Inhibition of lens epithelial cells by Fas-specific antibody activating Fas-Fas ligand system.
OBJECTIVE
To detect cell specific apoptosis factors, Fas and Fas ligand, and the common intracellular apoptosis modulators, interleukin-1 beta converting enzyme (ICE)-like protease (caspase 1), Bcl-2, Bcl-xL and Bax in lens epithelial cells (LEC) of human cataracts. To study the effects of Fas-stimulating monoclonal antibody on inhibition of LEC proliferation.


METHODS
Reverse-transcriptase-polymerase chain reaction (RT-PCR) was used to detect Fas, Fas ligand, caspase 1, Bcl-2, Bcl-xL and Bax, after cDNA was synthesized from the total RNA isolated from human cataractous LEC obtained by capsulotomy during cataract surgery. Fas-stimulating monoclonal antibody was added at the concentrations of 10, 30, 100, 300 and 1000 ng/ml to the incubation medium of human cataractous LEC; and the specimens were incubated for 24 h at 37 degrees C with 5% CO(2) circulation and 100% humidity. The specimens were then stained with Hoechst 33342, and the number of apoptotic cells was counted.


RESULTS
Fas, caspase 1, Bcl-2, Bcl-xL and Bax mRNA were detected by RT-PCR. Fas ligand mRNA was not detected by RT-PCR. At each concentration, Fas-stimulating monoclonal antibody significantly inhibited LEC proliferation.


CONCLUSIONS
Human cataractous LEC expressed mRNA of Fas and various modulators of apoptosis pathways. Fas-stimulating monoclonal antibody may have the potential to prevent posterior capsule opacification after cataract surgery by inhibiting LEC proliferation.
Nishi
O
O
Nishi Eye Hospital, Osaka, Nakamichi, Higashinari-ku, Japan okihior@nishi-ganka.or.jp
Nishi
K
K
Wada
K
K
Ohmoto
Y
Y
Akura
J
J
D000368:Aged; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D020170:Caspase 1 / Q000235:genetics / Q000378:metabolism; D002386:Cataract / Q000378:metabolism / Q000473:pathology; D002455:Cell Division / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D053222:Fas Ligand Protein; D006801:Humans; D007908:Lens, Crystalline / Q000187:drug effects* / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D008875:Middle Aged; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein; D019014:fas Receptor / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; C494826:BAX protein, human; C494811:BCL2L1 protein, human; C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein; D019014:fas Receptor; D020170:Caspase 1