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

Biochem J. 1991 Sep 1;278 ( Pt 2)(Pt 2):493-7. doi: 10.1042/bj2780493.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Chromatography, High Pressure Liquid
  • Cytokines / chemistry
  • Cytokines / isolation & purification*
  • Electrophoresis, Polyacrylamide Gel
  • Growth Substances / chemistry
  • Growth Substances / isolation & purification*
  • Inflammation / metabolism*
  • Interleukin-8 / chemistry
  • Molecular Sequence Data
  • Peritoneal Cavity / pathology
  • Rabbits
  • Sequence Homology, Nucleic Acid

Substances

  • Cytokines
  • Growth Substances
  • Interleukin-8