Fig 2: Ex vivo infection upregulates CD20 expression. Unstimulated PBMCs from three uninfected donors were infected with HIV strain NL4.3. Infection was monitored by simultaneous staining of HIV RNA using the RNA FISH-flow assay and the viral protein p24 at days 3, 4, and 5. a Gating strategy used to monitor HIV infection and expression of CD20dim. Previous sequential gates are represented in Supplementary Fig. 9. b Percentage of productively HIV-infected cells in CD3+ T cells (left panel) and in CD20dim or CD20- CD3+ T cells (right panel). c Expression of CD20dim in infected and uninfected CD3+ T cells (left panel) and in the total CD3+ T cell population (right panel). d Correlation between the proportion of infected cells within the CD3+ T cell population and CD20 expression (left panel) and the mean fluorescence intensity (MFI) of CD20 (right panel). e Percentage of HLA-DR and PD-1 in infected and uninfected cells expressing or not expressing CD20dim. f Proportion of CD20dim in infected cells expressing the CD4 cell receptor versus infected cells with marked downregulation of the CD4 receptor. In all panels, the mean and SEM value of three independent experiments is represented. In panel (d), Spearman’s nonparametric correlation coefficients and associated p values are shown. Data underlying this Figure are provided as Source Data file

Fig 3: T cell MHP is associated with functional changes upon activation. (A) T cell ??m of individuals whose cryopreserved PBMC samples were selected for in vitro activation. Black dot: type 1 diabetes patients. Open circles: healthy controls. (B) CD4+ T cells from cryopreserved PBMC of individuals with MHP (n = 6, including 5 type 1 diabetes patients and 1 healthy control) show higher intracellular IFN? staining after 72-hour stimulation with plate-bound anti-CD3 and anti-CD28, when compare to cells from individuals without MHP (n = 26, including 10 type 1 diabetes patients and 16 healthy controls). (C) Mitochondria-specific ROS produced by enriched total T cells after stimulating by plate-bound anti-CD3 and anti-CD28 for 24 hours, is correlated with the amount of IFN? secreted in the supernatant. (D) Activation-induced change of ??m is positively correlated with mt-specific ROS production after 24-hour stimulation in vitro. *P < 0.05, ****P < 0.0001, Mann-Whitney test (A and B). Linear regression (C and D).

Fig 4: T cells from patients with type 1 diabetes exhibited MHP. T cells from peripheral blood of type 1 diabetes patients (T1D; n = 29) exhibited a higher mitochondrial membrane potential (??m) when compared to T cells from healthy controls (HC; n = 38) in (A) CD4+, (B) CD8+ and (C) CD3+ subsets; (D) In an independent cohort, enriched total T cells from fresh blood of T1D patients (n = 39) showed higher ??m when compared to HC individuals (n = 34) or patients with type 2 diabetes (n = 20). T cell ??m was not correlated with (C) age or (D) HbA1c level. *P < 0.05, **P < 0.005, ****P < 0.001. Mann-Whitney test.

Fig 5: Activation-induced ATP and cell death. (A) T cells from type 1 diabetes patients (T?1D) have lower basal cellular ATP content compare to healthy controls (HC) but cellular ATP ?was? significantly increased after in vitro activation. Enriched total T cells were stimulated in vitro with plate-bound anti-CD3 and anti-CD28 or isotype antibodies for 24 hours. ATP content was detected in cell lysate and corrected by protein amount. The trend remains that T cells from type 1 diabetes patients have lower ATP content even though in vitro activation induces a significant increase of ATP in this group. (B) T cells from T1D patients and HC showed the same level of activation-induced apoptosis. After 24-hour activation with plate-bound anti-CD3 and anti-CD28, T cell apoptosis was identified by staining with Annexin-V and Propidium Iodide. Activation-induced apoptosis was calculated as mentioned in method session. (C) T cells with MHP showed the same activation-induced apoptosis as normal ??m T cells. (D) Activation-induced apoptosis was not correlated with T cell ??m. *P < 0.05, unpaired t test with Welch’s correction. **P < 0.01 Wilcoxon matched pair test for (A) and Mann-Whitney test for (B).