Proliferation of T cells from mice treated with ATG or isotype IgG in response to CD3 antibody stimulation. NOD mice were treated with ATG or isotype IgG twice with a 3-day interval. A, At day 3 post treatment, spleen cells were prepared and stimulated with anti-CD3 antibodies for 3 days, then 3H-thymidine (1 uCi/well) was added to the cultures for additional 16 hours. 3H-thymidine incorporation was measured by scintillation counting. Three mice were tested in each group. B, part of the above spleen cells was used to isolate CD4+ T cells. The purified CD4+ T cells were stimulated with anti-CD3 and anti-CD28 antibody coated beads for 3 days. Thereafter, 3H-thymidine (1 uCi/well) was added to the cultures for additional 16 hours. T cell proliferation was examined by the method described above. C, the production of IL-10, IFN-γ in the cultures from 3 animals in each group in B was measured by Luminex. The values for IL-10 and IFN-γ for treatment group and controls are (993.3±141.0 v.s. 366.7±96.1) and (1117.0±202.8 v.s. 3793±795.5), respectively. The difference between the two groups for IL-10 and IFN-γ by Student t test is significant with p value as of 0.0221 and 0.0253, respectively. An additional independent experiment was performed with the similar results.
Xia et al. BMC Immunology 2012 13:70 doi:10.1186/1471-2172-13-70