Ustekinumab (anti-IL-12/IL-23)

The addition of Ustekinumab (UST) significantly inhibits T follicular helper (TFH) cell differentiation in vitro.^[1]

Objective: TFH cell differentiation assay
Cells: naïve CD4+ T cells, T follicular helper (TFH) cells, PBMCs
Concentrations: 100 ng/mL
Incubation Time: 24 h
Method: The TFH cell differentiation assay is performed with freshly isolated PBMCs from healthy donors (n = 16). Naïve CD4+ T cells are isolated by using the human Naïve CD4+ T Cell Isolation Kit II. A total of 500,000 naïve CD4+ T cells are cultured in complete medium in the presence of 12.5 μL/mL anti-CD3/CD28 in a flat-bottom 48 well plate. After 24 hours of culture, cytokines are added and cells are incubated for additional 2.5 days. A total of 5 ng/mL IL-12 + 5 ng/mL TGFβ with and without 100 ng/mL UST and 100 ng/mL UST alone are used.
Reference: https://pubmed.ncbi.nlm.nih.gov/32679193

Ustekinumab can apply to humanized mice, peripheral blood and other related assays (Only for Reference)

Ustekinumab is generated via recombinant human IL-12 immunization of human Ig (hu-Ig) transgenic mice. Ustekinumab binds to the p40 subunit common to IL-12 and IL-23 and prevents their interaction with the IL-12 receptor β1 subunit of the IL-12 and IL-23 receptor complexes.^[2]