Introduction: In 2012, the Advisory Committee on Immunization Practices (ACIP) of the Centers for Disease Control and Prevention expanded their recommendation for the Tdap vaccination to include the antepartum period. Regardless of immunization history, the recommendation states that medical practitioners should administer the Tdap vaccination to every pregnant woman in each occurring pregnancy (Munoz, et al., 2014; Shakib, et al., 2013; Goldfarb, Little, Brown, Riley, 2014). Methods: To describe treatment practices and uptake of Tdap vaccine, a cross-sectional descriptive survey design was utilized. The purpose of survey is to gather information regarding prevalence, distribution, and interrelations of variables within a population (Polit & Beck). In this study, the survey questionnaire was conducted in an online format. Results: Of the six HBM questions included in the study, except for question four, the results of the chi-squared analysis suggest that any single measured dimension of the HBM cannot predict a health behavior, in this case receipt of the Tdap vaccination. The population is split regarding infants' perceived susceptibility to pertussis infection. Strong agreement to the benefit of vaccination was revealed. Question six regarded available information, although the majority were satisfied a significant percentage indicated a desire for more information. Discussion: This Doctorate of Nursing Practice project developed a survey based on the Health Belief Model with the intention of assessing perceived susceptibility, perceived severity, perceived benefits and perceived barriers to the health care preventative action of receiving the Tdap vaccination in the third trimester of pregnancy. Through in-depth literature review, consideration of the updated ACIP guidelines, and support of a developed theoretical framework, an eight-question survey was developed. The data examined in this project may serve to illustrate limitations in provider care that can be immediately improved upon, such as information sharing. The primary limitation of the study is in the sample size of 44 eligible survey responses and the uniform demographics of the population. Despite these limitations, the survey design may be extended to other populations of interest, with greater demographic variation for further study.

An aspartate transcarbamoylase (ATCase) gene from Bordetella pertussis was amplified by PCR and ligated into pT-ADV for expression in Escherichia coli. This particular ATCase (pyrB) was an inactive gene found adjacent to an inactive dihydroorotase (DHOase) gene (pyrC'). This experiment was undertaken to determine whether this pyrB gene was capable of expression alone or if it was capable of expression only when cotransformed with a functional pyrC'. When transformed into E. coli TB2 pyrB-, the gene did not produce any ATCase activity. The gene was then co-transformed into E. coli TB2 pyrB- along with a plasmid containing the pyrC' gene from Pseudomonas aeruginosa and assayed for ATCase activity. Negative results were again recorded.

Thesis (Ph. D.)--University of Virginia, 2001. / Includes bibliographical references (leaves 146-168). Also available online through Digital Dissertations.

Bordetella pertussis, the etiologic agent of whooping cough or pertussis, produces a multitude of virulence factors including a hemolysin. Virulent phase B. pertussis isolates are hemolytic, whereas avirulent isolates are not. Other investigations concerning B. pertussis adenylate cyclase toxin indicate it has hemolytic activity and is a member of the bacterial RTX toxin family. In an attempt to further characterize hemolysis by B. pertussis, a new B. pertussis gene was isolated which conferred a hemolytic phenotype on non-hemolytic E. coli. DNA sequencing of the putative B. pertussis hemolysin gene revealed it encoded a 27 kDa protein similar to HlyX, an FNR-like transcriptional regulator from Actinobacillus pleuropneumonia, which also confers hemolysis upon E. coli. No similarity to bacterial cytolysins was found. The B. pertussis transcriptional regulator-like gene and its encoded protein were named btr and BTR, respectively. BJB1, a BTR deficient B. pertussis strain was constructed. The btr::kan mutation was shown to have no effect on the production, or phenotypic modulation, of hemolysis by B. pertussis. BTR production was not regulated by the BvgA-S two component sensor-regulator. An FNR deficient E. coli, JRG1728 (Δfnr), was transformed with a btr recombinant plasmid pHLY1A. The B. pertussis btr gene complemented the FNR deficient E. coli to grow anaerobically on a non-fermentable carbon source. This suggested that BTR may function as a B. pertussis transcriptional regulator which responds to anoxic conditions.

No description available.

Whooping cough is a common childhood disease caused by infection with the bacteria Bordetella pertussis or Bordetella parapertussis. Although previously considered under control within developed countries due to widespread vaccination, whooping cough has undergone a resurgence in many developed countries, with recent outbreaks underscoring the need for new and more effective vaccines. Most crucially, there is a need for vaccines that can be administered during the neonatal period, to protect infants at their most susceptible, and vaccines that will provide long-lasting protection. Due to the functional differences of the neonatal immune system as compared to the adult, there are specific difficulties that must be overcome when attempting to create a vaccine that will be effective in the neonate. The objective of the current research was to examine the immunogenicity of several adjuvants, including CpG ODN, IDRP, and PP, in order to design a combined novel adjuvant platform that could be used with our pertussis vaccine antigen, PTd. After selection of each adjuvant component, we tested the ability of various adjuvant formulations to induce Th1 and Th2 humoral responses in both adult and neonatal mice. We found that a 1:2:1 ratio of CpG ODN: IDRP: PP was most effective, and that pre-complexation of the IDRP and CpG ODN components induced significantly higher Th1 (IgG2a) antibody titres than non-complexed vaccines. Our vaccine platform induced strong Th1 and Th2 antibody titres in both adult and neonatal BALB/c mice, with the immune response being of a mixed Th1/Th2 type. The Th1 type humoral response to our vaccine platform was significantly higher than that seen using current commercial vaccines such as Quadracel®, or when using the standard vaccine adjuvant alum. This Th1 antibody response was extremely long-lasting, with strong IgG2a titres being found up to 2 years post-vaccination. When examining the cell-mediated immune response in adult and neonatal BALB/c mice, a strong secretory IFN-g (Th1) response was present post-vaccination in the splenocytes of platform-vaccinated mice, with a large number of IFN-g secreting cells present. The IL-5 (Th2) response was found to be decreased in mice that received our novel vaccine as compared to mice vaccinated with Quadracel®, with no detectable cytokine secreted by stimulated splenocytes in vitro, and few to no IL-5 secreting cells visible through ELISPOT. To further improve our vaccine, a second antigen, pertactin (PRN), was added to the formulation. Upon live bacterial challenge, mice that received the two-antigen vaccine were completely protected against infection, and showed strong humoral response. This full protection and clearance was superior to the results seen using Quadracel®. Finally, the variability and cell-recruitment functions of the adjuvant platform were examined. The adjuvant platform successfully induced a strong mixed Th1 and Th2 humoral response when combined with the vaccine antigen HBsAg, with a significant increase in the Th1 (IgG2a) antibody response. Replacing the CpG ODN component of the adjuvant platform with Poly I:C through and using various immunization routes also resulted in an induction of IgG2a titres. Thus we have developed a novel vaccine formulation against B.pertussis that induces strong humoral and cell-mediated immune responses in both adult and neonatal mice, with these responses being both long-lasting and protective against infection. Our novel adjuvant platform itself has been shown to be adaptable for use with other vaccine antigens and through several routes of administration, and there is the possibility of adjusting the components while maintaining efficacy.

Typescript (photocopy). / Digitized by Kansas Correctional Industries

Background: Pertussis diagnosis may go unrecognized when other pathogens, such as respiratory syncytial virus (RSV) circulate. Methods: A prospective cross-sectional study was conducted in Lima, Peru from January 2009 to September 2010. A total of 596 children under 5 years old admitted with clinical diagnoses of acute respiratory infections were test for B. pertussis and RSV detection by polymerase chain reaction (PCR). Results: The pertussis toxin and IS481 genes were detected in 19.12% (114/596) of the cases and the respiratory syncytial viruses (RSV-A and RSV-B) were identified in 17.28% (103/596) of patients. Infants under 3 months old were the most frequently affected by this pathogens in 43% (49/114) and 35.9% (37/103) respectively. An increase of B. pertussis was observed from February to March and from October to November with a Seasonal index between 1.32-1.51 and 1.24-3.5 respectively. Conclusions: Epidemiologic surveillance for B. pertussis is essential in Peru, especially in children that could most benefit from the vaccine. B. pertussis should be suspected in infants hospitalized for acute respiratory symptoms for early treatment and prevent complications. / Tesis

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 5 uppsatser.

Bordetella pertussis, o agente etiológico da coqueluche ou tosse comprida, que estabelece a infecção através da fixação bacteriana no epitélio do trato respiratório superior. Os principais mediadores de adesão da bactéria são a toxina pertússica (PT) e a hemaglutinina filamentosa (FHA). A FHA é a adesina majoritária e contém pelo menos 4 domínios: porção Nterminal, domínio de reconhecimento de carboidrato (CRD) (FHA1141-1279), trinca de aminoácidos Arginina-Glicina-Ácido aspártico (RGD) (FHA1097-1099) e o sítio de ligação a heparina (domínio Hep) (FHA442-863). Neste trabalho, foi realizado a amplificação de duas regiões do domínio de ligação à heparina, as regiões MAL80 (FHA299-873) e HEP (FHA430-873). As regiões foram amplificadas, clonadas no vetor de expressão pAE, expressas utilizando a cepa BL21 SI de E. coli e purificadas. A proteína HEP purificada de E. coli possui baixa afinidade por heparina e não é capaz de aglutinar hemácias. A proteína recombinante HEP purificada foi utilizada para a produção de anticorpos. Através do experimento de ELISA foi demonstrado que o anti-soro anti-HEP é capaz de reconhecer além da região HEP, a região MAL80 e a proteína FHA. Estes resultados foram confirmados por experimentos de Western. Neste período também foi realizada a amplificação do domínio HEP de FHA e da subunidade S1 da toxina pertússica (PT) de B. pertussis através do método de TAP Express. Este método envolve duas reações de PCR e no final do processo é obtido um fragmento que contém uma região promotora (CMV), uma seqüência codificadora e uma região terminadora (SV40), que está pronto para ser introduzido e expresso em animais. De posse deste material e da proteína recombinante HEP, foi realizado o desafio intracerebral com Bordetella pertussis e através do monitoramento dos camundongos foi observado que nenhum dos candidatos testados foi capaz de proteger os animais contra B. pertussis. Foi realizado também a expressão do domínio Hep em lactobacilos, visando um possível sistema de imunizações de mucosas. Os anticorpos produzidos nos camundongos imunizados com a proteína HEP expressa em E. coli, lactobacilos e por vacina de DNA foram capazes de inibir a hemaglutinação promovida pela proteína FHA. / Bordetelfa pertussis, the agent of whopping cough, establishes infection by attaching to the ciliated epithelial cells of the respiratory tract. The bacterial adherence is mediated by pertussis toxin and filamentous hemagglutinin (FHA). FHA is the major adhesin of B. pertussis and displays multipie adherence activities. FHA contains four distin\'ct domains that exhibit specific affinities for different ligands or receptor, the amino-terminal end, the RGD triplet (FHA1097-1099), the lectin domain (FHA1141-1279) and the heparin-binding domain (FHA442-863). In this study, two overlapping regions of the heparin-binding domain, Mal80 (FHA299-873) and Hep (FHA442-873), were amplified by peR and subcloned in pAE expression vectors for E. coli. The fusion proteins in pAE were transformed in E. coli BL21 SI, induced with NaCI 0,3 M and purified using a nickel-charged metal chelating resin. The purified protein has low heparin affinity and does not have hemagglutination activity. The purified protein HEP was used to produce polyclonal antibodies in mouse. The anti-HEP antibodies are able to recognize the HEP, MAL80 and FHA proteins in ELISA and western assays, but anti-FHA only recognized the FHA protein. The genetically detoxified S1 subunit of pertussis toxin and Hep domain were amplified by the TAP Express method. There are two PCR reactions involved in the TAP processo At the end of the process the fragment of interest will carry a CMV promoter and a SV40 terminator and is ready to be introduced into animals or cell by transfection. Groups were immunized with proteins and/or DNA, challenged i.c. with a lethal dose of live Bordetelfa pertussis and the survival was monitored. No groups were protected against the challenge. The recombinant protein HEP were also expressed in Lactobacilfus aiming the development of potential mucosal vaccines. The polyclonal antibodies produce in mouse immunized with DNA and protein Hep expressed in E. coli and Lactobacillus were able to inhibition the FHA hemagglutination activity.