A role for apoptosis in the control of neutrophil homeostasis in the circulation: insights from CD18-deficient mice

Abstract

The control of neutrophil turnover in the circulation is a key event in homeostasis and inflammation. Using CD18- deficient (CD18 ^−/−) mice that show a 19-fold increase of blood neutrophil counts when compared with wild-type animals (CD18 ^+/+), we found that apoptosis of peripheral neutrophils was significantly reduced from 27.4% in the wild-type to 4.8% inCD18 ^−/− mice within 4 hours after isolation as measured by analysis of DNA content. This was confirmed by detecting CD16 expression, nuclear morphology, and internucleosomal DNA degradation. In contrast, no difference in apoptosis was observed in neutrophils derived from the bone marrow. Neutrophilia and delayed neutrophil apoptosis were also present inCD18 ^−/−/interleukin 6 (IL-6 ^−/−) double knockout mice. Moreover, plasma ofCD18 ^−/− mice was not able to delay apoptosis of CD18 ^+/+neutrophils and plasma ofCD18 ^+/+ mice did not augment apoptosis of CD18 ^−/−neutrophils. However,CD18 ^−/− neutrophils revealed an up-regulation of the antiapoptotic gene bcl-X_l and a down-regulation of the proapoptotic gene bax-α compared withCD18 ^+/+ neutrophils suggesting that this delayed apoptosis. Accordingly, down-regulation of Bax-α using antisense technique delayed apoptosis and prolonged neutrophil survival. The replacement of the hematopoietic system of CD18 ^+/+ mice by a 1:1 mixture of CD18 ^+/+ andCD18 ^−/− hematopoietic cells abolished the delay of apoptosis in peripheralCD18 ^−/− neutrophils and prevented neutrophilia. Altogether, this suggests that a delay of neutrophil apoptosis inCD18 ^−/− mice causes an alteration of neutrophil homeostasis, which may induce the massive increase of peripheral neutrophil counts. Thus, apoptosis seems to be critically involved in the control of neutrophil turnover in the circulation.