Effects of Serum on Superoxide Release from Single Pulmonary Alveolar Macrophages

Pulmonary inflammation is accompanied by increased microvascular permeability and leakage of plasma into alveolar spaces. Serum has been shown to affect superoxide (O2-) production from pulmonary alveolar macrophages (PAM) in vitro, and in some cases serum has been shown to be necessary for stimulation of PAM. An electro-optical method was used to measure O2- production from single PAM by measuring the reduction of nitroblue tetrazolium (NBT) to a diformazan precipitate (NBTH2) from videorecorded images of individual cells. PAM were stimulated by their initial attachment to culture dishes independent of serum. The addition of 2% serum to the culture medium increased NBTH2 production during attachment, whereas concentrations of 5 and 10% serum had no additional effect. In contrast, PAM incubated for 2 hrs could not be stimulated by phorbol myristate acetate (PMA) or zymosan particles to produce O2- unless serum was present during the incubation and/or stimulation; however, NBTH production was similar in all cases of serum addition. Also2, serum alone did not stimulate adherent PAM indicating that serum is necessary but not sufficient for stimulation of attached. PAM. These results suggest that changes in microcirculatory permeability may have a pronounced effect on superoxide production by PAM and that serum may help regulate or condition these cells during inflammation of the lung. (Supported by USBOM Gll35142.)