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Characterization of Murine Bronchoalveolar Macrophage Respiratory Burst: Comparison of Soluble and Particulate Stimuli
Author(s) -
Sugar Alan M.,
Field Keith G.
Publication year - 1988
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.44.6.500
Subject(s) - bronchoalveolar lavage , biology , macrophage , respiratory burst , respiratory system , particulates , immunology , lung , in vitro , biochemistry , medicine , ecology , anatomy
Stimulation of the respiratory burst of murine bronchoalveolar macrophages obtained by lung lavage was studied using four different stimuli and different assay conditions. One soluble stimulus, phorbol myristate acetate (PMA), two intracellular particles, zymosan and Blastomyces dermatitidis conida, and one extracellular particle, B. dermatitidis yeast, were incubated with either freshly obtained macrophages in suspension or 2‐ and 48‐hour macrophage monolayers. Suspension cultures were incubated with stimuli for 90 minutes and monolayers for 10 minutes before O 2 − was assayed. PMA did not elicit O 2 − production in macrophage suspensions or 2‐hour macrophage monolayers, but 48‐hour macrophage monolayers exhibited a 13‐fold increase above control values ( P = .0001). On the other hand, zymosan elicited an increase in O 2 − production in both suspensions and monolayers, although monolayers incubated for 48 hours produced almost fourfold more O 2 − than the other systems ( P = .025). Opsonization had no effect on the ability of zymosan to elicit respiratory burst B. dermatitidis conidia resulted in a two‐ to threefold increase in O 2 − production in macrophage suspensions and a five‐ to eightfold increase in 48‐hour monolayers, representing significantly less respiratory burst stimulation than with either zymosan or PMA. Similarly, B. dermatitidis yeasts demonstrated similar submaximal stimulation of O 2 − , 3–4 times that over control values, and again this was less than zymosan and PMA. We conclude that 1) freshly obtained murine bronchoalveolar macrophages do not respond to PMA with an increase in O 2 − production, but that responsiveness is evident after 48 hours of incubation in monolayers; 2) B. dermatitidis conidia and yeasts do not stimulate respiratory burst activity to the same degree as zymosan or PMA; and 3) opsonization of zymosan is not necessary for stimulation of the murine bronchoalveolar macrophage oxidative burst, confirming previous data that functional complement receptors are not present on these cells.