Monoclonal Autoantibody Against a Cryptic Epitope on Tissue-Adherent Low-Density Lipoprotein for Molecular Imaging in Atherosclerosis

Ramzi Y Khamis; Adam Hartley; Mikhail Caga-Anan; Samata S Pandey; Cinzia Marceddu; Chiari Kojima; Shang-Hung Chang; Joseph J Boyle; Jason L Johnson; Harry Björkbacka; Liang Guo; Aloke V Finn; Renu Virmani; Jan Nilsson; Dorian O Haskard

doi:10.1016/j.jcmg.2022.02.023

Monoclonal Autoantibody Against a Cryptic Epitope on Tissue-Adherent Low-Density Lipoprotein for Molecular Imaging in Atherosclerosis

Ramzi Y Khamis, Adam Hartley, Mikhail Caga-Anan, Samata S Pandey, Cinzia Marceddu, Chiari Kojima, Shang-Hung Chang, Joseph J Boyle, Jason L Johnson, Harry Björkbacka, Liang Guo, Aloke V Finn, Renu Virmani, Jan Nilsson, Dorian O Haskard^*

^*Corresponding author for this work

Bristol Medical School (THS)

Research output: Contribution to journal › Article (Academic Journal) › peer-review

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Abstract

BACKGROUND: Antibody-based constructs for molecular imaging and therapeutic delivery provide promising opportunities for the diagnosis and treatment of atherosclerosis.

OBJECTIVES: The authors aimed to generate and characterize immunoglobulin (Ig)G monoclonal autoantibodies in atherosclerosis for targeting of novel molecular determinants.

METHODS: The authors created hybridomas from an unimmunized low-density lipoprotein (LDL) receptor-deficient (Ldlr -/-) mouse and selected an IgG2b isotype autoantibody, LO9, for further characterization.

RESULTS: LO9 reacted well with native LDL bound to immobilized matrix components and less well to oxidized LDL. LO9 binding to immobilized native LDL was not neutralized by fluid-phase native LDL, indicating an adhesion-dependent epitope. The authors localized the epitope to a 20 amino-acid peptide sequence (P5) in the globular amino-terminus of apolipoprotein B. LO9 reacted with antigen in mouse atherosclerosis and in both human stable and ruptured coronary atherosclerosis. Furthermore, in vivo near-infrared fluorescence molecular tomographic imaging, and ex vivo confocal microscopy showed that intravenously injected LO9 localized beneath endothelium of the aortic arch in Ldlr -/- mice, in the vicinity of macrophages.

CONCLUSIONS: The authors believe LO9 is the first example of an IgG autoantibody that reacts with a native LDL epitope revealed by adherence to tissue matrix. Antibodies against adherent native LDL have potential as molecular targeting agents for imaging of and therapeutic delivery to atherosclerosis.

Original language	English
Pages (from-to)	1458-1470
Number of pages	13
Journal	JACC: Cardiovascular Imaging
Volume	15
Issue number	8
Early online date	13 May 2022
DOIs	https://doi.org/10.1016/j.jcmg.2022.02.023
Publication status	Published - 1 Aug 2022

Bibliographical note

Funding Information:
Dr Khamis has been funded by a British Heart Foundation (BHF) Intermediate Clinical Research Fellowship and has received a Wellcome Trust Clinical Research Fellowship (as part of the Wellcome Trust/GlaxoSmithKline Fellowship program). Prof Haskard has received Professorial Chair funding from the BHF. Dr Hartley has been funded by a Wellcome Trust Clinical Research Fellowship. Dr Kojima has received funding from the Japan Heart Foundation/Bayer Yakuhin Research Abroad Grant. Dr Guo and Dr Finn have been supported by a Leducq Foundation Grant. All other authors have reported that they have no relationships relevant to the contents of this paper to disclose.

Publisher Copyright:
© 2022 The Authors

Research Groups and Themes

Bristol Heart Institute

Keywords

Animals
Antibodies, Monoclonal
Atherosclerosis/metabolism
Autoantibodies/chemistry
Epitopes
Humans
Immunoglobulin G
Lipoproteins, LDL/chemistry
Mice
Molecular Imaging
Predictive Value of Tests

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Khamis, R. Y., Hartley, A., Caga-Anan, M., Pandey, S. S., Marceddu, C., Kojima, C., Chang, S.-H., Boyle, J. J., Johnson, J. L., Björkbacka, H., Guo, L., Finn, A. V., Virmani, R., Nilsson, J., & Haskard, D. O. (2022). Monoclonal Autoantibody Against a Cryptic Epitope on Tissue-Adherent Low-Density Lipoprotein for Molecular Imaging in Atherosclerosis. JACC: Cardiovascular Imaging, 15(8), 1458-1470. https://doi.org/10.1016/j.jcmg.2022.02.023

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title = "Monoclonal Autoantibody Against a Cryptic Epitope on Tissue-Adherent Low-Density Lipoprotein for Molecular Imaging in Atherosclerosis",

abstract = "BACKGROUND: Antibody-based constructs for molecular imaging and therapeutic delivery provide promising opportunities for the diagnosis and treatment of atherosclerosis.OBJECTIVES: The authors aimed to generate and characterize immunoglobulin (Ig)G monoclonal autoantibodies in atherosclerosis for targeting of novel molecular determinants.METHODS: The authors created hybridomas from an unimmunized low-density lipoprotein (LDL) receptor-deficient (Ldlr -/-) mouse and selected an IgG2b isotype autoantibody, LO9, for further characterization. RESULTS: LO9 reacted well with native LDL bound to immobilized matrix components and less well to oxidized LDL. LO9 binding to immobilized native LDL was not neutralized by fluid-phase native LDL, indicating an adhesion-dependent epitope. The authors localized the epitope to a 20 amino-acid peptide sequence (P5) in the globular amino-terminus of apolipoprotein B. LO9 reacted with antigen in mouse atherosclerosis and in both human stable and ruptured coronary atherosclerosis. Furthermore, in vivo near-infrared fluorescence molecular tomographic imaging, and ex vivo confocal microscopy showed that intravenously injected LO9 localized beneath endothelium of the aortic arch in Ldlr -/- mice, in the vicinity of macrophages. CONCLUSIONS: The authors believe LO9 is the first example of an IgG autoantibody that reacts with a native LDL epitope revealed by adherence to tissue matrix. Antibodies against adherent native LDL have potential as molecular targeting agents for imaging of and therapeutic delivery to atherosclerosis.",

keywords = "Animals, Antibodies, Monoclonal, Atherosclerosis/metabolism, Autoantibodies/chemistry, Epitopes, Humans, Immunoglobulin G, Lipoproteins, LDL/chemistry, Mice, Molecular Imaging, Predictive Value of Tests",

author = "Khamis, {Ramzi Y} and Adam Hartley and Mikhail Caga-Anan and Pandey, {Samata S} and Cinzia Marceddu and Chiari Kojima and Shang-Hung Chang and Boyle, {Joseph J} and Johnson, {Jason L} and Harry Bj{\"o}rkbacka and Liang Guo and Finn, {Aloke V} and Renu Virmani and Jan Nilsson and Haskard, {Dorian O}",

note = "Funding Information: Dr Khamis has been funded by a British Heart Foundation (BHF) Intermediate Clinical Research Fellowship and has received a Wellcome Trust Clinical Research Fellowship (as part of the Wellcome Trust/GlaxoSmithKline Fellowship program). Prof Haskard has received Professorial Chair funding from the BHF. Dr Hartley has been funded by a Wellcome Trust Clinical Research Fellowship. Dr Kojima has received funding from the Japan Heart Foundation/Bayer Yakuhin Research Abroad Grant. Dr Guo and Dr Finn have been supported by a Leducq Foundation Grant. All other authors have reported that they have no relationships relevant to the contents of this paper to disclose. Publisher Copyright: {\textcopyright} 2022 The Authors",

year = "2022",

month = aug,

day = "1",

doi = "10.1016/j.jcmg.2022.02.023",

language = "English",

volume = "15",

pages = "1458--1470",

journal = "JACC: Cardiovascular Imaging",

issn = "1936-878X",

publisher = "Elsevier Inc.",

number = "8",

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Khamis, RY, Hartley, A, Caga-Anan, M, Pandey, SS, Marceddu, C, Kojima, C, Chang, S-H, Boyle, JJ, Johnson, JL, Björkbacka, H, Guo, L, Finn, AV, Virmani, R, Nilsson, J & Haskard, DO 2022, 'Monoclonal Autoantibody Against a Cryptic Epitope on Tissue-Adherent Low-Density Lipoprotein for Molecular Imaging in Atherosclerosis', JACC: Cardiovascular Imaging, vol. 15, no. 8, pp. 1458-1470. https://doi.org/10.1016/j.jcmg.2022.02.023

TY - JOUR

T1 - Monoclonal Autoantibody Against a Cryptic Epitope on Tissue-Adherent Low-Density Lipoprotein for Molecular Imaging in Atherosclerosis

AU - Khamis, Ramzi Y

AU - Hartley, Adam

AU - Caga-Anan, Mikhail

AU - Pandey, Samata S

AU - Marceddu, Cinzia

AU - Kojima, Chiari

AU - Chang, Shang-Hung

AU - Boyle, Joseph J

AU - Johnson, Jason L

AU - Björkbacka, Harry

AU - Guo, Liang

AU - Finn, Aloke V

AU - Virmani, Renu

AU - Nilsson, Jan

AU - Haskard, Dorian O

N1 - Funding Information: Dr Khamis has been funded by a British Heart Foundation (BHF) Intermediate Clinical Research Fellowship and has received a Wellcome Trust Clinical Research Fellowship (as part of the Wellcome Trust/GlaxoSmithKline Fellowship program). Prof Haskard has received Professorial Chair funding from the BHF. Dr Hartley has been funded by a Wellcome Trust Clinical Research Fellowship. Dr Kojima has received funding from the Japan Heart Foundation/Bayer Yakuhin Research Abroad Grant. Dr Guo and Dr Finn have been supported by a Leducq Foundation Grant. All other authors have reported that they have no relationships relevant to the contents of this paper to disclose. Publisher Copyright: © 2022 The Authors

PY - 2022/8/1

Y1 - 2022/8/1

N2 - BACKGROUND: Antibody-based constructs for molecular imaging and therapeutic delivery provide promising opportunities for the diagnosis and treatment of atherosclerosis.OBJECTIVES: The authors aimed to generate and characterize immunoglobulin (Ig)G monoclonal autoantibodies in atherosclerosis for targeting of novel molecular determinants.METHODS: The authors created hybridomas from an unimmunized low-density lipoprotein (LDL) receptor-deficient (Ldlr -/-) mouse and selected an IgG2b isotype autoantibody, LO9, for further characterization. RESULTS: LO9 reacted well with native LDL bound to immobilized matrix components and less well to oxidized LDL. LO9 binding to immobilized native LDL was not neutralized by fluid-phase native LDL, indicating an adhesion-dependent epitope. The authors localized the epitope to a 20 amino-acid peptide sequence (P5) in the globular amino-terminus of apolipoprotein B. LO9 reacted with antigen in mouse atherosclerosis and in both human stable and ruptured coronary atherosclerosis. Furthermore, in vivo near-infrared fluorescence molecular tomographic imaging, and ex vivo confocal microscopy showed that intravenously injected LO9 localized beneath endothelium of the aortic arch in Ldlr -/- mice, in the vicinity of macrophages. CONCLUSIONS: The authors believe LO9 is the first example of an IgG autoantibody that reacts with a native LDL epitope revealed by adherence to tissue matrix. Antibodies against adherent native LDL have potential as molecular targeting agents for imaging of and therapeutic delivery to atherosclerosis.

AB - BACKGROUND: Antibody-based constructs for molecular imaging and therapeutic delivery provide promising opportunities for the diagnosis and treatment of atherosclerosis.OBJECTIVES: The authors aimed to generate and characterize immunoglobulin (Ig)G monoclonal autoantibodies in atherosclerosis for targeting of novel molecular determinants.METHODS: The authors created hybridomas from an unimmunized low-density lipoprotein (LDL) receptor-deficient (Ldlr -/-) mouse and selected an IgG2b isotype autoantibody, LO9, for further characterization. RESULTS: LO9 reacted well with native LDL bound to immobilized matrix components and less well to oxidized LDL. LO9 binding to immobilized native LDL was not neutralized by fluid-phase native LDL, indicating an adhesion-dependent epitope. The authors localized the epitope to a 20 amino-acid peptide sequence (P5) in the globular amino-terminus of apolipoprotein B. LO9 reacted with antigen in mouse atherosclerosis and in both human stable and ruptured coronary atherosclerosis. Furthermore, in vivo near-infrared fluorescence molecular tomographic imaging, and ex vivo confocal microscopy showed that intravenously injected LO9 localized beneath endothelium of the aortic arch in Ldlr -/- mice, in the vicinity of macrophages. CONCLUSIONS: The authors believe LO9 is the first example of an IgG autoantibody that reacts with a native LDL epitope revealed by adherence to tissue matrix. Antibodies against adherent native LDL have potential as molecular targeting agents for imaging of and therapeutic delivery to atherosclerosis.

KW - Animals

KW - Antibodies, Monoclonal

KW - Atherosclerosis/metabolism

KW - Autoantibodies/chemistry

KW - Epitopes

KW - Humans

KW - Immunoglobulin G

KW - Lipoproteins, LDL/chemistry

KW - Mice

KW - Molecular Imaging

KW - Predictive Value of Tests

U2 - 10.1016/j.jcmg.2022.02.023

DO - 10.1016/j.jcmg.2022.02.023

M3 - Article (Academic Journal)

C2 - 35926905

SN - 1936-878X

VL - 15

SP - 1458

EP - 1470

JO - JACC: Cardiovascular Imaging

JF - JACC: Cardiovascular Imaging

IS - 8

ER -

Monoclonal Autoantibody Against a Cryptic Epitope on Tissue-Adherent Low-Density Lipoprotein for Molecular Imaging in Atherosclerosis

Abstract

Bibliographical note

Research Groups and Themes

Keywords

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