Periodic Reporting for period 4 - FuncMAB (High-throughput single-cell phenotypic analysis of functional antibody repertoires)
Periodo di rendicontazione: 2022-08-01 al 2024-01-31
Vaccines have had a tremendously positive effect on individual and global health for decades. However, the understanding of their mechanism of action and induced immunological responses is still limited. The concept of vaccine-mediated protection and the quantitation thereof remain especially elusive. Indeed, protection is empirically assayed during clinical trials, and levels and thresholds are defined over time. Here, current testing methods for protection do not provide temporal, spatial, or analytical resolution to expose fundamental differences in the underlying mechanisms that ultimately mediate protection, and therefore the gained understanding is limited. Indeed, immune responses are highly dynamic, heterogeneous and their successful completion involves and needs many different interactions, cells, and functions throughout the organism. Individual cells are the functional units within any immune response, and their varying frequencies and degrees of activity shape and define the response. Therefore, cells from the immune system, their state, activation and ultimately functionality display high dynamic heterogeneity, and there is hence a need for quantitative high-throughput systems that allow for a dynamic, functional single-cell phenotyping, linking function to the individual cells.
Therefore, within this project, we aim to measure, understand, and exploit antibody-mediated vaccine-induced protection on the individual cell and antibody level. By doing so, we aim to not only measure and describe the functional antibody repertoires with single-cell resolution, but also to understand the influences that are introduced by varying vaccination strategies and measure the impact on the quality, quantity, and functionality of the humoral immune response. The overarching objective of this proposal is to quantitatively map antibody functions on the single-cell level, and to use these data sets to understand the selection mechanisms involved in their generation, evolution, and transfer to memory; and to finally exploit the measurement to screen for therapeutic candidates and to accelerate vaccine development.
Therefore, within this project, we aim to measure, understand, and exploit antibody-mediated vaccine-induced protection on the individual cell and antibody level. By doing so, we aim to not only measure and describe the functional antibody repertoires with single-cell resolution, but also to understand the influences that are introduced by varying vaccination strategies and measure the impact on the quality, quantity, and functionality of the humoral immune response. The overarching objective of this proposal is to quantitatively map antibody functions on the single-cell level, and to use these data sets to understand the selection mechanisms involved in their generation, evolution, and transfer to memory; and to finally exploit the measurement to screen for therapeutic candidates and to accelerate vaccine development.
The FuncMab project aimed to measure and understand antibody functionality to inform rational vaccine design. The project involved various investigations including quantifying the quality and quantity of functional antibody repertoires on the single-antibody level. We generated assays to analyze various parameters linked to monoclonal antibody functionality, ranging from their isotype, neutralization, binding, and complement deposition to various others. We focused on vaccination against infectious diseases.
In the next step, the researchers explored the impact of introduced changes in immunization on the antibody repertoire, providing insights into the interplay between the two parameters. These studies were done in a murine model system. Data compilation revealed key factors influencing the immune response to vaccination, such as antigenic dose and adjuvants. This work also provided deeper insights into complement activation processes due to antibody binding onto bacterial surfaces and investigated the influence of various agonists on the quality and quantity of the functional antibody repertoire induced by vaccination.
This understanding led to the establishment of criteria for rational vaccine design, resulting in patent applications and ongoing research on peptide mimetics, therapeutic antibodies, and analysis systems to dampen the impact of viral evolution with vaccination. In terms of dissemination of the results, the project led to several high-quality and impact papers in vaccine-related and interdisciplinary journals, talks at universities and conferences, and engagements with the public surrounding the topic of vaccination. The project is also linked to the creation of a start-up.
Overall, the project generated a variety of new measurement methods that can be applied to various fundamental and clinical problems, enhanced our understanding of antibody functionality, and provided insights for the rational design of vaccines.
In the next step, the researchers explored the impact of introduced changes in immunization on the antibody repertoire, providing insights into the interplay between the two parameters. These studies were done in a murine model system. Data compilation revealed key factors influencing the immune response to vaccination, such as antigenic dose and adjuvants. This work also provided deeper insights into complement activation processes due to antibody binding onto bacterial surfaces and investigated the influence of various agonists on the quality and quantity of the functional antibody repertoire induced by vaccination.
This understanding led to the establishment of criteria for rational vaccine design, resulting in patent applications and ongoing research on peptide mimetics, therapeutic antibodies, and analysis systems to dampen the impact of viral evolution with vaccination. In terms of dissemination of the results, the project led to several high-quality and impact papers in vaccine-related and interdisciplinary journals, talks at universities and conferences, and engagements with the public surrounding the topic of vaccination. The project is also linked to the creation of a start-up.
Overall, the project generated a variety of new measurement methods that can be applied to various fundamental and clinical problems, enhanced our understanding of antibody functionality, and provided insights for the rational design of vaccines.
We developed and used new, cutting-edge technologies to shed light on complex immune reactions, to first quantify the quality of the induced antibodies, secondly understand the mechanisms behind their generation, and lastly influence the immunological reaction upon vaccination. Our novel bioassays to functionally characterize secondary antibody functionalities are well beyond the state-of-art and allow us to study the antibody repertoire with unique quantitative resolution. The objective of this project was not only to develop new analytical approaches to be able to quantitatively map antibody functions with individual cell resolution but also to use these data sets to understand the underlying selection mechanisms involved in their generation, evolution, and transfer to the memory of the antibody repertoires. Therefore, the overarching objective of this proposal was to quantitatively map antibody functions on the single-cell level, to use these data sets to understand the selection mechanisms involved in their generation, evolution, and transfer to memory. While far from complete, these unique measurements gave novel insights into these mechanisms that are currently further investigated.