Cell-based ELISpot protocol to detect and quantify antigen-specific antibody-secreting cells in murine whole-organ
Christina Brunner1, Shannon R Menzel1, Franziska Rost1
1Division of Molecular Immunology, Department of Medicine 3 - Rheumatology and Immunology, Universitätsklinikum Erlangen, Nikolaus-Fiebiger Center, Friedrich-Alexander-University Erlangen-Nürnberg, 91054 Erlangen, Germany.
Insights
This study details a protocol for quantifying antibody-secreting cells (ASCs) using the enzyme-linked immunospot (ELISpot) assay. This method is crucial for understanding adaptive immunity in vaccine responses and disease research.
Area of Science:
- Immunology
- Cell Biology
Background:
- Antibody-secreting cells (ASCs) are critical components of the adaptive immune response.
- Understanding ASCs is vital for evaluating vaccine efficacy and studying immune-related diseases.
Purpose of the Study:
- To present a standardized protocol for quantifying ASCs.
- To enable accurate enumeration of ASCs for research applications.
Main Methods:
- Detailed protocol for the enzyme-linked immunospot (ELISpot) assay.
- Includes steps for cell sample preparation and ELISpot plate quantification.
Main Results:
- The ELISpot assay provides a reliable method for quantifying ASCs.
- The protocol is applicable to various research settings.
Conclusions:
- The presented ELISpot protocol offers a valuable tool for immunological research.
- Facilitates studies on adaptive immunity, vaccine responses, and ASC-associated pathologies.
Abstract:
Antibody-secreting cells (ASCs) are the hallmark of the adaptive immune response to vaccines or infections. Here, we present a protocol to quantify the number of ASCs using an enzyme-linked immunospot assay (ELISpot). We describe the preparation of cell samples, ELISpot plates, and their quantification. This assay has potential applications in vaccine studies and research on the adaptive immune system or ASC-associated diseases. For complete details on the use and execution of this protocol please, refer to Bierling et al.1.
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