Detection of the chemokine RANTES in cytokine-stimulated human dermal fibroblasts.
A novel family of structurally and functionally related polypeptides has recently been detected that are now referred to as chemokines. Within this family, a peptide with the acronym RANTES was shown to be chemotactic for memory T cells, monocytes, and eosinophilic and basophilic granulocytes, thus suggesting it plays an important role in chronic inflammatory and allergic diseases. Murine monoclonal antibodies as well as cDNA probes specific for human RANTES were raised and extensively characterized. With these antibodies, stimulated human dermal fibroblasts were shown to express intracellular RANTES peptide by immunocytochemistry. Furthermore, similar kinetics could be demonstrated in fibroblasts for both RANTES mRNA expression and secretion of RANTES peptide using Northern blot hybridization and sandwich-enzyme-linked immunosorbent assay, respectively. RANTES expression was induced upon stimulation with tumor necrosis factor-alpha as well as with interleukin-1 alpha and -beta in a concentration- and time-dependent manner. These results reinforce the role of both resident and circulating cells in the production and release of RANTES and their participation in inflammatory processes.
Sticherling
M
M
Department of Dermatology, University of Kiel, F.R.G.
Küpper
M
M
Koltrowitz
F
F
Bornscheuer
E
E
Kulke
R
R
Klinger
M
M
Wilhelm
D
D
Kameyoshi
Y
Y
Christophers
E
E
Schröder
J M
JM
D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D018946:Chemokine CCL5 / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D018925:Chemokines / Q000494:pharmacology*; D018076:DNA, Complementary / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000737:chemistry* / Q000187:drug effects; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis; D012867:Skin / Q000166:cytology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D018946:Chemokine CCL5; D018925:Chemokines; D018076:DNA, Complementary; D007375:Interleukin-1; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha