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Oxygen radical release from adhering polymorphonuclear leukocytes (PMN) has been implicated as an important feature of many vascular diseases. We developed a technique by which adherence and production of O2 radicals by PMN can be measured simultaneously. The technique combines the conventional nylon fiber assay for measuring adherence of PMN with concurrent scintillation counter measurement of chemiluminescence (CL) to assess O2 radical production by PMN. We found that adherence of PMN to nylon fiber is associated with increases in CL. Moreover, increases in CL appear to be dependent on generation of O2 radicals from PMN since they are not seen with PMN from a patient with chronic granulomatous disease (CGD) or in the presence of O2 radical scavengers, superoxide dismutase, or catalase. Furthermore, agents which increase the adherence of PMN to nylon fiber are associated with increases in CL. Use of this approach may facilitate simultaneous evaluation of adherence and O2 radical generation by PMN.

Chemiluminescence was used to determine opsonic requirements for interaction of Cryptococcus neoformans with human peripheral blood polymorphonuclear leukocytes (PMNLs) and monocytes (MNs). Peak chemiluminescence (mean 1 SD) was 172 1 25 x 103 for PMNLs and 50 1 13 x 103 for MNs with 20%0 pooled normal human serum vs. 10 1 3 x 103 for PMNLs and 6 1 1 x 103 for MNs without serum (P< 0.005 for each comparison). The nature of the serum requirement was investigated; alternative pathway serum complement components were necessary. The alternative pathway was activated directly by C. neoformans without the need for specific antibody. Chemiluminescence provided a quantitative and reproducible assay of the interaction of human phagocytes with C. neoformans and may be useful in studying opsonic requirements of other fungi. Previous investigations have suggested that heatlabile opsonins- most likely complement- are needed for the effective uptake of Cryptococcus neoformans by polymorphonuclear leukocytes (PMNLs), but the techniques that were used involved morphologic slide approaches, which are very difficult to evaluate and quantitate accurately