Purification and partial biologic characterization of a human lymphocyte-derived peptide with potent neutrophil-stimulating activity.
A novel neutrophil-activating peptide is detected in supernatants from mitogen-stimulated human T lymphocyte preparations. This chemotaxin was purified to apparent homogeneity by sequential Gel G-75 permeation chromatography, wide pore reversed phase (RP-8) HPLC, size exclusion HPLC, and reversed phase (RP-18) HPLC. Additional characterization of this lymphocyte-derived neutrophil-activating peptide (LYNAP) resulted in a single peak upon reversed phase HPLC and size exclusion HPLC. SDS-PAGE under nonreducing conditions revealed a single line at 10 kDa. LYNAP stimulated neutrophil chemotaxis (ED50 of 3 +/- 3 ng/ml), chemokinesis (ED50 of 2 +/- 2 ng/ml), and caused degranulation of cytochalasin B pretreated human neutrophils (ED50 of 20 ng/ml). In purified human monocytes, chemotactic responses to LYNAP at doses up to 100 ng/ml were absent, indicating nonidentity with a lymphocyte-derived monocyte chemotactic factor previously described by other workers. LYNAP shows biochemical and biologic similarities to a recently detected monocyte-derived neutrophil-activating peptide (MONAP). Moreover, desensitization experiments revealed cross-deactivation between LYNAP and MONAP, not, however, between these two chemotactic peptides and other well characterized polymorphonuclear leukocyte chemotaxins, e.g., C5a, FMLP, leukotriene B4, or platelet-activating factor. This finding points toward structure identity or homology of both chemotaxins, MONAP and LYNAP.
Schröder
J M
JM
Department of Dermatology, University of Kiel, Federal Republic of Germany.
Mrowietz
U
U
Christophers
E
E
D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002630:Chemotactic Factors / Q000096:biosynthesis / Q000302:isolation & purification* / Q000502:physiology; D002634:Chemotaxis, Leukocyte; D003429:Cross Reactions; D004305:Dose-Response Relationship, Drug; D005966:Glucuronidase / Q000378:metabolism; D006801:Humans; D016209:Interleukin-8; D008222:Lymphokines / Q000096:biosynthesis / Q000302:isolation & purification* / Q000502:physiology; D009000:Monocytes / Q000502:physiology; D009504:Neutrophils / Q000201:enzymology / Q000502:physiology*; D013601:T-Lymphocytes / Q000378:metabolism*
D002630:Chemotactic Factors; D016209:Interleukin-8; D008222:Lymphokines; D005966:Glucuronidase