Immunoglobulin Production
Purified B cells (1x106cells/ml) alone or with CD8
Treg at various ratios were activated with 2ug/ml of anti-human
CD40/TNFRSF5 antibody and 2.5ug/ml of CpG in AIM-V medium and cultuted
at 37°C for 14 days. Supernatant were collected and stored at −20°C
until assayed for detection of Immunoglobulins by ELISA. Immunoglobulin
levels of IgA, IgG, IgM were determined using appropriate human ELISA
kits (eBiosciences/Thermo Fisher, Waltham, MA) per manufacturer’s
protocol.
Statistical analysis was performed by paired student t test using Prism
7 software. A value of <0.05 is considered significant.