Journal: bioRxiv

Article Title: Aging restricts maturation of CXCL13 + T follicular helper cells in human immunity

doi: 10.64898/2026.04.03.715992

Figure Lengend Snippet: a, Schematics of key differences in Tfh cell maturation between humans (left) and mice (right). b, Gene expression signatures of all CD4 + T cell clusters from tonsillar T cell scRNA-seq. c, Volcano plot showing differential gene expression of GC-Tfh cells (left) vs. CXCL13 + GC-Tfh cells (right). d, Select subset-specific marker gene expression in Tfh, GC-Tfh, and CXCL13 + GC-Tfh cells. e, Schematic overview for assessment of CXCL13 production by non-, pre-, and GC-Tfh cells. f, Exemplary gating for non-Tfh (PD-1 - CXCR5 - ), pre-Tfh (PD-1 lo CXCR5 lo ), and GC-Tfh cells (PD-1 hi CXCR5 hi ). g, CXCL13 secretion by different sorted T cell subtypes cultured with or without B cells (n = 4). h, Schematic representation for CRISPR/Cas9-based editing of tonsil samples. i, CXCL13 ELISA of tonsillar cell culture supernatants for indicated transcription factor knockouts vs. scramble control (n = 14; ≤ 30 yo). Data were analyzed using two-way ANOVA with Šídák’s multiple comparison test ( g ) and one-way ANOVA with Dunnett’s multiple comparisons test ( i ). * P < 0.05, ** P < 0.01.

Article Snippet: Editing efficiencies ( Table S3 ) were calculated using the Inference of CRISPR Edits (ICE) analysis tool (Synthego Performance Analysis 2019, v3.0) to ensure target gene knockout.

Techniques: Gene Expression, Marker, Cell Culture, CRISPR, Enzyme-linked Immunosorbent Assay, Control, Comparison

Journal: bioRxiv

Article Title: Aging restricts maturation of CXCL13 + T follicular helper cells in human immunity

doi: 10.64898/2026.04.03.715992

Figure Lengend Snippet: a, Number of differentially expressed genes (DEGs, >50yo vs. <30yo) over pseudotime during Tfh cell development from naïve CD4 + T cells (left) to CXCL13 + GC-Tfh cells (right). False discovery rate (FDR) is set to < 0.05. b, Volcano plot showing differential gene expression of Tfh cells from older (left) vs. younger donors (right). c, Schematic representation for CRISPR/Cas9-based editing of naïve CD4 + T cells followed by T cell polarization and flow cytometric analysis. d, Expression of CXCR5, FOXP3, and PD-1 in naïve CD4 + T cells cultured under Tfh-polarizing conditions (dark grey) or non-polarizing conditions (light grey). e, Log2 fold change relative to matched scramble controls for Th1 (left) and Th17 (right) cell frequency in TF-knockouts under Th1- or Th17-polarizing conditions. f, Log2 fold change relative to matched scramble controls for Tfh cell frequency in TF-knockouts under Tfh-polarizing conditions (gated on CD4 + CD19 - FOXP3 - CXCR5 + PD-1 + cells). Data were analyzed using a mixed effects model and Dunnett’s multiple comparisons test ( f ), one sample t test ( e ). Error bars SEM ( e-f ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Editing efficiencies ( Table S3 ) were calculated using the Inference of CRISPR Edits (ICE) analysis tool (Synthego Performance Analysis 2019, v3.0) to ensure target gene knockout.

Techniques: Gene Expression, CRISPR, Expressing, Cell Culture