CD13 and CD10 expression of granulocytes as markers for the functioning of the immune system: quantification of the expression of membrane molecules using 1:1 labeled monoclonal antibodies and flow cytometry.

Abstract

HLA-DR expression on monocytes as a marker for the functioning of the immune system is known to be severely depressed in immunodeficiency. Up to now, other markers for the function of the immune system are scarce. In the peripheral blood of patients with open heart surgery the expression of the membrane peptidases neprilysin/CD10 and aminopeptidase N/CD13, was determined on granulocytes in comparison to the monocytic HLA-DR expression. We used the QuantiBRITE flow cytometry system, which yields an absolute antigen expression value (antibodies bound per cell) and may be useful in standardizing surface antigen expression analysis. This system makes use of a highly purified phycoerythrin-labeled antibody with a 1:1 fluorochrome-to-protein ratio, and multilevel calibrated beads with known absolute phycoerythrin fluorescence. Our results show that both membrane peptidases on granulocytes show a similar time-course of expression after heart surgery as do HLA-DR molecules on monocytes, with a decrease from days one to three and a subsequent recovery to normal values. In future analyses a possible relationship between the immunodeficiency of patients and a diminished expression of both membrane peptidases on granulocytes has to be investigated.