Nonspecific binding of antibodies to cell Fc receptors was assessed with a control isotype.
A third group of mice received 75 µg of a control isotype matched MAb 2C3 that binds to a Neisseria gonorrhoeae membrane protein.
Staining with a control isotype antibody is represented by a grey shaded peak in all plots.
The background staining was evaluated using a control isotype labeled with phycoerythrin (BD Bioscience Pharmingen) (data not shown).
Cells were pelleted, resuspended in PBS/10%, divided into three equal portions, and incubated with an E-cadherin antibody (1 1600), a control isotype equivalent antibody (anti-HA; 1 1600), or a secondary antibody Alexa-488 (1 400) in PBS/10% (1 hour at 4°C).
Mice receiving a control isotype Ab showed a significant decrease in disease burden compared with mice receiving control Th1 cells (Fig. 4B); however, mice receiving GSK3 inhibitor treated cells and anti-IL-10R Ab displayed an increase in disease burden compared with controls (Fig. 4B).
