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. 2022 Aug 18;83:104226. doi: 10.1016/j.ebiom.2022.104226

Figure 4.

Figure 4

Functional studies of CD8+ T cells in ABMR. Frozen PBMCs (n=24) from patients diagnosed with ABMR (biopsy group 3) was retrieved from our biocollection for functional studies. (A) Flow cytometry staining for CD45RA and CCR7 divides CD8+ T cells into 4 subpopulations: naïve (CD45RA+CCR7+), central memory (CM) (CD45RA-CCR7+), effector memory (EM) (CD45RA-CCR7-), and effector memory expressing CD45RA (TEMRA) (CD45RA+CCR7-). The percentage of these 4 subpopulations in CD28+CD8+ and CD28-CD8+ T cells are shown. (B) and (C) PBMCs were stimulated with plate-bound anti-CD3 mAb, soluble anti-CD28 mAb or IL-15, and plate-bound anti-CD16 mAb as indicated for 4 h. Cells were stained for surface CD8, CD28, and CD107a, followed by intracellular staining for TNFα and IFNγ and analyzed by flow cytometry. The percentage of cells double positive for TNFα and IFNγ (left) and positive for CD107a (right) among CD28+CD8+ (open circles) and CD28-CD8+ (filled circles) T cells are shown. (D) Frozen PBMCs (n=46) were also retrieved from patients with normal/subnormal biopsies (group 1) and stimulated as in (B) and (C). The functional responses of the CD28-CD8+ T cell subpopulation from patients with ABMR and normal/subnormal biopsies were compared (see also Supplementary Figure 3). *: p<0.05; **: p<0.01; ***:p<0.001; ****: p<0.0001.