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Relevant bibliographies by topics / Accelerating vaccine formulation development

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Academic literature on the topic 'Accelerating vaccine formulation development'

Author: Grafiati

Published: 4 June 2025

Last updated: 8 July 2025

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Journal articles on the topic "Accelerating vaccine formulation development"

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Ahl, Patrick L., Christopher Mensch, Binghua Hu, et al. "Accelerating Vaccine Formulation Development Using Design of Experiment Stability Studies." Journal of Pharmaceutical Sciences 105, no. 10 (2016): 3046–56. http://dx.doi.org/10.1016/j.xphs.2016.06.014.

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Kim, Jeremiah Y., Matthew G. Rosenberger, Nakisha S. Rutledge, and Aaron P. Esser-Kahn. "Next-Generation Adjuvants: Applying Engineering Methods to Create and Evaluate Novel Immunological Responses." Pharmaceutics 15, no. 6 (2023): 1687. http://dx.doi.org/10.3390/pharmaceutics15061687.

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Adjuvants are a critical component of vaccines. Adjuvants typically target receptors that activate innate immune signaling pathways. Historically, adjuvant development has been laborious and slow, but has begun to accelerate over the past decade. Current adjuvant development consists of screening for an activating molecule, formulating lead molecules with an antigen, and testing this combination in an animal model. There are very few adjuvants approved for use in vaccines, however, as new candidates often fail due to poor clinical efficacy, intolerable side effects, or formulation limitations. Here, we consider new approaches using tools from engineering to improve next-generation adjuvant discovery and development. These approaches will create new immunological outcomes that will be evaluated with novel diagnostic tools. Potential improved immunological outcomes include reduced vaccine reactogenicity, tunable adaptive responses, and enhanced adjuvant delivery. Evaluations of these outcomes can leverage computational approaches to interpret “big data” obtained from experimentation. Applying engineering concepts and solutions will provide alternative perspectives, further accelerating the field of adjuvant discovery.

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Koff, Wayne C. "Accelerating HIV vaccine development." Nature 464, no. 7286 (2010): 161–62. http://dx.doi.org/10.1038/464161a.

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He, Jason. "Accelerating Recombinant Protein Vaccine Development." Genetic Engineering & Biotechnology News 44, no. 1 (2024): 42–44. http://dx.doi.org/10.1089/gen.44.01.13.

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Larkin, Marilynn. "Seth Berkley—Accelerating AIDS vaccine development." Lancet Infectious Diseases 5, no. 1 (2005): 16–19. http://dx.doi.org/10.1016/s1473-3099(04)01249-6.

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Chiu, Christopher. "Novel immunological insights in accelerating RSV vaccine development." Vaccine 35, no. 3 (2017): 459–60. http://dx.doi.org/10.1016/j.vaccine.2016.11.069.

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Roordink, Danielle, Ann Williams, Bernard Fritzell, et al. "The TB vaccine development pathway – An innovative approach to accelerating global TB vaccine development." Tuberculosis 126 (January 2021): 102040. http://dx.doi.org/10.1016/j.tube.2020.102040.

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Koff, W. C., D. R. Burton, P. R. Johnson, et al. "Accelerating Next-Generation Vaccine Development for Global Disease Prevention." Science 340, no. 6136 (2013): 1232910. http://dx.doi.org/10.1126/science.1232910.

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Rahman, Mohammad Arif, and Marjorie Robert-Guroff. "Accelerating HIV vaccine development using non-human primate models." Expert Review of Vaccines 18, no. 1 (2018): 61–73. http://dx.doi.org/10.1080/14760584.2019.1557521.

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Alkaff, Ahmad Husein, Mutiara Saragih, Mochammad Arfin Fardiansyah, and Usman Sumo Friend Tambunan. "Role of Immunoinformatics in Accelerating Epitope-Based Vaccine Development against Dengue Virus." Open Biochemistry Journal 14, no. 1 (2020): 9–18. http://dx.doi.org/10.2174/1874091x02014010009.

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Dengue Fever (DF) has emerged as a significant public health problem of international concern with its high prevalence in the tropic and subtropical regions. Dengue Virus (DENV), which is the cause of DF, consists of four serotypes of antigenically distinct viruses. The immense variation and limited identity similarity at the amino acid level lead to a problematic challenge in the development of an efficacious vaccine. Fortunately, the extensively available immunological data, the advance in antigenic peptide prediction, and the incorporation of molecular docking and dynamics simulation in immunoinformatics have directed the vaccine development towards the rational design of the epitope-based vaccine. Here, we point out the current state of dengue epidemiology and the recent development in vaccine development. Subsequently, we provide a systematic review of our validated method and tools for B- and T-cell epitope prediction as well as the use of molecular docking and dynamics in evaluating epitope affinity and stability in the discovery of a new tetravalent dengue vaccine through computational epitope-based vaccine design.

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Dissertations / Theses on the topic "Accelerating vaccine formulation development"

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Ajmera, Ankur [Verfasser]. "Stable spray dried protein formulation and implementation in vaccine development / Ankur Ajmera." Kiel : Universitätsbibliothek Kiel, 2014. http://d-nb.info/1064906036/34.

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Li, Xinran. "Vaccine formulation development : towards addressing major limitations of vaccines that are adjuvanted with aluminum salts." Thesis, 2013. http://hdl.handle.net/2152/28737.

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Many vaccines require an adjuvant to induce a strong immune response. Aluminum–containing adjuvants have been approved by the United States Food and Drug Administration for human use for many years. There are two main aluminum-containing adjuvants, aluminum hydroxide and aluminum phosphate. Due to their favorable safety profile, aluminum-containing adjuvants have been widely used in human vaccines for decades. Many currently licensed and commercially available vaccines contain aluminum-containing adjuvants. However, aluminum-containing vaccine adjuvants suffer from two major limiting factors: (1) aluminum-containing adjuvants can only weakly or moderately potentiate antigen-specific antibody responses and are generally considered incapable of inducing cellular immune responses; (2) vaccines that contain aluminum-containing adjuvants require cold-chain refrigeration for storage and distribution, and may not be frozen, because freezing of the vaccine in dispersion causes irreversible coagulation that damages vaccines (e.g., loss in potency and stability). In this dissertation, the first limitation was addressed by reducing the size of the aluminum hydroxide from micrometers (3-10 micrometer) to nanometers of less than 200 nm, and the second limitation mentioned above was addressed by freeze-drying vaccines that contain aluminum salts as adjuvants into a dry powder using thin-film freeze-drying. In addition, using an improved experimental design, the vaccine adjuvant activities of nanoparticles of around 200 nm was compared to that of the nanoparticles of around 700 nm. The smaller 200 nm nanoparticles showed a more potent adjuvant activity than the larger nanoparticles. When dispersed in an aqueous medium, both aluminum hydroxide and aluminum phosphate are physically 1–20 micrometer particulates. There are data showing that particulate vaccine carriers of around 200 nm (or less) may be optimal in potentiating the immunogenicity of vaccines. Based on this finding, aluminum hydroxide nanoparticles of 112 nm were synthesized, and its adjuvant activity was compared to that of the traditional aluminum hydroxide adjuvant, which have particulates of 3-20 micrometer. Using ovalbumin and Bacillus anthracis protective antigen protein as model antigens, it was found that protein antigens adsorbed on the aluminum hydroxide nanoparticles induced stronger antigen-specific antibody responses than the same protein antigens adsorbed on the traditional aluminum hydroxide microparticles of around 9.3 µm. Importantly, the inflammation reactions induced by aluminum hydroxide nanoparticles in the injection sites were milder than that induced by microparticles. Simply reducing the particle size of the traditional aluminum hydroxide adjuvant in suspension from micrometers into nanometers represents a novel and effective approach to improve its potency. The second limitation was addressed by converting vaccines that contain an aluminum salt as an adjuvant from an aqueous dispersion into a dried powder using thin-film freeze-drying. There is evidence that aluminum-containing vaccines can be lyophilized to dry powders using high speed freezing methods. Thin-film freezing is a high speed freezing method with a freezing rate between 100 to 10,000 K/s, but the feasibility of using thin-film freeze-drying to freeze-dry vaccines that contain aluminum salts as adjuvants has not been tested before. In this dissertation, Using ovalbumin as a model protein antigen and aluminum hydroxide or aluminum phosphate as an adjuvant, it was confirmed that vaccines that are adjuvanted with aluminum hydroxide or aluminum phosphate can be freeze-dried with as low as 2% (w/v) of trehalose as a cryoprotectant by thin-film freeze-drying without causing vaccine aggregation while preserving the immunogenicity of the vaccine. Finally, the feasibility of using the thin-film freeze-drying method to freeze-drying vaccines that contain aluminum salts as adjuvants was further confirmed by drying a commercial aluminum salt-adjuvanted tetanus toxoid vaccine. Vaccines that contain aluminum salts as adjuvants may be converted to a dry powder using the thin-film freeze-drying method to avoid loss of potency due exposure to freezing conditions during transport and storage.<br>text

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Machado, Fátima Alexandra Meira. "Development of a liposomal formulation for peptide delivery to serve as vaccine against chronic myeloid leukemia." Master's thesis, 2013. http://hdl.handle.net/1822/29422.

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Dissertação de mestrado em Biophysics of Bionanosystems<br>The main goal of this work was to characterize and explore the potential of Dioctadecyldimethylammonium Chloride (DODAC) / Monoolein (MO) liposomes in a 1:2 proportion and identify the formulations that could be used in the development of an immunoprotective protocol for Chronic Myeloid Leukemia (CML). CML has long been recognized as one of the most responsive leukemic disorder to immunotherapy. CML is potent model for immune therapy in humans because there is a specific gene rearrangement, BCR/ABL, which product, P210bcr/abl, can be the target antigen. The loading of drugs into particles at the nanometer size range is a recognized technique for the optimization of controlled drug delivery. In its use in vaccines, liposomes have the advantage of being able to maintain antigens present in the organism for long enough to obtain an immune response. Different methods of preparation and distinct peptide/lipid molar ratios were used to prepare P210bcr/abl / DODAC:MO (1:2) nanoparticles. This thesis describes results for biophysical characterization of the peptide/lipid system, encapsulation efficiency and exposure of THP-1 cells to the nanoparticles. The lipid content was essential to achieve the desired nanoparticles. The highest lipid concentration showed higher encapsulation, however, a lower lipid content induced a more efficient cell response. The peptide/lipid system was capable of inducing a stronger cell response than the peptide by itself, emphasizing the potential of this system in vaccine development for the treatment of CML.<br>O objetivo principal deste trabalho foi caracterizar e explorar o potencial dos lipossomas de cloreto de Dioctadecildimetilamónio (DODAC) / Monooleina (MO) numa proporção de 1:2 e identificar as formulações que poderão ser usadas no desenvolvimento de um tratamento imunoprotetor para a Leucemia Mieloide Crónica (LMC). A LMC é desde há muito tempo conhecida como uma das desordens imunológicas mais responsivas à imunoterapia. A LMC é um poderoso modelo para imunoterapia em humanos devido à existência de um gene específico BCR/ABL, cujo produto, P210bcr/abl, pode ser usado como antigene-alvo. A incorporação de fármacos em partículas a uma escala nanométrica é uma técnica reconhecida para a optimização da entrega controlada de fármacos. No seu uso em vacinas, os lipossomas possuem a vantagem de ser capazes de manter os antigenes presentes no organismo o tempo suficiente para se obter uma resposta imune. Diferentes métodos de preparação e várias razões molares de péptido/lipido foram usadas para preparar nanoparticulas de P210bcr/abl / DODAC:MO(1:2). Esta tese descreve os resultados obtidos da caracterização biofísica do sistema péptido/lípido, eficiência de encapsulação e exposição das células THP-1 às nanopartículas. O conteúdo lipídico foi essencial para obter nanopartículas desejáveis. A concentração mais alta de lípido demonstrou maior eficiência de encapsulação, no entanto, uma concentração de lipído mais baixa mostrou-se mais eficiente em induzir uma resposta por parte das células. O sistema péptido/lipido foi capaz de induzir uma resposta mais forte do que o pétido por si só, enfatizando o seu potencial no desenvolvimento de uma vacina para o tratamento da LMC.

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Tantituvanont, Angkana. "Novel formulation : development of oral microparticulate non-viral DNA vaccine delivery system against infectious hematopoetic necrosis virus (IHNV) in Rainbow Trout, statistical design in matrix tablets formulation." Thesis, 2003. http://hdl.handle.net/1957/31818.

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This dissertation describes two different projects. The first is the development of an oral DNA vaccine delivery system for fish. A novel oral DNA vaccine delivery system was developed for Rainbow Trout by combining non-viral vectors (polycationic liposomes or polycationic polymer) to facilitate the DNA vaccine's uptake by cell membranes along with enteric-coated protection of the DNA embedded in microparticles to prevent DNA degradation in the gastrointestinal tract. Spray drying and spray coating bead techniques were employed in the preparation of the DNA vaccine microparticles. The spray drying technique allowed production of spherical shape enteric-coated microparticles with a particle size range of 0.18 to 20 ��m. Larger particle sizes of 40-50 mesh were obtained from the spray-coated bead technique. The resultant DNA vaccine microparticles were granulated with regular fish feed and given to fish to investigate the efficacy of the delivery system in providing protection against IHNV, and to demonstrate the ease of administration in fish. An in vivo fish trial experiment showed improvement in fish survival rate when fish were immunized with larger particle size DNA vaccine microparticles. Further research to find effective vector carriers for the DNA vaccine delivery system and to seek modifications of the delivery system that will still prevent the denaturation of plasmid DNA that will also facilitate membrane uptake of the DNA vaccine is needed in order to develop a safe, effective, and commercially viable vaccine to control the outbreak of IHNV. The second project of the dissertation is prediction of in vitro drug release profiles from a novel matrix tablet spray-coated with a barrier membrane using mathematical and statistical models. Tablets were prepared by direct compression followed by spray coating. The relationship of the amount of hydrophilic materials in the core tablets and barrier thickness on drug release mechanism was investigated using factorial design and regression analysis. Drug release characteristics were influenced and can be controlled by modifying the amount of hydrophilic materials in the core tablet and the barrier thickness. Mathematical equation generated from regression analysis of n-value, lag time, and percent drug release as a function of the amount of hydrophilic material and the amount of coating material applied can now be used as a tool for predicting and optimizing in vitro drug release from matrix tablets spray-coated with a barrier membrane.<br>Graduation date: 2003

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Book chapters on the topic "Accelerating vaccine formulation development"

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Lidgate, Deborah M., and Noelene E. Byars. "Development of an Emulsion-Based Muramyl Dipeptide Adjuvant Formulation for Vaccines." In Vaccine Design. Springer US, 1995. http://dx.doi.org/10.1007/978-1-4615-1823-5_12.

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Aatresh, Aishani, Nicole Lurie, and Richard Hatchett. "13 Accelerating Vaccine Development: The 100 Days Mission." In Principles and Practice of Emergency Research Response. Springer International Publishing, 2024. http://dx.doi.org/10.1007/978-3-031-48408-7_16.

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AbstractThis chapter explores the role of the Coalition for Epidemic Preparedness Innovations (CEPI) in global health preparedness and response. It specifically focuses on emergency research response and the 100 Days Mission—the goal of having vaccines ready for use with initial emergency authorization within 100 days from the identification of a novel pathogen with pandemic potential. If this can be accomplished, the world will have a better chance of containing or controlling future infectious disease outbreaks and averting global pandemics or at least minimizing their consequences. Meeting this timeline would require advances in scientific research, production technology, clinical trial readiness, governance and coordination, financing, and the policy ecosystems within which such preparedness and response efforts take place. A “second 100 days” must follow, with the goal of scaling up global vaccine production and delivering vaccines equitably to the world’s population and preventing excess morbidity and mortality as broadly as possible. Multiple countries have followed CEPI’s lead in developing plans consistent with the 100 Days Mission to accelerate vaccine development and delivery. The key to achieving such a goal is a rapid, coordinated research response to smaller outbreaks and epidemics. Smaller responses build the infrastructure and provide practice for testing products quickly in a new crisis, while revealing gaps and generating relevant data to strengthen a bidirectional relationship between preparedness and response.

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Wahome, Newton, John M. Hickey, David B. Volkin, and C. Russell Middaugh. "Formulation Studies During Preclinical Development of Influenza Hemagglutinin and Virus-Like Particle Vaccine Candidates." In Vaccine Design. Springer New York, 2016. http://dx.doi.org/10.1007/978-1-4939-3389-1_27.

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Schlegl, Robert, and Rainer Hahn. "Purification and Formulation: Silent but Important Players in Vaccine Development." In Development of Novel Vaccines. Springer Vienna, 2012. http://dx.doi.org/10.1007/978-3-7091-0709-6_7.

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Vermaak, Samantha, Samantha Sampson, and Helen McShane. "The VALIDATE Network: Accelerating Vaccine Development for Tuberculosis, Leishmaniasis, Melioidosis and Leprosy." In Vaccines for Neglected Pathogens: Strategies, Achievements and Challenges. Springer International Publishing, 2023. http://dx.doi.org/10.1007/978-3-031-24355-4_2.

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AbstractEstablished in June 2017, VALIDATE is an international network of researchers working to accelerate vaccine development for four neglected intracellular pathogens that cause significant mortality and morbidity globally: Mycobacterium tuberculosis, Leishmania spp., Burkholderia pseudomallei and Mycobacterium leprae. In 5 years, VALIDATE has grown to have more than 550 members from over 250 institutes across 72 countries and has had several successes and important outputs. This chapter discusses VALIDATE’s origins, achievements and future direction.

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Higgs, Elizabeth S., Sheri A. Dubey, Beth A. G. Coller, et al. "Accelerating Vaccine Development During the 2013–2016 West African Ebola Virus Disease Outbreak." In Current Topics in Microbiology and Immunology. Springer International Publishing, 2017. http://dx.doi.org/10.1007/82_2017_53.

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Chesko, James, Thomas Vedvick, and Steve Reed. "Development of Biophysical Assays to Better Understand Adjuvanted Vaccine Formulation Potency and Stability." In Novel Immune Potentiators and Delivery Technologies for Next Generation Vaccines. Springer US, 2012. http://dx.doi.org/10.1007/978-1-4614-5380-2_5.

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Higgs, Elizabeth S. "14 Accelerating Development of Therapeutics for Preparedness, Response, and a More Secure World." In Principles and Practice of Emergency Research Response. Springer International Publishing, 2024. http://dx.doi.org/10.1007/978-3-031-48408-7_17.

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AbstractThe accelerated development of therapeutics is essential to infectious disease preparedness and emergency response, just like vaccines and diagnostics. Vaccine research received more public attention than therapeutics during the COVID-19 pandemic. Still, existing drugs, monoclonal antibody cocktails, the combination drug nirmatrelvir/ritonavir (Paxlovid®), and other life-saving interventions for COVID-19 are the result of accelerated therapeutics research during the health emergency. This chapter outlines scientific, regulatory, and social factors that affect the entire therapeutics development process from “bench to bedside,” including preclinical and clinical research, regulatory review, manufacturing, and delivery of safe, effective therapeutics before and during an infectious disease outbreak with pandemic potential. There are both similarities and differences in the processes for vaccines and diagnostics. Still, in all three cases, emergency research response goals are the same: to save lives and avert suffering, accelerate the end of the outbreak, and develop measures to prevent and mitigate future outbreaks. Therapeutics preparedness and response for pathogens with epidemic and/or pandemic potential require innovation to: Correct market failures in therapeutics development, e.g., lack of commercial investment in therapeutics for diseases that affect few people or predominantly low-income populations. Accelerate preclinical development. Receive approvals from oversight bodies for trials in emergencies. Implement large-scale, well-designed randomized clinical studies following good participatory practice (GPP) guidelines, ideally structured by a master protocol so individual trials contribute to a statistically powerful whole. Obtain regulatory authorization or approval. Ensure that the products resulting from research are available to all those who may benefit. Throughout the process, multitasking, or simultaneously acting on steps normally done in sequence, is a key methodology to accelerate results without compromising ethical and scientific standards. A clear, practical vision of what a therapeutics preparedness state should be is outlined here. It will require political support and human and material resources to make it possible.

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Meppen, Malte, and Daniela Stranges. "QbD approach to formulation development for protein-based vaccines." In Practical Aspects of Vaccine Development. Elsevier, 2022. http://dx.doi.org/10.1016/b978-0-12-814357-5.00004-0.

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Smith, Lee C., and Paul Nelson. "Formulation design considerations and good practice for live attenuated vaccine development." In Practical Aspects of Vaccine Development. Elsevier, 2022. http://dx.doi.org/10.1016/b978-0-12-814357-5.00005-2.

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Conference papers on the topic "Accelerating vaccine formulation development"

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Marwiyah, Siti, and Vincensia Mutiara Rengganis. "Policy Formulation on Accelerating the Development of Electric Motor Vehicles." In International Conference For Democracy and National Resilience (ICDNR 2021). Atlantis Press, 2022. http://dx.doi.org/10.2991/assehr.k.211221.015.

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Nair, Vineet, Kritika Prakash, Michael Wilbur, et al. "ADVISER: AI-Driven Vaccination Intervention Optimiser for Increasing Vaccine Uptake in Nigeria." In Thirty-First International Joint Conference on Artificial Intelligence {IJCAI-22}. International Joint Conferences on Artificial Intelligence Organization, 2022. http://dx.doi.org/10.24963/ijcai.2022/712.

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More than 5 million children under five years die from largely preventable or treatable medical conditions every year, with an overwhelmingly large proportion of deaths occurring in under-developed countries with low vaccination uptake. One of the United Nations' sustainable development goals (SDG 3) aims to end preventable deaths of newborns and children under five years of age. We focus on Nigeria, where the rate of infant mortality is appalling. We collaborate with HelpMum, a large non-profit organization in Nigeria, to design and optimize the allocation of heterogeneous health interventions under uncertainty to increase vaccination uptake, the first such collaboration in Nigeria. Our framework, ADVISER: AI-Driven Vaccination Intervention Optimiser, is based on an integer linear program that seeks to maximize the cumulative probability of successful vaccination. Our optimization formulation is intractable in practice. We present a heuristic approach that enables us to solve the problem for real-world use-cases. We also present theoretical bounds for the heuristic method. Finally, we show that the proposed approach outperforms baseline methods in terms of vaccination uptake through experimental evaluation. HelpMum is currently planning a pilot program based on our approach to be deployed in the largest city of Nigeria, which would be the first deployment of an AI-driven vaccination uptake program in the country and hopefully, pave the way for other data-driven programs to improve health outcomes in Nigeria.

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Herzog, Jenna, Rebekah Fodale, Mohammad Alsager Alzayed, Elizabeth M. Starkey, and Rohan Prabhu. "Feeling the Distance: Exploring Novice Designers’ Perceptions of the Psychological Distance Towards and Empathy Induced by Problem Variations." In ASME 2023 International Design Engineering Technical Conferences and Computers and Information in Engineering Conference. American Society of Mechanical Engineers, 2023. http://dx.doi.org/10.1115/detc2023-114540.

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Abstract The accelerating depletion of natural resources has necessitated the design of environmentally sustainable engineering solutions. To meet this need for sustainable solutions, designers must actively incorporate considerations of environmental impact in their design decisions. Prior research suggests that the effects of climate change are often perceived to be psychologically distant, and this distance could inhibit individuals from actively engaging in environmentally sustainable behavior. Little research has investigated the impact of problem framing based on designers’ psychological distance on design performance. Furthermore, research suggests that empathy development could be an effective mechanism for bridging psychological distance. However, little research has assessed the utility of empathy-invoking problem framing in sustainable engineering design practice and education. Our aim in this study is to explore this research gap by comparing student designers’ perceptions of different problem formulations. Specifically, we tested the effects of variations in (1) the socio-spatial context and (2) the empathy focus of a similar design problem. The effects of these variations on the perceived psychological distance and empathy-invoking nature were tested through a 2 × 2 between-subjects experiment. From the results, we see that the variations in the problem formulation did not relate to either the perceived psychological distance or the perceived empathy-invoking nature. These findings suggest that issues related to environmental sustainability tend to be perceived similarly, despite differences in their socio-spatial context and empathy-invoking nature.

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Reports on the topic "Accelerating vaccine formulation development"

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Evans, Donald L., Avigdor Eldar, Liliana Jaso-Friedmann, and Herve Bercovier. Streptococcus Iniae Infection in Trout and Tilapia: Host-Pathogen Interactions, the Immune Response Towards the Pathogen and Vaccine Formulation. United States Department of Agriculture, 2005. http://dx.doi.org/10.32747/2005.7586538.bard.

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The objectives of the BARD proposal were to determine the mechanisms of nonspecific cytotoxic cells (NCC) that are necessary to provide heightened innate resistance to infection and to identify the antigenic determinants in Streptococcus iniae that are best suited for vaccine development. Our central hypothesis was that anti-bacterial immunity in trout and tilapia can only be acquired by combining "innate" NCC responses with antibody responses to polysaccharide antigens. These Objectives were accomplished by experiments delineated by the following Specific Aims: Specific aim (SA) #1 (USA) "Clone and Identify the Apoptosis Regulatory Genes in NCC"; Specific aim #2 (USA)"Identify Regulatory Factors that Control NCC Responses to S. iniae"; Specific aim #3 (Israel) "Characterize the Biological Properties of the S. iniae Capsular Polysaccharide"; and Specific aim #4 (Israel) "Development of an Acellular Vaccine". Our model of S. iniae pathogenesis encompassed two approaches, identify apoptosis regulatory genes and proteins in tilapia that affected NCC activities (USA group) and determine the participation of S.iniae capsular polysaccharides as potential immunogens for the development of an acellular vaccine (Israel group). We previously established that it was possible to immunize tilapia and trout against experimental S. difficile/iniaeinfections. However these studies indicated that antibody responses in protected fish were short lived (3-4 months). Thus available vaccines were useful for short-term protection only. To address the issues of regulation of pathogenesis and immunogens of S. iniae, we have emphasized the role of the innate immune response regarding activation of NCC and mechanisms of invasiveness. Considerable progress was made toward accomplishing SA #1. We have cloned the cDNA of the following tilapia genes: cellular apoptosis susceptibility (CAS/AF547173»; tumor necrosis factor alpha (TNF / A Y 428948); and nascent polypeptide-associated complex alpha polypeptide (NACA/ A Y168640). Similar attempts were made to sequence the tilapia FasLgene/cDNA, however these experiments were not successful. Aim #2 was to "Identify Regulatory Factors that Control NCC Responses to S. iniae." To accomplish this, a new membrane receptor has been identified that may control innate responses (including apoptosis) of NCC to S. iniae. The receptor is a membrane protein on teleost NCC. This protein (NCC cationic antimicrobial protein-1/ncamp-1/AAQ99138) has been sequenced and the cDNA cloned (A Y324398). In recombinant form, ncamp-l kills S. iniae in vitro. Specific aim 3 ("Characterize the Biological Properties of the S.iniae Capsular Polysaccharide") utilized an in- vitro model using rainbow trout primary skin epithelial cell mono layers. These experiments demonstrated colonization into epithelial cells followed by a rapid decline of viable intracellular bacteria and translocation out of the cell. This pathogenesis model suggested that the bacterium escapes the endosome and translocates through the rainbow trout skin barrier to further invade and infect the host. Specific aim #4 ("Development of an Acellular Vaccine") was not specifically addressed. These studies demonstrated that several different apoptotic regulatory genes/proteins are expressed by tilapia NCC. These are the first studies demonstrating that such factors exist in tilapia. Because tilapia NCC bind to and are activated by S. iniae bacterial DNA, we predict that the apoptotic regulatory activity of S. iniae previously demonstrated by our group may be associated with innate antibacterial responses in tilapia.

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Eldar, Avigdor, and Donald L. Evans. Streptococcus iniae Infections in Trout and Tilapia: Host-Pathogen Interactions, the Immune Response Toward the Pathogen and Vaccine Formulation. United States Department of Agriculture, 2000. http://dx.doi.org/10.32747/2000.7575286.bard.

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In Israel and in the U.S., Streptococcus iniae is responsible for considerable losses in various fish species. Poor understanding of its virulence factors and limited know-how-to of vaccine formulation and administration are the main reasons for the limited efficacy of vaccines. Our strategy was that in order to Improve control measures, both aspects should be equally addressed. Our proposal included the following objectives: (i) construction of host-pathogen interaction models; (ii) characterization of virulence factors and immunodominant antigens, with assessment of their relative importance in terms of protection and (iii) genetic identification of virulence factors and genes, with evaluation of the protective effect of recombinant proteins. We have shown that two different serotypes are involved. Their capsular polysaccharides (CPS) were characterized, and proved to play an important role in immune evasion and in other consequences of the infection. This is an innovative finding in fish bacteriology and resembles what, in other fields, has become apparent in the recent years: S. iniae alters surface antigens. By so doing, the pathogen escapes immune destruction. Immunological assays (agar-gel immunodiffusion and antibody titers) confirmed that only limited cross recognition between the two types occurs and that capsular polysaccharides are immunodominant. Vaccination with purified CPS (as an acellular vaccine) results in protection. In vitro and ex-vivo models have allowed us to unravel additional insights of the host-pathogen interactions. S. iniae 173 (type II) produced DNA fragmentation of TMB-8 cells characteristic of cellular necrosis; the same isolate also prevented the development of apoptosis in NCC. This was determined by finding reduced expression of phosphotidylserine (PS) on the outer membrane leaflet of NCC. NCC treated with this isolate had very high levels of cellular necrosis compared to all other isolates. This cellular pathology was confirmed by observing reduced DNA laddering in these same treated cells. Transmission EM also showed characteristic necrotic cellular changes in treated cells. To determine if the (in vitro) PCD/apoptosis protective effects of #173 correlated with any in vivo activity, tilapia were injected IV with #173 and #164 (an Israeli type I strain). Following injection, purified NCC were tested (in vitro) for cytotoxicity against HL-60 target cells. Four significant observations were made : (i) fish injected with #173 had 100-400% increased cytotoxicity compared to #164 (ii) in vivo activation occurred within 5 minutes of injection; (iii) activation occurred only within the peripheral blood compartment; and (iv) the isolate that protected NCC from apoptosis in vitro caused in vivo activation of cytotoxicity. The levels of in vivo cytotoxicity responses are associated with certain pathogens (pathogen associated molecular patterns/PAMP) and with the tissue of origin of NCC. NCC from different tissue (i.e. PBL, anterior kidney, spleen) exist in different states of differentiation. Random amplified polymorphic DNA (RAPD) analysis revealed the "adaptation" of the bacterium to the vaccinated environment, suggesting a "Darwinian-like" evolution of any bacterium. Due to the selective pressure which has occurred in the vaccinated environment, type II strains, able to evade the protective response elicited by the vaccine, have evolved from type I strains. The increased virulence through the appropriation of a novel antigenic composition conforms with pathogenic mechanisms described for other streptococci. Vaccine efficacy was improved: water-in-oil formulations were found effective in inducing protection that lasted for a period of (at least) 6 months. Protection was evaluated by functional tests - the protective effect, and immunological parameters - elicitation of T- and B-cells proliferation. Vaccinated fish were found to be resistant to the disease for (at least) six months; protection was accompanied by activation of the cellular and the humoral branches.

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