Increased cytotoxicity of soluble Fas ligand by fusing isoleucine zipper motif.
Fas (CD95) ligand (FasL) has the ability to induce apoptosis in Fas-expressing glioma cells by binding to Fas. Several molecular species have been designed to be soluble Fas ligands for therapeutic purposes. We successfully constructed a chimeric soluble FasL by fusing an isoleucine zipper motif for self-oligomerization and a FLAG sequence to the extracellular domain of the human Fas ligand (FIZ-shFasL). The cytotoxic effect of FIZ-shFasL on Jurkat cells was equivalent to that of membrane-bound FasL and approximately 10-fold stronger than that of agonistic anti-Fas antibody (CH-11). Flow cytometric analysis demonstrated that the differential Fas expression of human brain tumor cell lines partially correlated with levels of apoptosis through FIZ-shFasL. The upper limit of FIZ-shFasL for safe systemic administration to rat is estimated as below 2 microg/ml in plasma concentration. FIZ-shFasL could be applicable as a therapeutic agent for cancer.
Shiraishi
Tetsuya
T
Department of Neurosurgery, Faculty of Medicine, Saga University, Japan. tetsuya@csl.sony.co.jp
Suzuyama
Kenji
K
Okamoto
Hiroaki
H
Mineta
Toshihiro
T
Tabuchi
Kazuo
K
Nakayama
Kazuyuki
K
Shimizu
Yusuke
Y
Tohma
Junko
J
Ogihara
Takuo
T
Naba
Hiroyasu
H
Mochizuki
Hidenori
H
Nagata
Shigekazu
S
D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000633:toxicity; D017209:Apoptosis / Q000187:drug effects*; D001835:Body Weight / Q000187:drug effects; D045744:Cell Line, Tumor / Q000187:drug effects / Q000473:pathology; D002470:Cell Survival / Q000187:drug effects; D053222:Fas Ligand Protein; D005260:Female; D006801:Humans; D007532:Isoleucine / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics / Q000633:toxicity*; D019169:Jurkat Cells; D016350:Leucine Zippers; D008562:Membrane Glycoproteins / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics / Q000633:toxicity; D009929:Organ Size / Q000187:drug effects; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage* / Q000633:toxicity*; D012995:Solubility; D019014:fas Receptor / Q000378:metabolism*
D000970:Antineoplastic Agents; C505661:FASLG protein, human; D053222:Fas Ligand Protein; C505662:Faslg protein, rat; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D019014:fas Receptor; D007532:Isoleucine