In situ labeling of immune cells with iron oxide particles: an approach to detect organ rejection by cellular MRI - PubMed (original) (raw)

In situ labeling of immune cells with iron oxide particles: an approach to detect organ rejection by cellular MRI

Yijen L Wu et al. Proc Natl Acad Sci U S A. 2006.

Abstract

In vivo cell tracking by MRI can provide means to observe biological processes and monitor cell therapy directly. Immune cells, e.g., macrophages, play crucial roles in many pathophysiological processes, including organ rejection, inflammation, autoimmune diseases, cancer, atherosclerotic plaque formation, numerous neurological disorders, etc. The current gold standard for diagnosing and staging rejection after organ transplantation is biopsy, which is not only invasive, but also prone to sampling errors. Here, we report a noninvasive approach using MRI to detect graft rejection after solid organ transplantation. In addition, we present the feasibility of imaging individual macrophages in vivo by MRI in a rodent heterotopic working-heart transplantation model using a more sensitive contrast agent, the micrometer-sized paramagnetic iron oxide particle, as a methodology to detect acute cardiac rejection.

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Conflict of interest statement

Conflict of interest statement: No conflicts declared.

Figures

Fig. 1.

Fig. 1.

In vivo MRI of allograft hearts and lungs, 1 day after i.v. injection of MPIO particles. (A) Allograft heart on POD 5. (B and C) Allograft heart on POD 6. (D) Allograft lung on POD 6. (E) Isograft heart on POD 6. Shown with 156-μm in-plane resolution at 4.7 Tesla by using a Bruker Biospec AVANCE-DBX MRI instrument.

Fig. 2.

Fig. 2.

In vivo MRI of a rat allograft heart over time. MPIO particles are administered once on POD 3.5, and the same animal is imaged on POD 3.5 (A), 4.5 (B), and 5.5 (C), with 156-μm in-plane resolution at 4.7 Tesla by using a Bruker Biospec AVANCE-DBX MRI instrument.

Fig. 3.

Fig. 3.

Contrast patterns labeled with two different contrast agents. (A_–_C) In vivo MRI of macrophage accumulation on POD 6 of an allograft heart with MPIO-particle labeling (A), an isograft heart with MPIO-particle labeling (B), and an allograft heart with USPIO labeling (C), with 156-μm in-plane resolution at 4.7 Tesla. (D_–_F) MRM at 11.7 Tesla using a Bruker AVANCE-DBX MRI instrument with in-plane resolution of 40 μm of an allograft heart with MPIO-particle labeling (D), an isograft heart (E), and an allograft heart with USPIO labeling (F). All hearts used in the in vivo and ex vivo measurements were the same ones except those used in A and D.

Fig. 4.

Fig. 4.

Ex-vivo labeled macrophages. Light microscopy (A) and fluorescent microscopy (B) of ex vivo MPIO-labeled macrophages. (Scale bars: 50 μm.) (C) MRM at 11.7-Tesla using a Bruker AVANCE-DBX MRI instrument with in-plane resolution of 40 μm of gelatin phantoms containing isolated macrophages labeled with MPIO particles with different cell concentration.

Fig. 5.

Fig. 5.

Histological examination of allograft hearts on POD 6 after in vivo MPIO-particle labeling. (A_–_C) Optical micrograph (×200 magnification) of three neighboring 5-μm tissue sections of a POD 6 allograft heart stained with Perl’s Prussian blue for iron (A, blue; with pink background counterstaining), anti-rat ED1 for macrophage (B, brown; with blue background counterstaining), and hematoxylin/eosin staining (C) for tissue integrity. (D and E) Optical micrograph (×400 magnification) of two neighboring 5-μm tissue sections of another POD 6 allograft heart stained with Perl’s Prussian blue for iron (D, blue; with pink background counterstaining), and anti-rat ED1 for macrophage (E, brown; with blue background counterstaining). (F and G) Partial view of optical micrograph (×400 magnification) of two neighboring 5-μm tissue sections of a POD 6 allograft heart stained with Perl’s Prussian blue for iron (F, blue; with pink background counterstaining) and anti-rat ED1 for macrophage (G, brown; with blue background counterstaining). (Scale bars: 75 μm.) (H) Electron micrograph of an allograft heart harvested on POD 6 after in vivo MPIO-particle labeling. (Inset) Enlarged area. (Scale bar: 100 nm.)

Fig. 6.

Fig. 6.

Double fluorescence for MPIO particles and ED1. Shown are fluorescence microscopy of Dragon green for MPIOs (A) and anti-ED1 (red) immunofluorescence microscopy for microphages of the same field of view (B). The boxes select three regions for overlaying double fluorescence. The overlay images are enlarged on the Right.

Fig. 7.

Fig. 7.

Temporal progression of ED1+ cell infiltration. Optical micrograph (×400 magnification) of anti-rat ED1+ immunohistochemical staining sections of allograft hearts obtained on POD 3 (A), POD 4 (B), POD 5 (C), and POD 6 (D). The ED1+ cells appear brown and the background is counterstained blue.

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