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Increased circulating monocyte activation in patients with unstable coronary syndromes

Christian V. Zalai, MSc^* , M. Dean Kolodziejczyk, MSc^* , Linda Pilarski, PhD^||, Alexander Christov, PhD^*, Patric N. Nation, DVM^¶, Marita Lundstrom-Hobman, PhD, Wayne Tymchak, MD, FRCP(C), FACC^#, Vladimir Dzavik, MD, FRCP(C), FACC^{# **}, Dennis P. Humen, MD, FRCP(C), FACC^{# ||}, William J. Kostuk, MD, FRCP(C), FACC, George Jablonsky, MD, FRCP(C), FACC, Peter W. Pflugfelder, MD, FRCP(C), FACC, James E. Brown, MD, FRCP(C), FACC and Alexandra Lucas, MD, FRCP(C), FACC^{*,* #}

^* John P. Robarts Research Institute, University of Western Ontario, London, Ontario, Canada
Department of Microbiology and Immunology, University of Western Ontario, London, Ontario, Canada
Department of Medicine, University of Western Ontario, London, Ontario, Canada
Cardiovascular Research Group, University of Alberta, Edmonton, Alberta, Canada
^|| Department of Oncology, University of Alberta, Edmonton, Alberta, Canada
^¶ Department of Lab Animal Sciences, University of Alberta, Edmonton, Alberta, Canada
^# Department of Medicine, University of Alberta, Edmonton, Alberta, Canada
^** Department of Medicine, University of Toronto, Toronto, Ontario, Canada

View larger version (58K): [in a new window]   Figure 1 (A) Immunostaining for CD14, a monocyte-specific marker, demonstrated migration of mononuclear cell (MC) isolated from patients with unstable angina (UA) through a Matrigel-coated filter. (B) Bar graphs demonstrate a significant increase (p < 0.05) in the mean migration rates of MCs isolated from patients with UA on comparison with cells isolated from patients with stable angina or normal coronary arteries. No significant shifts in migration rates were detected for lymphocytes. (C) A significant increase in cellular migration rates was detected after treatment with lipopolysaccharide (LPS) for cells isolated from normal donors (p < 0.001) but not for cells isolated from patients with unstable coronary artery disease. Relative MC migration count was obtained by dividing MC cell counts at each LPS dose by untreated control counts. Results are presented as mean cell counts per high power field of all donor samples within a category ± SE. *p 0.05. AMI = acute myocardial infarction; CCS = Canadian Cardiovascular Society; L = lymphocyte; M = monocyte; N = normal; P = pore; SA = stable angina.

View larger version (42K): [in a new window]   Figure 2 Membrane fluidity assays: A significant increase in membrane fluidity is evident in pyrene fluorescence emission spectra recorded from labeled mononuclear cells (MC) after treatment with thrombin (A). Patients with unstable angina (UA) demonstrated a significantly altered membrane fluidity profile (B) on comparison with cells obtained from normal individuals (p < 0.01), patients with stable angina (p < 0.001), and patients with acute myocardial infarction (MI) (p < 0.05). Thrombin treatment resulted in a significant change in the measured membrane fluidity of cells obtained from normal individuals (p < 0.05), but did not significantly alter the fluidity of cells obtained from patients with UA (C). Saline-treated cells (striped bars) obtained from patients with UA show a similar profile to those thrombin-treated cells (black bars) obtained from normal individuals. Data represent the mean Iexc/Imon ratios ± SE. Cells were grouped according to the Canadian Cardiovascular Society guidelines. *p 0.05.

View larger version (44K): [in a new window]   Figure 3 Significant increases in serum levels of soluble CD14 (sCD14) (p < 0.01) were found in patients with unstable coronary artery disease as compared to normal donors and patients with acute myocardial infarction (MI). The level of sCD14 did not differ significantly from individuals with stable angina (p = 0.06), but did show a trend toward an increase. Data represent the mean level of sCD14 expressed in g/ml; error bars represent SE. UA = unstable angina.