Figure 2. Expression of β2-ARs on T cells regulates the T cell phenotype during allogeneic responses.

(A) Immune-related gene expression in WT or β2-AR^–/– CD4⁺ T cells sorted from spleen at day 14 after allo-HCT in the B6→BALB/c model transplanted with 3.5 × 10⁶ WT C57BL/6 TCD-BM alone or combined with 0.7 × 10⁶ WT C57BL/6 or β2-AR^–/– pan–T cells using NanoString nCounter Immunology Panel mRNA microarray (splenocytes pooled from 5–6 mice before CD4⁺ T cell sorting). (B) RORγt⁺T-bet⁺ frequencies in liver and spleen CD4⁺ T cells within single live H-2^b+H-2^d–CD45⁺CD3⁺ cells 7 and 14 days after allo-HCT. There are increased RORγt⁺T-bet⁺CD4⁺ T cells in recipients of β2-AR^–/– T cells. Representative plots are shown from 2 independent experiments (n = 5–6 in total). (C) T-bet⁺, Foxp-3⁺, IFN-γ⁺, IL-17⁺, and IL-10⁺ frequencies in CD4⁺ T cells within single live H-2^b+H-2^d–CD45⁺CD3⁺ cell populations from the spleen and liver at days 7 and 14, and the GI tract 14 days after allo-HCT. Data pooled from 2 individual experiments, total n = 5–6 per group. B6→BALB/c model using B6→BALB/c model transplanted with 3.5 × 10⁶ WT C57BL/6 TCD-BM alone or combined with 0.7 × 10⁶ WT C57BL/6 or β2-AR^–/– pan–T cells. CD4⁺T-bet⁺, CD4⁺IFN-γ⁺, CD4⁺IL-17⁺, and T-bet⁺Rorγt⁺CD4⁺ T cells were significantly increased in β2-AR^–/– versus WT T cells. CD4⁺Foxp-3⁺ and CD4⁺IL-10⁺ cells significantly increased in WT versus β2-AR^–/– T cells. (D) Plasma levels of inflammatory/inhibitory cytokines of irradiated BALB/c mice transplanted with TCD-BM plus WT or β2-AR^–/– T cells at 7 and 14 days. Plasma IFN-γ on days 7 and 14 and IL-17 levels on day 7 increased significantly, while IL-4 and IL-10 levels decreased significantly in β2-AR^–/– T cell recipients. There were no differences in plasma GM-CSF levels. For comparison of the means, an unpaired 2-tailed t test was used in B, C, and D. *P < 0.05, **P < 0.01, ***P < 0.001. Data are presented as median ± min to max.