Differential changes in gene expression in human neutrophils following TNF‐α stimulation: Up‐regulation of anti‐apoptotic proteins and down‐regulation of proteins involved in death receptor signaling

Responses of human neutrophils to TNF‐α are complex and multifactorial. Exposure of human neutrophils to TNF‐α in vitro primes the respiratory burst, delays apoptosis and induces the expression of several genes including chemokines, and TNF‐α itself. This study aimed to determine the impact of TNF‐α exposure on the expression of neutrophil genes and proteins that regulate apoptosis. Quantitative PCR and RNA‐Seq, identified changes in expression of several apoptosis regulating genes in response to TNF‐α exposure. Up‐regulated genes included TNF‐α itself, and several anti‐apoptotic genes, including BCL2A1, CFLAR (cFLIP) and TNFAIP3, whose mRNA levels increased above control values by between 4‐20 fold (n = 3, P < 0.05). In contrast, the expression of pro‐apoptotic genes, including CASP8, FADD and TNFRSF1A and TNFRSF1B, were significantly down‐regulated following TNF‐α treatment. These changes in mRNA levels were paralleled by decreases in protein levels of caspases 8 and 10, TRADD, FADD, TNFRSF1A and TNFRSF1B, and increased cFLIP protein levels, as detected by western blotting. These data indicate that when neutrophils are triggered by TNF‐α exposure, they undergo molecular changes in transcriptional expression to up‐regulate expression of specific anti‐apoptotic proteins and concomitantly decrease expression of specific proteins involved in death receptor signaling which will alter their function in TNF‐α rich environments.