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7. Microfluidic manufacturing and development of a liposomal vaccine for Clostridium difficile

Author

Forbes, Neil, Szita, Nicolas, and Perrie, Yvonne

Subjects
615.3
Abstract

Oral vaccines offer significant social and economic advantages over vaccines delivered by parenteral routes including higher patient compliance and ease of administration, allowing vaccines to be given by health workers without medical training. Furthermore, oral immunisation can generate both systemic and localised immune responses, making it ideal for generating effective protection against many enteric pathogens such as the gram-positive spore forming bacterium Clostridium difficile (C. difficile). However, design of an effective vaccine for oral administration remains a significant challenge. In order to reach the relevant sites for the induction of immunological responses, the vaccine must first traverse the hostile environment of the gastrointestinal tract (GIT). For subunit antigens, the acidic pH, degradative enzymes and bile salts present in the GIT can adversely impact their structural stability. This can limit the effectiveness of the immune response. In addition, the mucosal surface within the gut can impair the transport of the vaccine antigen and limit the accessibility to the underlying antigen presenting cells. In order to circumvent these barriers, delivery systems can be employed. Lipid-based delivery systems are commonly used to deliver therapeutics via the oral route of administration. Of the lipid based systems, liposomes have been widely investigated but tend to be used for parenteral delivery. Therefore, the aim of the work in this thesis was to explore the use of liposomes for the oral delivery of a C.difficile vaccine. Liposomes generally exhibit low toxicity, enhance interactions with biological membranes and can be incredibly versatile in regards to what compounds they can incorporate (whether they are encapsulated within the aqueous core, within the lipid bilayer or surface associated). However, the manufacture of liposomes has been limited to batch scale production which has inherent risk associated. By employing microfluidics as a manufacturing platform, in conjunction with tangential flow filtration for purification, scalable production can be achieved. Therefore, this method was employed for the formulation and manufacture of the liposomal vaccine systems. Key production parameters were identified and formulations were selected based upon their physicochemical characteristics and protein loading efficiency. Selected formulations were screened for immunological effectiveness on THP-1 macrophage-like cells regarding antigen processing ability and activation marker expression. Finally, a range of formulations were then administered orally to mice to determine in vivo efficacy. Enhanced antibody responses (IgG) could be observed when antigen was administered encapsulated within liposomal formulation DSPC:Chol (alongside the potent adjuvant cholera toxin) when compared to free antigen alone; however, poor localised IgA responses were observed for all liposomal formulations tested. The work within this thesis presents a platform for the rapid and scalable manufacture, purification and at-line analysis of liposomal formulations incorporating protein, de-risking the translation of liposomal based protein products from bench to large scale production.

Published
2020
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15. Uncovering the wider impact of COVID-19 measures on the lives of children with complex care needs and their families: A mixed-methods study protocol.

Author

Curran, Janet A., Lane, Jennifer, McCulloch, Holly, Keeping-Burke, Lisa, Johnson, Catie, Wong, Helen, Cassidy, Christine, McIsaac, Jessie-Lee, Lamptey, De-Lawrence, Clegg, Julie, Forbes, Neil, Breneol, Sydney, Sheriko, Jordan, Best, Shauna, Burgess, Stacy, Sinclair, Doug, Elliot Rose, Annette, Standing, Mary-Ann, Somerville, Mari, and King, Sarah

Subjects
CHILD care, COVID-19 pandemic, PUBLIC health, COVID-19, CHILD patients, ACCELERATED life testing
Abstract

Existing barriers to care were exacerbated by the development and implementation of necessary public health restrictions during the COVID-19 pandemic. Children with complex care needs and their families represent a small portion of the paediatric population, and yet they require disproportionately high access to services. Little is known about the impact of COVID-19 public health measures on this population. This study will generate evidence to uncover the wider impact of COVID-19 measures on the lives of children with complex care needs and their families in relation to policy and service changes. This multi-site sequential mixed methods study will take place across the Canadian Maritime provinces and use an integrated knowledge translation approach. There are two phases to this study: 1) map COVID-19 public health restrictions and service changes impacting children with complex care needs by conducting an environmental scan of public health restrictions and service changes between March 2020 and March 2022 and interviewing key informants involved in the development or implementation of restrictions and service changes, and 2) explore how children with complex care needs and their families experienced public health restrictions and service changes to understand how their health and well-being were impacted. [ABSTRACT FROM AUTHOR]

Published
2024
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22. Controlled production of lipopolysaccharides increases immune activation in Salmonella treatments of cancer.

Author

Howell, Lars M., Manole, Simin, Reitter, Alec R., and Forbes, Neil S.

Subjects
PRODUCTION increases, CANCER treatment, SALMONELLA, BINDING sites, CYTOKINE release syndrome
Abstract

Immunotherapies have revolutionized cancer treatment. These treatments rely on immune cell activation in tumours, which limits the number of patients that respond. Inflammatory molecules, like lipopolysaccharides (LPS), can activate innate immune cells, which convert tumour microenvironments from cold to hot, and increase therapeutic efficacy. However, systemic delivery of lipopolysaccharides (LPS) can induce cytokine storm. In this work, we developed immune‐controlling Salmonella (ICS) that only produce LPS in tumours after colonization and systemic clearance. We tuned the expression of msbB, which controls production of immunogenic LPS, by optimizing its ribosomal binding sites and protein degradation tags. This genetic system induced a controllable inflammatory response and increased dendritic cell cross‐presentation in vitro. The strong off state did not induce TNFα production and prevented adverse events when injected into mice. The accumulation of ICS in tumours after intravenous injection focused immune responses specifically to tumours. Tumour‐specific expression of msbB increased infiltration of immune cells, activated monocytes and neutrophils, increased tumour levels of IL‐6, and activated CD8 T cells in draining lymph nodes. These immune responses reduced tumour growth and increased mouse survival. By increasing the efficacy of bacterial anti‐cancer therapy, localized production of LPS could provide increased options to patients with immune‐resistant cancers. [ABSTRACT FROM AUTHOR]

Published
2024
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27. Parrots: Appropriate Pets or Best Not Bred?

Author

HESS, LAURIE, CLUBB, SUSAN, ECHOLS, M. SCOTT, FORBES, NEIL, HERNANDEZ, SONIA, HOOIMEIJER, JAN, JOYNER, LORAKIM, KLAPHAKE, ERIC, OROSZ, SUSAN, RICH, GREG, SCHNELLBACHER, RODNEY, and SPEER, BRIAN

Published
2016

31. Introduction

Author

Forbes, Neil, primary, Kurosawa, Takafumi, additional, and Wubs, Ben, additional

Published
2018
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37. British financial and economic policy towards Germany, 1931-39

Author

Forbes, Neil

Abstract

This thesis attempts to show the complexity and convoluted nature of Britain's financial and economic relations with Germany in the 1930s. A great variety of influences (often in competition with one another) acted upon policy formulation in Britain. This helped to create different, but seemingly equally valid, opinions as to what constituted the 'national interest'. This, in turn, tended to exacerbate the rivalry which existed between government officials. In particular, the Foreign Office showed extreme hostility to the attitude of the Governor of the Bank of England as the latter attempted to facilitate agreements between Britain and Germany. Such domestic constraints, combined with the economic pre-occupations of the period, prevented policy-makers from developing a clear picture of the nation's interests. A study of bank archives reveals that the extent of the influence of London banks in determining British official policy has been greatly exaggerated. In the face of competition from rival interests, the London banks could not secure any special consideration for their position under the Standstill Agreements from the Government. Thus while the formulation of policy, confronted with the problem of Nazi Germany, showed that political and economic considerations were linked, perhaps to a degree greater than ever before, the indiscriminate use of the term 'economic appeasement' to describe British policy tends to hinder rather than aid historical understanding.

Published
2023
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44. Cultural Heritage in a Changing World

Author

Borowiecki, Karol Jan, Fresa, Antonella, and Forbes, Neil

Subjects
Business & Economics / Management, Science / Earth Sciences, Social Science / Anthropology / Cultural & Social
Abstract

Cultural Heritage; Cultural Economics; Cultural Studies; Archaeology; Information Storage and Retrieval; Information Systems Applications (incl. Internet)

Published
2016

46. White paper on microbial anti-cancer therapy and prevention

Author

Forbes, Neil S., Coffin, Robert S., Deng, Liang, Evgin, Laura, Fiering, Steve, Giacalone, Matthew, Gravekamp, Claudia, Gulley, James L., Gunn, Hal, Hoffman, Robert M., Kaur, Balveen, Liu, Ke, Lyerly, Herbert Kim, Marciscano, Ariel E., Moradian, Eddie, Ruppel, Sheryl, Saltzman, Daniel A., Tattersall, Peter J., Thorne, Steve, Vile, Richard G., Zhang, Halle Huihong, Zhou, Shibin, and McFadden, Grant

Published
2018
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47. Brian Coles

Author

Forbes, Neil, primary, Payne‐Johnson, Catherine, additional, and Skerritt, Geoff, additional

Published
2022
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48. A Module to Foster Engineering Creativity: An Interpolative Design Problem and an Extrapolative Research Project

Author

Forbes, Neil S.

Abstract

This article describes a teaching module designed to enhance engineering creativity in an introductory chemical engineering course. The module includes an exercise to design column packing material, and an open-ended research project to describe the societal impact of chemical engineering. These assignments were created to illustrate the benefit of generating many ideas independent of analysis. The results of two years of survey-based assessment showed that these techniques improved students' confidence and ability with creative problems. (Contains 5 figures and 3 tables.)

Published
2008

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