Purification and amino acid analysis of two human glioma-derived monocyte chemoattractants

Teizo Yoshimura, E. A. Robinson, S. Tanaka, E. Appella, J. I. Kuratsu, E. J. Leonard

Research output: Contribution to journalArticle

381 Citations (Scopus)

Abstract

Two chemoattractants for human monocytes were purified to apparent homogeneity from the culture supernatant of a glioma cell line (U-105MG) by sequential chromatography on Orange A-Sepharose, an HPLC cation exchanger, and a reverse phase HPLC column. On SDS-PAGE gels under reducing or nonreducing conditions, the molecular masses of the two peptides glioma-derived chemotactic factor 1 and 2 were 15 and 13 kD, respectively. Amino acid composition of these molecules was almost identical, and differed from other cytokines that have been reported. The NH2 terminus of each peptide was apparently blocked. When tested for chemotactic efficacy, the peptides attracted ~30% of the monocytes added to chemotaxis chambers, at the optimal concentration of 10-9 M. Potency and efficacy were comparable with that of FMLP, which is often used as a reference attractant. The activity was chemotactic rather than chemokinetic. In contrast to their interaction with human monocytes, the pure peptides did not attract neutrophils. These pure tumor-derived chemoattractants can now be compared with attractants produced by normal cells and evaluated for their biological significance in human neoplastic disease.

Original languageEnglish
Pages (from-to)1449-1459
Number of pages11
JournalJournal of Experimental Medicine
Volume169
Issue number4
Publication statusPublished - 1989
Externally publishedYes

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Chemotactic Factors
Glioma
Monocytes
Amino Acids
Peptides
High Pressure Liquid Chromatography
Chemotaxis
Sepharose
Cations
Chromatography
Polyacrylamide Gel Electrophoresis
Neutrophils
Gels
Cytokines
Cell Line
Neoplasms

ASJC Scopus subject areas

  • Immunology

Cite this

Yoshimura, T., Robinson, E. A., Tanaka, S., Appella, E., Kuratsu, J. I., & Leonard, E. J. (1989). Purification and amino acid analysis of two human glioma-derived monocyte chemoattractants. Journal of Experimental Medicine, 169(4), 1449-1459.

Purification and amino acid analysis of two human glioma-derived monocyte chemoattractants. / Yoshimura, Teizo; Robinson, E. A.; Tanaka, S.; Appella, E.; Kuratsu, J. I.; Leonard, E. J.

In: Journal of Experimental Medicine, Vol. 169, No. 4, 1989, p. 1449-1459.

Research output: Contribution to journalArticle

Yoshimura, T, Robinson, EA, Tanaka, S, Appella, E, Kuratsu, JI & Leonard, EJ 1989, 'Purification and amino acid analysis of two human glioma-derived monocyte chemoattractants', Journal of Experimental Medicine, vol. 169, no. 4, pp. 1449-1459.
Yoshimura, Teizo ; Robinson, E. A. ; Tanaka, S. ; Appella, E. ; Kuratsu, J. I. ; Leonard, E. J. / Purification and amino acid analysis of two human glioma-derived monocyte chemoattractants. In: Journal of Experimental Medicine. 1989 ; Vol. 169, No. 4. pp. 1449-1459.
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