alp  (R&D Systems)

Journal: iScience

Article Title: Robust effector memory features of human T-bet hi B cells induced by repeated mRNA vaccination

doi: 10.1016/j.isci.2026.114804

Figure Lengend Snippet: RBD-specific T-bet hi B cells induced by multiple mRNA vaccinations are robustly affinity-matured and can rapidly differentiate into antibody-secreting cells (ASCs) (A) Gene set enrichment analysis (GSEA) showing that RBD-specific T-bet hi B cells are significantly enriched in gene sets related to effector functions and activation, compared with RBD-specific switched memory B cells. Dotted lines indicate false discovery rate (FDR) of 0.05. (B) Gating strategy for fluorescence-activated cell sorting (FACS) of T-bet hi and classical memory B cells. (C) Representative ASC frequencies among sorted T-bet hi and classical memory B cells at 48 h after in-vitro polyclonal stimulation with IL-2 and R848. (D) Summary of ASC frequencies after stimulation of sorted T-bet hi and classical memory B cells, showing more robust differentiation potentials of T-bet hi B cells. (E) Representative data from IgG ELISPOT with sorted memory B cell subpopulations. (F) Summary of Wuhan-Hu-1 RBD-specific IgG SFUs per 1000 total IgG SFUs normalized by RBD-specific B cell frequency of each subset between classical and T-bet hi B cells. (G) Summary of RBD-specific IgG SFUs per 1000 total IgG SFUs among stimulated classical memory, unstimulated classical memory and T-bet hi B cells. (H) Surrogate RBD neutralization capacity of antibodies secreted by memory B cell subsets, in terms of binding inhibition rates. (I) Heavy chain somatic hypermutation frequency of RBD-specific memory B cells 14 days after the 2 nd or 3 rd vaccination, showing significant and comparable affinity maturation of T-bet hi B cell. (J) LIBRA-seq scores calculated from RBD-tetramer ADT increases in both subpopulations following additional vaccine doses. Permutation test for GSEA terms in A, Wilcoxon rank-sum test for D–4H and Mann-Whitney U test for I and 4J, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; n.s., non-significant. Pooled data are represented as mean ± SD.

Article Snippet: ELISpot Flex: Human IgG (ALP) , MABTECH , Cat# 3850-2A.

Techniques: Activation Assay, Fluorescence, FACS, In Vitro, Enzyme-linked Immunospot, Neutralization, Binding Assay, Inhibition, MANN-WHITNEY

Journal: bioRxiv

Article Title: Duration of Initial Viremia Modulates Functional Properties of HIV-specific T Cell Receptors

doi: 10.64898/2026.01.29.702605

Figure Lengend Snippet: Breadth and magnitude of HIV-specific CTL responses in PBMCs, measured by IFN-g ELISPOT to five clade C HLA-B*58:01-restricted consensus epitopes (TW10, IW9, KW11, RY11, and HW9) in the 12 participants, measured during the acute phase of HIV-1 infection. Reported values represent spot-forming units (SFUs) after subtraction of background activity from triplicate negative control wells without peptide stimulation . Each column corresponds to an individual epitope, with response magnitudes depicted by a color scale. Numbers within each box indicate SFUs per 10 6 PBMCs. The intensity scale shown on the right denotes the strength of responses above background. Early and late treated persons are indicated by identifiers shown in red and blue boxes, respectively.

Article Snippet: IFN-gamma ELISPOT assays were performed according to the manufacturer’s instructions with the human IFN-gamma ELISPOT Basic Kit (ALP) (Cat. 3420-2 A, Mabtech).

Techniques: Enzyme-linked Immunospot, Infection, Activity Assay, Negative Control