Molecular cloning, functional characterization, and enzyme-linked immunosorbent assay of cynomolgus monkey Fas ligand.
Fas ligand (FasL) cDNAs were cloned and sequenced from cynomolgus, rhesus, and pig-tailed monkeys. The 840-bp cDNAs were identical among these three species of monkeys except for one nucleotide. The deduced 280 amino acids were completely identical and displayed 97% homology with human FasL (hFasL). Recombinant soluble FasL obtained from COS cells transfected with cynomolgus monkey FasL (cm-FasL) cDNA induced apoptosis in cells displaying human or cynomolgus monkey Fas-expressing cells. Several anti-human FasL monoclonal antibodies (mAbs) were able to neutralize the cytotoxic activity of monkey FasL, and a combination of mAbs was selected to obtain the most sensitive detection of monkey soluble FasL (sFasL) under sandwich enzyme-linked immunosorbent assay (ELISA). Plasma from normal monkey did not contain detectable levels of sFasL, whereas plasma from monkeys acutely infected with simian immunodeficiency virus (SIV) displayed increased levels of sFasL.
Kirii
Yasuyuki
Y
Research and Development Laboratories, Nippon Organon K.K., 1-5-90 Tomobuchi-cho, Miyakojima, Osaka 534-0016, Japan. Yasuyuki.Kirii@organon.jp
Inoue
Tomomi
T
Yoshino
Kohichiro
K
Kayagaki
Nobuhiko
N
Yagita
Hideo
H
Okumura
Ko
K
Shibata
Hiroaki
H
Yoshikawa
Yasuhiro
Y
Terao
Keiji
K
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D017209:Apoptosis; D015153:Blotting, Western; D019556:COS Cells; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary; D053222:Fas Ligand Protein; D000882:Haplorhini; D006801:Humans; D008562:Membrane Glycoproteins / Q000097:blood / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins; D017386:Sequence Homology, Amino Acid; D016097:Simian Acquired Immunodeficiency Syndrome / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D015302:Simian Immunodeficiency Virus; D013601:T-Lymphocytes / Q000473:pathology / Q000502:physiology; D014162:Transfection
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins