IAP in Crohn's disease

The study examines the effect of cellular inhibitor of apoptosis protein (cIAP) inhibition on monocyte and PBMC cytokine production and cell death in healthy and Crohn's disease subjects. PBMCs from healthy controls were exposed to 4h of doses ranging from 0.1 to 1.0 micromolar of SMAC mimetics (SM) LCL161 and BV6 and co-incubated with LPS. SM consistently and dose-dependently reduced mRNA production of TNF-a, IL6 and IL-1b as determined by standard qPCR. A similar effect was seen on protein expression of TNF-a as determined by ELISA. To determine if this reduction in cytokine production was secondary to cell death, standard flow cytometry using ANX5 antibodies and nuclear dye 7AAD was performed. PBMCs were exposed from 4-16 hours to SM and/or LPS. Overall, neither of the stimulations LPS, SM or combined LPS/SM induced cell death. To determine if the effect was specific to LPS-induced signaling, PBMCs from healthy controls and from patients with inactive Crohn’s disease were exposed to ..., PBMCs and monocytes were havested from healthy subjects and patients with Crohn's disease. Cells were then cultured and exposed to SMAC mimetics LCL161 or BV6 as indicated and/or inhibitors of upstream or downstream pathways (infliximab, zVAD, GSK 872, Nec1s, and NSA) as indicated. Cytokine production (TNF-a, IL-6, IL-1b) was measured by qPCR and ELISA (only TNF-a) as indicated. Cytotoxiciity was assessed by flow cytomertry using ANX5 antibodies and nuclear dye 7AAD as indicated. NFkB p65 and p-p65 expression was measured by standard Western blotting. Readouts were as follows: qPCR data normlized to LPS-only exposed cells, protein levels as pg/mL, flow cytometry data as % cells, and Western blot quantification as pixels (ImageJ). , XML, # IAP in Crohn's disease

### Brief summary of dataset contents, contextualized in experimental procedures and results.

The study examines the effect of cellular inhibitor of apoptosis protein (cIAP) inhibition on monocyte and PBMC cytokine production and cell death in heathy and Crohn's disease subjects. PBMCs from healthy controls were exposed to 4h of doses ranging from 0.1 to 1.0 micromolar of SMAC mimetics (SM) LCL161 and BV6 and co-incubated with LPS. SM consistently and dose-dependently reduced mRNA production of TNF-a, IL6 and IL-1b as determined by standard qPCR. A similar effect was seen on protein expression of TNF-a as determined by ELISA. To determine if this reduction in cytokine production was secondary to cell death, standard flow cytometry using ANX5 antibodies and nuclear dye 7AAD was performed. PBMCs were exposed from 4-16 hours to SM and/or LPS. Overall, neither of the stimulations LPS, SM or combined LPS/SM induced cell death. To determine if the effect was specif...