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Titolo:
CDNA CLONING OF GUINEA-PIG MONOCYTE CHEMOATTRACTANT PROTEIN-1 AND EXPRESSION OF THE RECOMBINANT PROTEIN
Autore:
YOSHIMURA T;
Indirizzi:
NCI,FREDERICK CANC RES & DEV CTR,IMMUNOPATHOL SECT,IMMUNOBIOL LAB,BLDG 560,ROOM 12-71 FREDERICK MD 21702
Titolo Testata:
The Journal of immunology
fascicolo: 11, volume: 150, anno: 1993,
pagine: 5025 - 5032
SICI:
0022-1767(1993)150:11<5025:CCOGMC>2.0.ZU;2-1
Fonte:
ISI
Lingua:
ENG
Soggetto:
BLOOD MONONUCLEAR LEUKOCYTES; AMINO-ACID ANALYSIS; CHEMOTACTIC PROTEIN; SPLEEN-CELLS; JE GENE; MCP-1; IDENTIFICATION; PURIFICATION; PRODUCT; RABBIT;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
34
Recensione:
Indirizzi per estratti:
Citazione:
T. Yoshimura, "CDNA CLONING OF GUINEA-PIG MONOCYTE CHEMOATTRACTANT PROTEIN-1 AND EXPRESSION OF THE RECOMBINANT PROTEIN", The Journal of immunology, 150(11), 1993, pp. 5025-5032

Abstract

Monocyte chemoattractant protein-1 (MCP-1) cDNA was cloned from guinea pig spleen cells stimulated with Con A. The cDNA comprised 647 bp with an open reading frame that encoded a 120 amino acid protein. The sequence similarity of the first 99 amino acids to human MCP-1 is 56%. Recombinant guinea pig MCP-1 was expressed in COS-7 cells, then purified by a three step procedure with orange A-agarose, carboxymethyl-HPLC,and reversed phase-HPLC. The purified protein was found around 25 kDaas a broad band on a polyacrylamide gel under reducing conditions. Guinea pig MCP-1 attracted about 34% of input human monocytes at 5 x 10(-9) M. Guinea pig peritoneal exudate macrophages migrated toward guinea pig MCP-1 dose dependently, but only 1% of input cells responded to guinea pig MCP-1 at its optimal concentration of 5 x 10(-9) M. Human MCP-1 attracted 1% of input guinea pig peritoneal exudate macrophages at its highest concentration of 2.5 x 10(-8) M. Neither human nor guinea pig MCP-1 attracted guinea pig peritoneal resident macrophages. These results suggest that monocytes lose responsiveness to MCP-1 after differentiating to macrophages. Finally, intradermal injection of the recombinant protein into guinea pigs caused marked macrophage infiltration. Cloned and expressed guinea pig MCP-1 will help in studying the role of MCP-1 in vivo.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 26/09/20 alle ore 14:41:50