mimmunМедицинская иммунологияMedical Immunology (Russia)1563-06252313-741XSPb RAACI10.15789/1563-0625-IDC-2594mimmun-2594Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESИнтеграционный диагностический критерий, оценивающий тяжесть течения COVID-19 и риск возникновения постковидного синдромаIntegrative diagnostic criterion for evaluation of COVID-19 severity and the risk of post-COVID syndromeНестероваИ. В.NesterovaI. V.

Нестерова И.В. – д.м.н., профессор, главный научный сотрудник отдела клинико-экспериментальной иммунологии и молекулярной биологии Центральной научно-исследовательской лаборатории; профессор кафедры клинической иммунологии, аллергологии и адаптологии факультета непрерывного медицинского образования Медицинского института

117513, Россия, Москва, Ленинский пр., 123, кв. 1

Тел.: 8 (916) 187-73-41

Nesterova I.V., PhD, MD (Medicine), Professor, Chief Research Associate, Department of Clinical and Experimental Immunology and Molecular Biology, Central Scientific Research Laboratory; Professor, Department of Clinical Immunology, Allergology and Adaptology

123 Leninsky Ave, Apt 1 Moscow 117513 Russian Federation

Phone: +7 (916) 187-73-41

 

inesterova1@yandex.ruАтажаховаМ. Г.AtazhakhovaM. G.

Атажахова М.Г. – аспирант кафедры клинической иммунологии, аллергологии и лабораторной диагностики ФПК и ППС

г. Краснодар

Atazhakhova M.G., Postgraduate Student, Department of Clinical Immunology, Allergology and Laboratory Diagnostics

Krasnodar

МатушкинаВ. А.MatushkinaV. A.

Матушкина В.А. – ассистент кафедры инфекционных болезней и эпидемиологии ФПК и ППС

г. Краснодар

Matushkina V.A., Assistant Professor, Department of Infectious Diseases and Epidemiology

Krasnodar

ТетеринЮ. В.TeterinYu. V.

Тетерин Ю.В. – аспирант кафедры клинической иммунологии, аллергологии и лабораторной диагностики ФПК и ППС

г. Краснодар

Teterin Yu.V., Postgraduate Student, Department of Clinical Immunology, Allergology and Laboratory Diagnostics

Krasnodar

ГородинВ. Н.GorodinV. N.

Городин В.Н. – д.м.н., доцент, заведующий кафедрой инфекционных болезней и эпидемиологии ФПК и ППС

г. Краснодар

Gorodin V.N., PhD, MD (Medicine), Associate Professor, Head, Department of Infectious Diseases and Epidemiology

Krasnodar

ЧудиловаГ. А.ChudilovaG. A.

Чудилова Г.А. – д.б.н., доцент, заведующая отделом клинико-экспериментальной иммунологии и молекулярной биологии Центральной научно-исследовательской лаборатории, профессор кафедры клинической иммунологии, аллергологии и лабораторной диагностики ФПК и ППС

г. Краснодар

Chudilova G.A., PhD, MD (Biology), Associate Professor, Head of the Department of Clinical and Experimental Immunology and Molecular Biology of the Central Scientific Research Laboratory, Professor of the Department of Clinical Immunology, Allergology and Laboratory Diagnostics

Krasnodar

ФГБОУ ВО «Кубанский государственный медицинский университет» Министерства здравоохранения РФ; ФГАОУ ВО «Российский университет дружбы народов имени Патриса Лумумбы»РоссияKuban State Medical University; P. Lumumba Peoples’ Friendship UniversityRussian FederationФГБОУ ВО «Кубанский государственный медицинский университет» Министерства здравоохранения РФРоссияKuban State Medical UniversityRussian Federation202409112022263545554Copyright © Нестерова И.В., Атажахова М.Г., Матушкина В.А., Тетерин Ю.В., Городин В.Н., Чудилова Г.А., 20242024Нестерова И.В., Атажахова М.Г., Матушкина В.А., Тетерин Ю.В., Городин В.Н., Чудилова Г.А.Nesterova I.V., Atazhakhova M.G., Matushkina V.A., Teterin Y.V., Gorodin V.N., Chudilova G.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.mimmun.ru/mimmun/article/view/2594

Резюме. Патофизиология тяжелой формы COVID-19 характеризуется изменением количества, фенотипа и функций нейтрофильных гранулоцитов (НГ). Среди эффекторных противовирусных механизмов НГ одними из наиболее важных являются нейтрофильные внеклеточные ловушки (NETs), но чрезмерное их образование усугубляет воспаление при остром респираторном дистресс-синдроме и способствует тромбозу микрососудов. Их обнаружение и количественная оценка могут иметь важное значение при различных формах течения COVID-19 для определения корреляции с исходом заболевания, оценки риска развития постковидного синдрома и, возможно, мониторинга будущей целевой терапии.

Цель исследования – разработать новый диагностический интеграционный критерий, позволяющий оценить тяжесть течения COVID-19 и риск развития осложнений в постковидном периоде, в том числе постковидного синдрома в периферической крови. Исследованы образцы периферической крови (ПК) 31 пациента с острым течением COVID-19 (среднетяжелого течения (n = 15) и тяжелого течения (n = 16)), 52 пациентов, выписанных из стационара после лечения COVID-19 тяжелой степени тяжести, в сроки от 30 до 60 дней, имеющие постковидный синдром (ПКС) и 100 условно здоровых добровольцев. Оценивались показатели общеклинического анализа крови (MicroCC-20Plus), в мазках ПК проводился подсчет лейкоцитарной формулы с учетом количества образованных NET и НГ, ушедших в патологический апоптоз. На основе полученных результатов рассчитывался интеграционный диагностический критерий по формуле:

$$ ИДК = \frac{\%\ неизменных\ НГ}{\%NET + \%НГ\ в\ апоптозе} $$

Показано снижение ИДК при среднетяжелом течении заболевания в 8,5 раза (p < 0,05), а при тяжелом течении – в 30 раз (p < 0,05), по сравнению со значениями в группе условно здоровых лиц. Также установлено, что у 88,5% пациентов с ПКС, перенесших SARS-CoV-2, в ПК не выявлено морфологически патологических измененных НГ. В то же время у 11,5% пациентов с ПКС отмечено появление NETs и клеток с патологическим апоптозом, при этом ИДК НГ-ПКС был в 8 раз меньше (p < 0,05), чем в группе сравнения и не отличался от показателей пациентов со среднетяжелым течением COVID-19 (p > 0,05), что диктует необходимость дальнейшего диспансерного наблюдения таких пациентов.

Полученные в настоящем исследовании данные свидетельствуют о том, что разработанный интеграционный диагностический критерий позволяет оценить как тяжесть течения COVID-19 в острый период, так и риск возникновения постковидного синдрома. Следует подчеркнуть, что выявленные при COVID-19 характерные изменения НГ можно легко идентифицировать в ПК и последовательно отслеживать по расчетному интегральному диагностическому критерию. Значительное снижение ИДК свидетельствует о сохраняющейся гиперактивации НГ и необходимости проведения таргетной иммунотерапии, направленной на модулирование дисфункций НГ.

Pathophysiology of severe COVID-19 is characterized by changes in the number, phenotype, and function of neutrophil granulocytes (NG). Among the effector antiviral mechanisms of NG, the neutrophil extracellular traps (NETs) are among the most important features. However, their excessive formation exacerbates inflammation in acute respiratory distress syndrome and contributes to microvascular thrombosis. Their detection and counting may be important in severity grading of COVID-19, for determining correlations with clinical outcome, assessing the risk of developing post-COVID syndrome, and, possibly, for monitoring future targeted therapy. Purpose of our study was to develop a new diagnostic integrative criterion to assess the severity of COVID-19 and the risk of complications in the post-COVID period, including post-COVID signs in peripheral blood. Peripheral blood (PB) samples were studied from 31 patients with acute COVID-19 of moderate (n = 15) and severe degrees (n = 16). Moreover, we observed 52 patients discharged from the hospital after severe COVID-19, with diagnosed post-COVID syndrome (PCS) over the period of 30 to 60 days, and 100 healthy volunteers. The parameters of routine blood counts (MicroCC-20Plus) were evaluated, the leukocyte formula was calculated in PC smears, taking into account the number of formed NETs, and NGs entering pathological apoptosis. Based on the obtained results, an integral diagnostic criterion was calculated using the formula:

$$ IDK = \frac{\%\ unchanged\ NG}{\%NET + \%NG\ in\ apoptosis} $$

A 8.5-fold decrease in IDK index (p < 0.05) was shown in the cases of moderate-severity course of the disease, and a 30-fold drop was seen in severe cases (p < 0.05) compared with appropriate values in the group of healthy individuals. It was also found that, in 88.5% of patients with PCS after the SARS-CoV-2 infection, no morphologically altered NG were detectable in PB samples. At the same time, in 11.5% of patients with PCS, we found NETs and cells with pathological apoptosis, whereas IDC of NG-PCS was 8 times less than in the comparison group, and did not differ from the parameters of patients with moderate COVID-19 (p > 0.05) thus requiring further dispensary observation of such patients. The data obtained in this study indicate that the developed integrative diagnostic criterion allows us to assess both the severity of COVID-19 over acute period, and the risk of post-COVID syndrome. It should be emphasized that the characteristic changes in NG detected in COVID-19 may be readily identified in PB and consistently monitored by the proposed integral diagnostic criterion. A significant decrease in IDC indicates the persisting hyper-activation of NG and a need for targeted immunotherapy aimed at modulating the NG dysfunction.

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The authors declare that there are no conflicts of interest present.