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In Vivo Immunogenicity Screening of Tumor-Derived Extracellular Vesicles by Flow Cytometry of Splenic T Cells
Published on: September 23, 2021
Assessment of cytoplasmic dsDNA for cellular immunogenicity
Isha Mondal1, Oishika Das2, Raymond Sun3
1University of California, San Francisco, San Francisco, California, United States; Duke University School of Medicine, Durham, NC, United States.
Insights
Detecting cytosolic double-stranded DNA (dsDNA) is key to understanding immunity, cancer, and autoimmune diseases. This review covers lab methods for identifying and quantifying cytosolic dsDNA in various samples.
Area of Science:
- Immunology
- Molecular Biology
- Biochemistry
Background:
- Cytosolic double-stranded DNA (dsDNA) plays a crucial role in innate immunity, infection, cancer, and autoimmune diseases.
- Accurate detection of cytosolic dsDNA is essential for advancing research in these fields.
Purpose of the Study:
- To provide a comprehensive overview of common laboratory methods for detecting cytosolic double-stranded DNA (dsDNA).
- To highlight the applications, limitations, and potential advancements of various dsDNA detection techniques.
Main Methods:
- Immunofluorescence microscopy for direct visualization of dsDNA localization.
- Enzyme-linked immunosorbent assays (ELISAs) and immunoblotting for indirect detection via associated proteins (e.g., cGAS).
- Fluorescent spectroscopy, biosensors, and nanotechnology for rapid, sensitive quantification and detection.
Main Results:
- Various methods offer different strengths in sensitivity, specificity, and throughput for dsDNA detection.
- Immunofluorescence provides spatial and dynamic information, while ELISAs and immunoblotting detect dsDNA indirectly.
- Spectroscopy, biosensors, and nanotechnology offer rapid and sensitive detection capabilities.
Conclusions:
- A range of established and emerging techniques are available for detecting cytosolic dsDNA.
- Method selection depends on specific research needs regarding sample type, sensitivity, and throughput.
- Advancements in biosensors and nanotechnology promise enhanced sensitivity for future dsDNA detection.
Abstract:
Detection of cytosolic double-stranded DNA (dsDNA) is critical for understanding its role in innate immunity, infection, cancer, and autoimmune diseases. Various methods have been developed to identify and quantify cytosolic dsDNA, each offering unique strengths in terms of sensitivity, specificity, and throughput. Immunofluorescence microscopy enables direct visualization of cytosolic dsDNA, providing insights into its localization and dynamics within cells. Enzyme-linked immunosorbent assays (ELISAs) and immunoblotting techniques detect dsDNA indirectly through associated proteins like cGAS. Fluorescent spectroscopy-based methods allow for rapid and specific quantification of dsDNA. Additionally, biosensors and nanotechnology-based approaches are emerging as novel tools for dsDNA detection with enhanced sensitivity. This methodological compendium provides an overview of the common lab bench methodologies, highlighting their applications, limitations, and potential advancements in the detection of cytosolic dsDNA in various sample forms, such as cells, sectioned tissue, and cellular cytosol extracts.
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