A flow cytometry-based proliferation assay for clinical evaluation of T-cell memory against SARS-CoV-2

Sara Lind Enoksson; Peter Bergman; Jonas Klingström; Fredrik Boström; Rui Da Silva Rodrigues; Malin Elisabeth Winerdal; Per Marits

doi:10.1016/j.jim.2021.113159

A flow cytometry-based proliferation assay for clinical evaluation of T-cell memory against SARS-CoV-2

J Immunol Methods. 2021 Dec:499:113159. doi: 10.1016/j.jim.2021.113159. Epub 2021 Sep 28.

Authors

Sara Lind Enoksson¹, Peter Bergman², Jonas Klingström³, Fredrik Boström⁴, Rui Da Silva Rodrigues⁴, Malin Elisabeth Winerdal¹, Per Marits⁵

Affiliations

¹ Department of Clinical Immunology and Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden; Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.
² Department of Infectious Diseases, The Immunodeficiency Unit, Karolinska University Hospital, Stockholm, Sweden; Department of Laboratory Medicine, Division of Clinical Microbiology, Karolinska Institutet, Stockholm, Sweden.
³ Center for Infectious Medicine, Department of Medicine Huddinge, Karolinska Institutet, Stockholm, Sweden.
⁴ Department of Clinical Immunology and Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden.
⁵ Department of Clinical Immunology and Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden; Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden. Electronic address: per.marits@regionstockhom.se.

Abstract

In general, the method of choice for evaluating immunity against Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is detection of antibodies against the virus in patient sera. However, this is not feasible in patients who do not produce antibodies, either due to a primary immunodeficiency or secondary to treatment with immunosuppressive drugs. Assessment of the antiviral T cell response is an alternative to serological tests, but most T cell assays are labor-intensive and unsuitable for a clinical routine laboratory. We developed a flow cytometry-based assay for T cell proliferative responses against SARS-CoV-2, based on the detection of blast transformation of activated cells. The assay was validated on previously SARS-CoV-2 infected individuals and healthy seronegative blood donors, displaying 74% sensitivity and 96% specificity for previous infection with SARS-CoV-2. The usefulness of the assay was demonstrated in a patient with common variable immunodeficiency with a history of COVID-19. The described T-cell assay is a clinically relevant complement to serology in the evaluation of cellular immunity against SARS-CoV-2, which can be emulated by any routine lab with flow cytometric competence.

Keywords: COVID-19; FASCIA; Flow cytometry; Immunity; SARS-CoV-2; T cell; Virus.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adult
Aged
Cell Proliferation
Female
Flow Cytometry*
Humans
Immunologic Memory / immunology*
Male
Middle Aged
SARS-CoV-2 / immunology*
T-Lymphocytes / immunology*
Young Adult