Перспективы использования технологической платформы ELISPOT в системе противоэпидемических мероприятий против новой коронавирусной инфекции COVID-19

Определение наличия Т-клеточного иммунного ответа к вирусу SARS-CoV-2 актуально как для диагностики заболевания у пациентов с наличием симптомов, так и для определения общего количества людей, перенесших данное заболевание, в том числе бессимптомно. Кроме того, данные тесты эффективны для оценки иммунного ответа после проведения вакцинации, а также оценки напряженности специфического иммунитета в группах риска и ранее переболевших. При этом среди методов оценки Т-клеточного иммунного ответа наиболее перспективным является тест ELISPOT на выброс интерферона гамма (IGRA) под действием специфических вирусных антигенов. В обзоре рассмотрены перспективы использования технологической платформы ELISPOT в клинико-лабораторной практике при работе с новой коронавирусной инфекцией COVID-19 с учетом особенностей иммунного ответа при данном заболевании. В качестве источников литературы использовались статьи, опубликованные в реферируемых журналах, препринты статей, размещенные на ресурсах arXiv, кроме того, использовались личные коммуникации авторов с ведущими иммунологами. Показано, что внедрение В- и Т-клеточного ELISPOT-тестов позволит осуществлять мониторинг иммунологического статуса пациентов и выбор тактики лечения, выявлять наиболее уязвимые группы населения, осуществлять комплексную оценку вакцинных препаратов на этапах разработки, клинических исследований и внедрения в практику. Обсуждаются вопросы сохранения Т-клеточного иммунитета в крови переболевших коронавирусными инфекциями HCoV, SARS, MERS, COVID-19 и преимущества Т-клеточного ELISPOT-теста перед серологическими тестами для эпидемиологической оценки распространенности новой коронавирусной инфекции и в клинических исследованиях вакцин против COVID-19. Биотехнологические компании имеют готовую технологическую платформу, легко адаптируемую под конкретный вид анализа и патогена, для разработки и промышленного производства наборов ELISPOT. Подтверждена необходимость разработки вакцин, стимулирующих как клеточный, так и гуморальный иммунные ответы, поднят вопрос о защитном потенциале перекрестного иммунитета, приобретенного до пандемии COVID-19.

1. Folegatti PM, Ewer KJ, Aley PK, Angus B, Becker S, Belij-Rammerstorfer S, et al. Safety and immunogenicity of the ChAdOx1 nCoV-19 vaccine against SARS-CoV-2: a preliminary report of a phase 1/2, single-blind, randomised controlled trial. Lancet. 2020;396(10249):467–78. https://doi.org/10.1016/S0140-6736(20)31604-4

2. Oi-Wing Ng, Adeline Chia, Anthony T Tan, Ramesh S Jadi, Hoe Nam Leong, Antonio Bertoletti, Yee-Joo Tan. Memory T cell responses targeting the SARS coronavirus persist up to 11 years post-infection. Vaccine. 2016;34(17):2008–14. https://doi.org/10.1016/j.vaccine.2016.02.063

3. Lehmann PV, Zhang W. Unique strengths of ELISPOT for T cell diagnostics. In: Kalyuzhny A, ed. Handbook of ELISPOT. Methods in Molecular Biology (Methods and Protocols). Vol. 792. Totowa, NJ: Humana Press; 2012. P. 3–23. https://doi.org/10.1007/978-1-61779-325-7_1

4. Umlauf BJ, Pinsky NA, Ovsyannikova IG, Poland GA. Detection of vaccinia virus-specific IFNγ and IL-10 secretion from human PBMCs and CD8+ T cells by ELISPOT. In: Kalyuzhny A, ed. Handbook of ELISPOT. Methods in Molecular Biology (Methods and Protocols). Vol. 792. Totowa, NJ: Humana Press; 2012. P. 199–218. https://doi.org/10.1007/978-1-61779-325-7_16

5. Saletti G, Çuburu N, Yang JS, Dey A, Czerkinsky C. Enzyme-linked immunospot assays for direct ex vivo measurement of vaccineinduced human humoral immune responses in blood. Nat Protoc. 2013;8(6):1073–87. https://doi.org/10.1038/nprot.2013.058

6. Kozlowski PA, Cu-Uvin S, Neutra MR, Flanigan TP. Comparison of the oral, rectal, and vaginal immunization routes for induction of antibodies in rectal and genital tract secretions of women. Infect Immun. 1997;65(4):1387–94.

7. Sundling C, Rönnberg C, Yman V, Asghar M, Jahnmatz P, Lakshmikanth T, et al. B cell profiling in malaria reveals expansion and remodelling of CD11c+ B cell subsets. JCI Insight. 2019;4(9):e126492. https://doi.org/10.1172/jci.insight.126492

8. Yue-Dan Wang, Wan-Yee Fion Sin, Guo-Bing Xu, Huang-Hao Yang, Tin-yau Wong, Xue-Wen Pang, et al. T-cell epitopes in severe acute respiratory syndrome (SARS) coronavirus spike protein elicit a specific T-cell immune response in patients who recover from SARS. J Virol. 2004;78(11):5612–8. https://doi.org/10.1128/JVI.78.11.5612-5618.2004

9. Jiang Gu, Encong Gong, Bo Zhang, Jie Zheng, Zifen Gao, Yanfeng Zhong, et al. Multiple organ infection and the pathogenesis of SARS. J Exp Med. 2005;202(3):415–24. https://doi.org/10.1084/jem.20050828

10. Rokni M, Ghasemi V, Tavakoli Z. Immune responses and pathogenesis of SARS-CoV-2 during an outbreak in Iran: comparison with SARS and MERS. Rev Med Virol. 2020;30(3):e2107. https://doi.org/10.1002/rmv.2107

11. Dawei Wang, Yimei Yin, Chang Hu, Xing Liu, Xingguo Zhang, Shuliang Zhou, et al. Clinical course and outcome of 107 patients infected with the novel coronavirus, SARS-CoV-2, discharged from two hospitals in Wuhan, China. Crit Care. 2020;24(1):188. https://doi.org/10.1186/s13054-020-02895-6

12. Li Tan, Qi Wang, Duanyang Zhang, Jinya Ding, Qianchuan Huang, Yi-Quan Tang, et al. Lymphopenia predicts disease severity of COVID-19: a descriptive and predictive study. Signal Transduct Target Ther. 2020;5(1):33. https://doi.org/10.1038/s41392-020-0148-4

13. Pak-Yin Lui, Lok-Yin Roy Wong, Cheuk-Lai Fung, Kam-Leung Siu, Man-Lung Yeung, Kit-San Yuen, et al. Middle East respiratory syndrome coronavirus M protein suppresses type I interferon expression through the inhibition of TBK1-dependent phosphorylation of IRF3. Emerg Microbes Infect. 2016;5(4):e39. https://doi.org/10.1038/emi.2016.33

14. Park A, Iwasaki A. Type I and type III interferons — induction, signaling, evasion, and application to combat COVID-19. Cell Host Microbe. 2020;27(6):870–8. https://doi.org/10.1016/j.chom.2020.05.008

15. Woo PCY, Lau SKP, Wong BHL, Kwok-hung Chan, Chung-ming Chu, Hoi-wah Tsoi, et al. Longitudinal profile of immunoglobulin G (IgG), IgM, and IgA antibodies against the severe acute respiratory syndrome (SARS) coronavirus nucleocapsid protein in patients with pneumonia due to the SARS coronavirus. Clin Diagn Lab Immunol. 2004;11(4):665–8. https://doi.org/10.1128/CDLI.11.4.665-668.2004

16. Fan Wu, Aojie Wang, Mei Liu, Qimin Wang, Jun Chen, Shuai Xia, et al. Neutralizing antibody responses to SARS-CoV-2 in a COVID-19 recovered patient cohort and their implications. medRxiv. 2020.03.30.20047365. https://doi.org/10.1101/2020.03.30.20047365

17. Chen Dong, Ling Ni, Fang Ye, Meng-Li Chen, Yu Feng, Yong-Qiang Deng, et al. Characterization of anti-viral immunity in recovered individuals infected by SARS-CoV-2. medRxiv. 2020.03.17.20036640. https://doi.org/10.1101/2020.03.17.20036640

18. Sekine T, Perez-Potti A, Rivera-Ballesteros O, Strålin K, Gorin J-B, Olsson A, et al. Robust T cell immunity in convalescent individuals with asymptomatic or mild COVID-19. Cell. 2020. https://doi.org/10.1016/j.cell.2020.08.017

19. Hong-Yi Zheng, Mi Zhang, Cui-Xian Yang, Nian Zhang, Xi-Cheng Wang, Xin-Ping Yang, et al. Elevated exhaustion levels and reduced functional diversity of T cells in peripheral blood may predict severe progression in COVID-19 patients. Cell Mol Immunol. 2020;17(5):541–3. https://doi.org/10.1038/s41423-020-0401-3

20. Braun J, Loyal L, Frentsch M, Wendisch D, Georg P, Kurth F, et al. Presence of SARS-CoV-2 reactive T cells in COVID-19 patients and healthy donors. medRxiv. 2020.04.17.20061440. https://doi.org/10.1101/2020.04.17.20061440

21. Shomuradova AS, Vagida MS, Sheetikov SA, Zornikova KV, Kiryukhin D, Titov A, et al. SARS-CoV-2 epitopes are recognized by a public and diverse repertoire of human T-cell receptors. medRxiv. 2020.05.20.20107813. https://doi.org/10.1101/2020.05.20.20107813

22. Pia L. SARS-CoV-2-reactive T cells in patients and healthy donors. Nat Rev Immunol. 2020;20(6):353. https://doi.org/10.1038/s41577-020-0333-2

23. Gaunt ER, Hardie A, Claas EC, Simmonds P, Templeton KE. Epidemiology and clinical presentations of the four human coronaviruses 229E, HKU1, NL63, and OC43 detected over 3 years using a novel multiplex real-time PCR method. J Clin Microbiol. 2010;48(8):2940–7. https://doi.org/10.1128/JCM.00636-10

24. Callow KA, Parry HF, Sergeant M, Tyrrell DA. The time course of the immune response to experimental coronavirus infection of man. Epidemiol Infect. 1990;105(2):435–46. https://doi.org/10.1017/s0950268800048019

25. Grifoni A, Weiskopf D, Ramirez SI, Mateus J, Dan JM, Moderbacher CR, et al. Targets of T cell responses to SARS-CoV-2 coronavirus in humans with COVID-19 disease and unexposed individuals. Cell. 2020;181(7):1489–501.e15. https://doi.org/10.1016/j.cell.2020.05.015

26. Peeples L. News feature: avoiding pitfalls in the pursuit of a COVID-19 vaccine. Proc Natl Acad Sci USA. 2020;117(15):8218–21. https://doi.org/10.1073/pnas.2005456117

27. Gallais F, Velay A, Wendling M-J, Nazon C, Partisani M, Sibilia J, et al. Intrafamilial exposure to SARS-CoV-2 induces cellular immune response without seroconversion. medRxiv. 2020.06.21.20132449. https://doi.org/10.1101/2020.06.21.20132449

28. Weiskopf D, Schmitz KS, Raadsen MP, Grifoni A, Okba NMA, Endeman H, et al. Phenotype and kinetics of SARS-CoV-2-specific T cells in COVID-19 patients with acute respiratory distress syndrome. Sci Immunol. 2020;5(48):eabd2071. https://doi.org/10.1126/sciimmunol.abd2071

29. Yan-Ying Fan, Zi-Tong Huang, Li Li, Man-Hui Wu, Tao Yu, Richard A Koup, et al. Characterization of SARS-CoV-specific memory T cells from recovered individuals 4 years after infection. Arch Virol. 2009;154(7):1093–9. https://doi.org/10.1007/s00705-009-0409-6

30. Greenbaum JA, Kotturi MF, Kim Y, Oseroff C, Vaughan K, Salimi N, et al. Pre-existing immunity against swine-origin H1N1 influenza viruses in the general human population. Proc Natl Acad Sci USA. 2009;106(48):20365–70. https://doi.org/10.1073/pnas.0911580106

31. Sridhar S, Begom S, Bermingham A, Hoschler K, Adamson W, Carman W, et al. Cellular immune correlates of protection against symptomatic pandemic influenza. Nat Med. 2013;19(10):1305–12. https://doi.org/10.1038/nm.3350

32. Wilkinson TM, Li CKF, Chui CSC, Huang AKY, Perkins M, Liebner JC, et al. Preexisting influenza-specific CD4+ T cells correlate with disease protection against influenza challenge in humans. Nat Med. 2012;18(2):274–80. https://doi.org/10.1038/nm.2612

33. Hancock K, Veguilla V, Xiuhua Lu, Zhong W, Butler EN, Hong Sun, et al. Cross-reactive antibody responses to the 2009 pandemic H1N1 influenza virus. N Engl J Med. 2009;361(20):1945–52. https://doi.org/10.1056/NEJMoa0906453

34. Gostic KM, Ambrose M, Worobey M, Lloyd-Smith JO. Potent protection against H5N1 and H7N9 influenza via childhood hemagglutinin imprinting. Science. 2016;354(6313):722–6. https://doi.org/10.1126/science.aag1322

35. Kissler SM, Tedijanto C, Goldstein E, Grad YH, Lipsitch M. Projecting the transmission dynamics of SARS-CoV-2 through the postpandemic period. Science. 2020;368(6493):860–8. https://doi.org/10.1126/science.abb5793

36. Chuan Qin, Luoqi Zhou, Ziwei Hu, Shuoqi Zhang, Sheng Yang, Yu Tao, et al. Dysregulation of immune response in patients with Coronavirus 2019 (COVID-19) in Wuhan, China. Clin Infect Dis. 2020;71(15):762–8. https://doi.org/10.1093/cid/ciaa248

37. Yufang Shi, Ying Wang, Changshun Shao, Jianan Huang, Jianhe Gan, Xiaoping Huang, et al. COVID-19 infection: the perspectives on immune responses. Cell Death Differ. 2020;27(5):1451–4. https://doi.org/10.1038/s41418-020-0530-3

38. Huang AT, Garcia-Carreras B, Hitchings MDT, Yang B, Katzelnick LC, Rattigan SM, et al. A systematic review of antibody mediated immunity to coronaviruses: antibody kinetics, correlates of protection, and association of antibody responses with severity of disease. medRxiv. 2020.04.14.20065771. https://doi.org/10.1101/2020.04.14.20065771

39. Fan Wu, Aojie Wang, Mei Liu, Qimin Wang, Jun Chen, Shuai Xia, et al. Neutralizing antibody responses to SARS-CoV-2 in a COVID-19 recovered patient cohort and their implications. medRxiv. 2020.03.30.20047365. https://doi.org/10.1101/2020.03.30.20047365

40. Thevarajan I, Thi H O Nguyen, Koutsakos M, Druce J, Caly L, van de Sandt CE, et al. Breadth of concomitant immune responses prior to patient recovery: a case report of non-severe COVID-19. Nat Med. 2020;26(4):453–5. https://doi.org/10.1038/s41591-020-0819-2

41. Bauer T, Jilg W. Hepatitis B surface antigen-specific T and B cell memory in individuals who had lost protective antibodies after hepatitis B vaccination. Vaccine. 2006;24(5):572–7. https://doi.org/10.1016/j.vaccine.2005.08.058

42. Melgaco JG, Morgado LN, Santiago MA, de Oliveira JM, Lewis-Ximenez LL, Hasselmann B, et al. A single dose of inactivated hepatitis A vaccine promotes HAV-specific memory cellular response similar to that induced by a natural infection. Vaccine. 2015;33(32):3813–20. https://doi.org/10.1016/j.vaccine.2015.06.099

43. Melgaço JG, Azamor T, Ano Bom APD. Protective immunity after COVID-19 has been questioned: what can we do without SARS-CoV-2-IgG detection? Cell Immunol. 2020;353:104114. https://doi.org/10.1016/j.cellimm.2020.104114

44. Pizzolla A, Wakim LM. Memory T cell dynamics in the lung during influenza virus infection. J Immunol. 2019;202(2):374–81. https://doi.org/10.4049/jimmunol.1800979