靶向超声成像技术无创定量检测心脏移植排斥反应中的颗粒酶 B。

Noninvasive quantification of granzyme B in cardiac allograft rejection using targeted ultrasound imaging.

机构信息

Shanghai Institute of Medical Imaging, Shanghai, China.

Department of Ultrasound, Zhongshan Hospital, Fudan University, Shanghai, China.

出版信息

Front Immunol. 2023 Jun 26;14:1164183. doi: 10.3389/fimmu.2023.1164183. eCollection 2023.

DOI:10.3389/fimmu.2023.1164183

PMID:37435082

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10331296/

Abstract

OBJECTIVE

Endomyocardial biopsy is the gold standard method for the diagnosis of cardiac allograft rejection. However, it causes damage to the heart. In this study, we developed a noninvasive method for quantification of granzyme B (GzB) by targeted ultrasound imaging, which detects and provides quantitative information for specific molecules, for acute rejection assessment in a murine cardiac transplantation model.

METHODS

Microbubbles bearing anti-GzB antibodies (MB) or isotype antibodies (MBcon) were prepared. Hearts were transplanted from C57BL/6J (allogeneic) or C3H (syngeneic) donors to C3H recipients. Target ultrasound imaging was performed on Days 2 and 5 post-transplantations. A pathologic assessment was performed. The expression of granzyme B and IL-6 in the heart was detected by Western blotting.

RESULTS

After MB injection, we observed and collected data at 3 and 6 min before and after the flash pulse. Quantitative analysis revealed that the reduction in peak intensity was significantly higher in the allogeneic MB group than in the allogeneic MB group and the isogeneic MB group at PODs 2 and 5. In the allogeneic groups, granzyme B and IL-6 expression levels were higher than those in the isogeneic group. In addition, more CD8 T cells and neutrophils were observed in the allogeneic groups.

CONCLUSION

Ultrasound molecular imaging of granzyme B can be used as a noninvasive method for acute rejection detection after cardiac transplantation.

摘要

目的

心肌内膜活检是诊断心脏移植物排斥反应的金标准方法。然而，它会对心脏造成损伤。在这项研究中，我们开发了一种通过靶向超声成像定量检测颗粒酶 B (GzB) 的非侵入性方法，该方法可检测和提供特定分子的定量信息，用于评估小鼠心脏移植模型中的急性排斥反应。

方法

制备携带抗 GzB 抗体（MB）或同种型抗体（MBcon）的微泡。将 C57BL/6J（同种异体）或 C3H（同基因）供体的心脏移植到 C3H 受体中。在移植后第 2 天和第 5 天进行靶向超声成像。进行病理评估。通过 Western blot 检测心脏中颗粒酶 B 和 IL-6 的表达。

结果

注射 MB 后，我们在闪光脉冲前后 3 分钟和 6 分钟观察并收集数据。定量分析显示，在 POD2 和 POD5 时，同种异体 MB 组的峰值强度降低幅度明显高于同种异体 MB 组和同基因 MB 组。在同种异体组中，颗粒酶 B 和 IL-6 的表达水平高于同基因组。此外，在同种异体组中观察到更多的 CD8 T 细胞和中性粒细胞。