Transcriptomic And Proteomic Analysis Of Cd4(+) T Cells To Identify Sex Differences In Angiotensin Ii Signaling Pathways

T cells are involved in hypertension pathogenesis in both males and postmenopausal females while premenopausal females are resistant to T cell-mediated Ang II-induced hypertension. The goals of this study were (1) to identify T cell specific proteomic pathways associated with postmenopausal susceptibility to hypertension (2) to identify T cell specific transcriptomic changes associated with premenopausal protection from hypertension. Proteomic analysis was performed on splenic CD4 + T cells isolated from premenopausal and postmenopausal females (VCD, 160 mg/kg/day i.p. 20d) following Ang II infusion (800 ng/kg/min 14d). 384 proteins from CD4 + T cells were identified as differentially expressed following Ang II infusion in premenopausal females. 285 proteins from CD4 + T cells were identified as differentially expressed between premenopausal and postmenopausal females following Ang II infusion. Gene ontology (GO) analysis of pre vs. postmenopausal proteins identified enriched pathways associated with RNA binding, chaperone activity and cellular stress responses. Transcriptomic changes were analyzed, via RNAseq, on isolated splenic CD4 + T cells from premenopausal females, with and without Ang II infusion. Thirty-four genes were identified as differentially expressed in CD4 + T cells following Ang II infusion. GO analysis of Ang II upregulated genes revealed an enrichment of five distinct molecular functions, including antioxidant activity. In a subsequent study to validate the RNAseq, we confirmed that Ang II increased CD4 + T cell mRNA expression of calprotectin (S100a8/S100a9), a calcium and zinc binding protein complex that contributes to antioxidant defense (S100a8: Con 1.0 ± 0.4 vs Ang II 5.0 ± 0.8*; S100a9: Con 1.0 ± 0.4 vs Ang II 6.0 ± 0.8*, *P<0.05 vs Con). Furthermore, we determined that Ang II did not increase calprotectin expression in mice lacking estrogen receptor α (ERKO) (S100a8: ERKO 0.1 ± 0.4 vs ERKO/Ang II 0.3 ± 0.7; S100a9: ERKO 0.2 ± 0.5 vs ERKO/Ang II 0.3 ± 0.6). The current studies demonstrate a role for estrogen in Ang II-induced T cell gene expression and signal transduction, and begin to elucidate the molecular mechanisms of female protection from T cell-mediated hypertension.