Recombinant Antibodies to an Oxidized Low-Density Lipoprotein Epitope Induce Rapid Regression of Atherosclerosis in Apobec-1 –/–/Low-Density Lipoprotein Receptor–/– Mice
Alexandru Schiopu, MD, PhD*,*,
Björn Frendéus, PhD ,
Bo Jansson, PhD,
Ingrid Söderberg, BSI*,
Irena Ljungcrantz, BSI*,
Zufan Araya, PhD,
Prediman K. Shah, MD, FACC ,
Roland Carlsson, PhD,
Jan Nilsson, MD, PhD* and
Gunilla Nordin Fredrikson, PhD*,
* Department of Clinical Sciences, Malmö University Hospital, Lund University, Malmö, Sweden
BioInvent International AB, Lund, Sweden
Atherosclerosis Research Center and Division of Cardiology, Cedars-Sinai Medical Center, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California
Department of Biomedical Laboratory Science, Malmö University, Malmö, Sweden.
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Figure 1 Antibody-Induced Regression of Plaque Area in the Descending Aorta Assessed by Oil Red O Staining
The values are expressed as the percentages of total plaque area per total area of the descending aorta. p values were calculated versus fluorescein isothiocyanate-8 (FITC-8). **p < 0.01 and ***p < 0.001 versus FITC-8.
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Figure 2 Recombinant Human Antibodies to MDA–apoB-100 Peptides Upregulate ABCA-1 Expression In Vivo and In Vitro
(A) ABCA-1 staining of aortic arch plaques from FITC-8–, IEI-E3–, and 2D03-treated mice. (B) The values represent the percentages of ABCA-1–stained area per total plaque area. (C) Flow cytometry histogram showing ABCA-1 expression of untreated (gray shaded peak), FITC-8–treated (black line overlay), and 2D03-treated (red line overlay) cells. (D) Mean ABCA-1 expression of untreated and FITC-8– and 2D03-treated cells. The values are expressed as percentages of mean fluorescent intensity of the untreated control. Scale bar = 100 µm. *p < 0.02 and **p < 0.01 versus FITC-8. ABCA-1 = adenosine triphosphate–binding cassette transporter A1; apoB-100 = apolipoprotein B-100; FITC-8 = fluorescein isothiocyanate-8; MDA = malondialdehyde.
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Figure 3 Macrophage and ABCA-1 Immunoreactivity Colocalize in Aortic Arch Plaques
(A and C) ABCA-1 and (B and D) macrophage staining of aortic arch plaques from 2 different mice treated with the 2D03 antibody and sacrificed at 29 weeks of age. Abbreviations as in Figure 2.
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Figure 4 Decreased Macrophage Content in Plaques From the Innominate Artery in the Antibody-Treated Groups
The values in the graph represent percentages of stained area per total plaque area. Scale bar = 100 µm. *p < 0.05 versus FITC-8. Abbreviations as in Figure 2.
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Figure 5 Antibodies to MDA–apoB-100 Peptides Block oxLDL-Induced Monocyte MCP-1 Production In Vitro
(A) MCP-1 levels in conditioned medium from monocytes incubated with titrated concentrations of IEI-E3 (shaded squares), 2D03 (solid squares), or FITC-8 control IgG1 (open squares) in the presence of oxLDL-containing human serum. The graph shows mean ± SD of triplicate values from a representative experiment. (B)
MCP-1 messenger ribonucleic acid (mRNA) relative to 18S mRNA levels in human monocytes incubated with 60 µg/ml 2D03, IEI-E3, or FITC-8 in the presence of oxLDL-containing human serum. Graphs show mean ± SD of triplicate values using cells obtained from 3 different donors (n = 9). ***p < 0.001 versus FITC-8. IgG = immunoglobulin G; MCP = monocyte chemoattractant protein; oxLDL = oxidized low-density lipoprotein; other abbreviations as in Figure 2.
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