Effect of IL-10 gene transmission on the PTg-stimulated splenocytes proliferation and Th1 cytokines production from experimental autoimminue thyroiditis rats1

ObjectiveTo investigate the effects of gene therapy with IL-10 on PTg-induced proliferation of splenocytes and Th1 cytokine production from PTg-stimulated splenocytes.MethodsEAT rats were divided into four groups: group A (PBS+PLL), group B (pORF+PLL), group C (pORFmIL10+PLL), and group D (pORFmIL10+MEM). The substances mixed with lipofectamine were injected into the thyroid tissues of rats on the 18th dday after immunization. The rats were sacrificed at the 8th week. In vitro proliferative responses to ConA and different concentration of PTg were measured by culturing 4×105 splenocytes pulsed with 18.5KBq of [3H] thymidine for the final 12h and then harvested for liquid scintillation counting. In vitro splenocytes were cultured with PTg (25 mg/L). Th1 cytokine IFN-γ, TNF-α and IL-2 were detected by ELISA.ResultsThe proliferative response to PTg was suppressed in group C, compared with that of group A and B (P < 0.05). The levels of IFN-γ, TNF-α and IL-2 in the supernatant of PTg-stimulated splenocytes were 3548.25 ± 779.47 pg/ml, 27.66±10.50 pg/ml and 3617.73±609.15 pg/ml, respectively, which were much lower in group C than those in group A and B (P < 0.01, P < 0.05, P < 0.001, respectively).ConclusionIL-10 gene transmission in thyroid tissues could inhibit PTg specific proliferation of splenocytes from EAT rats and the secretion of Th1 cytokines from PTg-stimulated splenocytes.