toxin Activity in Acellular Pertussis Vaccines

Whooping cough is caused by the bacterium Bordetella pertussis. There are currently two types of vaccines that can prevent the disease; whole cell vaccines (WCV) and acellular vaccines (ACV). The main virulence factor produced by the organism is pertussis toxin (PTx). This toxin is responsible for many physiological effects on the host, but it is also immunogenic and in its detoxified form is the main component of all ACVs. In producing toxoid for vaccines, it is vital to achieve a balance between sufficiently detoxifying PTx to render it safe while maintaining enough molecular structure that it retains its protective immunogenicity. To ensure that the first part of this balancing act has been successfully achieved, assays are required to accurately measure residual PTx activity in ACV products accurately. Quality control assays are also required to ensure that the detoxification procedures are robust and stable. This manuscript reviews the methods that have been used to achieve this aim, or may have the potential to replace them, and highlights their continuing requirement as vaccines that induce a longer lasting immunity are developed to prevent the re-occurrence of outbreaks that have been observed recently.

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A qPCR assay for Bordetella pertussis cells that enumerates both live and dead bacteria

10.1101/782177 ◽

2019 ◽

Author(s):

Stacy Ramkissoon ◽

Iain MacArthur ◽

Muktar Ibrahim ◽

Hans de Graaf ◽

Robert C. Read ◽

...

Keyword(s):

Causative Agent ◽

Bordetella Pertussis ◽

Whooping Cough ◽

Quantitative Measure ◽

Qpcr Assay ◽

Whole Cell ◽

Correlates Of Protection ◽

Immune Correlates ◽

Acellular Vaccines ◽

Future Utility

AbstractBordetella pertussis is the causative agent of whooping cough, commonly referred to as pertussis. Although the incidence of pertussis was reduced through vaccination, during the last thirty years it has returned to high levels in a number of countries. This resurgence has been linked to the switch from the use of whole-cell to acellular vaccines. Protection afforded by acellular vaccines appears to be short-lived compared to that afforded by whole cell vaccines. In order to inform future vaccine improvement by identifying immune correlates of protection, a human challenge model of B. pertussis colonisation has been developed. Accurate measurement of colonisation status in this model has required development of a qPCR-based assay to enumerate B. pertussis in samples that distinguishes between viable and dead bacteria. Here we report the development of this assay and its performance in the quantification of B. pertussis from human challenge model samples. This assay has future utility in diagnostic labs and in research where a quantitative measure of both B. pertussis number and viability is required.

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The relationship between mucosal immunity, nasopharyngeal carriage, asymptomatic transmission and the resurgence of Bordetella pertussis

F1000Research ◽

10.12688/f1000research.11654.1 ◽

2017 ◽

Vol 6 ◽

pp. 1568 ◽

Cited By ~ 14

Author(s):

Christopher Gill ◽

Pejman Rohani ◽

Donald M Thea

Keyword(s):

Bordetella Pertussis ◽

Mucosal Immunity ◽

Whooping Cough ◽

Critical Factor ◽

Nasopharyngeal Carriage ◽

Whole Cell ◽

The Us ◽

Acellular Vaccines ◽

The Uk ◽

The Relationship

The incidence of whooping cough in the US has been rising slowly since the 1970s, but the pace of this has accelerated sharply since acellular pertussis vaccines replaced the earlier whole cell vaccines in the late 1990s. A similar trend occurred in many other countries, including the UK, Canada, Australia, Ireland, and Spain, following the switch to acellular vaccines. The key question is why. Two leading theories (short duration of protective immunologic persistence and evolutionary shifts in the pathogen to evade the vaccine) explain some but not all of these shifts, suggesting that other factors may also be important. In this synthesis, we argue that sterilizing mucosal immunity that blocks or abbreviates the duration of nasopharyngeal carriage of Bordetella pertussis and impedes person-to-person transmission (including between asymptomatically infected individuals) is a critical factor in this dynamic. Moreover, we argue that the ability to induce such mucosal immunity is fundamentally what distinguishes whole cell and acellular pertussis vaccines and may be pivotal to understanding much of the resurgence of this disease in many countries that adopted acellular vaccines. Additionally, we offer the hypothesis that observed herd effects generated by acellular vaccines may reflect a modification of disease presentation leading to reduced potential for transmission by those already infected, as opposed to inducing resistance to infection among those who have been exposed.

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Pertussis

10.33442/vt2021311 ◽

2021 ◽

Author(s):

Carl Heinz Wirsing von König

Keyword(s):

Whooping Cough ◽

Severe Disease ◽

Newborn Infants ◽

School Entry ◽

Whole Cell ◽

Young Infants ◽

Combination Vaccines ◽

Adolescents And Adults ◽

Second Year ◽

Acellular Vaccines

The bacterium Bordetella pertussis causes disease by producing various virulence and adhesion factors, among them pertussis toxin (PT), filamentous hemagglutinin (FHA), pertactin (PRN) and agglutinogens (Agg), also called fimbriae (FIM) "Typical" pertussis or whooping cough starts with unspecific respiratory symptoms (catarrhal phase) followed by severe coughing spasms with whoops and vomiting (paroxysmal phase) and only after weeks or months disease severity slowly wanes (convalescent phase). "Atypical pertussis" with unspecific, long-lasting coughing episodes is seen in adolescents and adults; very young infants may die from apnoea. B. pertussis is transmitted by droplets, and neither infection nor vaccination produce long lasting protection. Macrolide antibiotics are given to patients and their contacts to reduce spread of the organism; however, antibiotics do NOT change the duration or course of the disease once symptoms are present. Whole cell pertussis vaccines (wP) consist of whole inactivated B. pertussis-cells, whereas acellular vaccines (aP) consist of one to five single components like PT, FHA, PRN or FIM. Pertussis vaccines are currently only available as combination vaccines with tetanus und diphtheria (DTP). Among these are DTwP; DTaP; TdaP; and various DTP-combinations with Hib, IPV, HBV vaccines. Whole cell pertussis (DTwP) combination vaccines are more reactogenic, whereas DTaP vaccines are generally well tolerated. Some DTwP had good efficacy/effectiveness (90%), it was low (40%) with others. Vaccine efficacy of DTaP vaccines ranges between 70% and 90%. As with most vaccines, efficiency is higher for severe disease. While pertussis vaccines did control clinical disease, protection is limited. Vaccination is recommended for all infants (three doses) worldwide with a booster in the second year of life. Many countries give additional doses at school entry and in adolescents, and some to adults. Vaccination of pregnant women effectively protects newborn infants and is increasingly recommended.

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Bordetella infection

Oxford Textbook of Medicine ◽

10.1093/med/9780198746690.003.0119 ◽

2020 ◽

pp. 1073-1076

Author(s):

Cameron C. Grant

Keyword(s):

Bordetella Pertussis ◽

Whooping Cough ◽

Severe Disease ◽

Diagnostic Methods ◽

Illness Onset ◽

Young Infants ◽

Adolescents And Adults ◽

Risk Of Exposure ◽

Acellular Vaccines ◽

Infectious Organism

Bordetella are small Gram-negative coccobacilli, of which Bordetella pertussis is the most important human pathogen. Bordetella pertussis is the cause of whooping cough, which remains one of the 10 leading causes of death among children less than five years old. Transmission of this highly infectious organism is primarily by aerosolized droplets. The preferred diagnostic methods are polymerase chain reaction detection from nasopharyngeal samples and serology (IgG antibodies to pertussis toxin). Macrolide antibiotics are recommended if started within four weeks of illness onset. Preventing severe disease in young children remains the primary goal, hence schedules consist of a three-dose infant series and subsequent booster doses. Acellular vaccines enable immunization schedules to include adolescents and adults. Acellular pertussis vaccine given to pregnant women reduces the risk of pertussis in young infants. Antibiotic prophylaxis is given when there is an infant at risk of exposure.

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Shortening the Lipid A Acyl Chains of Bordetella pertussis Enables Depletion of Lipopolysaccharide Endotoxic Activity

Vaccines ◽

10.3390/vaccines8040594 ◽

2020 ◽

Vol 8 (4) ◽

pp. 594

Author(s):

Jesús Arenas ◽

Elder Pupo ◽

Coen Phielix ◽

Dionne David ◽

Afshin Zariri ◽

...

Keyword(s):

Bordetella Pertussis ◽

Lipid A ◽

Whooping Cough ◽

Acyl Chain ◽

In Vitro Assays ◽

Whole Cell ◽

Whole Cells ◽

Acyl Chain Length ◽

Acyl Chains

Whooping cough, or pertussis, is an acute respiratory infectious disease caused by the Gram-negative bacterium Bordetella pertussis. Whole-cell vaccines, which were introduced in the fifties of the previous century and proved to be effective, showed considerable reactogenicity and were replaced by subunit vaccines around the turn of the century. However, there is a considerable increase in the number of cases in industrialized countries. A possible strategy to improve vaccine-induced protection is the development of new, non-toxic, whole-cell pertussis vaccines. The reactogenicity of whole-cell pertussis vaccines is, to a large extent, derived from the lipid A moiety of the lipopolysaccharides (LPS) of the bacteria. Here, we engineered B. pertussis strains with altered lipid A structures by expressing genes for the acyltransferases LpxA, LpxD, and LpxL from other bacteria resulting in altered acyl-chain length at various positions. Whole cells and extracted LPS from the strains with shorter acyl chains showed reduced or no activation of the human Toll-like receptor 4 in HEK-Blue reporter cells, whilst a longer acyl chain increased activation. Pyrogenicity studies in rabbits confirmed the in vitro assays. These findings pave the way for the development of a new generation of whole-cell pertussis vaccines with acceptable side effects.

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Overexpression of the RNA Polymerase Alpha Subunit Reduces Transcription of Bvg-Activated Virulence Genes inBordetella pertussis

Journal of Bacteriology ◽

10.1128/jb.182.2.529-531.2000 ◽

2000 ◽

Vol 182 (2) ◽

pp. 529-531 ◽

Cited By ~ 7

Author(s):

Nicholas H. Carbonetti ◽

Alla Romashko ◽

Teresa J. Irish

Keyword(s):

Rna Polymerase ◽

Virulence Factor ◽

Pertussis Toxin ◽

Bordetella Pertussis ◽

Virulence Genes ◽

Transcriptional Activator ◽

Alpha Subunit ◽

Terminal Domain

ABSTRACT Overexpression of the RNA polymerase alpha subunit inBordetella pertussis reduces expression of the virulence factor pertussis toxin. Here we show that this reduction is at the level of transcription, is reversed by overexpression of the transcriptional activator BvgA, and is dependent on the C-terminal domain of alpha.

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Complete Genome Sequence of Bordetella pertussis Pelita III, the Production Strain for an Indonesian Whole-Cell Pertussis Vaccine

Genome Announcements ◽

10.1128/genomea.00235-17 ◽

2017 ◽

Vol 5 (17) ◽

Author(s):

Yusuf Sofyan Efendi ◽

Dwi Susanti ◽

Erman Tritama ◽

Michelle Lueders Pasier ◽

Gilang Nadia Niwan Putri ◽

...

Keyword(s):

Genome Sequence ◽

Bordetella Pertussis ◽

Whooping Cough ◽

World Health ◽

Whole Genome Sequence ◽

Whole Genome ◽

Whole Cell ◽

Content Type ◽

Health Organization ◽

Production Strain

ABSTRACT PT Bio Farma, the sole World Health Organization-approved Indonesian vaccine producer, manufactures a whole-cell whooping cough vaccine (wP) that, as part of a pentavalent diphtheria-tetanus-pertussis/hepatitis B/Haemophilus influenzae b (DTP/HB/Hib) vaccine, is used in Indonesia and many other countries. We report here the whole-genome sequence for Bordetella pertussis Pelita III, PT Bio Farma’s wP production strain.

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G Proteins Gαi1/3Are Critical Targets for Bordetella pertussis Toxin-Induced Vasoactive Amine Sensitization

Infection and Immunity ◽

10.1128/iai.00971-13 ◽

2013 ◽

Vol 82 (2) ◽

pp. 773-782 ◽

Cited By ~ 7

Author(s):

Sean A. Diehl ◽

Benjamin McElvany ◽

Rajkumar Noubade ◽

Nathan Seeberger ◽

Brock Harding ◽

...

Keyword(s):

Pertussis Toxin ◽

Bordetella Pertussis ◽

Inflammatory Responses ◽

Whooping Cough ◽

Differential Susceptibility ◽

Inbred Strains ◽

Content Type ◽

Multiple Loci ◽

Inbred Strains Of Mice

ABSTRACTPertussis toxin (PTX) is an AB5-type exotoxin produced by the bacteriumBordetella pertussis, the causative agent of whooping cough.In vivointoxication with PTX elicits a variety of immunologic and inflammatory responses, including vasoactive amine sensitization (VAAS) to histamine (HA), serotonin (5-HT), and bradykinin (BDK). Previously, by using a forward genetic approach, we identified the HA H1receptor (Hrh1/H1R) as the gene in mice that controls differential susceptibility toB. pertussisPTX-induced HA sensitization (Bphs). Here we show, by using inbred strains of mice, F1hybrids, and segregating populations, that, unlike Bphs, PTX-induced 5-HT sensitivity (Bpss) and BDK sensitivity (Bpbs) are recessive traits and are separately controlled by multiple loci unlinked to 5-HT and BDK receptors, respectively. Furthermore, we found that PTX sensitizes mice to HA independently of Toll-like receptor 4, a purported receptor for PTX, and that the VAAS properties of PTX are not dependent upon endothelial caveolae or endothelial nitric oxide synthase. Finally, by using mice deficient in individual Gαi/oG-protein subunits, we demonstrate that Gαi1and Gαi3are the criticalin vivotargets of ADP-ribosylation underlying VAAS elicited by PTX exposure.

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