Department of Clinical Immunology and Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden; Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.
Department of Infectious Diseases, The Immunodeficiency Unit, Karolinska University Hospital, Stockholm, Sweden; Department of Laboratory Medicine, Division of Clinical Microbiology, Karolinska Institutet, Stockholm, Sweden.
J Immunol Methods. 2021 Dec;499:113159. doi: 10.1016/j.jim.2021.113159. Epub 2021 Sep 28.
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.
一般来说,评估对严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)免疫力的首选方法是检测患者血清中的病毒抗体。然而,对于那些由于原发性免疫缺陷或因免疫抑制药物治疗而不产生抗体的患者,这种方法是不可行的。评估抗病毒 T 细胞反应是血清学检测的替代方法,但大多数 T 细胞检测方法都很繁琐,不适合临床常规实验室。我们开发了一种基于流式细胞术的 SARS-CoV-2 特异性 T 细胞增殖反应检测方法,该方法基于激活细胞的细胞爆炸转化的检测。该检测方法在先前感染 SARS-CoV-2 的个体和健康血清阴性献血者中进行了验证,对先前 SARS-CoV-2 感染的敏感性为 74%,特异性为 96%。在一名患有 COVID-19 病史的常见可变免疫缺陷患者中证明了该检测方法的有用性。所描述的 T 细胞检测方法是评估针对 SARS-CoV-2 的细胞免疫的血清学的临床相关补充,可以通过具有流式细胞术能力的任何常规实验室来模拟。