Increased levels of granulocyte macrophage colony stimulating factor in hypercholesterolemic patients

Friday. 28 May 1999 Poster presentation: Inflammation, infection and cytokines in atherosclerosis

118 PI$

Age {yrs) SOP (mmHg)

GroupA (n = 18) Group B In = 201 controls In = 151

58.2±6.3 172±6.1 103±4.8 261.2 169.9 5:11.7 5:7.9 56.7±7.1 1705:6.8 105±4.2 218±12.3 138.3 ±8.7 54.3±5.9 125±4.3 88±3.8 208.5 133.2 5:14.8 5:91

DBP (mmHg)

TC" (mg/dl)

LDL-C" (mg/dl)

MCP-I"" MIP-Ia'"" (pg/mt) (pg/ml)

198.4 5:29.3 150.7 +33.2 116.2 +24.3

24.35:4.1 19.85:3.9 15.4±3.1

°p groupA vs B < 0.01. "°p group A vs B < 0.01.p group A vs controls < 0.001, p group B vs controls < 0.05. ' ' ' p groupA vs B > 0.05. p group A vs controls < 0.005, p group B vs controls < 0.05 in group A, MCP-I and MIP-la values were significantly correlated with LDL-C (r = 0.49, p < 0.05 and r = 0.56, p < 0.01, respectively). No correlation was found between smoking and C-C chemokine serum activity in two groups of patients. Conclusions: MCP-I and MIP-Ia serum levels are elevated in hypertensive patients, especially in those with significant hyperlipidemia. This elevation may reflect the initiation and progression of inflammatory atherosclerotic process triggered by arterial hypertension and enhanced by its coexistence with hyperlipidemia. INCREASED LEVELS OF G R A N U L O C Y T E M A C R O P H A G E C O L O N Y STIMULATING FACTOR IN H Y P E R C H O L E S T E R O L E M I C PATIENTS J. Parissis, K. Venetsanou, M. Kalantzi, D. Mentzikof, N. Ziras, K. Merkouris, M. Gikopoulos, M. Georgopoulou, S. Karas. Amalia Fleming

Hospital and St Anargiri Hospital. Athens. Greece Background: Hypercholesterolemia is considered a major risk factor for the development of atherosclerosis and induces an increased expression of cell adhesion molecules, such as ICAM-I, on the vascular endothelium. Furthermore, granulocyte macrophage-colony stimulating factor (GM-CSF) is a cytokine which may play an important role in multicellular events of atherosclerosis. This study investigates the impact of hypercholesterolemia on serum levels of GM-CSE Methods: Serum levels of GM-CSF and soluble ICAM-I (slCAM-I) were determined by commercially available Elisa tests in 20 hypercholesterolemic patients (TC: 278+18, LDL-C: 172+7, TG: 133+1 I, HDL-C: 41 + 3 mg/dl) with no clinical evidence of cardiovascular disease and in 15 gender -and age-matched normocholesterolemic subjects (TC: 196+7, LDL-C: 1055=6, TG: 128+9, HDL-C: 595=4 mg/dl). Patients did not receive any medications before blood sampling for measurements of serum GM-CSE Results: Hypercholesterolemic patients had higher levels of GM-CSF compared to controls (5.8+2.1 vs 2.1±0.9 pg/ml, p < 0.05). They also displayed increased serum levels of slCAM-I (236.15=19.7 vs 183.5±13.1 ng/ml, p < 0.01). A significant correlation was observed between GM-CSF levels and LDL-C values (r = 0.58, p < 0.01) as well as GM-CSF and slCAM-I levels (r = 0.78, p < 0.0011. Furthermore, sICAM-I levels were significantly correlated with LDL-C values (r = 0.69, p < 0.01 ). Conclusions: Hypercholesterolemia is associated with elevated serum levels of GM-CSE These results may be related with the endothelial dysfunction and the atherosclerotic inflammatory process induced by hypercholesterolemia. M Y O C A R D I A L INJURY M Y O C A R D I A L INJURY M A R K E R S AND ANTICARDIAC AUTO-ANTIBODIES. A PROSPECTIVE EVALUATION OF H U M O R A L A U T O I M M U N I T Y IN CORONARY H E A R T DISEASE G. Dangas, M.M. Konstadoulakis, A.M. Cohen, G.D. Kymionis, H. Kroubouzou, M.G. Toutouza, J.N. Nanas, A. Sadaniantz, C.I. Stefanadis, J.H. Chesebro, P.K. Toutouzas. Cardiology Research Foundation,

Washington, DC, Cardiovascular Institute, Mount Sinai School of Medicine, NY, Massachusetts General Hospital, Boston, MA. Brown University, Providence, R1, and National University of Athens. Greece Background: Development of cardiac auto-antibodies (AAb) has been described after surgical injury, but has unknown clinical implications. We hypothesized that ischemic myocardial injury may be in pa~ related to cardiac AAb, which may have developed from previous, possibly subclinical, exposure of intracellular cardiac proteins to the circulation. Methods: We investigated the development of lgG AAb against actin, myosin, and troponin after an acute clinical event, and their correlation

with CPK and Troponin-I (Tnl). Patients with prior myocardial infarction were excluded. We studied 33 consecutive hospitalized patients with acute coronary syndromes (ACS), and 25 controls, without evidence of coronary disease. Blood was drawn in-hospital, and at I and 3 months (mo) for Tnl, and AAb aginst actin, myosin, and troponin; CPK was measured only acutely. Since Tnl and CPK did not distribute normally, we used the non-parametric Wilcoxon z-test for comparisons of these parameters between AAb+ and A A b - patients. Results: In-hospital, 39% were actin AAb+, 44% myosin AAb+, and 27% troponin AAb+; in contrast, the respective AAb+ of the controls were 13%, 18%, 9% (p < 0.04 for AAb+ in ACS patients versus controls). At 1 month, 50% were actin+, 44% myosin+, and 33% troponin+, and no further change was observed at 3-month follow-up. Persistence of AAb+ at followup correlated with elevated Tnl in-hospital: z < 0.03 for both actin and myosin AAb+, at 1 and 3 months. Similarly, persistence of any AAb+ at I and 3 months occurred in patients with elevated in-hospital CPK (z < 0.01). Presence of a positive AAb in-hospital, correlated significantly with Tnl elevation at I month: z = 0.03 for myosin AAb+, z = 0.02 for troponin AAb+, z = 0.08 for actin AAb+. Persistence of AAb+ at 1 month correlated with Tnl elevation at 3 months: z = 0.04 for both actin and myosin AAb. Conclusions: Our findings indicate that myocardial injury is associated with presence of AAb against actin, myosin and troponin. Patients with elevated Tnl and CPK develop AAb at follow-up; this suggests a direct contribution of myocardial injury in the formation of AAb. Presence of AAb correlates with elevated Tnl at follow-up.

The stud), has been supported in part by grants from the National Institutes of Health (NIH), Bethesda. to the Mt. Sinai Clinical Research Center, New York, USA, and from the Central Health Council, Athens. Greece. A T H E R O G E N I C PROPERTIES OF HUMAN M O N O C Y T E S INDUCED BY THE CARBOXYL T E R M I N A L PROTEOLYTIC F R A G M E N T OF ALPHA-I -ANTITRYPSIN

S. Janciauskiene S. Lindgren. GastroenterrdoKv-Hepatology Division. Department of Medicine. Universi~, Hospital Malmo, 20502 Malmo, Sweden Backround: Atherosclerosis is characterized as a chronic inflammatory disease of the vessel wall. Following endothelial injury, monocytes attach to the subendothelium, penetrate into the vessel wall and form macrophage/foam cells by accumulating lipids. Release of various cytotoxic products from macrophages, such as interleukins, complement factor fragments, tumor necrosis factors, oxidized cholesterol, proteases and free radicals leads to further endothelial injury and the development of atheromatous plaques Alpha-l-antitrypsin (AAT) is serine proteinase inhibitor, acute phase protein produced by monocytes and is an important regulator of leukocyte elastase and proteinase-3. We have shown that the carboxyl terminal peptide (C36) resulting from proteinase cleavage of AAT polymerizes, and in its fibrillar form alters cellular cholesterol metabolism. To test for a possible link between the inflammation-generated C-36 peptide and cellular processes associated with atherogenesis, we studied the effects of the fibrillar form of this peptide at concentrations _> 2.5 Ixmol/L on human monocytes in culture. Methods: The following parameters were studied: 1251 -LDL binding, uptake and degradation, cholesterol ester formation, interleukin release (ELISA), cellular LDL receptor expression (using RT-PCR), glutathione reductase activity assay (spectrophotometry), CD36 expression (Western blot), AAT biosynthesis (metabolic labeling), monocyte morphology (light microscopy) Results: We found that fibrillar C-36 at concentrations > 5 I.tmol/L added into monocyte cultures for 24 hours increases LDL binding (from 138+17.5% to 499+32%, p < 0.001) and uptake (from 2 to 10-fold, p < 0.01) in a concentration dependent manner through transcriptional upregulation of LDL receptors Cup to 46+3.5%, p < 0.001) increase cholesterol ester formation by 75+ 9%, p < 0.01, induces cytokine production: IL-6 {8.6 fold), IL-I (3.8 fold) and TNFct (I0 fold) and ghitathione reductase activity (by 3--4 times) compared to controls, and upregulates AAT synthesis. The LDL scavenger receptor CD36 is also upregulated by fibrillar C-36, and native LDL in the the presence of C-36-activated monocytes is oxidized to a greater extent than with unactivated control monocytes. The majority of monocytes cultured for 24 hours in the presence of C-36 fibrils were transformed morphologically into macrophages. Conclusion: The stimulated biosynthesis of AAT during the chronic inflammatory process such as atherosclerosis might not only play a positive role as an inhibitor of proteases, but also a negative role as a protease substrate and a reservoir o f physiologically active degradation products. Our studies suggest that an imbalance in favor of the latter induces atherogenic

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