Fig. 4.
Association of p67phox with OpZ phagosomes. (A) Adherent PMNs from a normal control or patient HB with X-linked CGD, ingested OpZ for 0 to 15 minutes at 37°C as described above. Fixed cells were stained with pAbs to p67phox followed by secondary antibodies coupled to FITC and phagosomes were scored for p67phox using IFM. Data are the average of duplicate samples and 50 to 100 phagosomes were scored/sample/time. (•), control; (○), X-linked CGD. Note that p67phox was transiently recruited to OpZ phagosomes in X-linked CGD PMNs. (B) Adherent p47phox-deficient PMNs ingested OpZ for 0 to 15 minutes at 37°C before processing for IFM. Samples were double-stained to detect p67phox and p57 as described above. Data indicate the average ± SD of duplicate samples from a single experiment where 100 phagosomes/sample/time were scored. Comparable data were obtained using PMNs from an unrelated p47phox-deficient individual. (•), p57; (○), p67phox. By IFM, p47phox was undetectable (not shown).