Cellular immunity in adolescents and adults following acellular pertussis vaccine administration

Cell-mediated immune (CMI) responses to an acellular pertussis vaccine administered to 49 subjects, a subset of participants in the National Institutes of Health-funded adult acellular pertussis vaccine efficacy trial, were evaluated and compared with antibody responses to vaccine antigens. Levels of proliferation of and cytokine secretion from lymphocytes cultured in the presence of pertussis toxin, filamentous hemagglutinin, or pertactin were measured before vaccination and 1 month and 1 year after vaccination. Statistically significant increases in lymphocyte stimulation indices and cytokine secretion were noted at both 1 month and 1 year after vaccination. Brisk pertussis antigen-specific immunoglobulin G responses were also noted at 1 month after vaccination, but these responses had declined by nearly 50% at 1 year after vaccination. These studies clearly demonstrate that both cellular and humoral immune responses occur after the administration of acellular pertussis vaccines to adolescents and adults but that the CMI responses are of greater magnitude and longer duration. CMI responses may be a better correlate of long-term protection.     

Development and clinical testing of an acellular pertussis vaccine containing genetically detoxified pertussis toxin.

In 1924 Ramon described the inactivation of diphtheria toxin by formaldehyde treatment. This method allowed the introduction of mass vaccination against diphtheria and tetanus and opened the way to the inactivation of viruses by chemical treatment. In this review we describe the use of genetic manipulations for the inactivation of pertussis toxin. The toxin inactivated by this new method is an antigen superior to those obtained by chemical treatment and has been used to develop a new vaccine against whooping cough.     

Evolution of Bordetella pertussis in the acellular vaccine era in Norway, 1996 to 2019

European Journal of Clinical Microbiology & Infectious Diseases - We described the population structure of Bordetella pertussis (B. pertussis) in Norway from 1996 to 2019 and determined if...     

单磷酸脂质A佐剂对无细胞百日咳疫苗的免疫保护效果研究

目的 探讨单磷酸脂质A(MPLA)佐剂对无细胞百日咳疫苗(aP)的免疫保护效果影响.方法 aP中添加MPLA佐剂,在Balb/c小鼠上进行免疫以及感染保护实验.通过检测小鼠百日咳特异性IgG抗体及其分型抗体IgG1和IgG2a水平、感染后白细胞变化以及气管和肺组织细菌定植来进行免疫效果的评估.结果 aP+MPLA免疫的...     

Antibody decay after immunisation of health-care workers with an acellular pertussis vaccine

Antibody decay after a single dose of acellular pertussis vaccine containing 25 μg of pertussis toxin (PT), 25 μg of filamentous hemagglutin (FHA), and traces of pertactin (PRN) was monitored in health-care workers. Blood was sampled 4 weeks (n = 246), 1 year (n = 187), 2 years (n = 53), 3 years (n = 134), and 4 years (n = 37) after vaccination. IgG anti-PT, IgG anti-FHA and IgG anti-PRN were measured by ELISA. Peak median antibodies to PT, FHA, and PRN were 314, 785, and 84 EU/ml respectively. IgG anti-PT decreased to a median of 29% (76 EU/ml), 18% (64 EU/ml), 19% (58 EU/ml), and 20% (63 EU/ml) of the peak value after 1, 2, 3, and 4 years respectively. IgG anti-FHA decreased more slowly, but showed similar decay patterns. In German health-care workers antibodies to pertussis antigens decayed rapidly within the first year after vaccination, but remained stable after 2, 3, and 4 years. This observation may have implications for the timing of booster vaccinations in adults. Parts of this study were presented as a poster (Poster #129) during the 8th International Symposium “Saga of the genus Bordetella,” 7–10 November 2006 at the Institut Pasteur in Paris, France.     

Changes of the Swedish Bordetella pertussis population in incidence peaks during an acellular pertussis vaccine period between 1997 and 2004

In a surveillance programme undertaken from 1997 through 2004, Bordetella pertussis isolates and clinical information were collected after introduction of acellular pertussis vaccines (Pa) in 1996. Changes in the B. pertussis population were studied in three incidence peaks: 1999-2000, 2002 and 2004. Available isolates from 158 fully vaccinated children representing all of Sweden, plus 37 from the Gothenburg area 2003-2004, were analysed by pulsed-field gel electrophoresis (PFGE), serotyping and sequencing of the virulence factor genes pertussis toxin subunits 1 and 3 (ptxA, ptxC), pertactin (prn), tracheal colonisation factor (tcfA) and fimbria3 (fim3). Allele ptxA1 was found in all isolates. There was a statistically significant increasing trend in three out of five studied genes, ptxC, prn and tcfA, and for a fourth, Fim3, if Gothenburg strains were included. The PFGE profile BpSR11 appearing in the 1999-2000 peak dominated by >or=23% during the entire period, bringing with it the allele combination 1/2/2/2/B (ptxA1/ptxC2/prn2/tcfA2/fim3B). Other BpSR11-related profiles with the same allele combination and more than 82% similarity--BpSR5 in the 2002 peak and BpSR12 in the 2004 peak--appeared with an increasing trend. Although vaccination with Pa has reduced disease, new variants have emerged representing clones surviving in the immunized population.     

Pertussis-specific cell-mediated immunity in infants after vaccination with a tricomponent acellular pertussis vaccine.

The aim of this study was to investigate pertussis-specific cell-mediated immunity in infants vaccinated with a tricomponent acellular vaccine. Infants were investigated during a primary vaccination schedule from the third month of life to the sixth month as well as before and after a booster at 15 to 24 months. This is the first report of specific cell-mediated immune responses to pertussis-related antigens in infants below the age of 12 months. Our data show that the vaccine induces T-cell responses specific for the vaccine components, detoxified pertussis toxin, filamentous hemagglutinin, and pertactin, that increase progressively over the course of the vaccination schedule. In contrast to declining antibody titers, cell-mediated immune responses are stable over the postprimary to prebooster period. Vaccination results in a progressive increase in the number of T cells that express activation marker CD45RO preferentially on CD4-positive T cells after stimulation with pertussis antigens. Measurements of cytokine secretion profiles demonstrated a preferential induction of interleukin 2- and gamma interferon-producing T-helper 1 cells and only low production of interleukin 10. The observed persistence of the specific cell-mediated immunity may have a bearing on the protective mechanisms induced by pertussis vaccination. Cell-mediated immunity requires further study, particularly to improve our understanding of the persistence of protection afforded by vaccination up to the administration of booster doses.     

Assay of pertussis vaccine toxicity by a rat-paw-oedema method.

A method is described of using the production of oedema in the foot pad of rats as an index of the ability of pertussis vaccines to cause local reactions. At a suitable time after the subcutaneous injection of the vaccine into hind paw of the rat, the foot is excised and weighed. The technique is reproducible and most useful for the detection of oedema produced by pertussis-vaccine components sensitive to heating at 80 degrees C for 30 min. Substances in pertussis vaccine that produce rat-paw oedema gave maximal reactions at 4 and 12 h. but were best differentiated 9 h after injection.     

Lipopolysaccharide analogs improve efficacy of acellular pertussis vaccine and reduce type I hypersensitivity in mice

Pertussis is an infectious disease of the respiratory tract that is caused by the gram-negative bacterium Bordetella pertussis. Although acellular pertussis (aP) vaccines are safe, they are not fully effective and thus require improvement. In contrast to whole-cell pertussis (wP) vaccines, aP vaccines do not contain lipopolysaccharide (LPS). Monophosphoryl lipid A (MPL) and Neisseria meningitidis LpxL2 LPS have been shown to display immune-stimulating activity while exerting little endotoxin activity. Therefore, we evaluated whether these LPS analogs could increase the efficacy of the aP vaccine. Mice were vaccinated with diphtheria-tetanus-aP vaccine with aluminum, MPL, or LpxL2 LPS adjuvant before intranasal challenge with B. pertussis. Compared to vaccination with the aluminum adjuvant, vaccination with either LPS analog resulted in lower colonization and a higher pertussis toxin-specific serum immunoglobulin G level, indicating increased efficacy. Vaccination with either LPS analog resulted in reduced lung eosinophilia, reduced eosinophil numbers in the bronchoalveolar lavage fluid, and the ex vivo production of interleukin-4 (IL-4) by bronchial lymph node cells and IL-5 by spleen cells, suggesting reduced type I hypersensitivity. Vaccination with either LPS analog increased serum IL-6 levels, although these levels remained well below the level induced by wP, suggesting that supplementation with LPS analogs may induce some reactogenicity but reactogenicity considerably less than that induced by the wP vaccine. In conclusion, these results indicate that supplementation with LPS analogs forms a promising strategy that can be used to improve aP vaccines.     

A novel process for preparing an acellular pertussis vaccine composed of non-pyrogenic toxoids of pertussis toxin and filamentous hemagglutinin

A novel process for preparing non-pyrogenic toxoids of pertussis toxin (PT) and filamentous hemagglutinin (FHA) is described. The process consists of chromatographies on perlite then on hydroxylapatite. Purification yields for PT and FHA are 62 and 68%, respectively. The purification process takes advantage of the novel use of perlite (a filter aid) for the simultaneous purification of PT and FHA. The hydroxylapatite, in addition to removing the remaining contaminants, also concentrates the antigens. The resulting PT and FHA are approximately 95% pure, and are non-pyrogenic as judged by the rabbit pyrogen test. The purification process is simple, inexpensive, and does not use blood components or toxic substances. The mild conditions in which the PT and FHA are purified ensure the recovery of native protein. The purified PT and FHA are detoxified in the presence of glycerol using glutaraldehyde and formaldehyde, respectively, to produce antigenic components of an acellular pertussis vaccine. The final PT and FHA toxoids are immunogenic in guinea-pigs and have been shown to be protective in the mouse intracerebral challenge test.     

Immunoglobulin E response to pertussis toxin in whooping cough and after immunization with a whole-cell and an acellular pertussis vaccine

Immunoglobulin E antibodies to pertussis toxin (PT-IgE) were demonstrated in 15 of 23 (65%) patients with culture-confirmed pertussis. In 6 individuals there was a low-grade PT-IgE response after 6-9 weeks of disease and in 9 a rapid PT-IgE response, appearing 1-3 weeks after onset of symptoms. The PT-IgE antibody levels in immunized individuals were higher than in the non immunized. Following primary immunization of 23 children with a monovalent whole-cell pertussis vaccine (Burroughs-Wellcome, UK) or with an acellular pertussis vaccine (JNIH-6, Biken, Japan) a late low-grade PT-IgE response was found in 8 (35%). In 7/10 children previously immunized with the JNIH-6, a booster injection 16 months later with the same vaccine resulted in a rapidly appearing PT-IgE antibody response. In contrast, none of 13 children initially immunized with the monovalent whole-cell vaccine and then boostered with either this vaccine or JNIH-6 had detectable PT-IgE antibodies after the booster injection. The study shows that IgE-antibodies to pertussis toxin commonly appear in patients with whooping cough and that the kinetics and the magnitude of the response is influenced by previous exposure to the antigen. A PT-IgE response may also follow pertussis immunization.     

Reduction of antibody response to an 11-valent pneumococcal vaccine coadministered with a vaccine containing acellular pertussis components

In pneumococcal conjugate vaccines (PCVs), polysaccharide antigens are often conjugated to protein carriers related to other common vaccines. It is therefore important to test PCV interaction with other pediatric vaccines when administered simultaneously. We assessed the immune response to an 11-valent PCV conjugated to diphtheria and tetanus carriers (PncD/T11), administered concomitantly, but in separate sites, with a combined vaccine containing epitopes related antigenically to the carriers: polyribosylribitol phosphate-tetanus tox oid (PRP-T), diphtheria toxoid (DT), and tetanus toxoid (TT). In addition, these combinations contained inactivated poliovirus vaccine (IPV) and either whole-cell pertussis (wP) or acellular pertussis (aP) components. After coadministration of PncD/T11 with the combined vaccine containing wP (DTwP/IPV/PRP-T), the responses to all polysaccharides in the PncD/T11 were satisfactory. In contrast, when coadministered with an aP-containing combination (DTaP/IPV/PRP-T), the response to all seven pneumococcal conjugates to TT was significantly reduced after primary and booster immunization. The pneumococcal conjugates to DT were not significantly reduced after the primary series, but were somewhat reduced after booster. It is likely that some suppression of the tetanus-mediated response occurred even when the PncD/T11 was coadministered with wP, but this suppression was masked by the adjuvant effect of wP. By replacing wP with aP, this adjuvant effect was removed, unmasking the suppression of the tetanus-mediated response. With the increasing use of multiple aP-containing vaccines in infancy, novel approaches to adjuvants and carrier protein technology are likely to be required.     

Protective effect of acellular pertussis vaccines

Two acellular pertussis vaccines, a mono-component toxoid of pertussis toxin and a two-component vaccine containing both the toxoid and filamentous haemagglutinin, were used for primary immunization of infants 5 1/2 to 10 months of age in two clinical trials in Sweden. Over a follow-up period of 12 to 17 months, 37 children were exposed to pertussis in the household or a daycare centre. Only one child developed mild, culture-confirmed pertussis as opposed to the expected number of 30 to 33 cases based on an assumed attack rate of 80-90% in non-immunized children. These preliminary findings indicate that, given as primary immunization to infants, the vaccines provide protection against pertussis.     

Effect of different forms of adenylate cyclase toxin of Bordetella pertussis on protection afforded by an acellular pertussis vaccine in a murine model

Four recombinant forms of the cell-invasive adenylate cyclase toxin (CyaA) of Bordetella pertussis were compared for the ability to enhance protection against B. pertussis in mice when coadministered with an acellular pertussis vaccine (ACV). The four forms were as follows: fully functional CyaA, a CyaA form lacking adenylate cyclase enzymatic activity (CyaA*), and the nonacylated forms of these toxins, i.e., proCyaA and proCyaA*, respectively. None of these forms alone conferred significant (P > 0.05) protection against B. pertussis in a murine intranasal challenge model. Mice immunized with ACV alone showed significant (P < 0.05) reductions in bacterial numbers in the lungs after intranasal challenge compared with those for control mice. When administered with ACV, both CyaA and CyaA* further reduced bacterial numbers in the lungs of mice after intranasal challenge compared with those for ACV-immunized mice, but the enhanced protection was only significant (P < 0.05) with CyaA*. Coadministration of CyaA* with ACV caused a significant (P < 0.05) increase in immunoglobulin G2a antibody levels against pertactin compared with those in mice immunized with ACV alone. Spleen cells from mice immunized with ACV plus CyaA* secreted larger amounts of interleukin-5 (IL-5), IL-6, gamma interferon (IFN-gamma), and granulocyte-macrophage colony-stimulating factor (GM-CSF) than did cells from mice immunized with ACV plus CyaA or ACV alone after stimulation in vitro with a mixture of B. pertussis antigens. Spleen cells from mice immunized with ACV plus CyaA* also secreted larger amounts of IFN-gamma and GM-CSF than did cells from mice immunized with CyaA* alone after stimulation in vitro with CyaA*. Macrophages from mice immunized with ACV plus CyaA* produced significantly (P < 0.05) higher levels of nitric oxide than did macrophages from mice immunized with CyaA* alone, ACV alone, or ACV plus CyaA after stimulation in vitro with a mixture of B. pertussis antigens or heat-killed B. pertussis cells. These data suggest that the enhancement of protection provided by CyaA* was due to an augmentation of both Th1 and Th2 immune responses to B. pertussis antigens.     

A novel TLR2 agonist from Bordetella pertussis is a potent adjuvant that promotes protective immunity with an acellular pertussis vaccine

Bordetella pertussis causes whooping cough, a severe and often lethal respiratory infection in infants. A recent resurgence of pertussis has been linked with waning or suboptimal immunity induced with acellular pertussis vaccines (Pa) that were introduced to most developed countries in the 1990s because of safety concerns around the use of whole-cell pertussis vaccines (Pw). Pa are composed of individual B. pertussis antigens absorbed to alum and promote strong antibody, T helper type 2 (Th2) and Th17 responses, but are less effective at inducing cellular immunity mediated by Th1 cells. In contrast, Pw, which include endogenous Toll-like receptor (TLR) agonists, induce Th1 as well as Th17 responses. Here we report the identification and characterization of novel TLR2-activating lipoproteins from B. pertussis. These proteins contain a characteristic N-terminal signal peptide that is unique to Gram-negative bacteria and we demonstrate that one of these lipoproteins, BP1569, activates murine dendritic cells and macrophages and human mononuclear cells via TLR2. Furthermore, we demonstrated that a corresponding synthetic lipopeptide LP1569 has potent immunostimulatory and adjuvant properties, capable of enhancing Th1, Th17, and IgG2a antibody responses induced in mice with an experimental Pa that conferred superior protection against B. pertussis infection than an equivalent vaccine formulated with alum.     

Aerosolized vaccine as an unexpected source of false-positive Bordetella pertussis PCR results

When 13 of 13 nasal wash specimens from a single pediatrician's office tested positive for low quantities of Bordetella pertussis DNA, we suspected prelaboratory contamination. Investigation revealed that Pentacel and Adacel vaccines contain high copy numbers of B. pertussis DNA, which can be aerosolized, causing false-positive B. pertussis PCR results.     

A combined diphtheria, tetanus, five-component acellular pertussis, poliovirus and Haemophilus influenzae type b vaccine

Ideally, combination vaccines should not only be safe and effective, but also integrate smoothly into the vaccination schedule and provide advantages over the use of separately administered vaccines. Pentaceltrade mark (Sanofi Pasteur Ltd., Toronto, Canada), a combination vaccine first licensed in Canada and subsequently in other countries, is immunogenic against diphtheria, tetanus, pertussis, polio and Haemophilus influenzae type b when administered at 2, 4, 6 and 15-18 months of age. In published studies, the safety, immunogenicity and effectiveness of this combination vaccine were comparable with those of separately administered vaccines, with the advantage of a simplified dosing schedule.