Endotoxin induces the expression of macrophage inflammatory protein 1 alpha mRNA by rat alveolar and bone marrow-derived macrophages.
Macrophage inflammatory protein 1 alpha (MIP-1 alpha) is a newly described cytokine that is present in large amounts in the culture supernatant of an endotoxin-stimulated murine macrophage-like cell line (RAW 264.7). There is increasing information that suggests that this cytokine mediates acute neutrophilic inflammation, although the mechanism of mediation is unknown. Data examining the production and regulation of MIP-1 alpha by primary rat macrophages are lacking, and MIP-1 alpha has not been studied previously in an animal model of endotoxin-induced neutrophilic alveolitis. In this study, we performed Northern analysis of steady-state rat MIP-1 alpha mRNA using an oligonucleotide probe complementary to amino acids 4-13 of murine MIP-1 alpha. Our data demonstrate that rat alveolar and bone marrow-derived macrophages can be induced by in vitro endotoxin treatment to express a 1.1-kb MIP-1 alpha mRNA. Expression of the mRNA could be elicited by treatment with 0.1 to 10.0 micrograms/ml of endotoxin in vitro with peak steady-state levels detectable up to 9 h after adding endotoxin to the media. Alveolar macrophages recovered by whole lung lavage from endotoxin-treated rats expressed increased amounts of the mRNA homologous to MIP-1 alpha mRNA when treated in vitro with endotoxin. We also found that rat neutrophils could be induced by endotoxin in vitro to express the MIP-1 alpha mRNA. We were able to identify MIP-1 alpha in culture supernatant from endotoxin-stimulated rat alveolar and bone marrow-derived macrophages by immunoprecipitation with a specific goat anti-murine MIP-1 alpha.(ABSTRACT TRUNCATED AT 250 WORDS)
Christman
J W
JW
Department of Veterans Affairs, Vanderbilt University, Nashville, Tennessee 37232-2650.
Blackwell
T R
TR
Cowan
H B
HB
Shepherd
V L
VL
Rinaldo
J E
JE
D000818:Animals; D001345:Autoradiography; D001853:Bone Marrow / Q000502:physiology*; D001992:Bronchoalveolar Lavage Fluid; D002460:Cell Line; D002478:Cells, Cultured; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004731:Endotoxins / Q000494:pharmacology*; D004926:Escherichia coli; D007700:Kinetics; D019402:Macrophage Inflammatory Proteins; D008264:Macrophages / Q000187:drug effects / Q000502:physiology*; D016676:Macrophages, Alveolar / Q000187:drug effects / Q000502:physiology*; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D015846:Monokines / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D009504:Neutrophils / Q000502:physiology; D015345:Oligonucleotide Probes; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013462:Sulfur Radioisotopes; D013997:Time Factors
D054405:Chemokine CCL3; D054407:Chemokine CCL4; D016207:Cytokines; D004731:Endotoxins; D019402:Macrophage Inflammatory Proteins; D015846:Monokines; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D013462:Sulfur Radioisotopes; D008715:Methionine