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Impact of serum on γδ T cell expansion and phenotype. (A) Fold expansion comparison with and without 5% human AB serum in the culture medium, analyzed using the Wilcoxon matched pairs signed rank test, n=14. (B) Cellular composition assessed by flow cytometry throughout the expansion, illustrated for a representative donor. (C) γδ T cell phenotype determined by flow cytometry based on CD27 and CD45RA expression. Exemplary plots on day 13 and proportions throughout the culture for a representative donor. (D) Expression of activation markers on γδ T cells analyzed by flow cytometry. Šídák’s multiple comparisons test, n=15 for CD69, CD56, and HLA-DR; n=6 for NKG2D. (E) Expression of inhibitory and exhaustion markers on γδ T cells, analyzed by flow cytometry. Šídák’s multiple comparisons test, n=15. In the figures, the significance levels denoted by stars are as follows: *p < 0.05, ***p < 0.001, ****p < 0.0001, ns = non-significant.

Journal: Frontiers in Immunology

Article Title: Human serum influences functional plasticity and transcriptomic landscape of γδ T cells in vitro

doi: 10.3389/fimmu.2026.1722590

Figure Lengend Snippet: Impact of serum on γδ T cell expansion and phenotype. (A) Fold expansion comparison with and without 5% human AB serum in the culture medium, analyzed using the Wilcoxon matched pairs signed rank test, n=14. (B) Cellular composition assessed by flow cytometry throughout the expansion, illustrated for a representative donor. (C) γδ T cell phenotype determined by flow cytometry based on CD27 and CD45RA expression. Exemplary plots on day 13 and proportions throughout the culture for a representative donor. (D) Expression of activation markers on γδ T cells analyzed by flow cytometry. Šídák’s multiple comparisons test, n=15 for CD69, CD56, and HLA-DR; n=6 for NKG2D. (E) Expression of inhibitory and exhaustion markers on γδ T cells, analyzed by flow cytometry. Šídák’s multiple comparisons test, n=15. In the figures, the significance levels denoted by stars are as follows: *p < 0.05, ***p < 0.001, ****p < 0.0001, ns = non-significant.

Article Snippet: The following antibodies were used for cell surface staining and intracellular staining: anti-human CD107a REA803, anti-human CD14 REA599, anti-human CD19 REA675, anti-human CD27 REA499, anti-human CD3 REA613, anti-human CD45RA REA562, anti-human CD56 REA196, anti-human CD69 REA824, anti-human Granzyme B REA226, anti-human HLA/DR REA805, anti-human IFNγ 45-15, anti-human KIR2D REA1042, anti-human NKG2D REA797, anti-human PD-1 REA1165, anti-human Perforin REA1061, anti-human REA Control (I) REA293, anti-human REA Control (S) REA293, anti-human TCR Vδ1 REA173, anti-human TCR Vδ2 REA771, anti-human TCR γδ REA591, anti-human TIGIT REA1004, and anti-human TIM3 F38-2E2 (all from Miltenyi Biotec).

Techniques: Comparison, Flow Cytometry, Expressing, Activation Assay