Assesment of specific T-cell immunity to SARS-CoV-2 virus antigens in COVID-19 reconvalescents

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Abstract

Introduction. The development of the COVID-19 pandemic has stimulated the scientific research aimed at studying of the mechanisms of formation the immunity against SARS-CoV-2. Currently, there is a need to develop a domestic simple and cost-effective specific method suitable for monitoring of T-cell response against SARS-CoV-2 in reconvalescents and vaccinated individuals.

Aim: Development of a screening method for evaluation specific T-cell immunity against SARS-CoV-2.

Materials and methods. Total 40 individuals who had mild to moderate COVID-19 and 20 healthy volunteers who did not have a history of this disease were examined. The presence and levels of IgG and IgM antibodies to SARS-CoV-2 were identified in participant’s sera by ELISA using the diagnostic kits from JSC “Vector-Best” (Novosibirsk, Russian Federation). Antigenic stimulation of mononuclear cells was carried out on commercial plates with adsorbed whole-virion inactivated SARS-CoV-2 antigen (State Research Center of Virology and Biotechnology VECTOR Novosibirsk, Russian Federation). The concentration of IFN-γ was measured in ELISA using the test systems from JSC “Vector-Best” (Novosibirsk, Russian Federation). The immunophenotyping of lymphocytes was performed on a flow cytometer Cytomics FC500 (Beckman Coulter, USA). Statistical data processing was carried out using the Microsoft Excel and STATISTICA 10 software package.

Results. Stimulation of mononuclear cells isolated from the peripheral blood with whole-virion inactivated SARS-CoV-2 antigen fixed at the bottom of the wells of a polystyrene plate showed a significantly higher median response in terms of IFN-γ production in 40 people who had history of COVID-19 compared to 20 healthy blood donors (172.1 [34.3–575.1] pg/ml versus 15.4 [6.9–25.8] pg/ml, p < 0.0001).

There was no difference in median IFN-γ levels in supernatants collected from unstimulated mononuclear cells from COVID-19 reconvalescents and healthy donors (2.7 [0.4–11.4] pg/ml versus 0.8 [0.0–23.3] pg/ml, p < 0.05). The overall sensitivity and specificity of this method were 73% (95% CI 58–88%) and 100% (95% CI 100–100%), respectively, at a cut-off of 50 pg/ml.

Conclusion. The developed method for assessment of the cellular immune response to SARS-CoV-2 can be used as a screening method for monitoring the T-cell response in a population against a new coronavirus infection in recovered people.

About the authors

Maria S. Blyakher

G.N. Gabrichevsky Moscow Research Institute of Epidemiology and Microbiology

Email: maria.s.b@bk.ru
ORCID iD: 0000-0003-3480-6873

D. Sci. (Med.), Professor, Head Laboratory, Department of immunology

Russian Federation, 125212, Moscow

Irina M. Fedorova

G.N. Gabrichevsky Moscow Research Institute of Epidemiology and Microbiology

Email: vestnik-07@mail.ru
ORCID iD: 0000-0002-0335-2752

Cand. Sci. (Med.), leading researcher, Department of immunology

Russian Federation, 125212, Moscow

Elena A. Tulskaya

G.N. Gabrichevsky Moscow Research Institute of Epidemiology and Microbiology

Email: etul@mail.ru
ORCID iD: 0000-0003-1969-4009

Cand. Sci. (Biol.), leading researcher, Department of immunology

Russian Federation, 125212, Moscow

Ivan V. Kapustin

G.N. Gabrichevsky Moscow Research Institute of Epidemiology and Microbiology

Email: maria.s.b@bk.ru
ORCID iD: 0000-0001-6191-260X

Cand. Sci. (Med.), leading researcher, Department of immunology

Russian Federation, 125212, Moscow

Svetlana I. Koteleva

G.N. Gabrichevsky Moscow Research Institute of Epidemiology and Microbiology

Email: felileo@yandex.ru
ORCID iD: 0000-0003-1878-2234

к.м.н, в.н.с. отдела иммунологии

Russian Federation, 125212, Moscow

Zarema K. Ramazanova

G.N. Gabrichevsky Moscow Research Institute of Epidemiology and Microbiology

Email: rzarema@mail.ru
ORCID iD: 0000-0002-9314-3312

Cand. Sci. (Med.), leading researcher, Department of immunology

Russian Federation, 125212, Moscow

Evgeny E. Odintsov

G.N. Gabrichevsky Moscow Research Institute of Epidemiology and Microbiology

Email: rinocerus@yandex.ru
ORCID iD: 0000-0001-5895-2520

Cand. Sci. (Med.), leading researcher, Department of immunology

Russian Federation, 125212, Moscow

Svetlana V. Sandalova

G.N. Gabrichevsky Moscow Research Institute of Epidemiology and Microbiology

Email: s.sandalova74@mail.ru
ORCID iD: 0000-0002-4548-9888

Researcher, Department of immunology

Russian Federation, 125212, Moscow

Lidia I. Novikova

G.N. Gabrichevsky Moscow Research Institute of Epidemiology and Microbiology

Author for correspondence.
Email: vestnik-07@mail.ru
ORCID iD: 0000-0002-0307-4561

Cand. Sci. (Med.), Head Laboratory, Department of immunology

Russian Federation, 125212, Moscow

References

  1. Soresina A., Moratto D., Chiarini M., Paolillo C., Baresi G., Foca E., et al. Two X-linked agammaglobulinemia patients develop pneumonia as COVID-19 manifestation but recover. Pediatr. Allergy Immunol. 2020; 31(5): 565–9. https://doi.org/10.1111/pai.13263
  2. Mathew D., Giles J.R., Baxter A.E., Oldridge D.A., Greenplate A.R., Wu J.E., et al. COVID-19 patients reveals distinct immunotypes with therapeutic implications. Science. 2020; 369(6508): 1209. https://doi.org/10.1126/science.abc8511
  3. Peng Y., Mentzer A.J., Liu G., Yao X., Yin Z., Dong D., et al. Broad and strong memory CD4(+) and CD8(+) T cells induced by SARS-CoV-2 in UK convalescent individuals following COVID-19. Nat. Immunol. 2020; 21(11): 1336–45. https://doi.org/10.1038/s41590-020-0782-6
  4. Ni L., Ye F., Cheng M.L., Feng Y., Deng Y.Q., Zhao H., et al. Detection of SARS-CoV-2-specific humoral and cellular immunity in COVID-19 convalescent individuals. Immunity. 2020; 52(6): 971 7. https://doi.org/10.1016/j.immuni.2020.04.023
  5. Grifoni A., Weiskopf D., Ramirez S.I., Mateus J., Dan J.M., Moderbacher C.R., 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. https://doi.org/10.1016/j.cell.2020.05.015
  6. Braun J., Loyal L., Frentsch M., Wendisch D., Georg P., Kurth F., et al. SARS-CoV-2-reactive T cells in healthy donors and patients with COVID-19. Nature. 2020; 587(7833): 270–4. https://doi.org/10.1038/s41586-020-2598-9
  7. Gimenez E., Albert E., Torres I., Remigia M.J., Alcaraz M.J., Galindo M.J., et al. SARS-CoV-2-reactive interferon-gamma-producing CD8+ T cells in patients hospitalized with coronavirus disease. J. Med. Virol. 2021; 93(1): 375–82. https://doi.org/10.1002/jmv.26213
  8. Le Bert N., Tan A.T., Kunasegaran K., Tham C.Y.L., Hafezi M., Chia A., et al. SARS-CoV-2-specific T cell immunity in cases of COVID-19 and SARS, and uninfected controls. Nature. 2020; 584(7821): 457–62. https://doi.org/10.1038/s41586-020-2550-z
  9. Peng Y., Mentzer A.J., Liu G., Yao X., Yin Z., Dong D., et al. Broad and strong memory CD4+ and CD8+ T cells induced by SARS-CoV-2 in UK convalescent individuals following COVID-19. Nat. Immunol. 2020; 21(11): 1336–45. https://doi.org/10.1038/s41590-020-0782-6
  10. Thieme C.J., Anft M., Paniskaki K., Blazquez-Navarro A., Doevelaar A., Seibert F.S., et al. Robust T cell response toward spike, membrane, and nucleocapsid SARS-CoV-2 proteins is not associated with recovery in critical COVID-19 Patients. Cell Rep. Med. 2020; 1(6): 100092. https://doi.org/10.1016/j.xcrm.2020.100092
  11. Oja A.E., Saris A., Ghandour C.A., Kragten N.A.M., Hogema B.M., Nossent E.J., et al. Divergent SARS-CoV-2-specific T and B cell responses in severe but not mild COVID-19. bioRxiv. 2020; Preprint. https://doi.org/10.1101/2020.06.18.159202
  12. Auladell M., Jia X., Hensen L., Chua B., Fox A., Nguyen T.H.O., et al. Recalling the future: immunological memory toward unpredictable influenza viruses. Front. Immunol. 2019; 10: 1400. https://doi.org/10.3389/fimmu.2019.01400
  13. Robbiani D.F., Gaebler C., Muecksch F., Lorenzi J.C.C., Wang Z., Cho A., et al. Convergent antibody responses to SARS-CoV-2 in convalescent individuals. Nature. 2020; 584(7821): 437–42. https://doi.org/10.1038/s41586-020-2456-9
  14. Folegatti P.M., Ewer K.J., Aley P.K., 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
  15. Logunov D.Y., Dolzhikova I.V., Zubkova O.V., Tukhvatullin A.I., Schleblyakov D.V., Dzharullaeva A.S., et al. Safety and immunogenicity of an rAd26 and rAd5 vector-based heterologous prime-boost COVID-19 vaccine in two formulations: two open, non-randomised phase 1/2 studies from Russia. Lancet. 2020; 396(10255): 887–97. https://doi.org/10.1016/S0140-6736(20)31866-3
  16. Liu J., Li S., Liu J., Liang B., Wang X., Wang H., et al. Longitudinal characteristics of lymphocyte responses and cytokine profiles in the peripheral blood of SARS-CoV-2 infected patients. EBioMedicine. 2020; 55: 102763. https://doi.org/10.1016/j.ebiom.2020.102763
  17. Poteryaev D.A., Abbasova S.G., Ignat’eva P.E., Strizhakova O.M., Kolesnik S.V., Khamitov R.A. Assessment of T-cell immunity to SARS-CoV-2 in COVID-19 convalescents and vaccinated subjects, using TigraTest® SARS-CoV-2 ELISPOT kit. BIOpreparaty. Profilaktika, diagnostika, lechenie. 2021; 21(3): 178–92. https://doi.org/10.30895/2221-996X-2021-21-3-178-192 (in Russian)
  18. Youden W.J. Index for rating diagnostic tests. Cancer. 1950; 3(1): 32–5. https://doi.org/10.1002/1097-0142(1950)3:1<32:aid-cncr2820030106>3.0.co;2-3
  19. Solano C., Benet I., Clari M.A., Nieto J., de la Cámara R., López J., et al. Enumeration of cytomegalovirus-specific interferon gamma CD8+ and CD4+ T cells early after allogeneic stem cell transplantation may identify patients at risk of active cytomegalovirus infection. Haematologica. 2008; 93(9): 1434–6. https://doi.org/10.3324/haematol.12880
  20. de Araújo-Souza P.S., Hanschke S.C.H., Nardy A.F.F.R., Sécca C., Oliveira-Vieira B., Silva K.L., et al. Differential interferon-γ production by naive and memory-like CD8 T cells. J. Leukoc. Biol. 2020; 108(4): 1329–37. https://doi.org/10.1002/JLB.2AB0420-646R
  21. Murugesan K., Jagannathan P., Pham T.D., Pandey S., Bonilla H.F., Jacobson K., et al. Interferon-γ release assay for accurate detection of severe acute respiratory syndrome coronavirus 2 T-cell response. Clin. Infect. Dis. 2021; 73(9): 3130–2. https://doi.org/10.1093/cid/ciaa1537
  22. Fernández-González M., Agulló V., Padilla S., García J. A., García-Abellán J., Botella Á., et al. Clinical performance of a standardized severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) interferon-γ release assay for simple detection of T-cell responses after infection or vaccination. Clin. Infect. Dis. 2022; 75(1): e338–46. https://doi.org/10.1093/cid/ciab1021
  23. Echeverría G., Guevara Á., Coloma J., Ruiz A.M., Vasquez M.M., Tejera E., et al. Pre-existing T-cell immunity to SARS-CoV-2 in unexposed healthy controls in Ecuador, as detected with a COVID-19 interferon-gamma release assay. Int. J. Infect. Dis. 2021; 105: 21–5. https://doi.org/10.1016/j.ijid.2021.02.034
  24. Cibrián D., Sánchez-Madrid F. CD69: from activation marker to metabolic gatekeeper. Eur. J. Immunol. 2017; 47(6): 946–53. https://doi.org/10.1002/eji.201646837
  25. Chen Z.Y., Wang L., Gu L., Qu R., Lowrie D.B., Hu Z., et al. Decreased expression of CD69 on T Cells in tuberculosis infection resisters. Front. Microbiol. 2020; 11: 1901. https://doi.org/10.3389/fmicb.2020.01901
  26. Murata K., Hoshina T., Onoyama S., Tanaka T., Kanno S., Ishimura M., et al. Reduction in the number of Varicella-Zoster virus-specific T-Cells in immunocompromised children with varicella. Tohoku J. Exp. Med. 2020; 250(3): 181–90. https://doi.org/10.1620/tjem.250.181
  27. Muntasell A., Costa-Garcia M., Vera A., Marina-Garcia N., Kirschning C.J., López-Botet M. Priming of NK cell anti-viral effector mechanisms by direct recognition of human cytomegalovirus. Front. Immunol. 2013; 4: 40. https://doi.org/10.3389/fimmu.2013.00040
  28. Della Chiesa M., De Maria A., Muccio L., Bozzano F., Sivori S., Moretta L. Human NK cells and herpesviruses: mechanisms of recognition, response and adaptation. Front. Microbiol. 2019; 10: 2297. https://doi.org/10.3389/fmicb.2019.02297
  29. Varchetta S., Mele D., Oliviero B., Mantovani S., Ludovisi S., Cerino A., et al. Unique immunological profile in patients with COVID-19. Cell. Mol. Immunol. 2021; 18(3): 604–12. https://doi.org/10.1038/s41423-020-00557-9

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Copyright (c) 2023 Blyakher M.S., Fedorova I.M., Tulskaya E. ., Kapustin I.V., Koteleva S.I., Ramazanova Z.K., Odintsov E.E., Sandalova S.V., Novikova L.I.

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