Th17 T Cell Assay
Th17 T Cell Assay - Related Videos
10:15
Small RNA Transfection in Primary Human Th17 Cells by Next Generation Electroporation
0 Views •
Cited by 5 •
2017
Next generation electroporation is an efficient method for transfecting human Th17 cells with small RNAs to alter gene expression and cell behavior.
01:56
Inducing Paralysis in a Mouse Model via Transfer of Aquaporin-4-Specific Th17 Cells
0 Views •
2025
This video demonstrates the procedure of inducing paralysis by transferring activated Th17 cells targeting aquaporin-4 into a mouse model. In the central nervous system, the activated Th17 cells interact with aquaporin-4-expressing astrocytes, triggering an immune response that leads to tail and limb paralysis in the mouse.
12:59
Isolation and Th17 Differentiation of Naïve CD4 T Lymphocytes
0 Views •
Cited by 33 •
2013
With effector functions distinct from other T cell subsets, Th17 cells have been centrally implicated in inflammatory autoimmunity. This in vitro Th17 differentiation protocol provides a means to determine whether naïve CD4+ T lymphocytes can differentiate into Th17 cells, and to further examine their role in autoimmunity and host response.
07:46
In Vitro Differentiation of Naive CD4+ T Cells into Pathogenic Th17 Cells in Mouse
0 Views •
Cited by 1 •
2024
This protocol describes the differentiation of naïve CD4+ T cells into pathogenic Th17 cells in vitro. Specifically, when combined with a multi-parameter flow cytometry-based approach, 90% purity of pathogenic Th17 cells can be obtained from naïve CD4+ T cells using this differentiation method.
05:30
Clonogenic Assay: Adherent Cells
0 Views •
Cited by 228 •
2011
The applicability of the clonogenic assay for evaluating reproductive viability has been established for more than 50 years. Here we demonstrate the general procedure for performing the clonogenic assay with adherent cells.
FAQs
Antigen and mitogen stimulation answer different experimental questions. Antigen stimulation examines responses linked to a particular immune trigger, whereas mitogens provide a broader activation signal. Comparing these conditions can reveal whether IL-17A production depends on specific recognition or general T-cell activation. The selected stimulus therefore shapes how researchers interpret differences in Th17-associated responses.
IL-17A provides a functional indicator of Th17-associated activity because these cells are characterized by production of this cytokine. Measuring intracellular IL-17A identifies responding cells at the single-cell level, while measuring secreted IL-17A reflects the amount released into culture. Using either readout helps connect cellular activation with inflammatory and mucosal immune responses.
Culture conditions can either assess responses from existing CD4+ T cells or promote their differentiation toward a Th17 phenotype. This distinction is important when interpreting results: stimulated cultures examine functional responsiveness, whereas differentiation conditions examine how an experimental environment supports development of the response. Such comparisons help investigate mechanisms controlling Th17 activity and inflammation.
Flow cytometry with intracellular cytokine staining shows which cells produce IL-17A and can relate production to the CD4+ T-cell population. ELISA measures cytokine secretion in the culture sample, providing a bulk concentration rather than direct single-cell information. Selecting between them depends on whether the experiment prioritizes responding-cell frequency or overall cytokine release.
A typical workflow begins with CD4+ T cells, exposes them to an antigen or mitogen, and maintains them under conditions selected for response measurement or Th17 differentiation. Researchers then assess IL-17A expression or secretion using flow cytometry, intracellular cytokine staining, ELISA, or a related readout. The resulting measurements support comparisons among experimental conditions.
The assay is useful when investigators need to examine host responses to bacterial or fungal infection, particularly responses associated with mucosal defense and inflammation. It can also help define inflammatory or autoimmune mechanisms. By comparing IL-17A responses across conditions, researchers can characterize cellular function and examine how immune activity changes during disease-related experiments.
Researchers can compare stimulated or differentiated cultures in the presence and absence of an immunomodulatory treatment, then measure changes in IL-17A expression or secretion. Reduced or enhanced responses indicate that the treatment affects Th17-associated activity under the tested conditions. This approach supports investigation of treatment effects on inflammatory mechanisms and cellular immune function.