Characterization of a CD43/leukosialin-mediated pathway for inducing apoptosis in human T-lymphoblastoid cells.
The monoclonal antibody (mAb) J393 induces apoptosis in Jurkat T-cells. NH2-terminal amino acid sequence analysis identified the 140-kDa surface antigen for mAb J393 as CD43/leukosialin, the major sialoglycoprotein of leukocytes. While Jurkat cells co-expressed two discrete cell-surface isoforms of CD43, recognized by mAb J393 and mAb G10-2, respectively, only J393/CD43 signaled apoptosis. J393/CD43 was found to be hyposialylated, bearing predominantly O-linked monosaccharide glycans, whereas G10-2/CD43 bore complex sialylated tetra- and hexasaccharide chains. Treatment with soluble, bivalent mAb J393 killed 25-50% of the cell population, while concomitant engagement of either the CD3.TcR complex or the integrins CD18 and CD29 significantly potentiated this effect. Treatment of Jurkat cells with mAb J393 induced tyrosine phosphorylation of specific protein substrates that underwent hyperphosphorylation upon antigen receptor costimulation. Tyrosine kinase inhibition by herbimycin A diminished J393/CD43-mediated apoptosis, whereas inhibition of phosphotyrosine phosphatase activity by bis(maltolato)oxovanadium-IV enhanced cell death. Signal transduction through tyrosine kinase activation may lead to altered gene expression, as J393/CD43 ligation prompted decreases in the nuclear localization of the transcriptional regulatory protein NF-kappaB and proteins binding the interferon-inducible regulatory element. Since peripheral blood T-lymphocytes express cryptic epitopes for mAb J393, these findings demonstrate the existence of a tightly regulated CD43-mediated pathway for inducing apoptosis in human T-cell lineages.
Bristol-Myers Squibb Pharmaceutical Research Institute, Seattle, Washington 98121, USA.
T J
TJ
Brown
W W
WW
Shuford
W C
WC
Wang
S G
SG
Nadler
T S
TS
Bailey
H
H
Marquardt
R S
RS
Mittler
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000096:biosynthesis / Q000737:chemistry / Q000502:physiology*; D017209:Apoptosis; D001483:Base Sequence; D016227:Benzoquinones; D001665:Binding Sites; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002467:Cell Nucleus / Q000378:metabolism; D002846:Chromatography, Affinity; D004791:Enzyme Inhibitors / Q000494:pharmacology; D000939:Epitopes / Q000032:analysis; D005434:Flow Cytometry; D006020:Glycopeptides / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D019169:Jurkat Cells; D047029:Lactams, Macrocyclic; D051917:Leukosialin; D008213:Lymphocyte Activation; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism; D015345:Oligonucleotide Probes; D009844:Oligosaccharides / Q000737:chemistry; D010766:Phosphorylation; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011753:Pyrones / Q000494:pharmacology; D011809:Quinones / Q000494:pharmacology; D017828:Rifabutin / Q000031:analogs & derivatives; D012795:Sialoglycoproteins / Q000096:biosynthesis / Q000737:chemistry / Q000502:physiology*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000502:physiology*; D014157:Transcription Factors / Q000378:metabolism; D014638:Vanadates / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D016227:Benzoquinones; D004791:Enzyme Inhibitors; D000939:Epitopes; D006020:Glycopeptides; D047029:Lactams, Macrocyclic; D051917:Leukosialin; D016328:NF-kappa B; D015345:Oligonucleotide Probes; D009844:Oligosaccharides; D011753:Pyrones; D011809:Quinones; C465014:SPN protein, human; D012795:Sialoglycoproteins; D014157:Transcription Factors; D017828:Rifabutin; C074212:bis(maltolato)oxovanadium(IV); D014638:Vanadates; C020754:herbimycin; D011505:Protein-Tyrosine Kinases; D017027:Protein Tyrosine Phosphatases