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.