Premium
Further characterization of histamine releasing chemokines present in fractionated supernatants derived from human mononuclear cells
Author(s) -
KUNA P.,
REDDIGARI S. R.,
RUCINSKI D.,
KAPLAN A. P.
Publication year - 1996
Publication title -
clinical and experimental allergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.462
H-Index - 154
eISSN - 1365-2222
pISSN - 0954-7894
DOI - 10.1111/j.1365-2222.1996.tb00628.x
Subject(s) - histamine , chemokine , peripheral blood mononuclear cell , interleukin 8 , chemistry , elution , chromatography , cytokine , microbiology and biotechnology , immunology , biochemistry , biology , in vitro , endocrinology , receptor
Summary Background Histamine releasing factors (HRF) are members of the β chemokine family of cytokines and have been characterized using recombinant proteins. Mononuelcar cell and/or platelet supernatants have been shown to contain HRF and the initial void peak obtained using Mono Q anion exchange chromatography possesses such activity, as do two later peaks eluted from the column. Objective We wished to further characterize the activity present in the void peak and determine which of the chemokines present are responsible for the activity measured. Methods We fractionated the void peak obtained from Mono Q chromatography on Mono S. The elution profile of individual chemokities was determined and the fractions were assayed for histamine releasing capability. We used monospecific antisera to inhibit the activity and quantitate the contribution of each protein. Results The fractions contained MCP‐1/MCAF, CTAPIII/NAP‐2, IL8, and a small quantity of RANTES. About 90‐95% of the total histamine containing capability was attributable to MCP‐1/MCAF. There was a small contribution by CTAPIII/NAP‐2, and RANTES, and no activity associated with IL8. Conclusion MCP‐1/MCAF is the critical HRF present in the initial void peak obtained by anion exchange chromatography of supernatants derived from human mononuclear cells and platelets. The α chemokine CTAPIII/NAP‐2 has relatively weak activity and IL8 has none although they are prominent in this fraction and overlap with MCP‐1/MCAF. RANTES makes a minor contribution but most of it is eluted in a later peak.