Production and characterisation of a candidate hyper-immune serum for the replacement of the Bordetella pertussis mouse antiserum Biological Reference Preparation.

For acellular pertussis (aP) vaccines, the current European Pharmacopoeia (Ph. Eur.) monograph Pertussis vaccine (acellular, component, adsorbed) (1356) requires an immunogenicity assay in mice or guinea pigs to assess the potency of each lot of vaccine (Ph. Eur. general method 2.7.16. Assay of pertussis vaccine (acellular)). This biological assay, carried out on the final bulk of the vaccine lot, is based on the measurement of the specific antibody response to the 5 antigenic components (pertussis toxin (PT), Fimbrial haemagglutinin (FHA), pertactin (PRN) and Fimbriae 2 and 3 (FIM2/3)) that are present in the combined aP vaccines. In the mouse assay, serum antibody levels are measured by ELISA. The immunogenicity of a vaccine under test is estimated versus a homologous reference vaccine and a reference antiserum e.g. the first Ph. Eur. Biological Reference Preparation for Bordetella (B.) pertussis mouse anti-serum (BRP1), established in 1998, is used to normalise the titre of antibodies (expressed in ELISA Units (ELU)/mL). In anticipation of the depletion of BRP1 stocks, a project was launched in 2013 by the Biological Standardisation Programme (BSP) of the European Directorate for the Quality of Medicines & HealthCare (EDQM) in order to establish a new standardised reference serum. The project, referred to herein as BSP129, was conducted in 2 phases: 1) the production and characterisation of a mouse serum pool (using a multicomponent aP vaccine marketed in Canada similar to the vaccine used in the BRP1 production as immunogen) and of candidate BRP batches (cBRPs) and 2) an international collaborative study aimed at calibrating the cBRPs in terms of antibody levels against PT, FHA, PRN and FIM2/3. This article presents the design and results of the first phase of the collaborative study to establish the optimal conditions for immunisation and bleeding of mice in order to produce a large pool of hyper-immune serum against the 5 antigens. After the characterisation of this pool, cBRP pilot lots were manufactured by freeze-drying diluted solutions of the hyper-immune serum pool. The pilot lots were then characterised in two Official Medicines Control Laboratories (OMCLs) for their antibody contents against aP vaccine antigens using in-house ELISA (based on methods developed by 2 European vaccine manufacturers) and Multiplex Immunoassay (MIA) methods. The antibody titres recovered demonstrated that a dilution factor of 1/40 could be considered for the scaled-up manufacture of candidate reference preparations (cBRPs). Three batches (15 000 vials) of cBRP were manufactured and fully characterised. In light of the data obtained, and although titration results between the ELISA methods were sometimes discrepant, it was agreed that the establishment study (phase 2) could be launched. Real-time and accelerated stability studies were also included in the first study phase to document the stability of the cBRPs in freeze-dried form and after reconstitution and storage at -20°C±5°C. The results showed that the stability of the freeze-dried cBRPs at usual storage and shipment temperatures is acceptable and that reconstituted cBRP solutions are stable for 12 months at -20°C±5°C. It could therefore be recommended to freeze small aliquots of the 1 mL solution obtained by the reconstitution of one BRP vial in order to store them for use in separate assays. With the application of this strategy, the stocks of the BRP1 replacement batches should cover the needs of OMCLs and manufacturers for at least the next decade.

Establishment of Ph. Eur. Bordetella pertussis mouse antiserum Biological Reference Preparation batches 2, 3 and 4.

A project aimed at establishing replacement batches for the European Pharmacopoeia (Ph. Eur.) Biological Reference Preparation (BRP) Bordetella (B.) pertussis mouse antiserum was started in 2013 under the aegis of the Biological Standardisation Programme (BSP) of the European Directorate for the Quality of Medicines & HealthCare (EDQM). This BRP is used for the immunogenicity assay in mice to assess the potency of acellular pertussis (aP) vaccines as described in Ph. Eur. general method 2.7.16. Assay of pertussis vaccine (acellular). In a preliminary phase of the project (referred to herein as BSP129 phase 1) a hyper-immune serum pool was produced in mice using a combined aP vaccine as immunogen. This pool was used to generate 3 freeze-dried candidate (c) B. pertussis anti-mouse serum BRP batches (cBRP2, cBRP3 and cBRP4). After the pre-qualification that showed their suitability as candidate batches, an international collaborative study (BSP129 phase 2) was carried out in order to standardise these 3 batches against the current BRP1 in terms of anti-PT, -FHA, -PRN and -FIM2/3 antibody contents. For the sake of continuity with the standardisation of BRP1, the corresponding WHO standard (1RR 97/642) was introduced as a second reference for the calibration of the 3 candidate BRPs. Eleven laboratories took part in phase 2. Ten of them performed the ELISA method they use routinely for aP vaccine batch release and one laboratory performed the Multiplex Immunoassay (MIA) as an alternative test. Four participants titrated the antibodies against all 5 pertussis antigens, 5 participants determined the antibody content against 3 antigens (PT, FHA, PRN), one participant titrated the antibodies against PT and FHA antigens and one laboratory determined the antibody content for the PT antigen only. Details of all ELISA methods used were analysed to evaluate their impact on the calibration of the cBRPs. The variability of the results in relation to the nature and methodology of the tests appeared rather limited. Discrepant titres of cBRPs were measured depending on the reference used: the use of the 1RR induced an overestimation (in 8 out of 11 laboratories) and a large inter-laboratory variation in the calculated titres. Regardless of the reference used, equivalency between the calculated titres of cBRP2 and cBRP3 was observed, whilst cBRP4 had systematically lower titres for all antibodies against the 5 acellular pertussis vaccine components. Based on these observations, it was decided to establish the candidate BRP batches against BRP1 and to assign the following potencies based on the mean values determined through centrally calculated results of the calibration assays performed by ELISA in BSP129 phase 2: For cBRP2 and cBRP3 Anti-pertussis toxin: 37 ELISA Units (ELU) per vial Anti-filamentous haemagglutinin: 114 ELU per vial Anti-pertactin: 44 ELU per vial Anti-fimbrial agglutinogens (FIM2/3): 25 ELU per vial For cBRP4 Anti-pertussis toxin: 32 ELU per vial Anti-filamentous haemagglutinin: 98 ELU per vial Anti-pertactin 38 ELU per vial Anti-fimbrial agglutinogens (FIM2/3):23 ELU per vial In February 2018, BRP2, BRP3 and BRP4 were adopted by correspondence by the Ph. Eur. Commission.

Effectiveness of strategies to increase uptake of pertussis vaccination by new parents and family caregivers: A systematic review.

OBJECTIVES: Cocoon immunisation strategies involve administration of Bordetella pertussis containing vaccines to parents and family members who are in close contact with newborns. The objective of this systematic review was to evaluate the effectiveness of strategies to increase uptake of vaccination against Bordetella pertussis infection by parents and family caregivers of newborn children (< 3 months of age). DESIGN: A protocol driven systematic review was conducted between 2005 and February 2020. CINAHL, Medline, and Google Scholar databases were searched. SETTING: Inpatient maternity care units, ante-natal and post-natal clinics based in acute care or primary/community care contexts. PARTICIPANTS: (i) mothers, (ii) fathers and (iii) family caregivers or other regular household contacts of infants < 3 months of age. INTERVENTIONS: Health promotion interventions and immunisation clinics designed to promote "cocoon immunisation" against Bordetella pertussis infections of the newborn. MEASUREMENTS: Change in uptake of adult vaccination with a pertussis containing vaccine (dTpa or Tdap) by new parents and family caregivers. FINDINGS: Eight studies were included in this review. Strategies used to promote vaccination included: written and verbal education, promotional videos, provision of vaccine prescriptions and financial incentives, opportunistic vaccination of family members and population-based health promotional messaging. Six of the eight studies reported positive impacts on vaccination uptake. Four studies evaluating providing opportunistic immunisation during the obstetric admission reported statistically significant increases in maternal (+39% to +57%), paternal (+21% to +52%) and household members (+32%) vaccination rates. Targeted public health campaigns were also found to increase vaccination uptake but in isolation were insufficient to achieve vaccination of all household contacts. CONCLUSION: Promotion of pertussis vaccination to new parents and the provision of opportunistic vaccination during the obstetric admission or post-natal visit, was the most successful strategy to increase uptake of pertussis vaccination by family caregivers.

Collaborative study for the calibration of the replacement International Standard for pertussis toxin for use in histamine sensitisation and CHO cell clustering assays.

Pertussis toxin (PT) in its detoxified form is one of the major protective antigens in vaccines against Bordetella pertussis (whooping cough). Reference preparations of native PT are required for the quality control of pertussis vaccines. Stocks of the first WHO International Standard (IS) for PT (JNIH-5) were low and a replacement was required. One candidate material was donated by a vaccine manufacturer to NIBSC. It was formulated, lyophilised into sealed glass ampoules and coded 15/126. An international collaborative study assessed the suitability of this material to replace JNIH-5. Fourteen laboratories from 12 countries took part in the study. Eleven laboratories performed lethal murine histamine sensitisation assay (HIST), 14 performed Chinese Hamster Ovary (CHO) cell clustering assay. International Units (IU) were assigned to the material using these assays as they were used to assign units to JNIH-5. It was found that, unlike JNIH-5, the activities of 15/126 in HIST and CHO cell assays did not agree and therefore different unitage for each assay was assigned. The preparation 15/126 was established as the Second WHO IS for PT for HIST and CHO cell assays. It was assigned a unitage of 1,881 IU/ampoule in HIST and 680 IU/ampoule in the CHO cell clustering assay.

Calibration of pertussis toxin BRP batch 1 in a standardised CHO cell-based clustering assay.

The European Pharmacopoeia (Ph. Eur.) pertussis toxin (PT) Biological Reference Preparation (BRP) is used as a working standard for safety testing of acellular pertussis vaccines as prescribed in the Ph. Eur. monographs 1356 "Pertussis vaccine (acellular, component, adsorbed)" and 1595 "Pertussis vaccine (acellular, co-purified, adsorbed)". The BRP was calibrated in 2006 in the murine histamine sensitisation test (HIST) against the World Health Organization (WHO) 1st International Standard (IS) for PT. In recent years, there have been increasing efforts to replace the in vivo test with in vitro methods. The Chinese hamster ovary (CHO) cell clustering assay has been used for many years by manufacturers to monitor residual PT activity in detoxified non-adjuvanted bulks. More recently a standardised protocol has been developed for this assay and a PT reference preparation was needed. Due to low stocks, the WHO 1st International Standard for Pertussis Toxin (JNIH-5) needed to be replaced and therefore a joint study between the European Directorate for the Quality of Medicines & HealthCare (EDQM) and WHO was initiated to calibrate the PT BRP for the CHO clustering assay and to replace the IS. The collaborative study involved 14 laboratories from Europe, North America and Asia. The outcome of the study confirmed that the BRP is suitable for use as a reference preparation in the CHO clustering assay. The material was assigned a potency of 1360 IU per vial for the CHO clustering assay.

Transferability study of CHO cell clustering assays for monitoring of pertussis toxin activity in acellular pertussis vaccines.

Current regulations for acellular pertussis (aP) vaccines require that they are tested for the presence of residual or reversion-derived pertussis toxin (PTx) activity using the mouse histamine sensitisation test (HIST). Although a CHO cell clustering assay can be used by manufacturers to verify if sufficient inactivation of the substance has occurred in-process, this assay cannot be used at present for the final product due to the presence of aluminium adjuvants which interfere with mammalian cell cultures. Recently, 2 modified CHO cell clustering assays which accommodate for the adjuvant effects have been proposed as alternatives to the HIST. These modified assays eliminate the adjuvant-induced cytotoxicity either through dilution of the vaccine (called the Direct Method) or by introducing a porous barrier between the adjuvant and the cells (the Indirect Method). Transferability and suitability of these methods for testing of products present on the European market were investigated during a collaborative study organised by the European Directorate for the Quality of Medicines & HealthCare (EDQM). Thirteen laboratories participated in this study which included 4 aP-containing vaccines spiked by addition of PTx. This study also assessed the transferability of a standardised CHO cell clustering assay protocol for use with non-adjuvanted PTx preparations. Results showed that the majority of laboratories were able to detect the PTx spike in all 4 vaccines at concentrations of 4 IU/mL or lower using the Indirect Method. This sensitivity is in the range of the theoretical sensitivity of the HIST. The Direct Method however did not show the expected results and would need additional development work.

10 Practice Recommendations in Adult Vaccination Administration.

Although the provision of immunoprophylaxis to children has become routine in the practice of pediatric preventive care, the same is not true in adult primary care. Contributing to this problem is a lack of knowledge among providers of adult preventive care. This review aimed to bolster providers' understanding of adult vaccinations by highlighting changes in vaccination recommendations and addressing common knowledge gaps. This is not a comprehensive list of vaccination recommendations, but rather the "top 10" common misconceptions, advancements, and updates we have found in our reading of the vaccination literature and in our own experience in a training institution.

Hospitalizations and deaths due to pertussis in children from 1996 to 2013☆ / Internações e óbitos por coqueluche em crianças no período entre 1996 e 2013

ABSTRACT OBJECTIVES: To assess temporal trends of hospitalizations and deaths from pertussis in Brazilian children in the period of 1996-2013. METHODS: This was a descriptive ecological study of temporal trends, based on the DATASUS database. The number of hospitalizations and deaths from pertussis in children up to 19 years of age from January 1996 to December 2013 was obtained. Descriptive statistics were applied for data analysis. RESULTS: During the study period, a total of 19,047 hospital admissions from pertussis were recorded, of which 88.2% occurred in infants younger than 1 year. In the period 1996-2010, the mean annual number of admissions was 755, ranging from a maximum of 1179 in 2004 to a minimum of 400 in 2010. There was an increase of admissions in the last three consecutive years (2011, 2012, and 2013) with 1177, 2954 and 3589 hospitalizations, respectively. There were 498 deaths from pertussis throughout the study period, of which 96.8% occurred in children younger than one year. There was an increase in the number of deaths from pertussis in children in the years 2011, 2012, and 2013, with 40, 93, and 87 recorded deaths, respectively. The increase in hospitalizations and deaths from pertussis in children occurred in all regions of the country, with the highest increase observed in the Southeast, North and Northeast regions. CONCLUSIONS: There was a substantial increase in hospitalizations and deaths from pertussis in children for three consecutive years (2011, 2012, and 2013) in all Brazilian regions. The most affected age group was that of children younger than one year.

RESUMO OBJETIVOS: Avaliar a tendência temporal de internações e óbitos por coqueluche em crianças brasileiras de 1996 a 2013. MÉTODOS: Trata-se de um estudo ecológico descritivo de tendência temporal, baseado no banco de dados Datasus. Foram extraídos os números de internações e de óbitos por coqueluche em crianças até 19 anos de janeiro de 1996 a dezembro de 2013. A estatística descritiva foi aplicada para análise de dados. RESULTADOS: No período estudado foram registradas 19.047 internações por coqueluche, das quais 88,2% foram de lactentes menores de um ano. De 1996 a 2010, o número médio anual de internações foi de 755 e oscilou entre o máximo de 1.179 em 2004 e o mínimo de 400 em 2010. Houve um acréscimo de internações nos últimos três anos consecutivos (2011, 2012 e 2013), com 1.177, 2.954 e 3.589 registros, respectivamente. Ocorreram 498 óbitos por coqueluche em todo o período estudado, dos quais 96,8% eram menores de um ano. Houve acréscimo no número de óbitos por coqueluche em crianças em 2011, 2012 e 2013, com 40, 93 e 87 registrados, respectivamente. O aumento de internações e óbitos por coqueluche em crianças ocorreu em todas as regiões do país e houve maior acréscimo nas regiões Sudeste e Norte-Nordeste. CONCLUSÕES: Houve um aumento substancial de internações e de óbitos por coqueluche em crianças por três anos consecutivos (2011, 2012 e 2013) em todas as regiões brasileiras. A faixa etária mais atingida foi a de menores de um ano.

Comparison of Three Whole-Cell Pertussis Vaccines in the Baboon Model of Pertussis.

Pertussis is a highly contagious respiratory illness caused by the bacterial pathogen Bordetella pertussis. Pertussis rates in the United States have escalated since the 1990s and reached a 50-year high of 48,000 cases in 2012. While this pertussis resurgence is not completely understood, we previously showed that the current acellular pertussis vaccines do not prevent colonization or transmission following challenge. In contrast, a whole-cell pertussis vaccine accelerated the rate of clearance compared to rates in unvaccinated animals and animals treated with the acellular vaccine. In order to understand if these results are generalizable, we used our baboon model to compare immunity from whole-cell vaccines from three different manufacturers that are approved outside the United States. We found that, compared to clearance rates with no vaccine and with an acellular pertussis vaccine, immunization with any of the three whole-cell vaccines significantly accelerated the clearance of B. pertussis following challenge. Whole-cell vaccination also significantly reduced the total nasopharyngeal B. pertussis burden, suggesting that these vaccines reduce the opportunity for pertussis transmission. Meanwhile, there was no difference in either the duration or in B. pertussis burden between unvaccinated and acellular-pertussis-vaccinated animals, while previously infected animals were not colonized following reinfection. We also determined that transcription of the gene encoding interleukin-17 (IL-17) was increased in whole-cell-vaccinated and previously infected animals but not in acellular-pertussis-vaccinated animals following challenge. Together with our previous findings, these data are consistent with a role for Th17 responses in the clearance of B. pertussis infection.

In search of acceptable alternatives to the murine histamine sensitisation test (HIST): what is possible and practical?

The 'International Workshop on Alternatives to the Murine Histamine Sensitization Test for Acellular Pertussis Vaccines: In Search of Acceptable Alternatives to the Murine Histamine Sensitization Test (HIST): What is Possible and Practical?' was held on 4 and 5 March 2015 in London, United Kingdom. Participants discussed the results of the data generated from an international collaborative study (BSP114 Phase 2) sponsored by the European Directorate for the Quality of Medicines & Health Care (EDQM) to determine if a modified Chinese hamster ovary (CHO) cell-based clustering assay is a suitable alternative to replace HIST. Workshop participants agreed that protocol transferability demonstrated in the collaborative study indicates that a standardised CHO cell assay is adequate for measuring pure PTx in reference preparations. However, vaccine manufacturers would still need to demonstrate that the method is valid to detect or measure residual PTx in their specific adjuvanted products. The 2 modified CHO cell protocols included in the study (the Direct and the Indirect Methods) deserve further consideration as alternatives to HIST. Using the CHO cell assay, an in vitro alternative, for acellular pertussis (aP) vaccine batch release testing would reduce the number of animals used for aP vaccine safety testing. A strategic, stepwise adoption plan was proposed, in which the alternative test would be used for release purposes first, and then, once sufficient confidence in its suitable performance has been gained, its use would be extended to stability testing.

Hospitalizations and deaths due to pertussis in children from 1996 to 2013.

OBJECTIVES: To assess temporal trends of hospitalizations and deaths from pertussis in Brazilian children in the period of 1996-2013. METHODS: This was a descriptive ecological study of temporal trends, based on the DATASUS database. The number of hospitalizations and deaths from pertussis in children up to 19 years of age from January 1996 to December 2013 was obtained. Descriptive statistics were applied for data analysis. RESULTS: During the study period, a total of 19,047 hospital admissions from pertussis were recorded, of which 88.2% occurred in infants younger than 1 year. In the period 1996-2010, the mean annual number of admissions was 755, ranging from a maximum of 1179 in 2004 to a minimum of 400 in 2010. There was an increase of admissions in the last three consecutive years (2011, 2012, and 2013) with 1177, 2954 and 3589 hospitalizations, respectively. There were 498 deaths from pertussis throughout the study period, of which 96.8% occurred in children younger than one year. There was an increase in the number of deaths from pertussis in children in the years 2011, 2012, and 2013, with 40, 93, and 87 recorded deaths, respectively. The increase in hospitalizations and deaths from pertussis in children occurred in all regions of the country, with the highest increase observed in the Southeast, North and Northeast regions. CONCLUSIONS: There was a substantial increase in hospitalizations and deaths from pertussis in children for three consecutive years (2011, 2012, and 2013) in all Brazilian regions. The most affected age group was that of children younger than one year.

Suffer the Infants: A Severe Case of Pertussis in Oregon, 2012.

Pertussis remains a public health concern in Oregon, especially among young infants. The disease can be severe in this age group and is associated with a high inpatient cost. This report describes an Oregon infant who was hospitalized with pertussis for 90 days, required extracorporeal oxygenation for 43 days, suffered complications including stroke, and had hospital charges totaling $1.5 million. Pertussis morbidity among young infants argues for vaccination of women during each pregnancy and of infants beginning promptly at two months of age.

Estimation of the serial interval of pertussis in Dutch households.

Increasing incidence has led to the re-appearance of pertussis as a public health problem in developed countries. Pertussis infection is usually mild in vaccinated children and adults, but it can be fatal in infants who are too young for effective vaccination (≤3 months). Tailoring of control strategies to prevent infection of the infant hinges on the availability of estimates of key epidemiological quantities. Here we estimate the serial interval of pertussis, i.e., the time between symptoms onset in a case and its infector, using data from a household-based study carried out in the Netherlands in 2007-2009. We use statistical methodology to tie infected persons to probable infector persons, and obtain statistically supported stratifications of the data by person-type (infant, mother, father, sibling). The analyses show that the mean serial interval is 20 days (95% CI: 16-23 days) when the mother is the infector of the infant, and 28 days (95% CI: 23-33 days) when the infector is the father or a sibling. These time frames offer opportunities for early mitigation of the consequences of infection of an infant once a case has been detected in a household. If preventive measures such as social distancing or antimicrobial treatment are taken promptly they could decrease the probability of infection of the infant.

Identification of biomarkers to detect residual pertussis toxin using microarray analysis of dendritic cells.

In this study we aimed to identify genes that are responsive to pertussis toxin (PTx) and might eventually be used as biological markers in a testing strategy to detect residual PTx in vaccines. By microarray analysis we screened six human cell types (bronchial epithelial cell line BEAS-2B, fetal lung fibroblast cell line MRC-5, primary cardiac microvascular endothelial cells, primary pulmonary artery smooth muscle cells, hybrid cell line EA.Hy926 of umbilical vein endothelial cells and epithelial cell line A549 and immature monocyte-derived dendritic cells) for differential gene expression induced by PTx. Immature monocyte-derived dendritic cells (iMoDCs) were the only cells in which PTx induced significant differential expression of genes. Results were confirmed using different donors and further extended by showing specificity for PTx in comparison to Escherichia coli lipopolysaccharide (LPS) and Bordetella pertussis lipo-oligosaccharide (LOS). Statistical analysis indicated 6 genes, namely IFNG, IL2, XCL1, CD69, CSF2 and CXCL10, as significantly upregulated by PTx which was also demonstrated at the protein level for genes encoding secreted proteins. IL-2 and IFN-γ gave the strongest response. The minimal PTx concentrations that induced production of IL-2 and IFN-γ in iMoDCs were 12.5 and 25IU/ml, respectively. High concentrations of LPS slightly induced IFN-γ but not IL-2, while LOS and detoxified pertussis toxin did not induce production of either cytokine. In conclusion, using microarray analysis we evaluated six human cell lines/types for their responsiveness to PTx and found 6 PTx-responsive genes in iMoDCs of which IL2 is the most promising candidate to be used as a biomarker for the detection of residual PTx.

Acellular pertussis vaccines in China.

In China, whole-cell pertussis (Pw) vaccines were produced in the early 1960s and acellular pertussis (Pa) vaccines were introduced in 1995. Pa vaccines have now almost completely replaced Pw vaccines in the national immunization program. To strengthen the regulation of vaccines used in China, a vaccine lot release system was established in 2001 and Pa vaccines have been included in the system since 2006. This paper mainly described the current status of production and the quality control measures in place for Pa vaccines; and analyses quality control test data accumulated between 2006 and 2010.

Evaluation of an in vitro assay system as a potential alternative to current histamine sensitization test for acellular pertussis vaccines.

The histamine sensitization test (HIST) is a lethal test for batch release of acellular pertussis or its combination vaccines (ACV). Large numbers of animals have been used and it is difficult to standardize. Therefore there is an urgent need to develop an in vitro alternative to HIST. An in vitro test system has been developed as a potential alternative to HIST, to examine both the functional domains of PT based on a combination of enzyme coupled-HPLC (E-HPLC) and carbohydrate binding assays. We describe here an international collaborative study, which involved sixteen laboratories from 9 countries to assess the methodology transferability of the in vitro test system and its suitability for the testing of three different types of ACV products that are currently used worldwide. This study also evaluated further the relationship between the in vivo activity by HIST and the in vitro assay system. The results showed that the methodology of the E-HPLC and carbohydrate binding assays are transferable between laboratories worldwide and is suitable for the three types of ACV products included in the study. Although direct correlation between the in vitro assay system and the in vivo HIST (temperature reduction assay) for each individual vaccine lot cannot be established due to the large variation in the HIST results, the observation that the mean estimates of the in vitro and in vivo activities gave the same rank order of the three vaccine types included in the study is encouraging. The in vitro systems provide reproducible product specific profiles which supports their use as a potential alternative to the HIST.

The decline and resurgence of pertussis in the US.

Although the resurgence of pertussis in nations with long-standing vaccination programs has raised serious concerns about the effectiveness of current immunization policy, the epidemiology of resurgence remains poorly understood. We analyzed pertussis notifications in US states obtained from the National Notifiable Disease Surveillance System from 1951 to 2010 to explore the timing, spatial pattern and consistency of resurgence across the country. Here we show that resurgence occurred at different times in different states, spread out over a transition period of roughly three decades. Further, despite this spatial variation, broad patterns in pertussis epidemiology can be described by two dominant phases: (1) a period of decline ending in the mid-1970s, followed by (2) nationwide resurgence. Together, these patterns explain 89.7% of the variation in US case notifications between 1951 and 2005. This resurgence was interrupted, however, by a synchronized downturn in 2005 that continues to the present in many large states. The causes of these two transitions in pertussis epidemiology remain hotly debated, though our findings suggest that evolution of the Bordetella pertussis bacterium, loss of immunity and persistent transmission among adults, and demographic drivers are more probable explanations than changes in reporting or the introduction of acellular vaccines.

[Development of a time-resolved fluoroimmunoassay for detecting S1 subunit of pertussis toxin and its application].

OBJECTIVE: To develop a time-resolved fluoroimmunoassay (TRFIA) for detection of pertussis toxin (PT) S1 subunit for quality control of human PT vaccine. METHODS: A double antibody sandwich one-step method was used to establish the TRFIA for detecting PT S1 subunit in the vaccine. RESULTS: The sensitivity of c peptide analysis reached 2.5 ng/ml without cross-reactions with other antigens. This assay could be used in detecting S1 subunit in the vaccine. CONCLUSION: The TRFIA for detecting PT S1 subunit is simple, sensitive and rapid for quality control of the PT vaccine.

Evaluation of fourth international standard for whole cell pertussis vaccine.

Whole cell pertussis vaccine is still widely used in many countries. An International Standard is needed for its potency control. The Third International Standard for Pertussis Vaccine was prepared about 40 years ago and its replacement was recommended by the Expert Committee for Biological Standardisation (ECBS) of the WHO. Material in ampoules coded 94/532 was prepared as a candidate replacement and has been evaluated in international collaborative studies which consisted of two parts. The first part, to assess the suitability of the candidate standard by comparing it with the Second International Standard for Pertussis Vaccine (IS2) involved 14 laboratories in 11 countries. The second part to compare the candidate standard with the Third International Standard for Pertussis Vaccine (IS3) involved 16 laboratories in 14 countries. Since 1995 various other studies have included the international standards and the results of these are also considered in assessing likely continuity of the IU for potency of whole cell pertussis vaccine. The preparation in ampoules coded 94/532 was adopted by the WHO ECBS in October 2006 as the 4th International Standard for whole cell pertussis vaccine and assigned an activity of 40 IU per ampoule on the basis of the studies reported here.