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Detection of Atherosclerotic Plaques in the Rabbit Aorta Using Ultrasound Microbubbles Conjugated to Interleukin-18 Antibodies

Zhen-zhen Jiang, Xia-tian Liu, Cai-ye Ma, Cong He, Xing-yun Li, Chuan-lin Hou, Zu-sheng Cheng, Guo-yuan Xia

Department of Ultrasound, Shaoxing People’s Hospital (Shaoxing Hospital of Zhejiang University), Shaoxing, Zhejiang, China (mainland)

Med Sci Monit 2017; 23:5446-5454

DOI: 10.12659/MSM.907572

Available online:

Published: 2017-11-16


BACKGROUND: The purpose of the study was to investigate the ability of microbubbles (MBs) targeting interleukin-18 (IL-18) to detect plaques in a rabbit atherosclerotic plaque model.
MATERIAL AND METHODS: A rabbit atherosclerotic plaque model was established. The locations of the atherosclerotic plaques were verified by two-dimensional scanning and color Doppler flow imaging. An IL-18 antibody was conjugated to naked MBs (MBc) using the biotin-streptavidin conjugation method, resulting in the formation of MBIL-18. MBc and MBIL-18 were then used for contrast-enhanced ultrasound (CEUS) studies. The locations of CD34 and IL-18 within the plaques were determined by immunohistochemistry, and IL-18 expression levels in the plaques were determined by Western blot analysis. The relationships between IL-18 expression and the contrast intensity of the 2 MBs were analyzed.
RESULTS: MBc and MBIL-18 were both uniformly dispersed. Fluorescence microscopy and flow cytometry revealed that IL-18 was successfully conjugated to MBs. CEUS images showed that the intensity of the MBIL-18 signal was substantially enhanced and prolonged compared with that of the MBc signal. Immunohistochemistry showed that CD34 expression was significantly increased in the plaques and that IL-18 was mainly located in the inner parts and base of the atherosclerotic plaques. Western blot analysis revealed that IL-18 expression was higher in the plaque regions. Correlation analysis showed that IL-18 expression was correlated with the contrast intensity of MBIL-18 (r=0.903, P<0.05) but not with MBc (r=0.540, P>0.05).
CONCLUSIONS: MBs targeting IL-18 may be a novel, noninvasive method of diagnosing atherosclerotic plaques.

Keywords: Microbubbles, Plaque, Atherosclerotic, Ultrasonography, Doppler



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