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(Circulation. 1999;99:2858-2860.)
© 1999 American Heart Association, Inc.
Brief Rapid Communications |
Remnant Lipoprotein Levels in Fasting Serum Predict Coronary Events in Patients With Coronary Artery Disease
From the Department of Cardiovascular Medicine, Kumamoto University School of Medicine (K.K., H.D., K.T., H.K., H.S., Y.M., R.T., T.S., H.O., S.S., M.Y., H.Y.), Kumamoto, Japan, and Japan Immunoresearch laboratories (T.N., K.N.), Takasaki, Japan.
Correspondence to Kiyotaka Kugiyama, MD, PhD, Department of Cardiovascular Medicine, Kumamoto University School of Medicine, Honjo 1-1-1, Kumamoto City, Japan 860-8556. E-mail kiyo{at}gpo.kumamoto-u.ac.jp
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Abstract |
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Background—Remnant lipoproteins are atherogenic, but assays of remnants have not been available in routine clinical laboratories because of the lack of practical and validated methods. A simple and reliable method for such an assay, using an immunochemical approach, has recently been developed. This study prospectively examined whether remnant lipoprotein levels in fasting serum, measured by our method, may have prognostic value in patients with coronary artery disease (CAD).
Methods and Results—Remnant lipoprotein levels in fasting serum
were measured in 135 patients with CAD by an immunoaffinity mixed gel
containing anti-apolipoprotein (apo) A-1 and anti–apoB-100 monoclonal
antibodies. Patients were followed up for 
36 months until occurrence
of 1 of the following clinical coronary events: recurrent or
refractory angina pectoris requiring coronary
revascularization, nonfatal myocardial infarction,
or cardiac death. Kaplan-Meier analysis demonstrated a
significantly higher probability of developing coronary events
in patients with the highest tertile of remnant levels (>5.1 mg
cholesterol/dL; 75th percentile of distribution of remnant
levels) than in those with the lowest tertile of remnant levels (3.3
mg cholesterol/dL; 50th percentile of the distribution).
Higher levels of remnants were a significant and independent predictor
of developing coronary events in multivariate
Cox hazard analysis including the following covariates: extent
of coronary artery stenosis, age, sex, smoking,
hypertension, diabetes mellitus,
hypercholesterolemia, low HDL
cholesterol, and
hypertriglyceridemia.
Conclusions—Higher levels of remnant lipoproteins in fasting serum predict future coronary events in patients with CAD independently of other risk factors. Thus, measurement of fasting remnant levels, assessed by the current immunoseparation method, may be helpful in assessment of CAD risk.
Key Words: atherosclerosis • coronary disease • lipoproteins • prognosis • risk factors
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Introduction |
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Remnant lipoproteins, derived from VLDL and chylomicrons, are considered atherogenic.1 2 3 However, a simple and reliable method to isolate remnant lipoproteins has not been available because remnant lipoproteins have heterogeneous properties.4 Nakajima et al have recently developed a technique to isolate remnant-like lipoproteins using an immunoaffinity mixed gel containing anti-apolipoprotein (apo) A-1 and anti–apoB-100 monoclonal antibodies.4 5 6 7 8 This unique anti–apoB-100 monoclonal antibody has been shown to recognize apoB-100 in LDL and most VLDL but not in apoE-enriched VLDL.4 5 6 7 8 We and others have shown that this technique can isolate apoE-rich VLDL particles containing apoB-100 together with chylomicron remnants containing apoB-48, neither of which binds to the immunoaffinity gel.4 5 6 7 8 We7 8 further demonstrated that high levels of remnant lipoproteins, as assessed by this method, directly induced endothelial vasomotor dysfunction, an early sign of atherosclerosis, and were independently associated with the presence of myocardial infarction. In the present study, we prospectively examined whether remnant lipoprotein levels, measured by this method, may have predictive value for future coronary events in patients with coronary artery disease (CAD) and considered the possible validity of this assay in risk assessment of CAD.
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Methods |
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Study Patients
This study enrolled 147 consecutive patients with CAD (97 men and 50 women, ranging in age from 25 to 82 years [mean±SD, 65±9.7 years]) who underwent diagnostic cardiac catheterization for chest pain or ischemic changes in ECG in our hospital. All patients had angiographic documentation of organic stenosis of >70% of
1 major
coronary artery (single-vessel disease, 58 patients; 2-vessel
disease, 40 patients; 3-vessel disease, 37 patients; and left main
coronary artery disease, 12 patients). Written informed consent
was obtained from all patients before the study. This study was
conducted in agreement with guidelines approved by the ethics committee
at our institution.
Measurement of Lipoproteins
At the beginning of the study, venous blood was taken in all
patients after a 12-hour overnight fast. All patients ate a standard
meal (1900 kcal/d; 25% fat, 59% carbohydrate, and 16% protein) the
day before the sampling. Serum was stored at 4°C and was used for the
assays within 3 days after sampling. Remnant lipoproteins were isolated
by application of the fasting serum to the immunoaffinity mixed gel,
which contained anti–apoA-1 and anti–apoB-100 monoclonal antibodies
(Japan Immunoresearch Laboratories), and the unbound fraction
containing apoE-enriched lipoproteins was eluted with PBS, as described
in our previous reports.4 5 6 7 8 This assay takes 
2 hours
to complete. Cholesterol concentrations in the unbound
fraction were measured by the enzymatic method.7 8 Levels
of HDL cholesterol, LDL cholesterol, and
triglycerides in fasting serum were measured as described
previously.7
Follow-Up Study
After laboratory samples and angiographic data were obtained,
the 147 patients with CAD were followed up every month in the hospital
or with a clinic visit for 36 months until the occurrence of 1 of the
following clinical coronary events: recurrent or refractory
angina pectoris requiring coronary
revascularization by PTCA or CABG, nonfatal
myocardial infarction, or cardiac death. All patients received
standardized medical therapy. Time to first coronary event was
evaluated prospectively. Diagnosis of myocardial infarction was made by
chest pain, appearance of new Q wave on the ECG, and elevation of
creatinine kinase enzymes to more than twice the upper
limit of normal. Cause of death was determined from hospital
records.
Statistical Analysis
The Kaplan-Meier method (log-rank test) was applied in survival
analysis according to the levels of remnant lipoproteins. The
predictive value for coronary events during the follow-up
period was assessed by Cox proportional hazard analysis with
the following factors as categorical covariates: remnant levels,
stenosis of the left main coronary artery, number of
coronary arteries with stenosis, age (70 years), sex
(male), smoking history (defined as smoking 10 cigarettes/d for 10
years), hypertension (>140/90 mm Hg or taking antihypertensive
medication), diabetes mellitus (according to World Health Organization
criteria9 ), hypercholesterolemia
(>220 mg/dL or use of cholesterol-lowering medications),
low levels of HDL cholesterol (<35 mg/dL), and
hypertriglyceridemia (>150 mg/dL). In
these analyses, remnant levels were divided into tertiles that
were based on the 75th and 50th percentiles (5.1 and 3.3 mg/dL,
respectively) of the distribution of the fasting remnant levels in 250
consecutive patients hospitalized in the cardiology
section of the hospital, as described in our previous
report.7 When the number of coronary arteries with
stenosis was scored, stenosis of the left main
coronary artery was counted as 2-vessel disease. Statistical
significance was defined as P<0.05.
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Results |
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The distribution of fasting remnant levels in patients was skewed and shifted to lower levels, with a median level of 3.4 mg/dL (25th percentile, 2.0 mg/dL; 75th percentile, 5.4 mg/dL). The lipoproteins, isolated from fasting serum by the immunoaffinity mixed gel, consisted mainly of VLDL remnants and had only trace amounts of HDL, as shown in our previous analyses.4 5 6 7 8 Multivariate linear regression analysis (after log transformation) showed that remnant levels were independently correlated with triglyceride levels (partial regression coefficient, 0.480; P=0.001). Remnant levels were significantly higher in diabetic patients than in nondiabetic patients (n=44 and 103, respectively; P<0.01, Mann-Whitney U test).
Twelve patients were lost to follow-up. The remaining 135 patients were
followed up for 26.8±13.9 (mean±SD) months. Patients with the highest
tertile of remnant levels (39 patients) had 20 coronary events
(5 PTCA, 6 CABG, 3 myocardial infarctions, and 6 coronary
deaths) during the follow-up period, whereas patients in the lowest
tertile (56 patients) had 11 events (3 PTCA, 5 CABG, 1 myocardial
infarction, and 2 coronary deaths) (P<0.01 for
frequency of coronary events between the 2 subgroups by
2 test). Kaplan-Meier analysis
demonstrated a significantly higher probability of developing clinical
coronary events in patients with higher remnant levels, as
shown in the Figure
. In
univariate Cox proportional hazard model analysis,
higher levels of remnants (OR, 5.91; 95% CI, 2.0 to 17.2;
P<0.001, highest compared with lowest tertile),
stenosis of left main coronary artery (OR, 3.55; 95%
CI, 1.8 to 7.0; P<0.001), 3-vessel disease (OR, 2.96; 95%
CI, 1.5 to 6.0; P=0.002 compared with 1-vessel disease), and
diabetes mellitus (OR, 1.86; 95% CI, 1.1 to 3.4; P=0.04)
were significant predictors of clinical coronary events.
Multivariate Cox proportional hazard analysis
showed that only higher levels of remnants were a significant and
independent predictor of coronary events, as shown in the
Table. Higher remnant levels
remained a significant predictor of coronary events in
multivariate Cox analysis after addition of
high LDL-cholesterol levels (>130 mg/dL) into the
covariates (OR, 6.12; 95% CI, 2.1 to 15.2; P=0.001, highest
versus lowest tertile).
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Discussion |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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The present study demonstrated that higher levels of remnant lipoproteins in fasting serum predicted the development of clinical coronary events in patients with CAD independently of other risk factors. Thus, measurement of remnant levels by the current method may be helpful for CAD risk assessment in clinical laboratories. Previous reports showed that postprandial levels of triglyceride-rich lipoproteins may be a better predictor of the presence of CAD than fasting levels,1 but the assessment of postprandial levels of remnant lipoproteins is likely to be difficult for routine clinical examination because of postprandial variability of remnant levels in individuals, as well as requirements that the patient eat a standard meal and that multiple blood samples be collected over a prolonged period afterward. On the basis of our previous results7 showing that fasting remnant levels were closely correlated with postprandial remnant levels, remnant lipoprotein levels could be measured in the fasting blood as a reflection of abnormal postprandial remnant lipoproteins.
A previous in vitro experiment showed that remnant lipoproteins were taken up by macrophages and caused foam cell formation.10 Furthermore, high levels of remnant lipoproteins caused endothelial vasomotor dysfunction in human coronary arteries, as shown in our previous report.7 Remnant lipoproteins induce proinflammatory and prothrombogenic genes in cultured endothelial cells (H. Doi, MD, unpublished data, 1999). This atherothrombogenic role of remnant lipoproteins may result in the association of higher remnant levels with the increasingly high prevalence of future coronary events in patients with CAD, as observed in the present study.
Lipid-lowering drugs (especially fibric acid derivatives), dietary interventions, aerobic exercise, and obesity reduction might decrease remnant lipoprotein levels in hypertriglyceridemic patients. The measurement of remnant levels may also be useful for monitoring the therapeutic effect on levels of this atherogenic lipoprotein in these patients.
In conclusion, fasting remnant levels predicted future coronary events in patients with CAD independently of other risk factors. Thus, measurement of fasting remnant levels, as assessed by the current immunoseparation method, may be helpful in the assessment of CAD risk.
Received January 21, 1999; revision received March 24, 1999; accepted April 7, 1999.
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