Quantitation of tryptase, chymase, Fc epsilon RI alpha, and Fc epsilon RI gamma mRNAs in human mast cells and basophils by competitive reverse transcription-polymerase chain reaction

J Immunol. 1995 May 15;154(10):5472-80.

Abstract

Competitive reverse transcription-PCR assays developed for human tryptase, chymase, Fc epsilon RI alpha, and Fc epsilon RI gamma mRNA molecules were applied to the HMC-1 leukemic mast cell line, the KU812 leukemic basophil cell line, mast cells dispersed from lung and skin, and peripheral blood basophils. Relative amounts of alpha-tryptase and beta-tryptase mRNA were determined by analysis of BseAI digests of PCR products. Tryptase expression was highest in tissue-derived mast cells, lowest in basophils and KU812 cells, and intermediate in HMC-1 cells. beta-Tryptase mRNA predominated in HMC-1 and KU812 cells; mixtures of alpha- and beta-tryptase were found in tissue mast cells; and alpha-tryptase predominated in basophils. Chymase mRNA was more abundant in skin-derived (nearly all of the MCTC type) than lung-derived (variable amounts of MCTC and MCT cells) mast cells. Small amounts of chymase mRNA were detected in HMC-1 cells; none was found in basophils, in KU812 cells, or in the one preparation of 100% MCT cells derived from lung. Comparable amounts of Fc epsilon RI alpha and Fc epsilon RI gamma mRNA molecules were measured in basophils and tissue-derived mast cells, lesser amounts were detected in KU812 cells, and almost none was detected in HMC-1 cells. Thus, steady state levels of the granule and membrane resident molecules examined in our study are transcriptionally regulated in mast cells and basophils.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • Basophils / chemistry*
  • Basophils / enzymology
  • Basophils / immunology
  • Binding, Competitive
  • Cells, Cultured
  • Chymases
  • DNA, Complementary / analysis
  • Humans
  • Mast Cells / chemistry*
  • Mast Cells / enzymology
  • Mast Cells / immunology
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods
  • RNA, Complementary / analysis
  • RNA, Messenger / analysis
  • Receptors, IgE / analysis*
  • Serine Endopeptidases / analysis*
  • Tryptases

Substances

  • DNA, Complementary
  • RNA, Complementary
  • RNA, Messenger
  • Receptors, IgE
  • Serine Endopeptidases
  • chymase 2
  • Chymases
  • Tryptases