OBJECTIVE: To characterize the innate immune proteins expressed by human endometriosis cell line MD-E and begin to determine modulation of expression by hormones.DESIGN: This is an in vitro descriptive study.MATERIALS AND METHODS: Human endometriosis cell line MD-E maintained in RPMI 1640 10% heat inactivated fetal calf serum in a CO2 incubator were grown to confluence and split using standard trypsinization technique. In the hormone modulation studies, cells were washed with culture medium by centrifugation, suspended in fresh medium, and plated in 12 well plates. When near confluent the medium was replaced with fresh medium with/without hormone (25 ng/ml cetrotide) for 24 hours. Cells were then harvested by trypsinization and stained for LL-37 and aromatase. For IHC the trypsinized cells were washed with PBS, spotted on positively charged slides, air dried and fixed for 10 min in cold 4° acetone. The fixed cells were stained using mouse monoclonal or affinity purified rabbit polyclonal antibodies and Dako Mouse or Rabbit Envision Plus Staining kits according to their protocol. For negative controls, rabbit or mouse DAKO control reagents were subsituted for primary antibodies. All slides were processed and evaluated using Leitz microscope.Table 1Expression of Innate Immune Factors and Aromatase by MD-ELL-37AromataseHAD1HAD5HAD6HBD2cryopyrinControl++++++++++++++-- Open table in a new tab Glucocorticoid, progesterone, and estrogen receptors were positive. Aromatase was stongly expressed. There was no change in expression of LL-37 or aromatase by cetrotide.CONCLUSIONS: Endometrosis cells can express multiple innate immune proteins, which may play a role in the pathophysiology of endometriosis via influencing the inflammatory process. MD-E strongly expressed LL-37, which has been associated with ovarian cancer. We plan to continue our studies on the potential effects of steroid signaling (estrogen, progesterone, dexamethasone and letrozole) on these innate immune factors. OBJECTIVE: To characterize the innate immune proteins expressed by human endometriosis cell line MD-E and begin to determine modulation of expression by hormones. DESIGN: This is an in vitro descriptive study. MATERIALS AND METHODS: Human endometriosis cell line MD-E maintained in RPMI 1640 10% heat inactivated fetal calf serum in a CO2 incubator were grown to confluence and split using standard trypsinization technique. In the hormone modulation studies, cells were washed with culture medium by centrifugation, suspended in fresh medium, and plated in 12 well plates. When near confluent the medium was replaced with fresh medium with/without hormone (25 ng/ml cetrotide) for 24 hours. Cells were then harvested by trypsinization and stained for LL-37 and aromatase. For IHC the trypsinized cells were washed with PBS, spotted on positively charged slides, air dried and fixed for 10 min in cold 4° acetone. The fixed cells were stained using mouse monoclonal or affinity purified rabbit polyclonal antibodies and Dako Mouse or Rabbit Envision Plus Staining kits according to their protocol. For negative controls, rabbit or mouse DAKO control reagents were subsituted for primary antibodies. All slides were processed and evaluated using Leitz microscope. Glucocorticoid, progesterone, and estrogen receptors were positive. Aromatase was stongly expressed. There was no change in expression of LL-37 or aromatase by cetrotide. CONCLUSIONS: Endometrosis cells can express multiple innate immune proteins, which may play a role in the pathophysiology of endometriosis via influencing the inflammatory process. MD-E strongly expressed LL-37, which has been associated with ovarian cancer. We plan to continue our studies on the potential effects of steroid signaling (estrogen, progesterone, dexamethasone and letrozole) on these innate immune factors.