G. V. Borisevich, S. L. Kirillova, I. V. Shatokhina, V. Lebedev, S. S. Solov’ev, S. Syromyatnikova, N. Shagarova, N. Boyarskaya, N. G. Levkovich, D. A. Solyanik, A. F. Andrus, V. V. Rubtsov, V. T. Krotkov, V. Kulish, I. V. Surovyatkina, V. B. Kirillov, A. V. Koval’chuk, V. Pantyukhov, D. A. Kutaev, S. Borisevich
{"title":"恒河猴实验性感染严重急性呼吸系统综合征冠状病毒2型后细胞免疫的流式细胞术研究","authors":"G. V. Borisevich, S. L. Kirillova, I. V. Shatokhina, V. Lebedev, S. S. Solov’ev, S. Syromyatnikova, N. Shagarova, N. Boyarskaya, N. G. Levkovich, D. A. Solyanik, A. F. Andrus, V. V. Rubtsov, V. T. Krotkov, V. Kulish, I. V. Surovyatkina, V. B. Kirillov, A. V. Koval’chuk, V. Pantyukhov, D. A. Kutaev, S. Borisevich","doi":"10.21055/0370-1069-2022-3-53-60","DOIUrl":null,"url":null,"abstract":"Cellular immunity plays an important role in the pathogenesis and formation of protective immune defense against the SARS‑CoV‑2 virus.The aim of the work was to study the cellular immunity of rhesus monkeys applying flow cytometry after experimental infection with the SARS‑CoV‑2 virus.Materials and methods. Male rhesus monkeys were intranasally inoculated with the SARS‑CoV‑2 virus, Isolate B strain and hCoV-19/Russia/SP48-1226/2020 strain (abbreviated name U-2), at a dose of 5.0 lg PFU. Using flow cytometry, the levels of 21 populations/subpopulations of mononuclear cells in the peripheral blood of animals were determined before experimental infection with the pathogen and on day 14 after infection. SARS‑CoV‑2 coronavirus RNA was assessed using real-time polymerase chain reaction. Determination of the titer of virus-neutralizing antibodies to the SARS‑CoV‑2 virus in the blood sera of animals was conducted through neutralization test evaluating the ability to suppress negative colonies.Results and discussion. Infection with Isolate B strain culture has led to an increase in the relative content of total T-lymphocytes (p˂0.2), cytotoxic T-lymphocytes (p˂0.1), as well as monocytes expressing the early activation marker CD25 (p˂0.2). The decrease in levels has been observed for total B-lymphocytes (p˂0.2) and T-helper cells (p˂0.1). Infection with the U-2 strain culture revealed an increase in the relative content of monocytes expressing the early activation marker CD25 (p˂0.2). Thus, for the first time in the Russian Federation, flow cytometry was used to study the cellular immunity of rhesus monkeys before and after experimental infection with the SARS‑CoV‑2 virus. The obtained information can be used for studying the pathogenesis of SARS‑CoV‑2 infection, course, and outcome of the disease, and developing strategies for vaccination and treatment.","PeriodicalId":52264,"journal":{"name":"Problemy Osobo Opasnykh Infektsii","volume":" ","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2022-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"The Flow Cytometry Study of Cellular Immunity in Rhesus Monkeys after Experimental Infection with SARS CoV 2 Virus\",\"authors\":\"G. V. Borisevich, S. L. Kirillova, I. V. Shatokhina, V. Lebedev, S. S. Solov’ev, S. Syromyatnikova, N. Shagarova, N. Boyarskaya, N. G. Levkovich, D. A. Solyanik, A. F. Andrus, V. V. Rubtsov, V. T. Krotkov, V. Kulish, I. V. Surovyatkina, V. B. Kirillov, A. V. Koval’chuk, V. Pantyukhov, D. A. Kutaev, S. Borisevich\",\"doi\":\"10.21055/0370-1069-2022-3-53-60\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Cellular immunity plays an important role in the pathogenesis and formation of protective immune defense against the SARS‑CoV‑2 virus.The aim of the work was to study the cellular immunity of rhesus monkeys applying flow cytometry after experimental infection with the SARS‑CoV‑2 virus.Materials and methods. Male rhesus monkeys were intranasally inoculated with the SARS‑CoV‑2 virus, Isolate B strain and hCoV-19/Russia/SP48-1226/2020 strain (abbreviated name U-2), at a dose of 5.0 lg PFU. Using flow cytometry, the levels of 21 populations/subpopulations of mononuclear cells in the peripheral blood of animals were determined before experimental infection with the pathogen and on day 14 after infection. SARS‑CoV‑2 coronavirus RNA was assessed using real-time polymerase chain reaction. Determination of the titer of virus-neutralizing antibodies to the SARS‑CoV‑2 virus in the blood sera of animals was conducted through neutralization test evaluating the ability to suppress negative colonies.Results and discussion. Infection with Isolate B strain culture has led to an increase in the relative content of total T-lymphocytes (p˂0.2), cytotoxic T-lymphocytes (p˂0.1), as well as monocytes expressing the early activation marker CD25 (p˂0.2). The decrease in levels has been observed for total B-lymphocytes (p˂0.2) and T-helper cells (p˂0.1). Infection with the U-2 strain culture revealed an increase in the relative content of monocytes expressing the early activation marker CD25 (p˂0.2). Thus, for the first time in the Russian Federation, flow cytometry was used to study the cellular immunity of rhesus monkeys before and after experimental infection with the SARS‑CoV‑2 virus. 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引用次数: 0
摘要
细胞免疫在严重急性呼吸系统综合征冠状病毒2型的发病机制和保护性免疫防御的形成中发挥着重要作用。这项工作的目的是应用流式细胞术研究恒河猴在实验性感染严重急性呼吸系统综合征冠状病毒2型后的细胞免疫。材料和方法。雄性恒河猴鼻内接种SARS‑CoV‑2病毒,分离株B株和hCoV-19/Russia/SP48-1226/2020株(简称U-2),剂量为5.0 lg PFU。使用流式细胞术,在实验性感染病原体之前和感染后第14天测定动物外周血中21个单核细胞群/亚群的水平。使用实时聚合酶链式反应评估严重急性呼吸系统综合征冠状病毒2型的核糖核酸。通过中和试验评估抑制阴性菌落的能力,测定动物血清中针对严重急性呼吸系统综合征冠状病毒2型的病毒中和抗体的滴度。结果和讨论。用分离物B菌株培养物感染导致总T淋巴细胞(p 0.2)、细胞毒性T淋巴细胞(p 0.1)和细胞毒性T细胞的相对含量增加,以及表达早期激活标记CD25的单核细胞(p 0.2)。观察到总B淋巴细胞(p 0.2,流式细胞术用于研究恒河猴实验性感染SARS‑CoV‑2病毒前后的细胞免疫。所获得的信息可用于研究严重急性呼吸系统综合征冠状病毒2型感染的发病机制、病程和结果,以及制定疫苗接种和治疗策略。
The Flow Cytometry Study of Cellular Immunity in Rhesus Monkeys after Experimental Infection with SARS CoV 2 Virus
Cellular immunity plays an important role in the pathogenesis and formation of protective immune defense against the SARS‑CoV‑2 virus.The aim of the work was to study the cellular immunity of rhesus monkeys applying flow cytometry after experimental infection with the SARS‑CoV‑2 virus.Materials and methods. Male rhesus monkeys were intranasally inoculated with the SARS‑CoV‑2 virus, Isolate B strain and hCoV-19/Russia/SP48-1226/2020 strain (abbreviated name U-2), at a dose of 5.0 lg PFU. Using flow cytometry, the levels of 21 populations/subpopulations of mononuclear cells in the peripheral blood of animals were determined before experimental infection with the pathogen and on day 14 after infection. SARS‑CoV‑2 coronavirus RNA was assessed using real-time polymerase chain reaction. Determination of the titer of virus-neutralizing antibodies to the SARS‑CoV‑2 virus in the blood sera of animals was conducted through neutralization test evaluating the ability to suppress negative colonies.Results and discussion. Infection with Isolate B strain culture has led to an increase in the relative content of total T-lymphocytes (p˂0.2), cytotoxic T-lymphocytes (p˂0.1), as well as monocytes expressing the early activation marker CD25 (p˂0.2). The decrease in levels has been observed for total B-lymphocytes (p˂0.2) and T-helper cells (p˂0.1). Infection with the U-2 strain culture revealed an increase in the relative content of monocytes expressing the early activation marker CD25 (p˂0.2). Thus, for the first time in the Russian Federation, flow cytometry was used to study the cellular immunity of rhesus monkeys before and after experimental infection with the SARS‑CoV‑2 virus. The obtained information can be used for studying the pathogenesis of SARS‑CoV‑2 infection, course, and outcome of the disease, and developing strategies for vaccination and treatment.