Fig 2: Patch repair and membrane permeabilization are not sufficient to impair macrophages (A) BMDM were unchallenged or challenged with 500 HU/mL SLO WT, PFO or equivalent mass amount of SLO ML for 10 min at 37 °C in presence of FM1–43FX. Excess toxin and dye were washed away with PBS. Cells were then primed with 100 EU/mL LPS for 2 h and stained for intracellular TNFa as described in Fig. 1A. The percentage TNFa positive live cells in FM1–43Xlow and FM1–43Xhigh populations was measured by flow cytometry. (B) BMDM were challenged and stimulated as described in Fig. 2A. Cells were harvested, labeled with Ghost Red, stained for surface Lamp1, fixed, permeabilized and stained for intracellular TNFa. The percentage of TNFa positive live cells in Lamp1low and Lamp1high populations is shown. Graphs represent mean ± sem of three independent experiments.

Fig 3: Dysfunctional CD8+ T cells in BXD2 lupus mice cause viral hepatitis in LCMV infection.(A) MFI of CD38 in gp33+ CD8+ T cells of B6, diseased BXD2, or young nondiseased BXD2 mice 8 days after LCMV Armstrong infection. (B to D) Percentage of positive cells expressing CD107a (B), granzyme B (C), and IFN-? (D) in gp33+ CD8+ T cells of B6, diseased BXD2, or young nondiseased BXD2 mice 8 days after LCMV Armstrong infection. (E) LCMV viral load 8 days after LCMV Armstrong infection using quantitative PCR of glycoprotein gene normalized by tissue ACTB in liver, kidney, and lung of B6, diseased BXD2, or young nondiseased BXD2 mice. (F to H) Representative liver histology of B6 (F), diseased BXD2 (G), or young nondiseased BXD2 (H) mice 8 days after LCMV Armstrong infection. (I to K) Representative liver histology of diseased BXD2 mice 8 days after LCMV Armstrong infection showing focal lesion of necrosis (I) and portal and periportal inflammation (J and K). Data are means ± SD (A to D), mean ± SE (E); statistical analysis by two-tailed t test. ns = P > 0.05, *P < 0.05, **P < 0.01, ****P < 0.0001.

Fig 4: CD38 inhibition reverses mitochondrial defect and restores CD8+ T cell function in BXD2 lupus mice against LCMV infection.(A and B) Percentage of positive cells expressing CD107a (A) and granzyme B (B) in gp33+ CD8+ T cells 8 days after LCMV Armstrong infection from diseased BXD2 treated with control water or MK-0159. (C) Percentage of depolarized mitochondria in gp33+ CD8+ T cells 8 days after LCMV Armstrong infection from diseased BXD2 treated with control water or MK-0159. (D) LCMV viral load 8 days after LCMV Armstrong infection using qPCR of glycoprotein gene normalized by tissue ACTB in diseased BXD2 treated with control water or MK-0159. (E to G) Representative liver histology 8 days after LCMV Armstrong infection from diseased BXD2 treated with control water (E) or MK-0159 (F) and their respective hepatitis activity index (G). (H and I) Percentage of positive cells expressing CD107a (H) and granzyme B (I) in gp33+ CD8+ T cells 8 days after LCMV Armstrong infection from diseased BXD2 treated with control NP or NP-MK-0159. (J to M) Representative liver histology 8 days after LCMV Armstrong infection from diseased BXD2 treated with NP-control with the presentation of necrosis (J) or portal and periportal inflammation (K) or treated with NP-MK-0159 (L) and their respective hepatitis activity index (M). (N) LCMV viral load 8 days after LCMV Armstrong infection using qPCR of glycoprotein gene normalized by tissue ACTB in diseased BXD2 treated with NP-control or NP-MK-0159. (O) Percentage of depolarized mitochondria in gp33+ CD8+ T cells 8 days after LCMV Armstrong infection from diseased BXD2 treated with NP-control or NP-MK-0159. (P and Q) MFI of PINK1 (P) and LysoSensor Green (Q) in gp33+ CD8+ T cells 8 days after LCMV Armstrong infection from diseased BXD2 treated with NP-control or NP-MK-0159. Data are means ± SD; statistical analysis by two-tailed t test. *P < 0.05, **P < 0.01.

Fig 5: CTL serial conjugation and effector function in different tumor models.a Time-lapse sequence and migration track of one OT1 CTL killing seven MEC-1/OVA target cells sequentially within 11 h. Circles, CTL; cross, apoptotic target cell; scale bar, 20 µm. Representative example of a serial killer CTL derived from analysis of 43 CTL pooled from 8 independent experiments). b Lamp-1 expression at the surface of OT1 CTL after 24 h of 3D co-culture with MEC-1/OVA cells. Representative example from three independent experiments. c Lag phase until apoptosis of consecutive single interaction kills by the same CTL (43 CTL from 8 independent experiments). Red bars, median. p-Value, Kruskal–Wallis test. d Population survival of four antigen-dependent mouse and human target, and three control cell lines. Quantifications from three independent experiments for each cell line. B16F10, 98 cells; B16F10/OVA, 31 cells; MEC-1, 57 cells; MEC-1/OVA, 32 cells; BLM, 28 cells; MV3, 18 cells; MCF-7, 102 cells. e Inefficiency of inducing apoptosis by individual CTL contacts in OT1 and SMCY.A2 CTL models. Error bars, mean ± SD obtained from 104 MEC-1/OVA, 183 B16F10/OVA, 53 BLM, and 50 MV3 contacts from N = 5 (MEC-1/OVA, B16F10/OVA) or 3 (BLM, MV3) independent experiments per cell model. Source data are provided as a Source Data file.