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Targeted gold-coated iron oxide nanoparticles for CD163 detection in atherosclerosis by MRI.

Tarin C, Carril M, Martin-Ventura JL, Markuerkiaga I, Padro D, Llamas-Granda P, Moreno JA, García I, Genicio N, Plaza-Garcia S, Blanco-Colio LM, Penades S, Egido J - Sci Rep (2015)

Bottom Line: We have prepared a targeted probe based on gold-coated iron oxide nanoparticles vectorized with an anti-CD163 antibody for the specific detection of CD163 by MRI.Subsequently, the targeted probe was injected in 16 weeks old apoE deficient mice developing atherosclerotic lesions and the pararenal abdominal aorta was imaged by MRI.The accumulation of probe in the site of interest increased over time and the signal intensity decreased significantly 48 hours after the injection.

View Article: PubMed Central - PubMed

Affiliation: Laboratorio de Patología Vascular y Renal. IIS Fundación Jiménez Díaz, Universidad Autónoma. Av. Reyes Católicos 2, 28040, Madrid, Spain.

ABSTRACT
CD163 is a membrane receptor expressed by macrophage lineage. Studies performed in atherosclerosis have shown that CD163 expression is increased at inflammatory sites, pointing at the presence of intraplaque hemorrhagic sites or asymptomatic plaques. Hence, imaging of CD163 expressing macrophages is an interesting strategy in order to detect atherosclerotic plaques. We have prepared a targeted probe based on gold-coated iron oxide nanoparticles vectorized with an anti-CD163 antibody for the specific detection of CD163 by MRI. Firstly, the specificity of the targeted probe was validated in vitro by incubation of the probe with CD163(+) or (-) macrophages. The probe was able to selectively detect CD163(+) macrophages both in human and murine cells. Subsequently, the targeted probe was injected in 16 weeks old apoE deficient mice developing atherosclerotic lesions and the pararenal abdominal aorta was imaged by MRI. The accumulation of probe in the site of interest increased over time and the signal intensity decreased significantly 48 hours after the injection. Hence, we have developed a highly sensitive targeted probe capable of detecting CD163-expressing macrophages that could provide useful information about the state of the atheromatous lesions.

No MeSH data available.


Related in: MedlinePlus

CD163 detection in in vitro models.(A) Western blot of CD163 expression and α-actin (loading control) in THP-1 macrophages cell lysates with or without dexamethasone treatment for 24 hours. (B) Graph showing the normalized T2 values obtained from MRI images of human macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(h) and NP-IgG(h), n  = 3. All the replicates were normalized to the blank (cells that have not been incubated with any nanoparticles). The error bars shows the SEM. $p < 0.05 versus Blank; *p < 0.05, **p < 0.01 and ***p < 0.001 versus NP-CD163(+) (C) Representative MRI phantoms of human macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(h) and NP-IgG(h). (D) Immunodetection of CD163 expression in murine macrophages with or without dexamethasone treatment. Propidium iodide (PI) was used for nuclei staining. (E) Graph showing the normalized mean value obtained from MRI images of murine macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(m) and NP-IgG(m), n = 3. All the replicates were normalized to the blank (cells that have not been incubated with any nanoparticles). The error bars shows the SEM. $$p < 0.01 versus Blank; *p < 0.05 and **p < 0.01 versus NP-CD163(+); #p < 0.05 versus Ab 1:1. F) Representative MRI phantoms of murine macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(m) and NP-IgG(m).
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f2: CD163 detection in in vitro models.(A) Western blot of CD163 expression and α-actin (loading control) in THP-1 macrophages cell lysates with or without dexamethasone treatment for 24 hours. (B) Graph showing the normalized T2 values obtained from MRI images of human macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(h) and NP-IgG(h), n  = 3. All the replicates were normalized to the blank (cells that have not been incubated with any nanoparticles). The error bars shows the SEM. $p < 0.05 versus Blank; *p < 0.05, **p < 0.01 and ***p < 0.001 versus NP-CD163(+) (C) Representative MRI phantoms of human macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(h) and NP-IgG(h). (D) Immunodetection of CD163 expression in murine macrophages with or without dexamethasone treatment. Propidium iodide (PI) was used for nuclei staining. (E) Graph showing the normalized mean value obtained from MRI images of murine macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(m) and NP-IgG(m), n = 3. All the replicates were normalized to the blank (cells that have not been incubated with any nanoparticles). The error bars shows the SEM. $$p < 0.01 versus Blank; *p < 0.05 and **p < 0.01 versus NP-CD163(+); #p < 0.05 versus Ab 1:1. F) Representative MRI phantoms of murine macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(m) and NP-IgG(m).

Mentions: The incubation of cells that expressed (+) or not (−) CD163 (Fig. 2A,D) with the targeted or control probes showed that the NPs bearing the anti-CD163 antibody could selectively detect CD163 expressing macrophages (Fig. 2B-C, E-F). In both human and murine CD163 (+) cells incubated with NP-CD163, there was a significant decrease in the T2 values with respect to CD163 (−) macrophages incubated with the same probe (p < 0.05 either THP-1 or mouse peritoneal macrophages) (See Supplementary tables S1-S6 for statistical details). Likewise, T2 values were significantly lower than those for the same cells incubated with the NP-IgG control probe compared with NP-CD163 (p < 0.001 in THP-1 cell line and p < 0.01 in mouse peritoneal macrophages). Incubation with increasing amounts of free antibody blocked the targeting of CD163 by the NP-CD163 probe with a T2 values similar to that of cells not expressing CD163 or those that were incubated with control probe. This fact showed the specificity of the vectorized probe to target CD163 and excluded unspecific signal due to the probe.


Targeted gold-coated iron oxide nanoparticles for CD163 detection in atherosclerosis by MRI.

Tarin C, Carril M, Martin-Ventura JL, Markuerkiaga I, Padro D, Llamas-Granda P, Moreno JA, García I, Genicio N, Plaza-Garcia S, Blanco-Colio LM, Penades S, Egido J - Sci Rep (2015)

CD163 detection in in vitro models.(A) Western blot of CD163 expression and α-actin (loading control) in THP-1 macrophages cell lysates with or without dexamethasone treatment for 24 hours. (B) Graph showing the normalized T2 values obtained from MRI images of human macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(h) and NP-IgG(h), n  = 3. All the replicates were normalized to the blank (cells that have not been incubated with any nanoparticles). The error bars shows the SEM. $p < 0.05 versus Blank; *p < 0.05, **p < 0.01 and ***p < 0.001 versus NP-CD163(+) (C) Representative MRI phantoms of human macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(h) and NP-IgG(h). (D) Immunodetection of CD163 expression in murine macrophages with or without dexamethasone treatment. Propidium iodide (PI) was used for nuclei staining. (E) Graph showing the normalized mean value obtained from MRI images of murine macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(m) and NP-IgG(m), n = 3. All the replicates were normalized to the blank (cells that have not been incubated with any nanoparticles). The error bars shows the SEM. $$p < 0.01 versus Blank; *p < 0.05 and **p < 0.01 versus NP-CD163(+); #p < 0.05 versus Ab 1:1. F) Representative MRI phantoms of murine macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(m) and NP-IgG(m).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4663748&req=5

f2: CD163 detection in in vitro models.(A) Western blot of CD163 expression and α-actin (loading control) in THP-1 macrophages cell lysates with or without dexamethasone treatment for 24 hours. (B) Graph showing the normalized T2 values obtained from MRI images of human macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(h) and NP-IgG(h), n  = 3. All the replicates were normalized to the blank (cells that have not been incubated with any nanoparticles). The error bars shows the SEM. $p < 0.05 versus Blank; *p < 0.05, **p < 0.01 and ***p < 0.001 versus NP-CD163(+) (C) Representative MRI phantoms of human macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(h) and NP-IgG(h). (D) Immunodetection of CD163 expression in murine macrophages with or without dexamethasone treatment. Propidium iodide (PI) was used for nuclei staining. (E) Graph showing the normalized mean value obtained from MRI images of murine macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(m) and NP-IgG(m), n = 3. All the replicates were normalized to the blank (cells that have not been incubated with any nanoparticles). The error bars shows the SEM. $$p < 0.01 versus Blank; *p < 0.05 and **p < 0.01 versus NP-CD163(+); #p < 0.05 versus Ab 1:1. F) Representative MRI phantoms of murine macrophages CD163 (+) and CD163 (−) incubated with NP-CD163(m) and NP-IgG(m).
Mentions: The incubation of cells that expressed (+) or not (−) CD163 (Fig. 2A,D) with the targeted or control probes showed that the NPs bearing the anti-CD163 antibody could selectively detect CD163 expressing macrophages (Fig. 2B-C, E-F). In both human and murine CD163 (+) cells incubated with NP-CD163, there was a significant decrease in the T2 values with respect to CD163 (−) macrophages incubated with the same probe (p < 0.05 either THP-1 or mouse peritoneal macrophages) (See Supplementary tables S1-S6 for statistical details). Likewise, T2 values were significantly lower than those for the same cells incubated with the NP-IgG control probe compared with NP-CD163 (p < 0.001 in THP-1 cell line and p < 0.01 in mouse peritoneal macrophages). Incubation with increasing amounts of free antibody blocked the targeting of CD163 by the NP-CD163 probe with a T2 values similar to that of cells not expressing CD163 or those that were incubated with control probe. This fact showed the specificity of the vectorized probe to target CD163 and excluded unspecific signal due to the probe.

Bottom Line: We have prepared a targeted probe based on gold-coated iron oxide nanoparticles vectorized with an anti-CD163 antibody for the specific detection of CD163 by MRI.Subsequently, the targeted probe was injected in 16 weeks old apoE deficient mice developing atherosclerotic lesions and the pararenal abdominal aorta was imaged by MRI.The accumulation of probe in the site of interest increased over time and the signal intensity decreased significantly 48 hours after the injection.

View Article: PubMed Central - PubMed

Affiliation: Laboratorio de Patología Vascular y Renal. IIS Fundación Jiménez Díaz, Universidad Autónoma. Av. Reyes Católicos 2, 28040, Madrid, Spain.

ABSTRACT
CD163 is a membrane receptor expressed by macrophage lineage. Studies performed in atherosclerosis have shown that CD163 expression is increased at inflammatory sites, pointing at the presence of intraplaque hemorrhagic sites or asymptomatic plaques. Hence, imaging of CD163 expressing macrophages is an interesting strategy in order to detect atherosclerotic plaques. We have prepared a targeted probe based on gold-coated iron oxide nanoparticles vectorized with an anti-CD163 antibody for the specific detection of CD163 by MRI. Firstly, the specificity of the targeted probe was validated in vitro by incubation of the probe with CD163(+) or (-) macrophages. The probe was able to selectively detect CD163(+) macrophages both in human and murine cells. Subsequently, the targeted probe was injected in 16 weeks old apoE deficient mice developing atherosclerotic lesions and the pararenal abdominal aorta was imaged by MRI. The accumulation of probe in the site of interest increased over time and the signal intensity decreased significantly 48 hours after the injection. Hence, we have developed a highly sensitive targeted probe capable of detecting CD163-expressing macrophages that could provide useful information about the state of the atheromatous lesions.

No MeSH data available.


Related in: MedlinePlus