Failure of Nitro Blue Tetrazolium reduction in the phagocytic vacuoles of leukocytes in chronic granulomatous disease

Failure to trigger the oxidative metabolic burst by normal macrophages: possible mechanism for survival of intracellular pathogens.

“…Slides were incubated at 37°C for varying periods of time, rinsed in warmed saline, fixed in methanol, and counterstained with 0.2% safranin. The percentage of cells that contained formazan-stained organisms within phagocytic vacuoles was determined microscopically by counting 100-200 cells that contained organisms (24). Only cells that contained organisms that were stained deeply blue were scored as formazan positive.…”

Pyridine nucleotide-dependent superoxide production by a cell-free system from human granulocytes.

“…In view of the rather inconclusive nature of the evidence concerning defects in oxygen-metabolizing enzymes, it has been suggested that the metabolic lesion in CGD may involve an enzyme system physiologically connected to, but not identical with, the oxygen-consuming enzymes directly responsible for the respiratory burst (1). This possibility finds some support in studies showing differences in degranuation between normal and CGD cells (26,27).…”

“…In attempting to reproduce in a cell-free system one of the characteristic functional defects of CGD granulocytes, namely the failure of these cells to reduce NBT, these workers found instead active NBT reduction (not necessarily 02-mediated) by a cell-free preparation from CGD granulocytes that were unable to reduce NBT before their disruption (26). We report here similar observations, namely, that 02-production by lysates prepared from CGD cells was the same as its production by lysates from normal cells, notwithstanding our previous demonstration of a profound defect in 02-production by intact CGD granulocytes (10).…”

Immunoelectron Microscopy of Protein Kinase C in Resting and Phagocytosing Macrophages.

“…However, Brieland et al [3] reported that translocation of PKC is not necessary for the O-2 generation in macrophages. Microscopic analysis to visualize O-2 generated by human neutrophils was first reported by Nathan et al [22], and more recently by Hirai et al [11]. They showed that the cytochemical reaction is localized in phagocytic vacuoles and at areas of the plasma membrane attached to forein particle, but the free surface of the plasma membrane is completely devoid of reaction products.…”