Assessment of cell-mediated immune responses to SARS-CoV-2 in Syrian hamsters

Abstract

BACKGROUND: The pandemic caused by the SARS-CoV-2 virus at the end of 2019 remains to be a serious healthcare problem. Constant antigenic drift of the pathogen led to a decrease of licensed COVID-19 vaccines effectiveness. And the development of broad-spectrum vaccines with high effectiveness rate against evolutionarily divergent SARS-CoV-2 variants remains an urgent issue. Unlike virus-specific antibodies with limited spectrum of action, T-cell immunity has a wider cross-protective potential. Syrian hamsters are the most appropriate model for preclinical evaluation of new vaccine candidates, since these animals are susceptible to SARS-CoV-2 infection and show clinical symptoms of the disease. However, study of T-cell vaccine response in hamsters is complicated by the lack of available reagents and test systems for adequate assessment of the virus-specific cellular immunity levels after vaccination.

AIM: In this work, we report an optimized protocol of stimulation of Syrian hamsters’ immune cells with a live SARS-CoV-2 virus to assess virus-specific T-cell responses.

MATERIALS AND METHODS: Intranasal infection of animals with SARS-CoV-2 virus followed by stimulation of immune cells with different doses of whole live coronavirus and counting of IFNγ-producing cells by ELISpot method.

RESULTS: Stimulation of spleen and lung cells with SARS-CoV-2 at a dose 0.1 TCID₅₀/cell is the most optimal viral concentration for detecting maximum of cytokine-producing cells in SARS-CoV-2-infected animals. Stimulation of cells with whole virus revealed greater number of virus-specific cells compared to a stimulation with pools of SARS-CoV-2 lyophilized peptides (S and N proteins).

CONCLUSIONS: Overall, the new methodology allows assessment of the immunogenicity of COVID-19 T-cell vaccines more accurately in preclinical studies using the Syrian hamsters model.

Keywords

new coronavirus infection / SARS-CoV-2 / ELISpot / Syrian hamsters / IFNγ

References

[1] COVID-19 Dashboard by the Center for Systems Science and Engineering (CSSE) at Johns Hopkins University (JHU) [Internet]. Available from: https://coronavirus.jhu.edu/map.html. Accessed: May 1, 2022.

[2] COVID-19 Dashboard by the Center for Systems Science and Engineering (CSSE) at Johns Hopkins University (JHU) [Электронный ресурс]. Режим доступа: https://coronavirus.jhu.edu/map.html. Дата обращения: 01.05.2022.

[3] Zheng C, Shao W, Chen X, et al. Real-world effectiveness of COVID-19 vaccines: a literature review and meta-analysis. Int J Infect Dis. 2022;114:252–260. DOI: 10.1016/j.ijid.2021.11.0093

[4] Zheng C., Shao W., Chen X. et al. Real-world effectiveness of COVID-19 vaccines: a literature review and meta-analysis // Int. J. Infect. Dis. 2022. Vol. 114. P. 252–260. DOI: 10.1016/j.ijid.2021.11.0093.

[5] Tatsi EB, Filippatos F, Michos A. SARS-CoV-2 variants and effectiveness of vaccines: a review of current evidence. Epidemiol Infect. 2021;149:e237. DOI: 10.1017/S0950268821002430

[6] Tatsi E.B., Filippatos F., Michos A. SARS-CoV-2 variants and effectiveness of vaccines: a review of current evidence // Epidemiol. Infect. 2021. Vol. 149. P. e237. DOI: 10.1017/S0950268821002430

[7] Kedzierska K, Thomas PG. Count on us: T cells in SARS-CoV-2 infection and vaccination. Cell Rep Med. 2022;3(3):100562. DOI: 10.1016/j.xcrm.2022.100562

[8] Kedzierska K., Thomas P.G. Count on us: T cells in SARS-CoV-2 infection and vaccination // Cell Rep. Med. 2022. Vol. 3, No. 3. P. 100562. DOI: 10.1016/j.xcrm.2022.100562

[9] Zollner A, Watschinger C, Rössler A, et al. B and T cell response to SARS-CoV-2 vaccination in health care professionals with and without previous COVID-19. EBioMedicine. 2021;70:103539. DOI: 10.1016/j.ebiom.2021.103539

[10] Zollner A., Watschinger C., Rössler A. et al. B and T cell response to SARS-CoV-2 vaccination in health care professionals with and without previous COVID-19 // EBioMedicine. 2021. Vol. 70. P. 103539. DOI: 10.1016/j.ebiom.2021.103539

[11] Abbasi J. COVID-19 vaccine focused on T-Cell response promising in early trial. JAMA. 2022;327(2):115. DOI: 10.1001/jama.2021.24118

[12] Abbasi J. COVID-19 vaccine focused on T-Cell response promising in early trial // JAMA. 2022. Vol. 327, No. 2. P. 115. DOI: 10.1001/jama.2021.24118

[13] Cox JH, Ferrari G, Janetzki S. Measurement of cytokine release at the single cell level using the ELISPOT assay. Methods. 2006;38(4):274–282. DOI: 10.1016/j.ymeth.2005.11.006

[14] Cox J.H., Ferrari G., Janetzki S. Measurement of cytokine release at the single cell level using the ELISPOT assay // Methods. 2006. Vol. 38, No. 4. P. 274–282. DOI: 10.1016/j.ymeth.2005.11.006

[15] Bonifacius A, Tischer-Zimmermann S, Santamorena MM, et al. Rapid manufacturing of highly cytotoxic clinical-grade SARS-CoV-2-specific T cell products covering SARS-CoV-2 and its variants for adoptive T cell therapy. Front Bioeng Biotechnol. 2022;10:867042. DOI: 10.3389/fbioe.2022.867042

[16] Bonifacius A., Tischer-Zimmermann S., Santamorena M.M. et al. Rapid manufacturing of highly cytotoxic clinical-grade SARS-CoV-2-specific T cell products covering SARS-CoV-2 and its variants for adoptive T cell therapy // Front. Bioeng. Biotechnol. 2022. Vol. 10. P. 867042. DOI: 10.3389/fbioe.2022.867042

[17] Mak WA, Koeleman JGM, Ong DSY. Development of an in-house SARS-CoV-2 interferon-gamma ELISpot and plate reader-free spot detection method. J Virol Methods. 2022;300:114398. DOI: 10.1016/j.jviromet.2021.114398

[18] Mak W.A., Koeleman J.G.M., Ong D.S.Y. Development of an in-house SARS-CoV-2 interferon-gamma ELISpot and plate reader-free spot detection method // J. Virol. Methods. 2022. Vol. 300. P. 114398. DOI: 10.1016/j.jviromet.2021.114398

[19] Matyushenko V, Kotomina T, Kudryavtsev I, et al. Conserved T-cell epitopes of respiratory syncytial virus (RSV) delivered by recombinant live attenuated influenza vaccine viruses efficiently induce RSV-specific lung-localized memory T cells and augment influenza-specific resident memory T-cell responses. Antiviral Res. 2020;182:104864. DOI: 10.1016/j.antiviral.2020.104864

[20] Matyushenko V., Kotomina T., Kudryavtsev I. et al. Conserved T-cell epitopes of respiratory syncytial virus (RSV) delivered by recombinant live attenuated influenza vaccine viruses efficiently induce RSV-specific lung-localized memory T cells and augment influenza-specific resident memory T-cell responses // Antiviral. Res. 2020. Vol. 182. P. 104864. DOI: 10.1016/j.antiviral.2020.104864

[21] Fiolet T, Kherabi Y, MacDonald CJ, et al. Comparing COVID-19 vaccines for their characteristics, efficacy and effectiveness against SARS-CoV-2 and variants of concern: a narrative review. Clin Microbiol Infect. 2022;28(2):202–221. DOI: 10.1016/j.cmi.2021.10.005

[22] Fiolet T., Kherabi Y., MacDonald C.J. et al. Comparing COVID-19 vaccines for their characteristics, efficacy and effectiveness against SARS-CoV-2 and variants of concern: a narrative review // Clin. Microbiol. Infect. 2022. Vol. 28, No. 2. P. 202–221. DOI: 10.1016/j.cmi.2021.10.005

[23] Martinez-Flores D, Zepeda-Cervantes J, Cruz-Reséndiz A, et al. SARS-CoV-2 vaccines based on the spike glycoprotein and implications of new viral variants. Front Immunol. 2021;12:701501. DOI: 10.3389/fimmu.2021.701501

[24] Martinez-Flores D., Zepeda-Cervantes J., Cruz-Reséndiz A. et al. SARS-CoV-2 vaccines based on the spike glycoprotein and implications of new viral variants // Front. Immunol. 2021. Vol. 12. P. 701501. DOI: 10.3389/fimmu.2021.701501

[25] Swadling L, Maini MK. T cells in COVID-19 – united in diversity. Nat Immunol. 2020;21(11):1307–1308. DOI: 10.1038/s41590-020-0798-y

[26] Swadling L., Maini M.K. T cells in COVID-19 – united in diversity // Nat. Immunol. 2020. Vol. 21, No. 11. P. 1307–1308. DOI: 10.1038/s41590-020-0798-y

[27] Rong Y, Wang F, Liu J, et al. Clinical characteristics and risk factors of mild-to-moderate COVID-19 patients with false-negative SARS-CoV-2 nucleic acid. J Med Virol. 2021;93(1):448–455. DOI: 10.1002/jmv.26242

[28] Rong Y., Wang F., Liu J. et al. Clinical characteristics and risk factors of mild-to-moderate COVID-19 patients with false-negative SARS-CoV-2 nucleic acid // J. Med. Virol. 2021. Vol. 93, No. 1. P. 448–455. DOI: 10.1002/jmv.26242

[29] Dai L, Gao GF. Viral targets for vaccines against COVID-19. Nat Rev Immunol. 2021;21(2):73–82. DOI: 10.1038/s41577-020-00480-0

[30] Dai L., Gao G.F. Viral targets for vaccines against COVID-19 // Nat. Rev. Immunol. 2021. Vol. 21, No. 2. P. 73–82. DOI: 10.1038/s41577-020-00480-0

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