Connective tissue-activating peptide-III and its derivative, neutrophil-activating peptide-2, release histamine from human basophils.
Connective tissue-activating peptide-III (CTAP-III) is a 9 kd platelet alpha-granule-derived growth factor. It stimulates the synthesis of DNA, hyaluronic acid, glycosaminoglycans, and proteoglycan core protein in human fibroblasts. Human mononuclear cell-derived proteases have been previously demonstrated to digest the N-terminal 15 residues of CTAP-III (total, 85 residues) to produce neutrophil-activating peptide-2 (NAP-2). CTAP-III and NAP-2 belong to a class of proteins (platelet factor 4, interleukin-8/NAP-1, etc.) associated with inflammation and wound repair. In our efforts to purify human mononuclear cells and platelet-derived histamine-releasing factors, we had previously discovered that mixtures of CTAP-III and NAP-2 released histamine from human basophils. We have now developed simple protocols for the purification of CTAP-III and NAP-2, independently, from calcium ionophore (A23187)-stimulated platelet supernatants by affinity chromatography and have established their identity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and N-terminal sequence analysis. Each of these related histamine between 2 and 10 micrograms/ml, a range identical to that obtained with CTAP-III/NAP-2 mixtures that we reported earlier. Thus, our data suggest that CTAP-III and NAP-2 independently release histamine from human basophils in dose ranges similar to ranges required for fibroblast stimulation by each.
Department of Medicine, State University of New York, Stony Brook.
S R
SR
Reddigari
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Kuna
G F
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Miragliotta
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D
Kornfeld
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Baeza
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Castor
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Kaplan
D001491:Basophils / Q000187:drug effects* / Q000276:immunology; D002846:Chromatography, Affinity / Q000379:methods; D003238:Connective Tissue / Q000276:immunology*; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006632:Histamine / Q000032:analysis; D006636:Histamine Release / Q000187:drug effects*; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D010455:Peptides / Q000302:isolation & purification / Q000494:pharmacology*; D010983:Plateletpheresis / Q000379:methods; D001620:beta-Thromboglobulin
C061615:PPBP protein, human; D010455:Peptides; D001620:beta-Thromboglobulin; C009862:connective tissue-activating peptide; D006632:Histamine