The expression of genes modulating programmed cell death in normal human polymorphonuclear neutrophils.
Normal human polymorphonuclear neutrophils (PMN) have a short life and die in progression via apoptosis. In order to understand the molecular basis of PMN apoptosis, the expression of apoptosis-related (Fas, Fas-ligand, p53, and c-myc) and survival-related (bcl-2) genes was detected by flow cytometry, Western blot and reverse transcription-assisted polymerase chain reaction (RT-PCR). We found that Fas and Fas-ligand (FasL) were expressed on the surface of most of the cells. However, the disappearance of FasL was much faster than Fas after 24 h incubation. p53 and bcl-2 were also expressed in the cytoplasm of most of the cells. In contrast, the expression of c-myc was negligible in PMN. The addition of monoclonal anti-human Fas antibody (25 micrograms/ml) to PMN suspension enhanced whereas anti-FasL antibody (25 micrograms/ml) suppressed PMN apoptosis in 48 h incubation. These results suggest that the activation of Fas pathway induced by Fas-FasL interaction among PMNs is one of the mechanisms for spontaneous PMN apoptosis. Lack of proto-oncoprotein c-myc expression in PMN is responsible for their non-proliferative property and may aggravate the spontaneous apoptosis of the cells.
Hsieh
S C
SC
Department of Medicine, Veterans General Hospital, Taipei, Taiwan, Republic of China.
Huang
M H
MH
Tsai
C Y
CY
Tsai
Y Y
YY
Tsai
S T
ST
Sun
K H
KH
Yu
H S
HS
Han
S H
SH
Yu
C L
CL
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D017209:Apoptosis / Q000235:genetics*; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D053222:Fas Ligand Protein; D005434:Flow Cytometry; D015870:Gene Expression; D019254:Genes, bcl-2; D016259:Genes, myc; D016158:Genes, p53; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology; D051379:Mice; D009504:Neutrophils / Q000166:cytology* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D019014:fas Receptor / Q000235:genetics
D000911:Antibodies, Monoclonal; D017931:DNA Primers; C505661:FASLG protein, human; D053222:Fas Ligand Protein; C505658:Fasl protein, mouse; D008562:Membrane Glycoproteins; D019014:fas Receptor