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IDENTIFICATION OF A 2ND NEUTROPHIL-CHEMOATTRACTANT CYTOKINE GENERATED DURING AN INFLAMMATORY REACTION IN THE RABBIT PERITONEAL-CAVITY INVIVO - PURIFICATION, PARTIAL AMINO-ACID-SEQUENCE AND STRUCTURAL RELATIONSHIP TO MELANOMA-GROWTH-STIMULATORY ACTIVITY

JOSE, PJ; COLLINS, PD; PERKINS, JA; BEAUBIEN, BC; TOTTY, NF; WATERFIELD, MD; ... WILLIAMS, TJ; + view all (1991) IDENTIFICATION OF A 2ND NEUTROPHIL-CHEMOATTRACTANT CYTOKINE GENERATED DURING AN INFLAMMATORY REACTION IN THE RABBIT PERITONEAL-CAVITY INVIVO - PURIFICATION, PARTIAL AMINO-ACID-SEQUENCE AND STRUCTURAL RELATIONSHIP TO MELANOMA-GROWTH-STIMULATORY ACTIVITY. BIOCHEM J , 278 493 - 497.

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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 vitro will provide an opportunity to investigate the importance of their co-release in the inflammatory process.

Type:Article
Title:IDENTIFICATION OF A 2ND NEUTROPHIL-CHEMOATTRACTANT CYTOKINE GENERATED DURING AN INFLAMMATORY REACTION IN THE RABBIT PERITONEAL-CAVITY INVIVO - PURIFICATION, PARTIAL AMINO-ACID-SEQUENCE AND STRUCTURAL RELATIONSHIP TO MELANOMA-GROWTH-STIMULATORY ACTIVITY
Keywords:VASCULAR-PERMEABILITY, IN-111 NEUTROPHILS, INTERLEUKIN-8, ACCUMULATION, PRETREATMENT, MONOCYTES, INVITRO, CELLS, SKIN

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