Transient enrichment of p47phox on OpZ phagosomes in X-linked CGD neutrophils. (A) gp91phox is undetectable in X-linked CGD neutrophils. PMN proteins from four normal individuals (odd lanes) or three persons with X-linked CGD (AlS, MK, TB, even lanes) were resolved by SDS-PAGE and then immunoblotted with antibody to gp91phox as described in Materials and Methods. Individuals deficient in gp91phox also lacked detectable p22phox(data not shown). (B and C) Kinetics of p47phox association with OpZ phagosomes in X-linked CGD neutrophils. Adherent X-linked CGD PMNs ingested OpZ for 0 to 15 minutes before processing for IFM. Fixed cells were double-stained with MoAb to p57 and pAb to p47phoxfollowed by secondary antibodies coupled to Texas red or FITC, respectively. (B) Representative fluorescence data for X-linked CGD PMNs. Data shown are from MK. Note that p47phox was enriched on forming phagosomes, but was not retained on these structures once particle internalization was complete. Arrows, phagosomes. (C) Pooled data (mean ± SD) for PMNs isolated from six persons with X-linked CGD (TS, DS, MK, TB, AlS, and HB). All samples were assayed in duplicate or triplicate and at least 100 phagosomes were scored/sample/time. (•), p57; (○), p47phox.