Development and evaluation of a flow-cytometric assay of specific cell-mediated immune response in activated whole blood for the detection of cell-mediated immunity against varicella-zoster virus.
A new assay for the detection of specific cell-mediated immune (CMI) responses is described. Whole blood, diluted 1/10 in medium, was cultured in the presence or the absence of specific antigens. Results were assessed by flow cytometric analysis with or without immunophenotyping to detect proliferating lymphoblasts among cultured cells. Interferon-gamma, IL-10, and IL-5 in culture supernatants are measured by ELISAs. The assay was evaluated using samples from 37 VZV-antibody-positive children with a history of chickenpox and samples from 15 seronegative children without a history of chickenpox; it displayed a sensitivity of 95% and a specificity of 100% for the detection of varicella-zoster virus (VZV)-specific CMI. The intraassay and interassay variations of the new test were lower than with the conventional assay for CMI, detecting thymidine incorporation in peripheral blood mononuclear cells (PBMCs). Cytokines were detected in only 70% of cultures from VZV-antibody-positive subjects. The cytokine response was restricted to IFN-gamma in most cases. The Flow-cytometric Assay of Specific Cell-mediated Immune response in Activated whole blood (FASCIA) is a precise and accurate yet simple and convenient test that can be readily employed for the examination of single samples as well as for large-scale studies.
Svahn
A
A
Department of Virology, Swedish Institute for Infectious Disease Control, S-171 82, Solna, Sweden.
Linde
A
A
Thorstensson
R
R
Karlén
K
K
Andersson
L
L
Gaines
H
H
D000293:Adolescent; D000914:Antibodies, Viral / Q000097:blood*; D002644:Chickenpox / Q000097:blood / Q000276:immunology*; D002648:Child; D002675:Child, Preschool; D004279:DNA, Viral / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry / Q000379:methods*; D014645:Herpesvirus 3, Human / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007111:Immunity, Cellular / Q000276:immunology; D007223:Infant; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D016753:Interleukin-10 / Q000276:immunology / Q000378:metabolism; D015848:Interleukin-5 / Q000276:immunology / Q000378:metabolism; D012680:Sensitivity and Specificity; D013936:Thymidine / Q000378:metabolism
D000914:Antibodies, Viral; D004279:DNA, Viral; D015848:Interleukin-5; D016753:Interleukin-10; D007371:Interferon-gamma; D013936:Thymidine