Research Article EVALUATION OF NEUTROPHIL FUNCTION, OPSONISING CAPACITY AND LYMPHOCYTE PROLIFERATION FOR RISK OF DEVELOPING ISCHEMIC HEART DISEASE IN TYPE 2 DIABETES MELLITUS PATIENTS VIDYA BERNHARDT 1 , JANITA R. T. D'SOUZA 2 , ANUSHA SHETTY 4 , MANJULA SHANTARAM 1 , RAVI VASWANI 3 1 Department of Biochemistry, 2 Yenepoya Research Centre, 4 Department of General Medicine, 3 Received: 11 Jan 2012, Revised and Accepted: 12 Apr 2012 Yenepoya Medical College, Yenepoya University, Mangalore 575018, Karnataka, India Email: mirdad365@gmail.com ABSTRACT Background: Ischaemic heart disease (IHD) is a leading cause of mortality in type 2 diabetes mellitus (DM) patients. Elevated lipid profile levels may not always be associated with IHD. Increased reactive oxygen production, decreased chemotaxis, altered opsonization in DM patients alters phagocytosis. T and B lymphocyte proliferations are also found to be impaired. Objectives: Phagocytic index (PI), serum opsonization, T and B cell proliferation have been studied to evaluate their role as risk markers for developing type 2 DM mediated cardiac events. Methods: The patients were classified into three groups. Blood glucose, glycated haemoglobin (HbAlc) and ECG changes were taken into consideration when grouping. All the recruited patients had normal lipid profile and were not taking any lipid lowering drugs. Results: Both the study groups were compared to each other and the control. PI NO (without serum opsonisation) was decreased in group 1 & 2. Group 1 showed decreased PI (WBO, C3, IgG ). In group 2 forty patients had higher PI (WBO, C3, IgG), remaining 11 displayed values similar to group 2 indicating risk of developing IHD. T and B cell proliferation rate in groups 1 and 2 were decreased. The study cohort were grouped as Group 1 (DM-IHD) consisting of 33 patients with uncontrolled diabetes and ECG changes, Group 2 (DM) consisting of 51 patients with uncontrolled diabetes, without any ECG changes, both groups receiving antihypertensives and antidiabetic therapy. Group 3 (control) consisted of 30 healthy subjects with no history of DM or IHD and with normal fasting blood glucose levels and normal HBA1c values. Phagocytic index, opsonization with serum (WBO), IgG and C3 as well as T and B cell proliferation were assessed. Conclusions: PI can be used as a marker to predict IHD events in DM patients whose lipid values cannot predict IHD and can be used as part of an improved strategy for preventing atherosclerosis. Keywords: Ischaemic heart disease, Lymphocyte proliferation, Opsonisation, Phagocytic index, Type 2 Diabetes mellitus. INTRODUCTION Coronary artery disease (CAD) is a leading cause of mortality in patients with Diabetes Mellitus (DM) 1 . The various reported causes of atherosclerosis and CAD are smooth muscle cell proliferation induced by insulin stimulation that increases collagen synthesis within the vascular wall; increased accumulation of low density lipoproteins (LDL) 2 and its oxidized products 3 , premature senescence in vascular cells due to phenotypic changes in the vascular smooth muscle cell proliferation and adhesion molecule expression due to increased reactive oxygen species (ROS) 4 . The other reasons being decreased resorption of atherosclerotic plaques due to phagocytic dysfunction 5 . To prevent cardiovascular events in diabetic patients there is a need for simple markers that could identify patients with a risk of developing myocardial ischaemia. One of the commonly used predictor for coronary heart is the lipid profile 6 . Lipid profile may not always be elevated levels in DM led CAD patients indicating that not all DM patients with CAD develop an altered lipid profile. Thus lipid profile evaluation cannot always be used to predict cardiac events 7 Findings also suggest that polymorphonuclear leucocytes (PMNLs) from DM patients have an increased reactive oxygen metabolite production that may cause cell and tissue damage, opsonisation impairment (C3 and IgG) . Investigation into the role of other factors such as altered neutrophil phagocytic capacity, role of opsonins complement 3 (C3) and Immunoglobulin G (IgG) on phagocytic capacity and proliferation capacity of the T and B cells of the immune system may be helpful. 8 , decreased chemotaxis and decreased PMNLs phagocytic activity 9 which contributes to increased progression of atherosclerosis leading to CAD 10 . During opsonisation antigens are bound by antibody and/or complement molecules enhancing phagocytosis, important opsonins being IgG and C3 11 . Several lines of evidence suggest that PMNLs, which represent a major mechanism of innate immunity and inflammation which play a pivotal role in human atherosclerosis are altered in DM led CAD 12 . Peripheral PMNLs along with pro-inflammatory cytokines are found to be increased in the circulation in coronary artery diseases 13 . Patients with DM also display a significant increment in the basal levels of calcium in PMNLs and this is found to be associated with marked impairment in their phagocytosis 14 T and B lymphocyte proliferation rates are found to be impaired in DM . 15 . Common hypertensive drugs cause elevation in cell calcium, which reduces cell proliferation, mainly B cell proliferation along with PMNLs phagocytic function 16 . More over atherosclerosis being a chronic inflammatory disorder of the vessel wall, defective immune responses are known to influence disease progression 17 . We have studied the phagocytic function of neutrophils, serum opsonisation capacity, T and B cell proliferation in DM and DM Ischaemic heart disease (IHD) patients in order to assess these markers in predicting CAD events in DM patients. There is currently a great deal of interest in the role of immune system in vascular pathology. It is to be hoped that a better understanding of how immune response contributes to atherogenesis will suggest new targets for therapeutic intervention in cardiovascular disease. PATIENTS AND METHODS This prospective cohort study included 84 type 2 DM patients, with normal lipid profile and 30 healthy age and gender matched controls. The duration of the diabetes ranged from 3 to 7 years. Patients visiting on an outpatient basis to Tanvi Medical Center and Yenepoya Hospital, Mangalore, Karnataka, India were recruited for the study after obtaining their consent. The study was conducted after obtaining approval by the Yenepoya University Ethics Committee. Patients were sampled after fasting overnight, before taking any medication, after 10 – 15 minutes of resting in upright position. All the patients underwent a complete cardiac examination followed by International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 4, Suppl 3, 2012 A A c c a a d d e e m mi i c c S Sc c i i e e n n c c e e s s