Human lung mast cells: purification and characterization.

摘要: Detailed studies of the biochemistry and pharmacology mast cell-mediated inflammatory disorders have been hampered by inability to purify human cells. We now report techniques lung cells apparent homogeneity. The major purification steps are: 1) dispersion fragments into a single-cell suspension with enzyme combinations (pronase-chymopapain, collagenase-elastase); 2) partial countercurrent centrifugation elutriation (CCE); 3) affinity column chromatography. Enzymatic yielded suspensions congruent 10(6) per gram parenchyma in purities 1.2 9.7%. Dispersed responded comparably those parent challenge anti-human IgG pharmacologic agonists. Elutriation cell cell-enriched fractions up 70%. High purity were combined, passively sensitized purified penicillin (BPO)-specific IgE, BPO-affinity chromatography procedure. Post 29 +/- 3.5% increased 84 3% (range 65 98%) column. Short-term (24 hr) culture column-purified allowed adherence non-mast contaminants tissue plates, further increasing (up 100%). Purified intact functional as assessed dye exclusion, survival short-term culture, IgE-mediated histamine release, modulation release agonists adenosine, IBMX, prostaglandin E2, fenoterol.