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Further Characterization of Histamine Releasing Chemokines Present in Fractionated Supernatants Derived From Human Mononuclear Cells

Further Characterization of Histamine Releasing Chemokines Present in Fractionated Supernatants Derived From Human Mononuclear Cells

P Kuna, S R Reddigari, D Rucinski, A P Kaplan

Clin Exp Allergy. 1996 Aug;26(8):926-33.

PMID: 8877158

Abstract:

Background:
Histamine releasing factors (HRF) are members of the beta chemokine family of cytokines and have been characterized using recombinant proteins. Mononuclear 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 chemokines 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 alpha 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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42413095	MCP-1/MCAF human			Price

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