Identification of an N-terminally truncated form of the chemokine RANTES and granulocyte-macrophage colony-stimulating factor as major eosinophil attractants released by cytokine-stimulated dermal fibroblasts.
Eosinophil (Eo) granule proteins and, rarely, intact Eos represent a characteristic histopathologic feature of the dermal part of affected tissue in atopic dermatitis and some allergic reactions. Dermal fibroblasts are a rich source of cytokines and inflammatory mediators; therefore, we have investigated whether these cells release Eo chemoattractants when stimulated with different stimuli. Eo-chemotactic activity was detected after stimulation of cells with TNF-alpha and IL-1, but not when phorbol ester, PHA, or medium alone was used. Biochemical characterization of Eo-chemotactic activity in supernatants of NF-alpha-stimulated cells revealed both heparin-binding and nonbinding activity. HPLC purification with subsequent N-terminal sequencing and mass spectrometric analysis showed that the heparin-binding Eo-chemotactic peak corresponded to the chemokine [Tyr-RANTES]66 that also contained [Ser-RANTES]68 as contaminant, whereas the nonheparin-binding activity was identified as granulocyte-macrophage CSF (GM-CSF) by the use of neutralizing Abs. [Tyr-RANTES]66 was found to show identical behavior in the chemotaxis assay system with respect to potency and efficacy as natural [Ser-RANTES]68. These findings support the hypothesis that dermal fibroblasts can play an important role in the recruitment of Eo by release of the chemokine RANTES together with GM-CSF.
Noso
N
N
Department of Dermatology, University of Kiel, Federal Republic of Germany.
Sticherling
M
M
Bartels
J
J
Mallet
A I
AI
Christophers
E
E
Schröder
J M
JM
D000595:Amino Acid Sequence; D002478:Cells, Cultured; D018946:Chemokine CCL5 / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology*; D002631:Chemotactic Factors, Eosinophil / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D003470:Culture Media; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis / Q000494:pharmacology*; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology; D008297:Male; D008969:Molecular Sequence Data; D012867:Skin / Q000166:cytology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D018946:Chemokine CCL5; D002631:Chemotactic Factors, Eosinophil; D003470:Culture Media; D007375:Interleukin-1; D014409:Tumor Necrosis Factor-alpha; D016178:Granulocyte-Macrophage Colony-Stimulating Factor