In vivo imaging of the inflammatory receptor CD40 after cerebral ischemia using a fluorescent antibody
In vivo imaging of the inflammatory receptor CD40 after cerebral ischemia using a fluorescent antibody
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DOI：
10.1161/strokeaha.107.509844
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发表时间：
2008-10-01
期刊：
影响因子：
8.3
通讯作者：
Wunder, Andreas
中科院分区：
文献类型：
--
作者：
Klohs, Jan;Graefe, Michael;Wunder, Andreas
背景和目的-脑部炎症是中风的一个标志，它与组织损伤和修复有关。将这些过程具体可视化的成像技术是非常理想的。本研究利用近红外荧光染料Cy5.5(Cy5.5-CD40MAb)标记的荧光单抗，探讨CD40受体在小鼠脑缺血后能否无创、特异地显影。小鼠静脉注射Cy5.5-CD40MAb或对照Cy5.5-IgGMAb。注射化合物后进行非侵入性和体外近红外荧光成像。结果：在注射Cy5.5-CD40MAb的野生型小鼠中，仅在注射Cy5.5-CD40MAb的野生型小鼠中检测到明显高于对侧的荧光强度，而在注射Cy5.5-CD40MAb的CD40缺陷小鼠中或注射Cy5.5-IgGMAb的野生型小鼠中未检测到明显的荧光强度。体外近红外荧光显示，只有注射了Cy5.5-CD40MAb的野生型小鼠在缺血区内有强烈的荧光。在这些小鼠的大脑中，单平面照明显微镜显示了血管和实质的分布，共聚焦显微镜显示了注射Cy5.5-CD40MAb的实质荧光与激活的小胶质细胞和缺血区的血源性细胞部分共存。结论--研究表明，CD40靶向的荧光抗体能够利用光学技术对脑缺血后炎症受体CD40进行特异性的非侵入性检测。
Background and Purpose - Brain inflammation is a hallmark of stroke, where it has been implicated in tissue damage as well as in repair. Imaging technologies that specifically visualize these processes are highly desirable. In this study, we explored whether the inflammatory receptor CD40 can be noninvasively and specifically visualized in mice after cerebral ischemia using a fluorescent monoclonal antibody, which we labeled with the near-infrared fluorescence dye Cy5.5 (Cy5.5-CD40MAb).Methods - Wild-type and CD40-deficient mice were subjected to transient middle cerebral artery occlusion. Mice were either intravenously injected with Cy5.5-CD40MAb or control Cy5.5-IgGMAb. Noninvasive and ex vivo near-infrared fluorescence imaging was performed after injection of the compounds. Probe distribution and specificity was further assessed with single-plane illumination microscopy, immunohistochemistry, and confocal microscopy.Results - Significantly higher fluorescence intensities over the stroke-affected hemisphere, compared to the contralateral side, were only detected noninvasively in wild-type mice that received Cy5.5-CD40MAb, but not in CD40-deficient mice injected with Cy5.5-CD40MAb or in wild-type mice that were injected with Cy5.5-IgGMAb. Ex vivo near-infrared fluorescence showed an intense fluorescence within the ischemic territory only in wild-type mice injected with Cy5.5-CD40MAb. In the brains of these mice, single-plane illumination microscopy demonstrated vascular and parenchymal distribution, and confocal microscopy revealed a partial colocalization of parenchymal fluorescence from the injected Cy5.5-CD40MAb with activated microglia and blood-derived cells in the ischemic region.Conclusions - The study demonstrates that a CD40-targeted fluorescent antibody enables specific noninvasive detection of the inflammatory receptor CD40 after cerebral ischemia using optical techniques.