THE INFLUENCE OF INSULIN-DEPENDENT DIABETES-MELLITUS (IDDM) DURATION ON SUPEROXIDE ANION AND HYDROGEN-PEROXIDE PRODUCTION BY POLYMORPHONUCLEAR NEUTROPHILS

We address the question whether oxygen metabolism of polymorphonuclear neutrophils (PMN) is influenced by disease duration in patients with insulin-dependent diabetes mellitus (IDDM). PMN were isolated from pat ients with IDDM of various durations and from healthy controls. We mea sured PMN production of superoxide anions (O-2(-)) by cytochrome c red uction (see Babior, B.M. et al. (1973) J. Clin. Invest. 52, 741-746) a nd PMN production of hydrogen peroxide (H2O2) by phenol red oxygenatio n (see Pick, E. (1980) J. Immunol. Methods 38, 161-169) in three group s of IDDM patients subdivided according to disease duration (group A: IDDM less than 10 years; group B: IDDM of 10-15 years; group C: IDDM o f more than 15 years) and in control healthy subjects (group H). Unsti mulated O-2(-) production in all IDDM patients was not statistically d ifferent from control values (A: 4.3 +/- 0.4 nmol/10(6) PMN per 30 min ; B: 4.1 +/- 0.2 nmol/10(6) PMN per 30 min; C: 4.9 +/- 0.9 nmol/10(6) PMN per 30 min; and H: 3.5 +/- 0.2 nmol/10(6) PMN per 30 min, respecti vely). In contrast, stimulated O-2(-) production was significantly low er in both patients with 10-15 years, and patients with more than 15 y ears, duration of IDDM than in controls (B: 25.7 +/- 2.5 nmol/10(6) PM N per 30 min; C: 21.1 +/- 3.4 nmol/10(6) PMN per 30 min and H: 42.2 +/ - 1.1 nmol/10(6) PMN per 30 min, respectively) correlating with diseas e duration (r = -0.44. P < 0.033). The stimulated O-2(-) production in patients with less than 10 years duration of IDDM (A: 35.7 +/- 1.9 nm ol/10(6) PMN per 30 min) was slightly lower than in controls. H2O2 pro duction of unstimulated PMN (A: 4.0 +/- 0.5 nmol/10(6) PMN per 30 min; B: 4.4 +/- 0.8 nmol/10(6) PMN per 30 min and C: 4.4 +/- 1.0 nmol/10(6 ) PMN per 30 min, respectively) was much higher than those in controls . In contrast, stimulated H2O2 production did not differ statistically from the value noticed in healthy subjects. The results obtained migh t indicate that production of H2O2 by unstimulated cells is increased in diabetic patients while generation of O-2(-) by stimulated neutroph ils is markedly impaired, suggesting that toxic oxygen species product ion might be influenced by disease duration.