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

Yunjie Jin; Peng Gao; Lifei Liang; Yuhang Wang; Jiawei Li; Jiyan Wang; Jiangang Hou; Cheng Yang; Xiaolin Wang

doi:10.3389/fimmu.2023.1164183

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

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

Authors

Yunjie Jin^{1

2}, Peng Gao³, Lifei Liang^{4

5}, Yuhang Wang⁶, Jiawei Li^{4

5}, Jiyan Wang^{4

5}, Jiangang Hou³, Cheng Yang^{4

5

7}, Xiaolin Wang¹

Affiliations

¹ Shanghai Institute of Medical Imaging, Shanghai, China.
² Department of Ultrasound, Zhongshan Hospital, Fudan University, Shanghai, China.
³ Department of Urology, Huashan Hospital, Fudan University, Shanghai, China.
⁴ Department of Urology, Zhongshan Hospital, Fudan University, Shanghai, China.
⁵ Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
⁶ Shanghai Medical College, Fudan University, Shanghai, China.
⁷ Zhangjiang Institute of Fudan University, Shanghai, China.

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) in vivo 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_Gzb) 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_Gzb group than in the allogeneic MB_con group and the isogeneic MB_con 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.

Keywords: cardiac transplantation; contrast enhanced ultrasound (CEUS); granzyme B (GzmB); rejection; ultrasound imaging.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Allografts
Animals
Diagnostic Imaging*
Granzymes
Interleukin-6*
Mice
Ultrasonography

Substances

Granzymes
Interleukin-6

Grants and funding

This study was supported by the National Natural Science Foundation of China (82170765 to CY), Shanghai Municipal Key Clinical Specialty (shslczdzk05802), Project ELITE: A Special Supportive Program for Organ Transplantation by COTDF (2019JYJH05 to CY), and the 2019 Shanghai Youth Talent Development Program (to CY).