Increased expression of CR3 (C3bi receptor) on neutrophils in human inflammatory skin reactions

To help determine whether the neutrophils (PMN) found in skin inflammatory reactions are activated, we have compared the expression of the C3bi receptor (CR3) on such cells with that on autologous blood PMN in 10 pollen-sensitive subjects. Using skin chambers overlying denuded blister bases we collected PMN at 2 or 4 hr at sites of challenge with pollen antigen or buffer solution. These cells and PMN in autologous blood were incubated with monoclonal anti-CR3 antibody and the expression of CR3 was measured by indirect fluorescence and flow cytometry. Significantly more PMN were found at antigen than at buffer sites at 2 hr (7.02 +/- 0.45 X 10(5) vs 0.71 +/- 0.25 X 10(5] and at 4 hr (2.2 +/- 0.57 X 10(6) vs 5.47 X 10(5). The mean CR3 expressions on PMN at antigen and buffer sites were similar (117 +/- 7.4 vs 118 +/- 9.0); both were significantly greater than on blood PMN (17.6 +/- 1.5; P less than 0.005). PMN from both sites could be stimulated further in vitro with formyl-methionyl-leucyl-phenylalanine (FMLP) to express more CR3 to a level even greater than in FMLP-stimulated blood PMN (155 +/- 11 and 157 +/- 12, respectively, vs 108 +/- 7 in blood PMN). The incubation of blood PMN with the noncellular component of the chamber fluid led to a moderate (28-100%) increase in CR3 expression, but far less than the CR3 expression on the chamber fluid PMN themselves. Since surface CR3 is thought to be an activation marker important in PMN adhesion, these findings may be important in understanding the emigration of PMN in skin inflammatory reactions.