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. 2023 Feb 25;26(3):106269. doi: 10.1016/j.isci.2023.106269

Figure 2.

Figure 2

Frequencies and phenotypes of pigtail macaque tissue Vγ9Vδ2 T cells

Vγ9Vδ2 T cell frequencies and phenotypes were evaluated in peripheral blood mononuclear cells (PBMC), inguinal lymph nodes/tracheobronchial lymph nodes (ILN/TBLN), mesenteric lymph nodes (MLN), spleen, thymus, lung, liver, small intestine (SI), and large intestine (LI).

(A) Representative Vγ9Vδ2 T cell frequencies in paired cryopreserved PBMC and tissue samples from an adult male pigtail macaque. CD3+ Vγ9Vδ2 T cells were identified based on the flow cytometry gating strategy illustrated in Figure 1A. FACS plots show Vγ9Vδ2 T cell (magenta) and bulk CD3+ T cells (black).

(B) Frequencies of CD28+CD95− (naive; TN), CD28+CD95+ (central memory; TCM), CD28−CD95+ (effector memory; TEM), and CD28−CD95− Vγ9Vδ2 T cells in peripheral blood and tissue sites.

(C) Frequencies of CD26 and NKG2A expression of peripheral blood- and tissue-derived Vγ9Vδ2 T cells.

(D) Representative histograms and frequencies of CD69 expression on peripheral blood- and tissue-derived Vγ9Vδ2 T cells.

(E) Representative histograms and frequencies of TNFα and IFNγ expression in peripheral blood- and tissue-derived Vγ9Vδ2 T cells, following a 16 h in vitro stimulation with HMB-PP. In vitro stimulation was performed as described in the method details. Cytokine production frequencies were calculated following background subtraction using paired unstimulated control samples.

(F) Representative histograms and median fluorescence intensities (MFI) of granzyme B (GzmB) expression in peripheral blood- and tissue-derived Vγ9Vδ2 T cells at steady state. Additional tissue phenotyping data presented in Figure S1.

Each point on the graphs represents an individual animal for each tissue sample. Lines and error bars indicate median and interquartile range. All analysis was performed on cryopreserved samples (n = 2–6 animals depending on tissue availability and cell recovery, from 2 to 5 independent experiments). Statistics assessed by Kruskal-Wallis test with Dunn’s multiple comparisons correction (Aa dn D–F). ∗p < 0.05, ∗∗p < 0.01.