Journal: The Journal of Infectious Diseases
Article Title: Functional Diversity of Cytomegalovirus–Specific T Cells Is Maintained in Older People and Significantly Associated With Protein Specificity and Response Size
Figure Lengend Snippet: The functional T-cell response composition in young and older people appears to be very similar. Peripheral blood mononuclear cells (PBMCs) from cytomegalovirus (CMV)–positive donors were stimulated overnight with 19 CMV protein–derived overlapping peptide-pools. Activated T cells and their functional composition were analyzed by flow cytometry, including the activation markers, degranulation (CD107), CD154, interleukin 2 (IL-2), tumor necrosis factor (TNF), and interferon γ (IFN-γ). Responses from the United Kingdom cohort are shown for the 2 most frequently targeted proteins for each T-cell subset. A , Nonoverlapping subsets exhibiting 5, 4, 3, or 2 parallel functions or just 1 single function are shown as a proportion of the specifically activated T cells (in order of decreasing polyfunctionality). A significant difference between the age groups is observed for UL-83–specific CD4 + T cells with 2 functions, but, on the whole, subset distributions appear to be very similar. B , The functional dominance of each activation marker in a given response is assessed by the proportion of activated cells expressing it. Some variability between proteins was revealed, but there was no significant differences between the age groups. There were 5 end points evaluated, with the significance threshold set at a P value of ≤ .01. P values were determined by the Mann–Whitney test with Bonferroni correction for multiple end points.
Article Snippet: Antibodies We used the following fluorescence-conjugated monoclonal antibodies and staining reagents: anti-CD3-v500, anti-CD8-allophyocyanine (APC)-H7, anti-CD27-phycoerythrine (PE), interleukin 2 (IL-2)-fluorescein isothiocyanate (FITC), tumor necrosis factor α (TNF-α)-Alexa 700, and CD107a-APC (all BD Biosciences, Oxford, United Kingdom); anti-CD4-peridinin chlorophyll (PerCP), anti-interferon γ (IFN-γ) PE-cyanine 7, anti-CD154 Pacific-Blue (BioLegend, Cambridge, United Kingdom), anti-CD45RA-ECD (Beckman Coulter, United Kingdom), and Yellow live-dead stain (Invitrogen, Paisley, United Kingdom).
Techniques: Functional Assay, Derivative Assay, Flow Cytometry, Cytometry, Activation Assay, Marker, Expressing, MANN-WHITNEY