Journal: BMC Cancer

Article Title: HLA class II-restricted T cell epitopes in public neoantigens of ESR1 and PIK3CA in breast cancer

doi: 10.1186/s12885-025-13992-6

Figure Lengend Snippet: Summary of the ELISPOT assay of healthy donors. A Representative data of the ELISPOT assay for ES#2_E380Q, PIK#5_E545A, PIK#8_H1047L, and PIK#9_H1047Y, measured by IFN-γ (red spots) and IL-2 (blue spots). The numbers denote the spot count for IFN-γ (red). NC, negative control. TNTC: too numerous to count. B , C The ELISPOT profiles for DRB4*01:03 positive donors ( B ) and DP5 positive donors ( C ). The data for peptide pairs that showed penetrance ≥ 0.4 and g15 ratio > 2.0 are presented. HLA alleles of each donor are displayed on the top. Positive ELISPOT responses (IFN-γ) are highlighted in red. Numbers indicate the positive responses/total number of experiments (range of spot counts). The penetrance represents the number of positive donors/numbers of total donors (in red letters). The peptides with %Rank < 10 (NetMHCIIpan-4.1) is highlighted in yellow, and those with a g15 ratio > 2.0 (MHC-density assay) is highlighted in light blue. NA: not analyzed (shadowed in gray). TNTC: too numerous to count

Article Snippet: The ELISPOT assay was conducted using the human interferon-gamma (IFN-γ)/IL-2 double-color ELISPOT kit (Cellular Technology Limited, Cleveland, OH, USA), following the manufacturer’s instructions.

Techniques: Enzyme-linked Immunospot, Negative Control

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: RSC Advances

Article Title: A novel phosphoester-based cationic co-polymer nanocarrier delivers chimeric antigen receptor plasmid and exhibits anti-tumor effect †

doi: 10.1039/c8ra02133c

Figure Lengend Snippet: The mPEG-bPEI-PEBP-DNA transfected Jurkat cells were mixed with CD19-K562 cells ((A) bright field panel, (B) green fluorescence panel, (C) red fluorescence panel, scale bar = 200 μm); after incubation for 48 h, the IFN-γ (D) and IL-2 (E) expression was detected by ELISA assay; the cytotoxic effects were measured at different E/T ratios (2 : 1, 4 : 1, 8 : 1) by the LDH assay (F). Data are expressed as mean ± sd. ** P < 0.01, * P < 0.05.

Article Snippet: To further study the biologic effects of mPEG-bPEI-PEBP-DNA transfected Jurkat cells (transfected Jurkat cells), secretion of T-helper type I cytokines, interferon-γ (IFN-γ) and interleukin-2 (IL-2), was measured using commercially available ELISA reagents (Human IL-2 ELISA Kit, Human IFN-γ ELISA Kit, Becton, Dickinson and Company).

Techniques: Transfection, Fluorescence, Incubation, Expressing, Enzyme-linked Immunosorbent Assay, Lactate Dehydrogenase Assay