Figure 4. Expressions of 5-HT and SERT in ×Aa-T and DC as well as the effects of synthetic 5-HT on the proliferative response of ×Aa-T cells.
(A) ×Aa-T cells (4 × 105 cells/well) were co-cultured with DCs (2 × 104 cells/well, pretreated with MMC) in the presence and absence of Aa (107 fixed bacteria/mL/well). In addition, Fluoxetine or Desipramine (1 μM) was applied to the co-culture. After 3 days, the supernatants were collected for the measurement of 5-HT. *, higher than control co-culture with Aa (Student’s t test, P < 0.05), #, there is significant difference between the culture with and without Aa (Student’s t test, P < 0.05). (B) mRNA for SERT expressed in DCs was monitored using RT-PCR. Total RNA isolated from DCs incubated with or without LPS for 24 hours was subjected to RT-PCR using PCR primer set specific to SERT orβ-actin. As a positive control, brain tissue isolated from normal C57BL/6 mice was used. (C) Effects of synthetic 5-HT on the proliferative response of ×Aa-T cells in the co-culture with DC were evaluated. ×Aa-T cells (4 × 105 cells/well) were co-cultured with DCs (2 × 104 cells/well, pretreated with MMC) in the presence and absence of Aa (107 fixed bacteria/mL/well). Synthetic 5-HT (1 and 10 μM) was applied to the co-culture. [3H] thymidine (0.5 μCi) was applied to each well during the last 16 hours of a total 4-day culture. #, there is a significant difference between the culture with 5-HT (10 μM) and without 5-HT (Student’s t test, P < 0.05)