Abstract
Objective:
Treatment with human umbilical cord mesenchymal stem cells (hUC-MSCs) attenuated the clinical manifestations of systemic lupus erythematosus (SLE). We investigated the metabolic mechanism whereby hUC-MSCs modify CD4+ T cell cytokine secretion in lupus.
Methods:
The study enrolled 30 untreated lupus patients and 20 sex, age, and body mass index matched healthy controls (HCs). CD4+ T cells were isolated by magnetic sorting, and stimulated with anti-CD3/CD28. The hUC-MSCs treatment (MSCT) groups were coculturing hUC-MSCs to CD4+ T cells from moderate and severe SLE (SLE-MS) groups for 72 h at ratios of 1:25 (T1), 1:10 (T2), and 1:5 (T3). Cytokine concentration and proliferation of the CD4+ T cells were measured by Luminex liquid chip assay and cell counting kit-8, respectively. Glucose metabolic capacity was measured by Seahorse real-time metabolic analysis. The role of hUC-MSCs on cytokine secretion was analyzed by transcriptome sequencing. Glucose enzymes levels and HSP90AA1/PI3K/AKT pathway activity were analyzed by real-time quantitative PCR and western blot. The CD4+ T cell subsets were detected by flow cytometry.
Results:
Compared with HCs, the enhanced glycolysis and mitochondrial oxygen consumption of SLE-CD4+ T cells were positively associated with disease activity. Treatment with hUC-MSCs proportionally decreased glucose metabolism and proliferation of SLE-CD4+ T cells. The hUC-MSCs treatment significantly diminished supernatant concentrations of interferon-γ, tumor necrosis factor-α, interleukin (IL)-4, and IL-17 in SLE-MS group, as well as inhibited HSP90AA1 in the glucose-activated PI3K-AKT pathway. In animal experiment, the systemic administration of hUC-MSCs and inhibition of HSP90AA1 resulted in a reduction of glucose metabolites, enzymes, pro-inflammatory factor levels, and HSP90AA1/PI3K/AKT signaling pathway activity.
Conclusions:
The hUC-MSCs treatment inhibited overactive glucose metabolism of SLE-CD4+ T cells. HSP90AA1 in the PI3K-AKT pathway induced by the glucose metabolism may be involved in the anti-inflammatory function of hUC-MSCs treatment.
Keywords:
CD4+ T cell; HSP90AA1; Lupus; PI3K‐AKT pathway; cytokine; glucose metabolism; mesenchymal stem cell.