Further characterization of histamine releasing chemokines present in fractionated supernatants derived from human mononuclear cells.
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.


CONCLUSIONS
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.
Kuna
P
P
Department of Medicine, SUNY-Stony Brook, Health Sciences Center 11794-8160, USA.
Reddigari
S R
SR
Rucinski
D
D
Kaplan
A P
AP
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001667:Binding, Competitive / Q000276:immunology; D014408:Biomarkers, Tumor; D002458:Cell Fractionation; D002474:Cell-Free System / Q000737:chemistry / Q000276:immunology; D018932:Chemokine CCL2 / Q000097:blood / Q000276:immunology / Q000494:pharmacology; D018946:Chemokine CCL5 / Q000097:blood / Q000276:immunology / Q000494:pharmacology; D018925:Chemokines / Q000097:blood / Q000737:chemistry*; D002852:Chromatography, Ion Exchange; D006636:Histamine Release / Q000187:drug effects*; D006801:Humans; D007106:Immune Sera / Q000494:pharmacology; D016209:Interleukin-8 / Q000097:blood / Q000276:immunology / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism*; D008222:Lymphokines / Q000037:antagonists & inhibitors / Q000097:blood / Q000737:chemistry*; D010455:Peptides / Q000097:blood / Q000276:immunology / Q000494:pharmacology; D000090324:Tumor Protein, Translationally-Controlled 1; D001620:beta-Thromboglobulin
D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D018932:Chemokine CCL2; D018946:Chemokine CCL5; D018925:Chemokines; D007106:Immune Sera; D016209:Interleukin-8; D008222:Lymphokines; C061615:PPBP protein, human; D010455:Peptides; D000090324:Tumor Protein, Translationally-Controlled 1; D001620:beta-Thromboglobulin; C009862:connective tissue-activating peptide