ASSAY – Th17
HUMAN CELL LINE BASED
Upon antigen processing and maturation the dendritic cell-MHC Class II complex interacts with CD4 T cells with a TCR activation response that leads to the differentiation of T-helper (Th) cells. One of the pro-inflammatory differentiation of Th cells is the Th17 that is associated with the production of IL-17, IL-22, IL-21 and IL-26. Cytokines that contribute to Th17 development are TGFbeta, IL-6, IL-21 and IL-23. The transcription factors STAT3, RORgamma and RORalpha are key factors in the Th17 differentiation.
In vitro Th17 DIFFERENTIATION ASSAY
Our in-house human T cell assay is adapted to differentiate Th17 cells from TALL-1 cells. On stimulation (e.g. with PMA) these cells produce IL-17 and IL-22 which are quantified by intracellular flow cytometry or ELISA. This assay can be performed in 96-well plates to test the effect of compounds on Th17 differentiation and activation (e.g. inhibitory effect) and on the viability of Th17 cells. The assay can be adapted to be used with primary T cells.
The Th17 pathway is important in studies of various therapeutic areas and biological pathways:
- Tumour biology and oncology
- Cardiovascular disease
- Tissue damage and cell death
In recent years the need to develop and source human cell based Th17 assay has increased.
Cell Line Based Approach
We use myeloid and lymphocytic cell lines for various immune response assays such as dendritic cell maturation, T cell activation, phagocytosis, etc. We are among the few groups who can generate and demonstrate the use of dendritic cells and macrophages (including M1 and M2) from monocytic cell lines and T cells in activation studies. Conventionally dendritic cells are generated from primary monocytes or bone marrow and T cells or monocytes are separated from primary PBMCs from human donors. The use of cell lines in the initial stage offers a consistent reference assay prior to using primary human cells. Primary human cells used in the initial stage can be variable, expensive, time consuming, and often inconclusive.