Dysfunctional HDL and progression of atherosclerosis in HIV-1-infected and -uninfected adults
© Kelesidis et al; licensee BioMed Central Ltd. 2013
Received: 17 December 2012
Accepted: 28 February 2013
Published: 5 March 2013
HDL function rather than absolute level may be a more accurate indicator for risk of developing atherosclerosis. Dysfunctional HDL has increased redox activity and reduced antioxidant properties, but it is unknown whether abnormal HDL function is associated with progression of atherosclerosis in HIV-1-infected subjects.
We retrospectively measured serum HDL function in 91 subjects from a prospective 3-year study of carotid artery intima-media thickness (CIMT), which enrolled triads of risk factor-matched persons that were HIV-1-uninfected (n=36) or HIV-1+ with (n=29) or without (n=26) protease inhibitor (PI)-based therapy for ≥ 2 years. HDL function was assessed using a biochemical assay that measures the oxidation of dihydrorhodamine 123 (DHR oxidation rate, DOR), in which higher DOR readout corresponds to dysfunctional HDL phenotype.
There were no significant associations between DOR and HIV-1 infection. In univariate analysis of 55 HIV-1-infected subjects, greater waist circumference and lower serum HDL were significantly associated with higher baseline levels of DOR (p=0.01). These subjects had significant increases in levels of DOR over time (3 years) that were associated with white race (p=0.03), higher nadir CD4 count (p<0.001), and lower baseline CIMT (p<0.001). Lower baseline HDL levels, but not function of HDL (p>0.1) (DOR), were significantly associated (p=0.02) with progression of CIMT.
In a small matched cohort study of HIV-1-infected subjects who had a low cardiovascular risk profile, HDL function changed over time and was independently associated with anthropometric parameters of obesity but not with progression of CIMT.
KeywordsHIV-1 High density lipoprotein (HDL) HDL function Dysfunctional HDL Redox activity Atherosclerosis
High-density lipoprotein (HDL) function rather than absolute level may be a more accurate indicator for risk of developing atherosclerosis . Systemic inflammation may involve the conversion of HDL to a dysfunctional form that is no longer cardioprotective and contributes to the risk of coronary disease [2, 3]. Dysfunctional HDL has increased redox activity  and reduced antioxidant properties that may contribute to atherogenesis [5, 6]. We have recently shown that HIV-1-infected (HIV-1+) subjects have dysfunctional HDL [4, 7, 8]. However, no study has directly explored the relationships between dysfunctional HDL, HIV-1 infection and progression of atherosclerosis as well as predictors of dysfunctional HDL in HIV-1 infection.
Using a recently developed biochemical assay that quantifies the redox activity of HDL based on the oxidation rate of the fluorogenic probe dihydrorhodamine 123 (DOR)  and serum specimens from a previously reported clinical study, we examined the longitudinal association of DOR with the progression of atherosclerosis, as evaluated by measurement of carotid artery intima-media thickness (CIMT), in HIV-1-infected (HIV-1+) and -uninfected (controls) individuals. Since residual immune activation despite antiretroviral treatment may contribute to the presence of dysfunctional HDL in HIV-1+ persons  and progression of atherosclerosis , we also investigated possible associations between DOR and biomarkers of macrophage activation, such as soluble CD14 (sCD14) and biomarkers of bacterial translocation such as lipopolysaccharide (LPS) .
The current study is a subset analysis of samples obtained from a prospective, matched cohort study  in which subjects were enrolled as risk factor-matched triads of HIV-1+ individuals with viremia < 500 RNA copies/ml with (n=29) or without (n=26) use of PI therapy, and HIV-1-uninfected (control) individuals (n=36) from 41 triads.
Presence of metabolic syndrome, CIMT and baseline variables (fasting glucose, lipids, insulin, cardiovascular disease-related measurements, CD4+ T cell counts and HIV RNA levels, high-sensitivity C-reactive protein, serum sCD14, serum LPS) have previously been determined . Stored samples from baseline and week 144 (or 96 if week 144 was missing) were assayed. HDL was isolated using ultracentrifugation and the DOR of each sample was determined as previously  and was normalized (nDOR) as ratio to the DOR of a control HDL isolated from pooled serum from healthy subjects (see Additional file 1).
Matched analyses comparing the combined HIV-1+ group to the controls (N=32) assessed the effect of HIV-1 infection on nDOR. Progression of atherosclerosis was evaluated as yearly rate of change in CIMT (ΔCIMT); and CIMT progression (previously defined as yearly rate of change ≥ 12.2 μm/yr) . Mixed models regression with triad as a random effect and repeated measures regression evaluated associations of baseline measurements of nDOR and yearly rates of change in the outcome, respectively, with baseline covariates. SAS 9.2 (SAS Institute Inc., Cary, NC, USA) was used for all statistical analyses.
HIV-1+ subjects had more metabolic abnormalities than the control subjects
As reported previously the HIV-1+ subjects in this cohort had significantly (p<0.05) higher presence of metabolic syndrome, higher levels of total cholesterol, triglycerides and non-HDL cholesterol and insulin levels, serum sCD14 and similar levels of serum LPS compared to the control subjects . Due to matching the groups were similar with respect to age, sex, race, body mass index (BMI), LDL and HDL levels .
HIV-1 infection was not significantly associated with baseline HDL function, but HDL function changed significantly over time within the HIV-1+ subjects
Summary of normalized to control (pooled HDL sample from all study subjects) DOR (nDOR) results [Median (IQR)] by group and of parameters that were significantly associated with nDOR and normalized Yearly Rate of Change in nDOR (ΔnDOR) (see Table 2 )
Not HIV (N=36)
1.64 (1.47, 2.02)
1.62 (1.44, 1.96)
1.60 (1.46, 1.95)
1.65 (1.44, 1.96)
1.65 (1.51, 2.07)
Week 96/144 nDOR
1.80 (1.50, 2.00)
1.80 (1.45, 1.92)
1.79 (1.45, 1.83)
1.82 (1.45, 2.29)
1.82 (1.53, 2.14)
Yearly rate of change in nDOR
0.009 (-0.055, 0.108)
0.005 (-0.048, 0.110)
-0.004 (-0.055, 0.101)
0.059 (-0.031, 0.118)
0.009 (-0.059, 0.102)
0.94; 0.58; 0.29; 0.653
P Value for ΔnDOR within group 4
White non-Hispanic race
Body mass index (kg/m2)
Waist circumference (cm) ≥ 90 6
Waist/Hip ratio 6
Baseline CIMT (μm)
Nadir CD4+ T-cells ≤ 200 (cells/mm3) 7
HDL Cholesterol < 35 (mg/dl)
Insulin mU/l 8
Lower baseline HDL levels and anthropometric parameters of obesity were independently associated with HDL function
Univariate associations of nDOR with baseline variables
Parameter estimate (95% CI)
Parameter estimate (95% CI)
Baseline nDOR (normalized ratio).
HDL Cholesterol (ref: ≥35 mg/dl)
0.357 (0.114, 0.601)
0.365 (0.029, 0.701)
Body mass index (per 1 kg/m 2 increase)
0.032 (-0.005, 0.069)
Waist circumference (ref: <90 cm)
Waist/Hip ratio (per 0.1 unit increase)
0.212 (0.035, 0.388)
0.215 (-0.019, 0.448)
Yearly rate of change in nDOR (normalized ratio/yr).
0.156 (-0.018, 0.329)
0.067 (-0.001, 0.144)
White vs Non-White
0.119 (0.033, 0.206)
Body mass index (per 1 kg/m 2 increase)
-0.010 (-0.020, 0.0001)
Reported Nadir CD4 (ref: <200 cells/mm 3 )
0.083 (0.039, 0.128)
Baseline CIMT (per 1 μm decrease)
0.474 (0.229, 0.719)
HDL Cholesterol (ref: ≥35 mg/dl)
-0.055 (-0.109, -0.001)
Insulin (per 10 mU/l decrease)
0.120 (0.056, 0.184)
LPS (per 100 pg/ml decrease)
0.005 (0.002, 0.008)
Changes in HDL function over time were associated with baseline HDL, race, nadir CD4 count, and CIMT in HIV-1+ subjects
Because there were significant changes in nDOR over time within the HIV-1+ group, we sought factors that were associated with yearly rate of change in HDL function (ΔnDOR) in HIV-1+ subjects. Similar to the univariate analysis (Table 2), multivariate analysis in HIV-1+ subjects showed a positive ΔnDOR was associated with lower baseline CIMT (parameter estimate 0.45, 95% CI: 0.23-0.67; p<0.001), white race (estimate 0.12, 95% CI: 0.01-0.23; p=0.03), baseline HDL ≥35 mg/dl (estimate 0.07, 95% CI: 0.02-0.12; p=0.007), and nadir CD4 >200 cells/mm3 (estimate 0.08, 95% CI: 0.04-0.12; p<0.001). In the control subjects, lower baseline LPS (p=0.03) and decreased baseline insulin (p=0.002) were associated with a positive ΔnDOR in multivariate analysis. There were too few non-white subjects to determine the effect of race on ΔnDOR. Finally, increased rate of change in nDOR in HIV-1+ subjects was associated with favorable metabolic and immunologic parameters with higher baseline HDL levels, in contrast to baseline nDOR, which was associated with lower HDL levels. Further larger prospective studies will be required to explore these findings and determine predictors of both baseline HDL function and changes in HDL function over time.
HDL function was not associated with progression of subclinical atherosclerosis in HIV-1+ subjects
Considering the previously described role of HDL function in progression of atherosclerosis in HIV-1-uninfected subjects, we investigated the possible relationship between HDL function and progression of atherosclerosis in HIV-1+ subjects. In a prior multivariate analysis in the HIV-1+ subjects, baseline HDL <35 mg/dl (p=0.04) and higher levels of baseline sCD14 (p=0.04) and LPS (p=0.003) were associated with a positive ΔCIMT . In univariate analysis in HIV-1+ subjects, baseline nDOR was not associated with baseline CIMT (estimate: 53.2; 95% CI: (-16.6-123; p=0.13), ΔCIMT (estimate: -7.5; 95% CI: (-15.6- 0.7; p=0.07), and CIMT progression (estimate: 0.8; 95% CI: 0.2-3.6; p=0.81). It is possible that different factors influence the presence of plaque and CIMT, and thus the relationship of HDL function with different measures of atherosclerosis  requires further examination in larger studies.
The interaction of the HDL redox activity with levels of cholesterol may be important in HIV-1+ subjects
The redox activity of HDL may depend on the interplay between HDL and LDL and the concentrations of both HDL and LDL and their ratio (LDL/HDL) [3, 4]. In HIV-1+ subjects, the baseline interaction model of nDOR*(LDL/HDL), including HDL, LDL/HDL, and nDOR*HDL, nDOR*(LDL/HDL) was significantly associated with baseline CIMT (estimate: 120; 95% CI: 15.4-225; p=0.03), but not with CIMT progression (OR: 0.5; 95% CI: 0.06-4.5; p=0.55), and ΔCIMT (estimate: -3.5; 95% CI: -14.2-7.3; p=0.52). Thus, the interaction of the HDL redox activity with levels of cholesterol may be taken into consideration when studying the association of dysfunctional HDL with atherosclerosis in HIV-1+ subjects who are known to have dyslipidemia and dysfunctional HDL [4, 7, 12].
HDL redox activity was not associated with biomarkers of macrophage activation in HIV-1+ subjects
Residual immune activation despite effective antiretroviral therapy could contribute to the presence of dysfunctional HDL in HIV-1 infection [8, 13, 14]. However, despite the significantly higher baseline levels of sCD14, a marker of macrophage activation, in HIV-1+ compared to control subjects  in our small study, we did not detect any significant associations between nDOR and sCD14 (p>0.1). Larger studies are needed to elucidate the role of dysfunctional HDL in HIV-1-induced immune activation.
Our study had limited statistical power to detect small differences in HDL redox activity between groups, especially in the context of the limitations of the DHR assay (see Additional file 1). Moreover, despite preliminary data from in vitro studies that support a role of oxidized HDL in atherosclerosis , the significance of the nDOR, which is a biochemical and not a physiological parameter, in studying atherosclerosis in vivo, needs to be validated in large-scale human studies.
Despite these limitations, our study is among the first attempts to measure HDL function in HIV-1+ and controls longitudinally and define their possible associations with progression of subclinical atherosclerosis. Thus, this study may set the basis for further studies on the role of HDL function in HIV-1 infection given that standard clinical lipid profile testing may not be an adequate measurement of the risk for atherosclerosis.
Body mass index
Carotid artery intima-media thickness
Dihydrorhodamine 123 oxidation rate
Yearly rate of change in CIMT
Yearly rate of change in nDOR
High Density Lipoprotein
Human Immunodeficiency virus-1
Low Density Lipoprotein
This work was supported by the National Institute of Allergy and Infectious Diseases (AI068634 and AI056933), and UCLA AIDS Institute and the UCLA Center for AIDS Research (AI28697). Partial funding for laboratory work was provided by the University of Washington’s CVD and Metabolic Complications of HIV/AIDS Data Coordinating Center (5R01HL095126).
- Navab M, Reddy ST, Van Lenten BJ, Anantharamaiah GM, Fogelman AM: The role of dysfunctional HDL in atherosclerosis. J Lipid Res. 2009, 50 (Suppl): S145-S149.PubMed CentralPubMedGoogle Scholar
- McMahon M, Grossman J, FitzGerald J, Dahlin-Lee E, Wallace DJ, Thong BY, Badsha H, Kalunian K, Charles C, Navab M, Fogelman AM, Hahn BH: Proinflammatory high-density lipoprotein as a biomarker for atherosclerosis in patients with systemic lupus erythematosus and rheumatoid arthritis. Arthritis Rheum. 2006, 54: 2541-2549. 10.1002/art.21976View ArticlePubMedGoogle Scholar
- Navab M, Reddy ST, Van Lenten BJ, Fogelman AM: HDL and cardiovascular disease: atherogenic and atheroprotective mechanisms. Nat Rev Cardiol. 2011, 8: 222-232. 10.1038/nrcardio.2010.222View ArticlePubMedGoogle Scholar
- Kelesidis T, Currier JS, Huynh D, Meriwether D, Charles-Schoeman C, Reddy ST, Fogelman AM, Navab M, Yang OO: A biochemical fluorometric method for assessing the oxidative properties of HDL. J Lipid Res. 2011, 52: 2341-2351. 10.1194/jlr.D018937PubMed CentralView ArticlePubMedGoogle Scholar
- Navab M, Ananthramaiah GM, Reddy ST, Van Lenten BJ, Ansell BJ, Fonarow GC, Vahabzadeh K, Hama S, Hough G, Kamranpour N, Berliner JA, Lusis AJ, Fogelman AM: The oxidation hypothesis of atherogenesis: the role of oxidized phospholipids and HDL. J Lipid Res. 2004, 45: 993-1007. 10.1194/jlr.R400001-JLR200View ArticlePubMedGoogle Scholar
- Navab M, Ananthramaiah GM, Reddy ST, Van Lenten BJ, Ansell BJ, Hama S, Hough G, Bachini E, Grijalva VR, Wagner AC, Shaposhnik Z, Fogelman AM: The double jeopardy of HDL. Ann Med. 2005, 37: 173-178. 10.1080/07853890510007322View ArticlePubMedGoogle Scholar
- Kelesidis T, Reddy ST, Huynh D, Meriwether D, Fogelman AM, Navab M, Yang OO: Effects of lipid-probe interactions in biochemical fluorometric methods that assess HDL redox activity. Lipids Health Dis. 2012, 11: 87- 10.1186/1476-511X-11-87PubMed CentralView ArticlePubMedGoogle Scholar
- Kelesidis T, Yang OO, Currier JS, Navab K, Fogelman AM, Navab M: HIV-1 infected patients with suppressed plasma viremia on treatment have pro-inflammatory HDL. Lipids Health Dis. 2011, 10: 35- 10.1186/1476-511X-10-35PubMed CentralView ArticlePubMedGoogle Scholar
- Kelesidis T, Kendall MA, Yang OO, Hodis HN, Currier JS: Biomarkers of Microbial Translocation and Macrophage Activation: Association With Progression of Subclinical Atherosclerosis in HIV-1 Infection. J Infect Dis. 2012, 206: 1558-1567. 10.1093/infdis/jis545PubMed CentralView ArticlePubMedGoogle Scholar
- Crowe SM, Westhorpe CL, Mukhamedova N, Jaworowski A, Sviridov D, Bukrinsky M: The macrophage: the intersection between HIV infection and atherosclerosis. J Leukoc Biol. 2010, 87: 589-598. 10.1189/jlb.0809580PubMed CentralView ArticlePubMedGoogle Scholar
- Patel PJ, Khera AV, Jafri K, Wilensky RL, Rader DJ: The anti-oxidative capacity of high-density lipoprotein is reduced in acute coronary syndrome but not in stable coronary artery disease. J Am Coll Cardiol. 2011, 58: 2068-2075. 10.1016/j.jacc.2011.08.030View ArticlePubMedGoogle Scholar
- Currier JS: Update on cardiovascular complications in HIV infection. Top HIV Med. 2009, 17: 98-103.PubMedGoogle Scholar
- Almeida CA, Price P, French MA: Immune activation in patients infected with HIV type 1 and maintaining suppression of viral replication by highly active antiretroviral therapy. AIDS Res Hum Retroviruses. 2002, 18: 1351-1355. 10.1089/088922202320935429View ArticlePubMedGoogle Scholar
- French MA, King MS, Tschampa JM, da Silva BA, Landay AL: Serum immune activation markers are persistently increased in patients with HIV infection after 6 years of antiretroviral therapy despite suppression of viral replication and reconstitution of CD4+ T cells. J Infect Dis. 2009, 200: 1212-1215. 10.1086/605890View ArticlePubMedGoogle Scholar
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