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. Supported by grants from the Swedish Medical Research Council; Swedish Heart-Lung Foundation; Crafoord Foundation; Royal Physiographic Society; Albert Påhlsson Foundation, Malmö University Hospital Foundation; Lundström Foundation; Heart Foundation; and the United Hostesses at Cedars-Sinai
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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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