Immunogenicity and protective efficacy of an anti-Streptococcus pyogenes vaccine candidate in multiple animal species

Streptococcus pyogenes, also known as Group A Streptococcus (GAS) has been associated with a range of diseases from the mild pharyngitis and pyoderma to more severe invasive infections such as streptococcal toxic shock. GAS also causes a number of non-suppurative post-infectious diseases such as rheumatic fever, rheumatic heart disease and glomerulonephritis. The large extent of GAS disease burden necessitates the need for a prophylactic vaccine that could target the diverse GAS emm types circulating globally. Anti-GAS vaccine strategies have focused primarily on the GAS M-protein, an extracellular virulence factor anchored to GAS cell wall. As opposed to the hypervariable N-terminal region, the C-terminal portion of the protein is highly conserved among different GAS emm types and is the focus of a leading GAS vaccine candidate, J8-DT/alum. The vaccine candidate J8-DT/alum was shown to be immunogenic in mice, rabbits and the non-human primates, hamadryas baboons. Similar responses to J8-DT/alum were observed after subcutaneous and intramuscular immunization with J8-DT/alum, in mice and in rabbits. Further assessment of parameters that may influence the immunogenicity of J8-DT demonstrated that the immune responses were identical in male and female mice and the use of alum as an adjuvant in the vaccine formulation significantly increased its immunogenicity, resulting in a long-lived serum IgG response. Contrary to the previous findings, the data in this thesis indicates that a primary immunization with J8-DT/alum (50ƒÊg) followed by a single boost is sufficient to generate a robust immune response in mice. As expected, the IgG response to J8- DT/alum was a Th2 type response consisting predominantly of the isotype IgG1 accompanied by lower levels of IgG2a. Intramuscular vaccination of rabbits with J8-DT/alum demonstrated that an increase in the dose of J8-DT/alum up to 500ƒÊg does not have an impact on the serum IgG titers achieved. Similar to the immune response in mice, immunization with J8-DT/alum in baboons also established that a 60ƒÊg dose compared to either 30ƒÊg or 120ƒÊg was sufficient to generate a robust immune response. Interestingly, mucosal infection of naive baboons with a M1 GAS strain did not induce a J8-specific serum IgG response. As J8-DT/alum mediated protection has been previously reported to be due to the J8- specific antibody formed, the efficacy of J8-DT antibodies was determined in vitro and in vivo. In vitro opsonization and in vivo passive transfer confirmed the protective potential of J8-DT antibodies. A reduction in the bacterial burden after challenge with a bioluminescent M49 GAS strain in mice that were passively administered J8-DT IgG established that protection due to J8-DT was mediated by antibodies. The GAS burden in infected mice was monitored using bioluminescent imaging in addition to traditional CFU assays. Bioluminescent GAS strains including the ‘rheumatogenic’ M1 GAS could not be generated due to limitations with transformation of GAS, however, a M49 GAS strain was utilized during BLI. The M49 serotype is traditionally a ‘nephritogenic’ serotype associated with post-streptococcal glomerulonephritis. Anti- J8-DT antibodies now have been shown to be protective against multiple GAS strains such as M49 and M1. This study evaluated the immunogenicity of J8-DT/alum in different species of experimental animals in preparation for phase I human clinical trials and provided the ground work for the development of a rapid non-invasive assay for evaluation of vaccine candidates

Island Dreaming: Applied Epidemiology in the Pacific Region

This bound volume describes four significant public health problems in Australia and the Pacific Island Countries of Fiji and American Samoa. The four main epidemiological components are: 1) Australian vaccine preventable disease epidemiological review series: varicella-zoster virus infections, 1998–2015. The review was conducted to assess the impact of the national varicella immunisation program and provide a baseline for monitoring the impact of the national herpes zoster immunisation program. The national varicella immunisation program led to significant reductions in varicella. In Australia, the burden of herpes zoster is substantial, and high quality and timely surveillance will be crucial to assess the impact of the national herpes zoster immunisation program. 2) Investigation into increased lymphogranuloma venereum (LGV) in New South Wales, Australia. LGV is a sexually transmitted infection (STI) caused by L1-L3 serovars of chlamydia, and can lead to irreversible complications. LGV is notifiable condition in New South Wales (NSW). Following a noticeable increase in number of LGV notifications, I conducted a retrospective case series of all cases diagnosed between 1 January 2016 and 31 March 2017. During this period, all reported cases were among men who have sex with men. This chapter examines factors contributing to increase in LGV cases in NSW in 2016. It also describes the challenges associated with investigating STI outbreaks in NSW. 3) An evaluation of an early warning alert and response system (EWARS in a Box) implemented after Cyclone Winston, Fiji 2016. The World Health Organization recommends implementation of early warning systems for timely disease surveillance and early detection of outbreaks during humanitarian emergencies. This chapter describes the EWARS system, and its usefulness at timely monitoring of communicable diseases trends during a national health emergency. Findings include strengths and limitations of the system at conducting surveillance, along with practical recommendations for improving surveillance using EWARS. 4) Identifying residual transmission of lymphatic filariasis in post-mass drug administration surveillance phase: Comparing school-based versus community-based surveys – American Samoa, 2016. This study compares the effectiveness of two cross-sectional survey designs, a school-based and a community-based survey, for assessing transmission of lymphatic filariasis. Under the Global Programme for Elimination of Lymphatic Filariasis, American Samoa conducted seven rounds of mass drug administration (MDA) from 2000-2006. The World Health Organization recommends systematic post-MDA surveillance for epidemiological assessment of recent lymphatic filariasis transmission. Finger prick blood samples were collected from study participants to measure the prevalence of circulating filarial antigen (CFA). I recruited 1143 grade 1 and 2 school students from 29 elementary schools. For the community survey, 30 out of 70 villages were randomly selected, from which 2507 community members were recruited. The school survey was cheaper and logistically easier to implement. The estimated CFA prevalence by school survey was 0.7%, and was significantly lower than the community survey (6.2%). The community survey was more effective for collecting information required for identifying residual transmission of lymphatic filariasis. Both surveys provided evidence of ongoing lymphatic filariasis transmission in American Samoa

Comparison of immunochromatographic test (ICT) and filariasis test strip (FTS) for detecting lymphatic filariasis antigen in American Samoa, 2016

Circulating filarial antigen (Ag) prevalence, measured using rapid point-of-care tests, is the standard indicator used for monitoring and surveillance in the Global Program to Eliminate Lymphatic Filariasis. In 2015, the immunochromatographic test (ICT) was replaced with the filariasis test strip (FTS), which has higher reported sensitivity. Despite differences in sensitivity, no changes in recommended surveillance targets were made when the FTS was introduced. In 2016, we conducted lymphatic filariasis surveys in American Samoa using FTS, which found higher Ag prevalence than previous surveys that used ICT. To determine whether the increase was real, we assessed the concordance between FTS and ICT results by paired testing of heparinised blood from 179 individuals (63% FTS-positive). ICT had 93.8% sensitivity and 100% specificity for identifying FTS-positive persons, and sensitivity was not associated with age, gender, or presence of microfilariae. Based on these findings, if ICT had been used in the 2016 surveys, the results and interpretation would have been similar to those reported using FTS. American Samoa would have failed Transmission Assessment Survey (TAS) of Grade 1 and 2 children with either test, and community prevalence would not have been significantly different (4.1%, 95% CI, 3.3–4.9% with FTS vs. predicted 3.8%, 95%, CI: 3.1–4.6% with ICT)

Lymphatic Filariasis in 2016 in American Samoa: Identifying Clustering and Hotspots Using Non-Spatial and Three Spatial Analytical Methods

Background: American Samoa completed seven rounds of mass drug administration from 2000–2006 as part of the Global Programme to Eliminate Lymphatic Filariasis (LF). However, resurgence was confirmed in 2016 through WHO-recommended school-based transmission assessment survey and a community-based survey. This paper uses data from the 2016 community survey to compare different spatial and non-spatial methods to characterise clustering and hotspots of LF. Method: Non-spatial clustering of infection markers (antigen [Ag], microfilaraemia [Mf], and antibodies (Ab [Wb123, Bm14, Bm33]) was assessed using intra-cluster correlation coefficients (ICC) at household and village levels. Spatial dependence, clustering and hotspots were examined using semivariograms, Kulldorf’s scan statistic and Getis-Ord Gi* statistics based on locations of surveyed households. Results: The survey included 2671 persons (750 households, 730 unique locations in 30 villages). ICCs were higher at household (0.20–0.69) than village levels (0.10–0.30) for all infection markers. Semivariograms identified significant spatial dependency for all markers (range 207–562 metres). Using Kulldorff’s scan statistic, significant spatial clustering was observed in two previously known locations of ongoing transmission: for all markers in Fagali’i and all Abs in Vaitogi. Getis-Ord Gi* statistic identified hotspots of all markers in Fagali’i, Vaitogi, and Pago Pago-Anua areas. A hotspot of Ag and Wb123 Ab was identified around the villages of Nua-Seetaga-Asili. Bm14 and Bm33 Ab hotspots were seen in Maleimi and Vaitogi-Ili’ili-Tafuna. Conclusion: Our study demonstrated the utility of different non-spatial and spatial methods for investigating clustering and hotspots, the benefits of using multiple infection markers, and the value of triangulating results between method

Information seeking behaviors of individuals impacted by COVID-19 international travel restrictions: an analysis of two international cross-sectional studies

Access to accurate information during a crisis is essential. However, while the amount of information circulating during the COVID-19 pandemic has increased exponentially, finding trustworthy resources has been difficult for many, including those affected by international travel restrictions. In this study, we examined the information-seeking behaviors of individuals seeking to travel internationally during the COVID-19 pandemic. We also explored perceptions regarding the value of resources in supporting understanding of COVID-19 travel restriction-related information. Two online cross-sectional surveys targeting four groups were conducted. The groups targeted were: (1) citizens and permanent residents stranded abroad; (2) individuals separated from their partners; (3) individuals separated from immediate families; and (4) temporary visa holders unable to migrate or cross international borders. In total, we analyzed 2,417 completed responses, and a further 296 responses where at least 75% of questions were completed. Findings suggest that social media groups (78.4%, 1,924/2,453), specifically Facebook (86.6%, 2,115/2,422) were the most useful or most used information resource for these groups. Some significant information seeking behavior differences across age and gender were also found. Our study highlights the diversity in information needs of people impacted by COVID-19 travel restrictions and the range of preferred channels through which information is sought. Further, it highlights which challenges hold legitimacy in their target audiences' eyes and which do not. Policymakers may use these results to help formulate more nuanced, consumer-tailored—and hence likely more acceptable, trusted, and impactful—communication strategies as part of future public health emergencies

Field epidemiology in action: an Australian perspective of epidemic response to the Rohingya health emergencies in Cox’s Bazar, Bangladesh

Approximately one million Rohingya persons who fled waves of violence in Myanmar at different times, the latest being 25 August 2017, now live in two coastal districts in Bangladesh; Cox’s Bazar and Bandarban (1). In makeshift shelters made from bamboo and tarpaulin in camps sprawling through rough terrains, the Rohingya live in conditions of overcrowding, poor sanitation, high malnutrition and, on arrival, extremely low vaccination coverage (1-3).BK, JEM and MXT were supported by Australian Government Research Training Program (RTP) Scholarships

Potential strategies for strengthening surveillance of lymphatic filariasis in American Samoa after mass drug administration: reducing ‘number needed to test’ by targeting older age groups, hotspots, and household members of infected persons

Under the Global Programme to Eliminate Lymphatic Filariasis (LF), American Samoa conducted mass drug administration (MDA) from 2000–2006. Despite passing Transmission Assessment Surveys (TAS) in 2011/2012 and 2015, American Samoa failed TAS-3 in 2016, with antigen (Ag) prevalence of 0.7% (95%CI 0.3–1.8%) in 6–7 year-olds. A 2016 community survey (Ag prevalence 6.2% (95%CI 4.4–8.5%) in age ≥8 years) confirmed resurgence. Using data from the 2016 survey, this study aims to i) investigate antibody prevalence in TAS-3 and the community survey, ii) identify risk factors associated with being seropositive for Ag and anti-filarial antibodies, and iii) compare the efficiency of different sampling strategies for identifying seropositive persons in the post-MDA setting. Antibody prevalence in TAS-3 (n = 1143) were 1.6% for Bm14 (95%CI 0.9–2.9%), 7.9% for Wb123 (95%CI 6.4–9.6%), and 20.2% for Bm33 (95%CI 16.7–24.3%); and in the community survey (n = 2507), 13.9% for Bm14 (95%CI 11.2–17.2%), 27.9% for Wb123 (95%CI 24.6–31.4%), and 47.3% for Bm33 (95%CI 42.1–52.6%). Multivariable logistic regression was used to identify risk factors for being seropositive for Ag and antibodies. Higher Ag prevalence was found in males (adjusted odds ratio [aOR] 3.01), age ≥18 years (aOR 2.18), residents of Fagali’i (aOR 15.81), and outdoor workers (aOR 2.61). Ag prevalence was 20.7% (95%CI 9.7–53.5%) in households of Ag-positive children identified in TAS-3. We used NNTestav (average number needed to test to identify one positive) to compare the efficiency of the following strategies for identifying persons who were seropositive for Ag and each antibody: i) TAS of 6–7 year-old children, ii) population representative surveys of older age groups, and iii) targeted surveillance of subpopulations at higher risk of being seropositive (older ages, householders of Ag-positive TAS children, and known hotspots). For Ag, NNTestav ranged from 142.5 for TAS, to <5 for households of index children. NNTestav was lower in older ages, and highest for Ag, followed by Bm14, Wb123 and Bm33 antibodies. We propose a multi-stage surveillance strategy, starting with population-representative sampling (e.g. TAS or population representative survey of older ages), followed by strategies that target subpopulations and/or locations with low NNTestav. This approach could potentially improve the efficiency of identifying remaining infected persons and residual hotspots. Surveillance programs should also explore the utility of antibodies as indicators of transmission

Potential use of antibodies to provide an earlier indication of lymphatic filariasis resurgence in post–mass drug ad ministration surveillance in American Samoa

Background: Under the Global Programme to Eliminate Lymphatic Filariasis (LF), American Samoa conducted 7 rounds of mass drug administration (MDA) between 2000 and 2006. The territory passed transmission assessment surveys (TASs) in 2011 (TAS-1) and 2015 (TAS-2). In 2016, the territory failed TAS-3, indicating resurgence. This study aims to determine if antibodies (Abs) may have provided a timelier indication of LF resurgence in American Samoa. Methods: We examined school-level antigen (Ag) and Ab status (presence/absence of Ag- and Ab-positive children) and prevalence of single and combined Ab responses to Wb123, Bm14, and Bm33 Ags at each TAS. Pearson chi-square test and logistic regression were used to examine associations between school-level Ab prevalence in TAS-1 and TAS-2 and school-level Ag status in TAS-3. Results: Schools with higher prevalence of Wb123 Ab in TAS-2 had higher odds of being Ag-positive in TAS-3 (odds ratio [OR] 24.5, 95% confidence interval [CI] 1.2–512.7). Schools that were Ab-positive for WB123 plus Bm14, Bm33, or both Bm14 and Bm33 in TAS-2 had higher odds of being Ag-positive in TAS-3 (OR 16.0–24.5). Conclusion: Abs could provide earlier signals of resurgence and enable a timelier response. The promising role of Abs in surveillance after MDA and decision making should be further investigated in other settings

Genetic epidemiology of lymphatic filariasis in American Samoa after mass drug administration

Over 892 million people in 48 countries are at risk of infection by nematodes that cause lymphatic filariasis. As part of the Global Programme to Eliminate Lymphatic Filariasis, mass drug administration is distributed to communities until surveillance indicates infection rates are below target prevalence thresholds. In some countries, including American Samoa, lymphatic filariasis transmission persists despite years of mass drug administration and/or has resurged after cessation. Nothing is known about the population genetics of Wuchereria bancrofti worms in Polynesia, or whether local transmission is persisting and/or increasing due to inadequate mass drug administration coverage, expansion from residual hotspots, reintroduction from elsewhere, or a combination. We extracted DNA from microfilariae on blood slides collected during prevalence surveys in 2014 and 2016, comprising 31 pools of five microfilariae from 22 persons living in eight villages. We sequenced 1104 bp across three mitochondrial markers (ND4, COI, CYTB). We quantified parasite genetic differentiation using variant calls and estimated haplotypes using principal components analysis, F-statistics, and haplotype networks. Of the variants called, all but eight were shared across the main island of Tutuila, and three of those were from a previously described hotspot village, Fagali’i. Genotypic data did not support population genetic structure among regions or villages in 2016, although differences were observed between worms collected in Fagali’i in 2014 and those from 2016. Because estimated haplotype frequency varied between villages, these statistics suggested genetic differentiation, but were not consistent among villages. Finally, haplotype networks demonstrated American Samoan sequence clusters were related to previously published sequences from Papua New Guinea. These are, to our knowledge, the first reports of W. bancrofti genetic variation in Polynesia. The resurgent parasites circulating on the main island of American Samoa represent a single population. This study is the first step towards investigating how parasite population structure might inform strategies to manage resurgence and elimination of lymphatic filariasis

An outbreak investigation of paediatric severe acute respiratory infections requiring admission to intensive care units - Fiji, May 2016

Introduction Influenza-associated severe acute respiratory infections (SARI) are a major contributor to global morbidity and mortality. In response to a cluster of SARI cases and deaths in pregnant women, with two deceased cases testing positive for influenza A(H1N1)pdm09, an investigation was initiated to determine whether there was an increase of paediatric SARI cases admitted to divisional hospital intensive care units in Fiji in may 2016 compared to May 2013-2015. Methods Retrospective case finding was conducted at the paediatric intensive care units (PICUs) in Fiji's three divisional hospitals. Data were collected from 1 January 2013 to 26 May 2016. Cases were identified using a list of clinical diagnoses compatible with SARI. Results A total of 632 cases of paediatric SARI with complete details were identified. The median age of cases was 6 months (Interquartile range: 2-14 months). Children aged less than 5 years had a higher rate of paediatric SARI requiring admission to a divisional hospital PICU in May 2016 compared to May 2013-2015 (Incidence rate ratio: 1.7 [95% CI: 1.1-2.6]). This increase was not observed in children aged 5-14 years. The case-fatality ratio was not significantly different in 2016 compared to previous years. Conclusion The investigation enabled targeted public health response measures, including enhanced SARI surveillance at divisional hospitals and an emergency influenza vaccination campaign in the Northern Division