Identification of two neutrophil chemotactic peptides produced by porcine alveolar macrophages.
We have purified to homogeneity two distinct 10-kDa proteins with potent chemotactic activity for neutrophils from porcine alveolar macrophages incubated for 24 h with Escherichia coli endotoxin (lipopolysaccharide (LPS), 10 micrograms/ml). Neutrophil chemotactic activity in alveolar macrophage supernatants was concentrated by adsorption to SP-Sephadex, and purified by cation exchange and reversed phase high performance liquid chromatography. The first peptide, alveolar macrophage chemotactic factor (AMCF)-I, had chemotactic activity for both porcine and human neutrophils. The chemotactic activity for porcine neutrophils was detectable at 3 x 10(-10) M, peaked at 3 x 10(-8) M, and was comparable to that of zymosan-activated porcine serum. Segmental instillation of AMCF-I into porcine lungs caused marked neutrophil accumulation at 4 h in both bronchoalveolar lavage fluid and in lung tissue. The second peptide, AMCF-II, was active at 1.4 x 10(-9) M for porcine neutrophils, but it was less active for human polymorphonuclear neutrophils than was AMCF-I. Oligonucleotide probes to regions of the N-terminal sequences of AMCF-I and AMCF-II hybridized to mRNA recovered from LPS-stimulated alveolar macrophages. The N-terminal sequences and amino acid compositions indicate that AMCF-I and AMCF-II are distinct proteins, but that both have homologies with a family of peptide chemoattractants produced by human blood monocytes and platelets. Thus, alveolar macrophages stimulated with LPS produce two distinct 10-kDa cytokines with potent chemotactic activity for neutrophils. This indicates that there are two different peptide pathways by which alveolar macrophages can recruit neutrophils into the lung.
Goodman
R B
RB
Medical Research Service, Seattle Veterans Administration Medical Center, Washington.
Forstrom
J W
JW
Osborn
S G
SG
Chi
E Y
EY
Martin
T R
TR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002630:Chemotactic Factors / Q000302:isolation & purification* / Q000378:metabolism; D002634:Chemotaxis, Leukocyte; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid; D004247:DNA; D016209:Interleukin-8; D007975:Leukotriene B4 / Q000032:analysis; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000378:metabolism*; D011650:Pulmonary Alveoli / Q000166:cytology; D013552:Swine; D015054:Zymosan / Q000494:pharmacology
D002630:Chemotactic Factors; D016209:Interleukin-8; D008070:Lipopolysaccharides; C068711:alveolar macrophage-chemotactic factor-II; D007975:Leukotriene B4; D004247:DNA; D015054:Zymosan