Makarov Igor, Mayrina Sofya, Makarova Taiana, Karonova Tatiana, Starshinova Anna, Kudlay Dmitry, Mitrofanova Lubov
Almazov National Medical Research Centre, 197341 St. Petersburg, Russia.
Department of Pharmacology, Institute of Pharmacy, I.M. Sechenov First Moscow State Medical University, 119435 Moscow, Russia.
Diagnostics (Basel). 2023 Jun 29;13(13):2212. doi: 10.3390/diagnostics13132212.
The clinical manifestation study of post-acute sequelae of SARS-CoV-2 infection (PASC) has shown a lack of knowledge regarding its morphology and pathogenesis. The aim of this research was to investigate morphological manifestations of PASC in the myocardium.
The study included 38 patients requiring endomyocardial biopsy (EMB) during the post-acute phase of coronavirus infection and a control group including patients requiring EMB prior to the SARS-CoV-2 pandemic. The patients' clinical and laboratory data were analyzed. Histological examination and immunohistochemistry (IHC) of the myocardial tissue was conducted with antibodies to CD3, CD68, HLA-DR, MHC1, C1q, VP1 enteroviruses, spike protein SARS-CoV-2, Ang1, von Willebrand factor (VWF), and VEGF. The morphometric analysis included counting the mean number of inflammatory infiltrate cells per mm and evaluating the expression of SARS-CoV-2 spike protein, HLA-DR, MHC1, C1q, Ang1, VWF, and VEGF using a scoring system. If the expression of SARS-CoV-2 spike protein was >3 points, an additional IHC test with antibodies to ACE2, CD16 as well as RT-PCR testing of the myocardial tissue were performed. For two patients, immunofluorescence tests of the myocardial tissue were performed using antibody cocktails to SARS-CoV-2 spike protein/CD16, SARS-CoV-2 spike protein/CD68, CD80/CD163. The statistical data analysis was carried out using the Python programming language and libraries such as NumPy, SciPy, Pandas, and Matplotlib.
The study demonstrated a significant increase in the number of CD68+ macrophages in the myocardium of PASC patients compared to patients who did not have a history of COVID-19 ( = 0.014 and = 0.007 for patients with and without myocarditis, respectively), predominantly due to M2 macrophages. An increase in the number of CD68+ macrophages was more frequently observed in patients with shorter intervals between the most recent positive SARS-CoV-2 PCR test and the time of performing the EMB (r = -0.33 and r = -0.61 for patients with and without myocarditis, respectively). The expression scores of Ang1, VEGF, VWF, and C1q in PASC patients did not significantly differ from those in EMB samples taken before 2019.
The myocardium of PASC patients demonstrated a significant increase in the number of CD68+ macrophages and a decrease in the expression of markers associated with angiopathy. No evidence of coronavirus-associated myocarditis was observed in any PASC patient.
严重急性呼吸综合征冠状病毒2感染后急性后遗症（PASC）的临床表现研究表明，人们对其形态和发病机制缺乏了解。本研究的目的是调查PASC在心肌中的形态学表现。
该研究纳入了38例在冠状病毒感染急性期需要进行心内膜心肌活检（EMB）的患者，以及一个对照组，该对照组包括在严重急性呼吸综合征冠状病毒2大流行之前需要进行EMB的患者。分析了患者的临床和实验室数据。使用针对CD3、CD68、HLA-DR、MHC1、C1q、肠道病毒VP1、严重急性呼吸综合征冠状病毒2刺突蛋白、血管生成素1（Ang1）、血管性血友病因子（VWF）和血管内皮生长因子（VEGF）的抗体对心肌组织进行组织学检查和免疫组织化学（IHC）检测。形态计量分析包括计算每毫米炎症浸润细胞的平均数，并使用评分系统评估严重急性呼吸综合征冠状病毒2刺突蛋白、HLA-DR、MHC1、C1q、Ang1、VWF和VEGF的表达。如果严重急性呼吸综合征冠状病毒2刺突蛋白的表达>3分，则使用针对血管紧张素转换酶2（ACE2）和CD16的抗体进行额外的IHC检测，并对心肌组织进行逆转录聚合酶链反应（RT-PCR）检测。对两名患者，使用针对严重急性呼吸综合征冠状病毒2刺突蛋白/CD16、严重急性呼吸综合征冠状病毒2刺突蛋白/CD68、CD80/CD163的抗体混合物对心肌组织进行免疫荧光检测。使用Python编程语言以及NumPy、SciPy、Pandas和Matplotlib等库进行统计数据分析。
研究表明，与没有冠状病毒病（COVID-19）病史的患者相比，PASC患者心肌中CD68+巨噬细胞数量显著增加（有心肌炎和无心肌炎的患者分别为P = 0.014和P = 0.007），主要是M2巨噬细胞。在最近一次严重急性呼吸综合征冠状病毒2聚合酶链反应（PCR）检测呈阳性与进行EMB的时间间隔较短的患者中，更常观察到CD68+巨噬细胞数量增加（有心肌炎和无心肌炎的患者分别为r = -0.33和r = -0.61）。PASC患者中Ang1、VEGF、VWF和C1q 的表达评分与2019年之前采集的EMB样本中的评分没有显著差异。
PASC患者的心肌显示CD68+巨噬细胞数量显著增加，与血管病变相关的标志物表达降低。在任何PASC患者中均未观察到冠状病毒相关心肌炎的证据。
