7,12-Dimethylbenz[a]anthracene (DMBA)

7,12-Dimethylbenz[a]anthracene (DMBA) is an immunosuppressor as well as a potent organ-specific carcinogen.

In splenocytes, which is isolated from mice, lymphocyte function is completely suppressed by the treatment with 50 μM DMBA. The susceptibility of splenocytes isolated from mEH-null mice is lower than from wildtype mice on the immunosuppression activity of DMBA.^[1]

The metabolic activation of DMBA in the spleen plays a key role in the splenic immunotoxicity, mEH-null mice are more resistant to DMBA-induced cytotoxicity as well as immunosuppression.^[1]

• Cell lines

  Splenocytes

• Concentrations

  10 μM, 20 μM, 50 μM

• Incubation Time

  --

• Method

  Spleens are harvested and placed in a sterile phosphatebuffered saline. Cell suspensions are prepared by gentle dissociation using glass slides. Debris is separated from cells by filtration and cell suspensions are centrifuged at 250 x g for 5 min. Cells are seeded in a volume of 100 µl in 96-well plates. Mitogens were added in a volume of 10 µl at final concentrations of 25 µg/ml for LPS and 5 µg/ml for PHA, and then the cells were incubated in 5% CO2 at 37°C for 3 days. The cells were harvested onto filter strips using a harvester and [3 H]-thymidine incorporation was determined by liquid scintillation. For mitogen-induced lymphocyte proliferation assays after DMBA treatment in vitro, cell preparations from untreated mice were simultaneously exposed to mitogen (LPS) and DMBA.

• Animal Models

  mEH-null mice

• Dosages

  30 mg/kg, 100 mg/kg

• Administration

  i.p.