Human glioblastoma cells produce 77 amino acid interleukin-8 (IL-8(77)).
The production of interleukin 8 (IL-8), a neutrophil chemotactic factor, and its amino acid sequence were examined in glioblastoma cell lines in vitro. Neutrophil chemotactic activity was demonstrated in 9 conditioned media of 15 human glioblastoma cell lines. Tumor necrosis factor (TNF)-alpha stimulated secretion of the activity in 7 lines and induced secretion in 4 other lines. ELISA quantification disclosed that the conditioned media contained interleukin 8 (IL-8) in an amount equivalent to the chemotactic activity. The IL-8 secretion increased with the stimulation by TNF-alpha. Northern blot analysis and the RT-PCR method confirmed expression of mRNA in the glioblastoma cells and its augmentation by TNF-alpha and/or IL-beta. Reversed-phase HPLC following ion-exchange chromatography revealed that the chemotactic activity was a single peptide, which was determined to be IL-8 by the retention time and ELISA. Furthermore, amino acid analysis disclosed that a major part of the glioblastoma-cell derived IL-8 peptide was 77 amino acid IL-8 (IL-8(77); with the N-terminal sequence AVLPRSAKELRCQCI-).
Tada
M
M
Department of Neurosurgery, Hokkaido University School of Medicine, Sapporo, Japan.
Suzuki
K
K
Yamakawa
Y
Y
Sawamura
Y
Y
Sakuma
S
S
Abe
H
H
van Meir
E
E
de Tribolet
N
N
D000595:Amino Acid Sequence; D015152:Blotting, Northern; D001932:Brain Neoplasms / Q000378:metabolism*; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D002853:Chromatography, Liquid; D016207:Cytokines / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D005909:Glioblastoma / Q000378:metabolism*; D006801:Humans; D016209:Interleukin-8 / Q000031:analogs & derivatives* / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000187:drug effects; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D014407:Tumor Cells, Cultured
D016207:Cytokines; D016209:Interleukin-8; D012333:RNA, Messenger; C083743:interleukin 8 (77)