Identification of a second neutrophil-chemoattractant cytokine generated during an inflammatory reaction in the rabbit peritoneal cavity in vivo. Purification, partial amino acid sequence and structural relationship to melanoma-growth-stimulatory activity.
The intraperitoneal injection of zymosan in the rabbit results in the generation of an inflammatory exudate containing oedema-forming and chemoattractant activities. Previous studies demonstrated the early appearance of the complement fragment C5a, followed by the generation of two mediators related to the cytokine interleukin-8 that were separable by cation-exchange h.p.l.c. N-Terminal amino acid sequencing identified one of these mediators as rabbit interleukin-8. This paper describes the purification of the second cytokine by cation-exchange, gel-filtration and reversed-phase h.p.l.c. The purified material had both oedema-forming and chemoattractant activity when assayed in rabbit skin in vivo. On SDS/PAGE a single 6-8 kDa band was observed and N-terminal amino acid sequencing of the reduced and alkylated protein positively identified 36 amino acids. This sequence revealed the rabbit homologue of melanoma-growth-stimulatory activity. The identification of these two cytokines in vivo will provide an opportunity to investigate the importance of their co-release in the inflammatory process.
Jose
P J
PJ
Department of Applied Pharmacology, National Heart and Lung Institute, London, U.K.
Collins
P D
PD
Perkins
J A
JA
Beaubien
B C
BC
Totty
N F
NF
Waterfield
M D
MD
Hsuan
J
J
Williams
T J
TJ
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D016207:Cytokines / Q000737:chemistry / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D006133:Growth Substances / Q000737:chemistry / Q000302:isolation & purification*; D007249:Inflammation / Q000378:metabolism*; D016209:Interleukin-8 / Q000737:chemistry; D008969:Molecular Sequence Data; D010529:Peritoneal Cavity / Q000473:pathology; D011817:Rabbits; D012689:Sequence Homology, Nucleic Acid
D016207:Cytokines; D006133:Growth Substances; D016209:Interleukin-8