title: Development of an immune function assay by measuring intracellular adenosine triphosphate (iATP) levels in mitogen-stimulated CD4+ T lymphocytes.
abstract: We developed an immune function assay for monitoring CD4+ T cells activity based on changes in intracellular adenosine triphosphate (iATP) levels after phytohemagglutinin (PHA) stimulation. Blood samples were obtained from 40 healthy subjects and 30 RTRs and incubated with 5 µg/mL of PHA for 15-18 hr at 37°C and 5% CO2. Afterward, the CD4+ T cells were separated by antibody-coated magnetic beads and lysed. Then, iATP content in unstimulated and stimulated conditions was measured by luciferin-luciferase reaction using a log-log standard curve. The iATP levels showed significant increase in CD4+ T cells in both healthy persons (mean: 550 ± 142 ng/mL vs. 109 ± 54 ng/mL) and RTRs (mean: 394 ± 160 ng/mL vs. 52 ± 37 ng/mL) after PHA stimulation (P < 0.001). However, the iATP production in RTRs was significantly lower than that in healthy individuals; both prior to and after stimulation with PHA (P < 0.001). No gender-specific difference in iATP production was observed between women and men subjects. This rapid and low-cost assay reflects the degree of immune cell function through assessment of CD4+ T cells activation. Thus, it can be used for evaluation of immune system status in immunodeficient individuals as well as in immunosuppressed transplant recipients who needs drug adjustment.
authors.affiliation: a Immunology Research Center (IRC) , Iran University of Medical Sciences , Tehran , Iran.|b Department of Immunology , School of Medicine, Tehran University of Medical Sciences , Tehran , Iran.
authors.forename: Hadi
authors.initials: H
authors.lastname: Naderi
authors.affiliation: a Immunology Research Center (IRC) , Iran University of Medical Sciences , Tehran , Iran.
authors.forename: Alireza
authors.initials: A
authors.lastname: Najafi
authors.affiliation: a Immunology Research Center (IRC) , Iran University of Medical Sciences , Tehran , Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Khoshroo
authors.affiliation: a Immunology Research Center (IRC) , Iran University of Medical Sciences , Tehran , Iran.
authors.forename: Nader
authors.initials: N
authors.lastname: Tajik
mesh_terms: D000255:Adenosine Triphosphate / Q000097:blood* / Q000276:immunology*; D000328:Adult; D015496:CD4-Positive T-Lymphocytes / Q000737:chemistry* / Q000276:immunology*; D005260:Female; D006801:Humans; D007111:Immunity, Cellular / Q000276:immunology; D007118:Immunoassay; D008297:Male; D008875:Middle Aged; D010835:Phytohemagglutinins / Q000276:immunology; D055815:Young Adult
chemical_list: D010835:Phytohemagglutinins; D000255:Adenosine Triphosphate
keywords: CD4+ T cells; immune function assay; immunodeficiency; intracellular adenosine triphosphate (iATP); renal transplantation