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Immunostimulatory Agent Evaluation: Lymphoid Tissue Extraction and Injection Route-Dependent Dendritic Cell Activation
Published on: September 16, 2018
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Detergent wash improves vaccinated lymph node handling ex vivo.
Alexander G Ball1, Maura C Belanger2, Rebecca R Pompano3
1Department of Microbiology, Immunology and Cancer Biology, University of Virginia, Charlottesville, USA; Carter Immunology Center, University of Virginia, Charlottesville, USA.
Journal of Immunological Methods
|December 17, 2020
Summary
A new digitonin wash method improves the slicing of lymph nodes (LNs) for studying adaptive immunity. This technique preserves tissue viability and function, enabling better analysis of immune responses to vaccines.
Area of Science:
- Immunology
- Cell Biology
- Tissue Engineering
Background:
- Live ex vivo lymph node (LN) slices are crucial for studying adaptive immunity dynamics.
- Reactive LNs from vaccinated animals often detach during slicing due to lipid accumulation, hindering analysis.
Purpose of the Study:
- To develop a method to prevent LN detachment during slicing.
- To assess if a mild detergent wash improves LN adhesion to agarose matrices without compromising tissue viability or function.
Main Methods:
- A brief digitonin wash was applied to reactive LNs before embedding in agarose.
- Lipid droplet removal was assessed visually.
- LN loss during slicing was quantified.
- Tissue viability and immune functions (particle capture, antigen processing, cytokine secretion) were evaluated.
Main Results:
- The digitonin wash significantly reduced LN loss during slicing from 13-75% to 0-25%.
- Lipid droplets were visibly removed from the LN surface.
- The wash did not substantially impact LN viability or key immune functions.
Conclusions:
- A simple digitonin wash effectively prevents detachment of reactive LNs during ex vivo slicing.
- This method preserves tissue integrity and function, facilitating the study of adaptive immune responses.
- The technique supports ex vivo analysis of immunotherapies and vaccine responses.
