Quantitative assessment of rabbit alveolar macrophage function by chemiluminescence

Rabbit alveolar macrophages (RAM) were cultured for 24 hr with concentrations ranging from 3 to 12 micrograms/ml of vanadium oxide (V2O5), a known cytotoxic agent, or with high-molecular-weight organic by-products from coal gasification processes. After culture the cells were harvested and tested for functional capacity using three types of indicators: (1) luminol-amplified chemiluminescence (CL), which quantitatively detects photon emission due to respiratory burst activity measured in a newly designed instrument with standardized reagents; (2) the reduction of nitro blue tetrazolium-saturated polyacrylamide beads, a semiquantitative measure of respiratory burst activity; and (3) phagocytic efficiency, defined as percentage of cells incorporating immunoglobulin-coated polyacrylamide beads. Chemiluminescence declined linearly with increasing concentrations of V2O5 over the dose range tested. Dye reduction and phagocytic efficiency similarly decreased with increasing V2O5 concentration, but were less sensitive indicators of functional impairment than CL as measured by the amount required to reduce the response to 50% of untreated cells. The effect of coal gasification condensates on RAM function varied, but in general these tests also indicated that the CL response was the most sensitive indicator.