Figure 5.

Aldesleukin induces cytotoxicity and pro-inflammatory mediator release in CD8⁺ T cell/primary human hepatocyte co-cultures. Primary human CD8⁺ T cells were stimulated with aldesleukin or activation controls (act I, CD3/CD28 + rhIL-2 activation; act II, PHA-L+ rhIL-2 activation) and co-incubated with primary human hepatocytes (PHHs). (A) For cytotoxicity assessment, CD8⁺ T cells were co-cultured with hepatocytes for 5 d. Lactate dehydrogenase (LDH) activity was determined via colorimetric assay and samples were referred to the lysis control. (B-E) For quantification of released proteins, 3 days pre-stimulated CD8⁺ T cells were added onto hepatocytes for 2 days. Proteins were quantified via (B) flow cytometry or via (C-E) proximity extension assay. Data are shown as mean ± SEM. N = 4 (B-E) and N = 5 (A) biological replicates were evaluated. Per measurement, one technical replicate was acquired. For statistical analysis, one-way ANOVA with Dunnett´s multiple comparison test was used. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 indicate significant differences between aldesleukin-treated or activated and vehicle-treated samples. CCL3, Chemokine (C-C motif) ligand 3; CXCL6, Chemokine (C-X-C motif) ligand 6; L, Triton-X-lysed PHHs; MCP-1, monocyte chemoattractant protein 1; NPX, Normalized Protein eXpression; untr., untreated PHH monocultures.