The Journal of Heart and Lung Transplantation ( IF 8.9 ) Pub Date : 2021-10-25 , DOI: 10.1016/j.healun.2021.10.010 Rafaela V P Ribeiro 1 , Terrance Ku 2 , Aizhou Wang 1 , Layla Pires 1 , Victor H Ferreira 2 , Vinicius Michaelsen 1 , Aadil Ali 1 , Marcos Galasso 1 , Sajad Moshkelgosha 1 , Anajara Gazzalle 1 , Mads G Jeppesen 3 , Mette M Rosenkilde 4 , Mingyao Liu 1 , Lianne G Singer 2 , Deepali Kumar 2 , Shaf Keshavjee 5 , John Sinclair 6 , Thomas N Kledal 3 , Atul Humar 2 , Marcelo Cypel 5
Background
Transmission of latent human cytomegalovirus (HCMV) via organ transplantation with post-transplant viral reactivation is extremely prevalent and results in substantial adverse impact on outcomes. Therapies targeting the latent reservoir within the allograft to mitigate viral transmission would represent a major advance. Here, we delivered an immunotoxin (F49A-FTP) that targets and kills latent HCMV aiming at reducing the HCMV reservoir from donor lungs using ex-vivo lung perfusion (EVLP).
Methods
HCMV seropositive human lungs were placed on EVLP alone or EVLP + 1mg/L of F49A-FTP for 6 hours (n = 6, each). CD14+ monocytes isolated from biopsies pre and post EVLP underwent HCMV reactivation assay designed to evaluate viral reactivation capacity. Off-target effects of F49A-FTP were studied evaluating cell death markers of CD34+ and CD14+ cells using flow cytometry. Lung function on EVLP and inflammatory cytokine production were evaluated as safety endpoints.
Results
We demonstrate that lungs treated ex-vivo with F49A-FTP had a significant reduction in HCMV reactivation compared to controls, suggesting successful targeting of latent virus (76% median reduction in F49A-FTP vs 15% increase in controls, p = 0.0087). Furthermore, there was comparable cell death rates of the targeted cells between both groups, suggesting no off-target effects. Ex-vivo lung function was stable over 6 hours and no differences in key inflammatory cytokines were observed demonstrating safety of this novel treatment.
Conclusions
Ex-vivo F49A-FTP treatment of human lungs targets and kills latent HCMV, markedly attenuating HCMV reactivation. This approach demonstrates the first experiments targeting latent HCMV in a donor organ with promising results towards clinical translation.
中文翻译:
使用一种新型的基于趋化因子的免疫毒素对人类供体肺中的巨细胞病毒进行离体治疗
背景
通过器官移植传播潜伏性人巨细胞病毒(HCMV) 并伴有移植后病毒再激活的情况极为普遍,并对结果造成重大不利影响。针对同种异体移植物内的潜在储库以减轻病毒传播的疗法将代表重大进步。在这里,我们提供了一种免疫毒素 (F49A-FTP),它靶向并杀死潜伏的 HCMV,旨在使用离体肺灌注 (EVLP) 减少供体肺中的 HCMV 储库。
方法
将 HCMV 血清反应阳性的人肺置于单独的 EVLP 或 EVLP + 1mg/L 的 F49A-FTP 上 6 小时(每个n = 6 )。从 EVLP 前后活检中分离的CD14+ 单核细胞进行了 HCMV 再激活测定,旨在评估病毒再激活能力。研究了 F49A-FTP 的脱靶效应,使用流式细胞术评估 CD34+ 和 CD14+ 细胞的细胞死亡标志物。EVLP 的肺功能和炎性细胞因子的产生被评估为安全终点。
结果
我们证明,与对照组相比,用 F49A-FTP 离体治疗的肺在 HCMV 再激活方面有显着减少,表明成功靶向潜伏病毒(F49A-FTP 中位数减少 76%,而对照组增加 15%,p = 0.0087)。此外,两组之间靶细胞的细胞死亡率相当,表明没有脱靶效应。离体肺功能在 6 小时内保持稳定,并且未观察到关键炎症细胞因子的差异,证明了这种新型治疗的安全性。
结论
人肺的离体 F49A-FTP 治疗靶向并杀死潜伏的 HCMV,显着减弱 HCMV 再激活。这种方法展示了第一个针对供体器官中潜伏的 HCMV 的实验,对临床转化有希望的结果。