来自新冠疫情四个阶段患者的心肌比较研究。

Comparative Study of the Myocardium of Patients from Four COVID-19 Waves.

作者信息

Mitrofanova Lubov Borisovna, Makarov Igor Aleksandrovich, Gorshkov Andrey Nikolaevich, Runov Andrey Leonidovich, Vonsky Maxim Sergeevich, Pisareva Maria Mikhailovna, Komissarov Andrey Borisovich, Makarova Taiana Alekseevna, Li Qingli, Karonova Tatiana Leonidovna, Konradi Alexandra Olegovna, Shlaykhto Evgeny Vladimirovich

机构信息

Almazov National Medical Research Centre, St. Petersburg 197341, Russia.

Smorodintsev Research Institute of Influenza, St. Petersburg 197376, Russia.

出版信息

Diagnostics (Basel). 2023 May 7;13(9):1645. doi: 10.3390/diagnostics13091645.

DOI:10.3390/diagnostics13091645

PMID:37175037

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10178873/

Abstract

BACKGROUND

Few studies have compared COVID-19 patients from different waves. This study aims to conduct a clinical and morphological analysis of patients who died from COVID-19 during four waves.

METHODS

The study involved 276 patients who died from COVID-19 during four waves, including 77 patients in the first wave, 119 patients in the second wave, and 78 patients in the third wave. We performed a histological examination of myocardium samples from autopsies and additionally analyzed the samples by PCR. We conducted immunohistochemistry of the myocardium for 21 samples using antibodies against CD3, CD45, CD8, CD68, CD34, Ang1, VWF, VEGF, HLA-DR, MHC1, C1q, enteroviral VP1, and SARS-CoV-2 spike protein. We also did immunofluorescent staining of three myocardial specimens using VP1/SARS-CoV-2 antibody cocktails. Further, we ran RT-ddPCR analysis for 14 RNA samples extracted from paraffin-embedded myocardium. Electron microscopic studies of the myocardium were also performed for two samples from the fourth wave.

RESULTS

Among the 276 cases, active myocarditis was diagnosed in 5% (15/276). Of these cases, 86% of samples expressed VP1, and individual cells contained SARS-CoV-2 spike protein in 22%. Immunofluorescence confirmed the co-localization of VP1 and SARS-CoV-2 spike proteins. ddPCR did not confidently detect SARS-CoV-2 RNA in the myocardium in any myocarditis cases. However, the myocardium sample from wave IV detected a sub-threshold signal of SARS-CoV-2 by qPCR, but myocarditis in this patient was not confirmed. Electron microscopy showed several single particles similar to SARS-CoV-2 virions on the surface of the endothelium of myocardial vessels. A comparison of the cardiovascular complication incidence between three waves revealed that the incidence of hemorrhage (48 vs. 24 vs. 17%), myocardial necrosis (18 vs. 11 vs. 4%), blood clots in the intramural arteries (12 vs. 7 vs. 0%), and myocarditis (19 vs. 1 vs. 6%) decreased over time, and CD8-T-killers appeared. Immunohistochemistry confirmed the presence of endotheliitis in all 21 studied cases.

CONCLUSIONS

This study compared myocardial damage in patients who died during three COVID-19 waves and showed a decrease in the incidence of endotheliitis complications (thrombosis, hemorrhage, necrosis) and myocarditis over time. However, the connection between myocarditis and SARS-CoV-2 infection remains unproven.

摘要

背景

很少有研究对不同疫情波次的新冠病毒病（COVID-19）患者进行比较。本研究旨在对在四个疫情波次中死于COVID-19的患者进行临床和形态学分析。

方法

本研究纳入了276例在四个疫情波次中死于COVID-19的患者，其中第一波77例，第二波119例，第三波78例。我们对尸检获取的心肌样本进行了组织学检查，并通过聚合酶链反应（PCR）对样本进行了额外分析。我们使用抗CD3、CD45、CD8、CD68、CD34、血管生成素1（Ang1）、血管性血友病因子（VWF）、血管内皮生长因子（VEGF）、人类白细胞抗原-DR（HLA-DR）、主要组织相容性复合体1（MHC1）、补体C1q、肠道病毒VP1和严重急性呼吸综合征冠状病毒2（SARS-CoV-2）刺突蛋白的抗体，对21个心肌样本进行了免疫组织化学检测。我们还使用VP1/SARS-CoV-2抗体混合物对三个心肌标本进行了免疫荧光染色。此外，我们对从石蜡包埋心肌中提取的14个RNA样本进行了逆转录-数字液滴聚合酶链反应（RT-ddPCR）分析。还对第四波的两个样本进行了心肌的电子显微镜研究。

结果

在276例病例中，5%（15/276）被诊断为活动性心肌炎。在这些病例中，86%的样本表达VP1，22%的单个细胞含有SARS-CoV-2刺突蛋白。免疫荧光证实了VP1和SARS-CoV-2刺突蛋白的共定位。ddPCR在任何心肌炎病例的心肌中均未可靠检测到SARS-CoV-2 RNA。然而，第四波的心肌样本通过定量聚合酶链反应（qPCR）检测到SARS-CoV-2的亚阈值信号，但该患者的心肌炎未得到证实。电子显微镜显示心肌血管内皮表面有几个类似于SARS-CoV-2病毒粒子的单个颗粒。对三个疫情波次之间心血管并发症发生率的比较显示，出血（48%对24%对17%）、心肌坏死（18%对11%对4%）、壁内动脉血栓（12%对7%对0%）和心肌炎（19%对1%对6%）的发生率随时间下降，并且出现了CD8-T杀伤细胞。免疫组织化学证实在所有21例研究病例中均存在内皮炎。