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

Triggered fragmentation in gravitationally unstable discs: forming fragments at small radii

We carry out three dimensional radiation hydrodynamical simulations of gravitationally unstable discs using to explore the movement of mass in a disc following its fragmentation. Compared to a more quiescent state before it fragments, the radial velocity of the gas increases by up to a factor of ~2-3 after fragmentation. While the mass movement occurs both inwards and outwards, the inwards motion can cause the inner spirals to be sufficiently dense that they may become unstable and potentially fragment. Consequently, the dynamical behaviour of fragmented discs may cause subsequent fragmentation at smaller radii after an initial fragment has formed in the outer disc.Comment: Submitted to the conference proceedings of: Instabilities and Structures in Proto-Planetary Disks. 5 pages; 4 figure

Non-convergence of the critical cooling timescale for fragmentation of self-gravitating discs

We carry out a resolution study on the fragmentation boundary of self-gravitating discs. We perform three-dimensional Smoothed Particle Hydrodynamics simulations of discs to determine whether the critical value of the cooling timescale in units of the orbital timescale, beta_{crit}, converges with increasing resolution. Using particle numbers ranging from 31,250 to 16 million (the highest resolution simulations to date) we do not find convergence. Instead, fragmentation occurs for longer cooling timescales as the resolution is increased. These results suggest that at the very least, the critical value of the cooling timescale is longer than previously thought. However, the absence of convergence also raises the question of whether or not a critical value exists. In light of these results, we caution against using cooling timescale or gravitational stress arguments to deduce whether gravitational instability may or may not have been the formation mechanism for observed planetary systems.Comment: Accepted for publication by MNRAS Letters. 6 pages, 3 figure

Large grains can grow in circumstellar discs

We perform coagulation & fragmentation simulations to understand grain growth in T Tauri & brown dwarf discs. We present a physically-motivated approach using a probability distribution function for the collision velocities and separating the deterministic & stochastic velocities. We find growth to larger sizes compared to other models. Furthermore, if brown dwarf discs are scaled-down versions of T Tauri discs (in terms of stellar & disc mass, and disc radius), growth at the same location with respect to the outer edge occurs to similar sizes in both discs.Comment: Submitted to the conference proceedings of the IAU Symposium 299 - Exploring the formation and evolution of planetary systems. 2 pages; 2 figure

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

On the gap-opening criterion of migrating planets in protoplanetary disks

We perform two-dimensional hydrodynamical simulations to quantitatively explore the torque balance criterion for gap-opening (as formulated by Crida et al. 2006) in a variety of disks when considering a migrating planet. We find that even when the criterion is satisfied, there are instances when planets still do not open gaps. We stress that gap-opening is not only dependent on whether a planet has the ability to open a gap, but whether it can do so quickly enough. This can be expressed as an additional condition on the gap-opening timescale versus the crossing time, i.e. the time it takes the planet to cross the region which it is carving out. While this point has been briefly made in the previous literature, our results quantify it for a range of protoplanetary disk properties and planetary masses, demonstrating how crucial it is for gap-opening. This additional condition has important implications for the survival of planets formed by core accretion in low mass disks as well as giant planets or brown dwarfs formed by gravitational instability in massive disks. It is particularly important for planets with intermediate masses susceptible to Type III-like migration. For some observed transition disks or disks with gaps, we expect that estimates on the potential planet masses based on the torque balance gap-opening criterion alone may not be sufficient. With consideration of this additional timescale criterion theoretical studies may find a reduced planet survivability or that planets may migrate further inwards before opening a gap.Comment: Accepted by ApJ, 22 pages, 13 figures, 6 table