Ex Vivo T Cell Function Analysis
Spleen and lymph node cells are restimulated with antigen to analyze cytokine production of T cells in vitro. This can be performed either as a separate study, or the end of an EAE study.
Spleens or lymph nodes are collected and cell suspensions plated in 96-well plates with multiple concentrations of the selected antigen in triplicate.
After 72 hours of culture, supernatants are collected and analyzed for cytokines using Luminex, ELISA, or CBA.
Analysis is performed on the supernatant from each well.
Typical results
C57BL/6 mice were immunized with MOG35-55/CFA, or with MOG35-55/CFA followed by pertussis toxin injection. Eleven (11) or 27 days later, spleen or lymph node cells were restimulated with MOG35-55 for 72 hours. Cytokine production was compared across a range of MOG35-55 concentrations (0.0, 0.7, 2.2, 6.6, and 20.0 µg/mL) and analyzed using a cytometric bead assay (CBA) in accordance with the manufacturer's instructions.
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