Journal: Frontiers in Immunology

Article Title: Differences in Humoral and Cellular Vaccine Responses to SARS-CoV-2 in Kidney and Liver Transplant Recipients

doi: 10.3389/fimmu.2022.853682

Figure Lengend Snippet: Principal component analysis (PCA) of all the assays after second vaccination. (A) Analysis included antibodies against anti-SARS-CoV-2, anti-SARS-CoV-2 variants, and common coronaviruses, and IFN-γ and IL-2 T cell responses. The axes represent the two variables, among all those generated by PCA, that ranked first in terms of proportion of assay variability. The variables are represented by arrows; blue arrows = Fluorospot assays, orange arrows = anti-SARS-CoV-2 serological assays, yellow = serological assays for other coronaviruses. The angle between arrows represents the correlation between assays: assays with the same direction have a correlation coefficient of 1, those with opposite directions have a correlation coefficient of -1. Those that are perpendicular to each other have a correlation coefficient of 0. The patients are represented by data points with individual point size proportional to the quality of representation in the bi-dimensional plot. The plot is based on data after two vaccinations only (T2). The categorical variable type-of-solid-organ-transplantation (i.e. kidney vs liver) is added to the plot as a supplementary variable to visualize how the pattern of correlated variables and cloud of data points are distributed between types of solid organ transplantation. The colored ellipses represent the 95% confidence ellipses of the scatter around overall assay mean of each group (liver = blue or kidney = red). (B) Correlation between the five principal components extracted from PCA and the original variables (assays: T cell reactivity and IgG against SARS-CoV2 antigens and variants and other coronaviruses, including common coronaviruses: HCoV229E-S1, HCoVHKU1-S1, HCoVNL63S-S1, HCoVOC43-S1). As shown by the legend in the rightmost column, the correlation is represented by a color gradient as follows: blue for positive correlation, red for negative correlation, and white for no correlation. The correlation coefficient is represented by a circle, the diameter of which is proportional to the strength of the correlation. The variability explained by the principal components Dim.1 to Dim.5 was 67.7, 8.7, 5.6, 3.8, and 3.5%, respectively (not shown).

Article Snippet: The readout was performed following the manufacturer’s instructions for the Human IFN-γ/IL-2 Double-Color FluoroSpot kit and spots were counted using an automated ImmunoSpot Analyzer Professional System (both from Cellular Technology Ltd.).

Techniques: Generated, Transplantation Assay

Journal: Molecular Medicine Reports

Article Title: Continuous expression of CD83 on activated human CD4 + T cells is correlated with their differentiation into induced regulatory T cells

doi: 10.3892/mmr.2015.3796

Figure Lengend Snippet: Time-dependent expression of CD83, proliferation and the production of IL-2 and IFN-γ in CD4 + T cells. Following purity identification, the purified CD4 + T cells were either (A) directly stained with PE-labeled CD83 or (B–D) stimulated with anti-CD3/CD28 for 1–3 days, followed by staining with PE-labeled CD83. All samples were analyzed by flow cytometry and the typical flow cytometric histograms indicating the percentages of CD83-positive cells are shown. (E) Additionally, cell proliferation at days 1, 2 and 3 were determined using Cell Counting Kit 8. (F) The levels of IL-2 and IFN-γ in the supernatants from 1-, 2- and 3-day cultures of activated CD4 + T cells were analyzed by ELISA. Values are expressed as the mean ± standard deviation of three independent experiments ( ** P<0.01; * P<0.05). PE, phycoerythrin; CD, cluster of differentiation; IFN, interferon; IL, interleukin; neg.con, negative control.

Article Snippet: The levels of IL-2 and IFN-γ in CD4 + T-cell culture supernatants were measured in duplicate for each of the serial aliquots using a Human IL-2 Quantikine ELISA kit (cat. no. D2050) and Human IFN-γ Quantikine ELISA kit (cat. no. DIF50), purchased from R&D Systems (Minneapolis, MN, USA), according to the manufacturer's instructions.

Techniques: Expressing, Purification, Staining, Labeling, Flow Cytometry, Cell Counting, Enzyme-linked Immunosorbent Assay, Standard Deviation, Negative Control