The recent rise to prominence of clinically beneficial immune-based therapies, particularly for cancer patients, has put critical emphasis on the need for validated bio-assays measuring functional correlates of the immune response. The ELISpot assay is one the most commonly used immune-monitoring assays for basic and translational research in immunology, enabling the detection of rare cells and RNA transcripts. The assay represents an ideal platform for immuno-biomarker assessment, particularly in vaccine development, immune-oncology therapeutics, and diagnostics. However, performance characteristics can be greatly influenced by a variety of factors, including sample handling/processing, cell recovery and viability post-cryopreservation, operator technique/execution, and the use of manual or automated spot enumeration and analysis. In this webinar, we will address the technical and methodological requirements for the reliable use of ELISpot assays, introduce important quality control measures, discuss essential assay qualification parameters, and provide advice for proper interpretation of results obtained by automated ELISpot imaging systems.
Overcoming Sample-related Challenges for Elispot and Other Functional T cell Assays To Produce More Reliable Immune Monitoring
Immune monitoring is an integral part of immunological research and translational applications in the fields of cancer, infectious disease, autoimmunity, transplantation and others. The functional assessment of immune cells heavily relies on the availability of adequate amounts of intact samples. In this live webinar with Q&A, you’ll hear Elispot expert Dr. Sylvia Janetzki (ZellNet Consulting) describe the latest improvements made to strategies for obtaining and handling patient samples to conserve sample integrity. Various specific topics will be discussed, including obtaining blood and tissue samples; processing, storage and transport of samples; handling of samples in multi-center trials; and tools to preserve stability and functionality of samples including re-setting circulating T cell functionality to a tissue-like state.
Strategies for enhancing Elispot/Fluorospot testing in research and clinical trials
Elispot assays are powerful tools for monitoring immune response, whether desired (as in the case of immunotherapies) and undesired (immunotoxicity). Not only do Elispot assays provide functional cytokine secretion data on single cells, but they are also sensitive, easy, robust and compatible with high-throughput testing. Recent studies have revealed the importance of sample integrity and strategies for maintaining functionality of the cells being analyzed. In this live webinar with Q&A, you’ll hear Elispot expert Dr. Sylvia Janetzki (ZellNet Consulting) describe the latest improvements made to technical aspects of the assay, including sample preparation solutions, specialized materials and reagents, and enhancements in defining positive Elispot responses. Dr. Janetzki will also show examples of polyfunctional, fluorescent Elispot analysis of T cells, B cells and macrophages/monocytes and examples of successful applications of Elispot in clinical trials.
Using imaging flow cytometry to study dendritic cell biology and function.
Dendritic cells (DCs) are the most efficient antigen-presenting cells in the immune system. Upon recognition of pathogens, DCs become activated and migrate to lymphoid structures where they present relevant antigens to lymphocytes. The uptake of pathogens and the mechanisms leading to proper processing and presentation of relevant antigens are crucial to an immune response.
In this webinar, Dr. Juan Garcia-Vallejo and Dr. Edith Janssen discuss how they have used imaging flow cytometry to gain a greater understanding of DC biology and function.
Before they begin Robert Smith-McCollum of $ will give a brief introduction to imaging flow cytometry.
Dr. Garcia-Vallejo will then talk about his research into the C-type lectin receptors (CLRs) expressed by DCs and the mechanisms leading to CLR-mediated internalization, intracellular routing and antigen processing. He will describe a series of imaging flow cytometry-based methods he developed to study antigen/receptor internalization as well as tracking of antigens/receptors through different intracellular compartments.
Finally Dr. Janssen will outline her studies comparing dendritic cell functionality in primary dendritic cells from normal mice and mice susceptible to various autoimmune diseases as well as primary and cultured DCs from healthy human donors. She will discuss how imaging flow cytometry can be used to assess the type and mechanism of endocytosis as well as the trafficking of endocytosed materials in phagosomes.