Human lung mast cells: Purification and characterization

E. S. Schulman, D. W. MacGlashan, S. P. Peters, R. P. Schleimer, H. H. Newball, L. M. Lichtenstein

Research output: Contribution to journalArticlepeer-review

Abstract

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

Original languageEnglish (US)
Pages (from-to)2662-2667
Number of pages6
JournalJournal of Immunology
Volume129
Issue number6
StatePublished - Dec 1 1982

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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