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During two successive years the phagocytic capacity of the granulocytes of 800 children from areas with environmental pollution and with clean air was examined and compared. To this end, the luminol chemiluminescence during phagocytosis of opsonized zymosan was measured in whole blood. In all areas the phagocytic activity of the granulocytes of boys was higher than that of girls. In area with environmental pollution it was higher than in clean-air areas. The cells disposition towards phagocytosis was lower in clean-air areas than in areas with an environmental burden. Boys showed a more rapid increase in phagocytosis than girls. Between both years of examination distinct differences were observed in the level of chemiluminescence. The phagocytic defense systems of children who grew up in areas rated as having an environmental burden, showed a distinct change in activity as opposed to that of children from so-called clean-air areas. It could be proved that the phagocytic defense system of clinically healthy children reacts to environmental irritants with a change of its activity state.

Granulocytes (PMNs) were harvested from 16 normal subjects by filtration leukapheresis (FL) in which the filter loading time (FLT) ranged between 30 min and 4 h, and the rate of filter perfusion was 60 ml whole blood x min-1. The function of PMNs eluted from filters with or without the aid of manual tapping (MT) at each of the various FLTs was assessed in vitro by tests of chemotaxis, phagocytosis and bactericidal capacity. The results indicated that the function of PMNs in vitro was compromised by eluting filters with MT, and this appeared to be independent of the FLT. PMNs collected without MT were nearly normal relative to the control regardless of the FLT.

Inoculation of mature gerbils with BCG gave protection to subsequent infection with B. divergens when inoculated by the intracardiac and intraperitoneal routes, the latter showing a dose dependent relationship. BCG vaccination was most effective in immature gerbils (less than 4 weeks old), which are innately resistant to B. divergens. Vaccination of gerbils with killed Propionesbacterium acne and zymosan A failed to elicit a protective response, which contrasts conspicuously with rodent babesia studies. Incubation of B. divergens-infected gerbil blood with hydrogen peroxide produced parasite inhibition only at the highest concentration and treatment of parasitized gerbils with the oxidative radical inducer, alloxan monohydrate, gave equivocal results so it is evident that, unlike Plasmodium spp., B. divergens is not significantly susceptible to the action of reactive oxygen forms.

Chemiluminescence (CL) occurs as a consequence of phagocytosis of bacteria or inert particles by human polymorphonuclear neutrophils. This phenomenon appears to be related to the production of singlet oxygen and/or electronically excited carbonyl groups which relax with light emission. We report that human monocytes also produce CL after phagocytosis of opsonized bacteria, fungi, or zymosan particles. Monocytes produce approximately one-third of the CL produced by polymorphonuclear neutrophils for all three particles, reflecting possible metabolic differences between cells of these types. Addition of exogenous superoxide dismutase reduced the peak production of CL by 65% for cells of both types, whereas raising the temperature of the cultures from 25 to 37 C approximately doubled the peak CL responses.

N-formylmethionyl (F-Met) peptides, when added alone to macrophages or polymorphonuclear leukocytes (PMN), were found to induce a chemiluminescent response of shorter duration than that produced by the commonly employed particulate stimulant, zymosan. The cellular nature of F-Met peptide-induced chemiluminescence was indicated by its dependence on cell concentration, and by its inhibition by cell disruption, heat inactivation, or previous maximal stimulation by the peptides. Comparison of PMN and macrophages from different species showed that the maximal chemiluminescent response seen in the dose-response curve of F-Met- Phe was different in different cell types. Chemiluminescence reached highest values in human PMN, it was intermediate in guinea pig macrophages and PMN, and in rabbit PMN; but it was nonexistent in rabbit alveolar macrophages and very low in rabbit peritoneal macrophages. A definite relationship was observed between peptide structure and chemiluminescent activity. Met-Phe, F- Met and Phe were inactive even at millimolar concentrations, while F-Met-Phe caused chemiluminescence at micromolar concentrations. Four active peptides were tested in guinea pig, rabbit, and human PMN, and in guinea pig alveolar and peritoneal macrophages. The relative activity of these peptides was the same in all cells studied, e.g. F-Met-Leu-Phe >> F-Met-Phe > F-Met-Val > F- Met-Ala. The values of ED50 for each peptide were also comparable to previously reported ED50 values of these peptides in inducing lysosomal enzyme release. These results were seen both in the presence and absence ofthe chemiluminescent oxidant indicator, luminol. Low concentrations of superoxide dismutase (10 mug/ml) completely inhibited chemiluminescence caused by the F-Met peptides, suggesting the involvement of 0(2)(-) or O(2)(-)-derived compounds in this response. Sodium azide, an inhibitor of peroxidase reactions, had either no effect or a slight inhibitory effect on chemiluminescence. However, when the extracellular release of lysosomal enzymes was induced by cytochalasin B, an azide- inhibitable enhancement of chemiluminescence was seen in PMN, but not in macrophages. This effect appears to be correlated with the presence of granule-associated myeloperoxidase. Although azide-inhibitable peroxidases could be a potential source of light, they did not appear to be a significant contributor in these experiments. Based on these results and on those of previous investigators, we postulate that the F-Met-peptides stimulate 0(2)(-) production in addition to stimulating lysosomal enzyme release and chemotaxis. The similar structure- activity relationship which appears to exist for these processes may indicate that they are all initiated by a single receptor mechanism. Since F-Met peptides are formed in bacteria it is likely that their actions represent an important physiologic response.

The terminal complement complex (TCC) was quantified in sera from patients with a genetic deficiency of C8 alpha-gamma or C8 beta. The individual sera contained only trace amounts of TCC compared with a normal serum pool. The content of TCC increased after mixing the two sera, which was consistent with reconstitution of C8 activity. Only a moderate increase in TCC was obtained after zymosan activation of the individual sera, whereas activation of the mixture resulted in high amounts of TCC. C8 was demonstrated in the TCC of both deficient sera. These results may indicate that functional C8 is present in trace amounts despite the genetic deficiency, and that the terminal pathway may function to some extent although not enough to be detectable in less sensitive assays.