Monitoring of polymorphonuclear leukocyte functions in diabetes mellitus--a comparative study of conventional...

In this study neutrophil (PMN) phagocytic capacity was investigated using a conventional radiometric ingestion assay (IN) in comparison with PMN respiratory burst activity assessed by luminol-enhanced chemiluminescence (LCL) in response to phorbolesters and LCL induction during phagocytosis of opsonized Staphylococcus aureus (STLCL) in diabetes mellitus and healthy controls. PMN ingestion was measured with 3H-thymidine-labelled S. aureus in a kinetic radiometric assay. LCL and STLCL were assessed in a parallel detecting microtitre-plate luminometer (MTP-Reader). PMN of diabetic subjects showed a highly significant reduction of peak LCL in response to PMA as well as during phagocytosis of S. aureus (STLCL) compared to non-diabetic controls (p < 0.001 respectively). PMN ingestion in diabetic patients (51.8 +/- 4.6%) was significantly reduced compared to controls (78.3 +/- 6.2%) (p < 0.01). The in vitro data displayed impaired PMN oxidative burst activity at glucose concentrations > or = 13.8 mmol/L, whereas PMN IN was significantly reduced at glucose levels > or = 27.75 mmol/L. The control group showed a positive correlation of peak LCL response and IN (p < 0.05) but not of STCL and IN; in diabetic patients this was also true, but did not reach statistical significance. The data obtained in this study clearly demonstrated impaired PMN respiratory burst activity and markedly reduced phagocytic PMN functions in diabetic patients ex vivo and in vitro as measured by LCL and by ingestion of 3H-thymidine-labelled S. aureus suggesting inhibitory effects of elevated glucose concentrations on various PMN-functions, which might be of clinical importance concerning altered host defence.