Quantifying T Cell Proliferation Assay

For T cell proliferation assays, MACS sorted cDC from rhMPV (-WT or -ΔG) and mock-infected mice (10⁵ cells/well) were cultured for 5 days with CD4+ T cells from DO11.10 mice (2x10⁵ cells/well) in 96-well round bottom microtiter plates. cDC were loaded with 10 μg/mL of OVA peptide (323–339, Genscript, Piscataway, NJ) 2h prior to co-culture with T-cells. The CD4+ T cells were tagged with CFSE prior to culture and after 5 days of co-culture, cells were collected, washed and analyzed by FACS for T-cell proliferation, as determined by increase in CFSE low positive cells (36 (link)).