Identification of lymphocyte subgroups with flow cytometry in COVID-19 patients

Year 2022, , 1183 - 1189, 20.07.2022

İlhami Berber , Nurcan Kırıcı Berber , Ahmet Sarıcı , Harika Gözükara Bağ , Soykan Biçim , Burhan Turgut , Furkan Çağan , Mehmet Ali Erkurt , Ayşe Uysal , Nihal Sümeyye Ulutaş , Emin Kaya , İrfan Kuku

https://doi.org/10.32322/jhsm.1129894

Abstract

Objective: We aimed to determine lymphocyte subgroups and activation status of flow cytometry in COVID-19 patients and examine their relationship with disease stage and length of hospital stay.
Material and Method: Forty patients were analyzed in this study and compared with the age and sex-matched 40 healthy controls. COVID-19 patients have split as early and advanced-stage diseases. Flow cytometry assay was performed to determine the counts of lymphocyte subsets and activation status. Total lymphocyte count was calculated and CD45 (cluster of differentiation), CD3, CD4, CD8, CD19, CD27, CD38, CD56, CD57, and IgD were studied on lymphocyte gate. T helper / T cytotoxic rates and length of hospital stay were recorded.
Results: The patients' CD3(+)CD4(+) ( T helper) count and CD27 expression on T cells counts were significantly lower, and CD57 expression on CD3(+)CD8(+) T cytotoxic cells were significantly higher (p<0.05) than the control group. When the patients were divided into early and advanced stages, it was observed that CD38 expression on T cells was significantly lower in advanced-stage patients (p< 0.05) Total lymphocyte count and CD3(+) T lymphocyte count were negatively correlated with the duration of hospitalization as statistically significant (p <0.05).
Conclusion: Our data showed that the SARS-CoV-2 primarily affects T lymphocytes. It was thought that this effect occurred by impairment of development and activation of T lymphocytes. There are some discordances among the studies on T lymphocytes in the literature.

Keywords

COVID-19, flow cytometry, lymphocytes, SARS-CoV-2, T cells

Project Number

YOK

References

• Li Q, Guan X, Wu P, et al. Early transmission dynamics in Wuhan, China, of novel coronavirus-infected pneumonia. N Engl J Med 2020; 382: 1199-207.
• Pal M, Berhanu G, Desalegn C, Kandi V. Severe acute respiratory syndrome coronavirus-2(SARS-CoV-2): an update. Cureus 2020; 12: e7423.
• World Health Organization Press Conference. The World Health Organization (WHO) Has Officially Named the Disease Caused by the Novel Coronavirus as COVID-19. Available online: URL: https://www.who.int/ emergencies/diseases/novel-coronavirus-2019 (accessed on 18 May 2020).
• Chan JF, Yuan S, Kok KH, et al. A familial cluster of pneumonia associated with the 2019 novel coronavirus indicating person-to-person transmission: a study of a family cluster. Lancet 2020; 395: 514-23.
• Zhu N, Zhang D, Wang W, et al. A Novel coronavirus from patients with pneumonia in China. N Engl J Med 2020; 382: 727-33.
• Rodriguez-Morales AJ, Cardona-Ospina JA, Gutiérrez-Ocampo E, et al Clinical, laboratory and imaging features of COVID-19: A systematic review and meta-analysis. Travel Med Infect Dis 2020; 34: 101623.
• Weiliang C, Li S, Lin C, et al. Clinical features, and laboratory inspection of novel coronavirus pneumonia (COVID-19) in Xiangyang, Hubei. medRxiv. URL: https://www.medrxiv.org/content/10.1101/2020.02.23.20026963v1
• Gao Y, Li T, Han M, et al. Diagnostic utility of clinical laboratory data determinations for patients with the severe COVID-19. J Med Virol. 2020; 92: 791-96.
• Lauer SA, Grantz KH, Bi Q, et al. The incubation period of coronavirus disease 2019 (COVID-19) from publicly reported confirmed cases: estimation and application. Ann Intern Med 2020; 172: 577-82.
• Eubank S, Eckstrand I, Lewis B, et al. Impact of Non-pharmaceutical Interventions (NPIs) to Reduce COVID-19 Mortality and Healthcare Demand. Bull Math Biol 2020; 82: 52.
• Lederman S, Yellin MJ, Krichevsky A, et al. Identification of a novel surface protein on activated CD4+ T cells that induces contact-dependent B cell differentiation(help). J Exp Med 1992; 175: 1091-101.
• Buchan SL, Rogel A, Al-Shamkhani A. The immunobiology of CD27 and OX40 and their potential as targets for cancer immunotherapy. Blood 2018; 131: 39–48.
• Kared H, Martelli S, Ng TP, et al. CD57 in human natural killer cells and T-lymphocytes. Cancer Immunol Immunother 2016; 65: 441-52.
• Lino VA, Santos SM, Bittencourt HN, et al. Quantification of CD8(+)CD38(+) T lymphocytes by flow cytometry does not represent a good biomarker to monitor the reactivation of cytomegalovirus infection after allogeneic hematopoietic stem cell transplantation. Rev Bras Hematol Hemoter 2011; 33: 268-73.
• Jiang Y, Wei X, Guan J, et al. COVID-19 pneumonia: CD8+ T and NK cells are decreased in number but compensatory increased in cytotoxic potential. Clin Immunol 2020; 218: 108516.
• Kazancioglu S, Yilmaz FM, Bastug A, et al. Lymphocyte subset alteration and monocyte CD4 expression reduction in patients with severe COVID-19. Viral Immunol 2021; 34: 342-51.
• Wang F, Nie J, Wang H, et al. Characteristics of peripheral lymphocyte subset alteration in COVID-19 pneumonia. J Infect Dis 2020; 221: 1762-9.
• Almeida M, Cordero M, Almeida J, et al. CD38 on peripheral blood cells: the value of measuring CD38 expression on CD8 T-cells in patients receiving highly active anti-retroviral therapy. Clin Appl Immunol Rev 2002; 2: 307-20.
• Kang CK, Han GC, Kim M, et al. Aberrant hyperactivation of cytotoxic T-cell as a potential determinant of COVID-19 severity. Int J Infect Dis 2020; 97: 313-21.
• Mazzoni A, Salvati L, Maggi L, et al. Impaired immune cell cytotoxicity in severe COVID-19 is IL-6 dependent. J Clin Invest 2020; 130: 4694-703.
• Huang W, Berube J, McNamara M, et al. Lymphocyte subset counts in COVID-19 patients: a meta-analysis. Cytometry Part A 2020; 97: 772-6.