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Isolation and Th17 Differentiation of Naïve CD4 T Lymphocytes
Published on: September 26, 2013
Measurement of interleukin-17
1M.D. Anderson Cancer Center, Houston, Texas.
Current Protocols in Immunology
|April 25, 2008
Summary
This study details methods for generating and detecting T helper 17 (T(H)17) cells, which produce IL-17 and drive inflammation. These protocols aid in understanding T(H)17 cell regulation and monitoring inflammatory diseases.
Area of Science:
- Immunology
- Cell Biology
Background:
- Naive CD4+ T cells differentiate into effector T helper (T(H)) cells upon antigenic stimulation.
- T(H)1 cells mediate cell-mediated immunity via IFN-gamma; T(H)2 cells handle extracellular pathogens and allergies via IL-4, IL-5, and IL-13.
- A novel T(H) subset, T(H)17 cells, produces the proinflammatory cytokine IL-17, regulating inflammatory responses.
Purpose of the Study:
- To describe protocols for the in vitro differentiation of mouse IL-17-expressing T cells.
- To outline methods for detecting IL-17-expressing T cells using intracellular cytokine staining.
- To detail the measurement of IL-17 secretion in culture supernatants via ELISA.
Main Methods:
- In vitro differentiation of mouse T cells under defined culture conditions.
- Intracellular cytokine staining for IL-17 detection in T cells.
- Enzyme-linked immunosorbent assay (ELISA) for quantifying IL-17 in supernatants.
Main Results:
- Established protocols for generating and quantifying IL-17-producing T cells in vitro.
- Demonstrated the utility of these methods for studying T(H)17 cell differentiation.
- Validated techniques for detecting IL-17 in biological samples.
Conclusions:
- In vitro generation of T(H)17 cells facilitates research into their differentiation and regulation.
- Detection methods for IL-17 are crucial for monitoring inflammatory diseases.
- These protocols support the assessment of therapeutic interventions for inflammatory conditions.
