Related Experiment Video
Updated: Aug 9, 2026
Detection and Isolation of Viable Mouse IL-17-Secreting T Cells
Published on: December 18, 2008
Detection of intracellular interleukin-10 by flow cytometry
B Kreft1, G G Singer, C Diaz-Gallo
1Brigham and Women's Hospital, Department of Medicine, Boston, MA 02115.
Insights
This study introduces a method to detect intracellular cytokines, like interleukin-10 (IL-10), by permeabilizing cell membranes. This technique allows for the localization and measurement of cytokine expression within cells, overcoming previous limitations.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Cytokine detection traditionally relies on measuring secreted molecules.
- Intracellular cytokine expression remains largely unquantified due to accessibility challenges.
- Existing methods lack the ability to localize cytokine production within cells.
Purpose of the Study:
- To develop and validate a method for detecting intracellular cytokines.
- To overcome the limitations of measuring only secreted cytokine molecules.
- To enable the study of intracellular cytokine expression in cell lines.
Main Methods:
- Cell membrane permeabilization using digitonin.
- Intracellular cytokine localization using specific antibodies.
- Detection and analysis via flow cytometry.
- Application in the HT-2 T cell line for interleukin-10 (IL-10) detection.
Main Results:
- Successfully detected intracellular interleukin-10 (IL-10) in HT-2 T cells.
- Demonstrated that membrane permeabilization is essential for intracellular cytokine detection.
- Confirmed the feasibility of localizing and quantifying intracellular cytokines.
Conclusions:
- Permeabilization of the cell membrane is a viable strategy for detecting intracellular cytokines.
- The developed flow cytometry technique allows for ready detection of intracellular cytokine expression.
- This method expands the possibilities for studying cytokine biology at the intracellular level.
Abstract:
Detection of cytokines is limited to measurements of the secreted molecule. To circumvent this problem we permeabilized the cell membrane with digitonin and localize cytokine expression using antibodies by flow cytometry. In this report we demonstrate that we can detect specific intracellular interleukin-10 (IL-10) in the HT-2 T cell line only after membrane permeabilization. With this technique intracellular cytokines are readily detectable.
