HRF of 30 kDa: evidence for active synthesis.

A variety of mediators are involved in the pathophysiology of a number of clinical conditions. Among them histamine-releasing factors (HRF) act as secretagogue for basophils and mast cells to cause cell degranulation and histamine release. Studies on the kinetics of HRF production by activated mononuclear cells (MNC) have suggested an active synthesis of this cytokine. Using a [35S]-metabolic cell labeling method, we first analyzed the capacity of MNC from patients with allergic rhinitis to ragweed to actively synthesize HRF. We then found that stimulation of MNC with allergen promoted a synthesis of large quantities of proteins, including a protein of about 30 kDa. Secondly, we developed a method for the enrichment of HRF molecules from crude supernatants of ragweed-stimulated MNC using ion exchange and gel filtration chromatography. We observed that the yield of HRF activity is seen just before the chymotrypsinogen marker. The estimated molecular weight was 30-35 kDa.