Journal: Experimental & Molecular Medicine

Article Title: Discovery of intrahepatic CD103 + cDC1/CD8 + T RM protective immune axis against acetaminophen-induced acute liver injury

doi: 10.1038/s12276-025-01565-3

Figure Lengend Snippet: a , b The frequency of hepatic CD69 + T RM cells in WT APAP mice ( n = 3 per group per time point) ( a ) and in WT and Batf3-KO mice 24 h after APAP administration ( n = 3 per group) ( b ). c The frequency of Ki67 + cells in hepatic CD69 + KLRG1 − CD44 + CD8 + T RM cells in APAP mice. n = 4 per group. d The population of adoptively transferred OT-I T cells and of OT-I T RM cells in the liver of recipient mice after APAP administration. n = 3 per group. e The frequency of CD69 + T RM cells was assessed when APAP mice were pretreated with blocking antibodies against IL-12, IL-15 and IL-18. n = 4 per group. f The ALT level was assessed in APAP mice pretreated with IL-15 blocking antibodies (α-CD122). n = 3 per group per time point. g The frequency of IL-15Rα + cells in the hepatic DC1s and DC2s of APAP mice. n = 3 per group. h The frequency of T RM cells in hepatic CD8 + T cells when CD8 + T cells were cultured in the presence of IL-15. n = 4 per group. i The frequency of T RM cells among CD8 + T cells when hepatic CD8 + T cells were cocultured with LPS-stimulated hepatic DC1s of APAP mice in the presence of IL-15 blocking antibodies. n = 4 per group. Ordinary one-way ANOVA with Dunnett for posttest in a and e , unpaired two-tailed Student’s t -test in b – d and g – i and two-way ANOVA with Bonferroni for posttest in f were used to measure significance. The error bars indicate mean ± s.e.m. ns, not significant. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: For IL-12, IL-15, IL-18 and CCL2 neutralizing, mice were i.p. injected with 200 μg per mouse of anti-IL-12 antibody (clone R1-5D9), anti-CD122 (IL-15Rβ) antibody (clone TM-Beta1), anti-IL-18 antibody (clone YIGIF74-1G7) and anti-CCL2 antibody (clone 2H5) (all from BioXCell).

Techniques: Blocking Assay, Cell Culture, Two Tailed Test

Journal: Experimental & Molecular Medicine

Article Title: Discovery of intrahepatic CD103 + cDC1/CD8 + T RM protective immune axis against acetaminophen-induced acute liver injury

doi: 10.1038/s12276-025-01565-3

Figure Lengend Snippet: a Based on the public single-cell RNA sequencing dataset from liver tissues of healthy donors and patients with APAP-ALI ( GSE223581 ), the relative expression levels of IL-15 were assessed between APAP patients and healthy donors, and the correlation between IL-15 level and CD69 + T RM population among the CD8 + T cells was evaluated. b Using the same dataset, the frequency of CD14⁺ monocytes among total liver immune cells (left) and the populaton of FCGR3A⁺ inflammatory monocytes among the CD14 + monocytes in the liver (right) were assessed between healthy donors and patients with APAP-ALI. For inflammatory monocytes, the ratio of FCGR3A >0.1 was assessed in the CD14 >0.1 population of monocytes. c The mRNA levels of IL-15 / IL-15RA were assessed by quantitative real-time PCR in LPS-activated hepatic DC1s isolated from noncancerous healthy tissues of resected liver specimens obtained from patients with HCC and other liver diseases. n = 3 per group. d , e The frequency of human hepatic CD8 + T RM cells when hepatic CD8 + T cells were cocultured with LPS-treated hepatic DC1s ( d ), in the presence of IL-15 blocking antibodies (α-CD122) ( e ). f The human hepatic inflammatory monocytes were cocultured with human hepatic T RM cells at a different ratio, and the frequency of annexin V + monocytes was assessed. n = 4 per group. Unpaired two-tailed Student’s t -test in a and b , multiple unpaired t -test in c and ordinary one-way ANOVA with Dunnett for the posttest f were used to measure significance. The error bars indicate mean ± s.e.m. ns, not significant. * P < 0.05.

Article Snippet: For IL-12, IL-15, IL-18 and CCL2 neutralizing, mice were i.p. injected with 200 μg per mouse of anti-IL-12 antibody (clone R1-5D9), anti-CD122 (IL-15Rβ) antibody (clone TM-Beta1), anti-IL-18 antibody (clone YIGIF74-1G7) and anti-CCL2 antibody (clone 2H5) (all from BioXCell).

Techniques: RNA Sequencing, Expressing, Real-time Polymerase Chain Reaction, Isolation, Blocking Assay, Two Tailed Test

Journal: Skeletal Muscle

Article Title: Skeletal muscle interleukin 15 promotes CD8 + T-cell function and autoimmune myositis

doi: 10.1186/s13395-015-0058-2

Figure Lengend Snippet: Skeletal muscle cells express IL-15/IL-15Rα protein complex in response to TNF-α and IFN-γ stimulation. a Expression of Il15 and Il15ra mRNA during C2C12 myoblast differentiation. Samples were collected before (0) and 2, 4, and 6 days after differentiation induction. b Expression of Il15 and Il15ra mRNA in C2C12 myotubes treated with TNF-α (10 ng/ml), IFN-γ (10 ng/ml), TNF-α + IFN-γ (TNF + IFN, 10 ng/ml each), or without cytokine ( con ) for 1, 2, 3, 6, 12, and 24 h. c Expression of Il15 and Il15ra mRNA in primary myotubes treated with TNF-α (5 ng/ml), IFN-γ (5 ng/ml), TNF + IFN (5 ng/ml each), or without cytokine ( con ) for 24 h. d Expression of IL-15/IL-15Rα complex protein in C2C12 myoblasts and myotubes treated with TNF-α (1 or 10 ng/ml), IFN-γ (1 or 10 ng/ml), TNF + IFN (1 or 10 ng/ml each), or without cytokine ( con ) for 24 h. e Expression of IL-15/IL-15Rα complex protein in primary myotubes treated with TNF + IFN (5 ng/ml each) or vehicle ( con ) for 24 h. f Expression of IL-15/IL-15Rα complex protein in skeletal muscle in vivo. TNF-α plus IFN-γ (1 mg each/injection) or PBS ( con ) was injected into the quadriceps muscles of mice three times at 4-h intervals. The injected muscles were collected 16 h after the last injection. Total RNA was isolated and analyzed by qPCR ( a–c ). Protein lysate and cell culture medium were collected and measured by ELISA ( d–f ). Data in ( a–c ) were triplicates and representative of two independent experiments with similar result. Data in ( d, e ) were pooled from three independent experiments. Data in ( f ) was pooled from three mice in each group. Data are mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, in comparison to “0” or “con”

Article Snippet: C2C12 cells were stained with biotin-conjugated antibodies against IL-15 (Cat. No. 500-P173Bt, PeproTech), IL-15Rα (Cat. No. BAF551, R&D), IL-15Rβ (CD122, clone: TM-b1, eBioscience), and γ c (CD132, Cat. No. 554470, BD Biosciences) then incubated with APC-conjugated streptavidin (BD Biosciences).

Techniques: Expressing, In Vivo, Injection, Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Skeletal Muscle

Article Title: Skeletal muscle interleukin 15 promotes CD8 + T-cell function and autoimmune myositis

doi: 10.1186/s13395-015-0058-2

Figure Lengend Snippet: IL-15 is presented on the surface of skeletal muscle cells. a Expression of IL-15 and IL-15Rα on C2C12 myoblasts. Cells were treated with TNF-α and IFN-γ (TNF + IFN) or without cytokine ( Control ) for 24 h and then stained with anti-IL-15 and IL-15Rα antibodies for flow cytometry analysis. “ Unstained ” indicates cells without antibody staining. Data are representative of three independent experiments with similar results. b Quantification of IL-15 bound by cell surface IL-15Rα on C2C12 myotubes. C2C12 myotubes were first treated with TNF-α and IFN-γ for 24 h, incubated with acid glycine buffer to dissociate IL-15 from cell surface IL-15Rα, and then re-treated with exogenous IL-15. Cell lysates from each step were collected and the amount of IL-15/ IL-15Rα complex was measured by ELISA. Data are mean ± SEM of triplicates and representative of two independent experiments with similar results. *** p < 0.001

Article Snippet: C2C12 cells were stained with biotin-conjugated antibodies against IL-15 (Cat. No. 500-P173Bt, PeproTech), IL-15Rα (Cat. No. BAF551, R&D), IL-15Rβ (CD122, clone: TM-b1, eBioscience), and γ c (CD132, Cat. No. 554470, BD Biosciences) then incubated with APC-conjugated streptavidin (BD Biosciences).

Techniques: Expressing, Staining, Flow Cytometry, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Skeletal Muscle

Article Title: Skeletal muscle interleukin 15 promotes CD8 + T-cell function and autoimmune myositis

doi: 10.1186/s13395-015-0058-2

Figure Lengend Snippet: Skeletal muscle IL-15 contributes to the progression of autoimmune myositis. a The expression of Il15 mRNA in various tissues of wt ( n = 3), Il15 f/f ( n = 5), and ACTA-Il15 −/− ( n = 5) mice was detected by qPCR. WAT white adipose tissue. Data are mean ± SEM. *** p < 0.001. b The amount of IL-15/IL-15Rα complex protein in the serum of Il15 f/f and ACTA-Il15 −/− mice ( n = 9–10) was measured by ELISA. Data are mean ± SEM. c Comparison of memory CD8 + T cell (mCD8) and NK cell (NK) level in the peripheral blood among wt, Il15 −/− , Il15 f/f , and ACTA-Il15 −/− mice ( n = 5 in each group). Fold change was calculated by normalizing the percentage of indicated cell type in mutant mice to that in wt mice. mCD8 were H57 + CD19 − CD8 + CD44 hi CD122 hi , and NK were H57 − CD19 − NK1.1 + . Data are mean ± SEM. *** p < 0.001. d Mononuclear cell infiltration in the quadriceps muscles of Syt7 −/− Il15 f/f and Syt7 −/− ACTA-Il15 −/− mice 14 days after C protein immunization ( Left ). Mononuclear cell infiltration was found in the endomysium ( black square ), perimysium ( black arrowhead ), and perivascular region ( white arrowhead ). Focal lymphatic invasion of myofibers were observed in C-protein-immunized Syt7 −/− ACTA-Il15 −/− mice as indicated by the white arrowhead ( Middle ). The histopathology scores were compiled in the right panel with each symbol representing one mouse ( Right ). Scale bar in left = 100 μm; middle = 25 μm. ** p < 0.01. e Expression of mononuclear cell markers, Cd4 , Cd8 , and F4/80 , mRNA in the quadriceps muscles of C-protein-immunized mice measured by qPCR. Each symbol representing one mouse. * p < 0.05, ** p < 0.01. f Expression of immune-relevant genes mRNA in the quadriceps muscles of C-protein-immunized mice measured by qPCR. Each symbol representing one mouse. * p < 0.05, ** p < 0.01

Article Snippet: C2C12 cells were stained with biotin-conjugated antibodies against IL-15 (Cat. No. 500-P173Bt, PeproTech), IL-15Rα (Cat. No. BAF551, R&D), IL-15Rβ (CD122, clone: TM-b1, eBioscience), and γ c (CD132, Cat. No. 554470, BD Biosciences) then incubated with APC-conjugated streptavidin (BD Biosciences).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Mutagenesis, Histopathology

Journal: PLoS Pathogens

Article Title: Human Herpesvirus 8 (HHV8) Sequentially Shapes the NK Cell Repertoire during the Course of Asymptomatic Infection and Kaposi Sarcoma

doi: 10.1371/journal.ppat.1002486

Figure Lengend Snippet: (A) Summary graphs of statistical dot plots with medians (horizontal black bars) showing expression (mean fluorescence intensity, MFI) of NKp30, NKp46, CD161, DNAM-1 and NKG2D receptors on gated CD3−CD56+ NK cells in the different study groups. Cells were analyzed on a FACSCalibur flow cytometer. P values<0.05 after adjustment for multiple comparisons (Kruskall-Wallis test with Dunn's post test) are indicated. * P<0.01, ** P<0.005, *** P<0.0001. (B) Box and whisker plots showing the median and 25–75 th percentiles of NKG2D expression in healthy controls (n = 35), patients with active KS (n = 10, all classical HIV- KS) and patients with resolved KS (n = 35, including 21 classical HIV- KS and 14 HIV+ KS). Horizontal bars indicate the minimal and maximal values. ** P<0.01, *** P<0.001. (C) Representative flow cytometry analysis in healthy controls (upper panel), asymptomatic HHV8 carriers (middle panel) and patients with active classical KS (lower panel). Control isotypes are shown as dotted lines.

Article Snippet: Cells were incubated for 20 min at 4°C with combinations of the following antibodies: FITC-conjugated anti-CD3; PE-conjugated anti-CD56, anti-NKp30, anti-CD158e1e2 (KIR3DL1/S1), anti-CD158i (KIR2DS4); APC-conjugated anti-CD56, anti-CD158ah (KIR2DL1/S1), anti-CD158bbj (KIR2DL2/L3/S2), PE-Cy7-conjugated anti-CD56, Pacific Blue-conjugated anti-CD3 (all from Beckman Coulter); PE-conjugated anti-CD3; PercP-conjugated anti-CD3, APC-conjugated anti-NKp46, anti-CD161; FITC-conjugated anti-CD94, anti-DNAM-1 (BD Pharmingen); PE-conjugated anti-NKG2D (eBioscience); FITC-conjugated anti-CD122 (IL-15Rβ) APC-conjugated anti-NKG2A, anti-NKG2C, anti-CD132 (IL-15RγR&D Systems).

Techniques: Expressing, Fluorescence, Flow Cytometry, Whisker Assay

Journal: Experimental & Molecular Medicine

Article Title: Discovery of intrahepatic CD103 + cDC1/CD8 + T RM protective immune axis against acetaminophen-induced acute liver injury

doi: 10.1038/s12276-025-01565-3

Figure Lengend Snippet: a The absolute number of liver immune cells in WT and KO mice 24 h after APAP. n = 3 per group. b The ALT level of WT and KO mice administrated with anti-CCL2 antibody before APAP. n = 3 per group per time point. c The relative CCR2 expression levels of liver immune cells analyzed by flow cytometry. n = 4 per group. d The frequency of Ly6C hi monocytes in the liver of WT and KO APAP mice pretreated with anti-CCL2 neutralizing antibodies. n = 3 per group. e The absolute cell number of Ly6C hi monocytes and neutrophils in normal and ABXs-treated mice without APAP. n = 3 per group. f The absolute cell number of Ly6C hi monocytes in normal and ABXs-treated mice after APAP administration. n = 3 per group per time point. g The concentration of CCL2 in the liver of ABXs-pretreated APAP mice. n = 4 per group per time point. h The concentration of CCL2 in the liver of ABXs-pretreated WT mice 6 h after intraperitoneal injection of PGN. n = 3 per group. i The ALT level of ABXs-mice adoptively transferred with hepatic Ly6C hi monocytes before APAP. n = 3 per group per time point. Ordinary one-way ANOVA with Dunnett in a and Tukey in d for the posttest, two-way ANOVA with Dunnett in b and Bonferroni in f , g and i for the posttest and unpaired two-tailed t -test in e and h were used to measure significance. The error bars indicate mean ± s.e.m. The black, blue and red asterisks represent the differences by time point compared with WT, Batf3-KO and Rag1-KO mice, respectively. ns, not significant. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: For IL-12, IL-15, IL-18 and CCL2 neutralizing, mice were i.p. injected with 200 μg per mouse of anti-IL-12 antibody (clone R1-5D9), anti-CD122 (IL-15Rβ) antibody (clone TM-Beta1), anti-IL-18 antibody (clone YIGIF74-1G7) and anti-CCL2 antibody (clone 2H5) (all from BioXCell).

Techniques: Expressing, Flow Cytometry, Concentration Assay, Injection, Two Tailed Test

Journal: Experimental & Molecular Medicine

Article Title: Discovery of intrahepatic CD103 + cDC1/CD8 + T RM protective immune axis against acetaminophen-induced acute liver injury

doi: 10.1038/s12276-025-01565-3

Figure Lengend Snippet: a , b The correlation between the ALT levels and the number of Ly6C hi monocytes ( a ) and the frequency of TNFα + or IL-6 + Ly6C hi monocytes ( b ) in the liver of APAP mice. c The CCL2 concentrations in the liver and CCR2 expression on the hepatic Ly6C hi monocytes of WT and KO APAP mice. n = 3 per group. d The correlation between the number of Ly6C hi monocytes and the frequency of Ki67 + cells in the liver of WT and KO APAP mice. e The apoptotic cell death of Ly6C hi monocytes in the liver of WT and KO APAP mice were assessed after annexin V and FVD staining. n = 3 per group. f The correlation between the number of Ly6C hi monocytes and the frequency of annexin V + cells in the liver of WT and KO APAP mice. g The frequency of annexin V + Ly6C hi monocytes in the liver and the spleen of APAP mice. n = 3 per group. h The cytotoxicity of hepatic CD8 + T cells and NK cells of APAP mice on hepatic Ly6C hi monocytes as a target cells. n = 3 per group. i The apoptosis of Ly6C hi monocytes ( n = 6 per group) and hepatocytes ( n = 3 per group) in the coculture with hepatic CD8 + T cells of APAP mice. Unpaired one-way ANOVA with Dunnett’s multiple comparisons test in c and e , unpaired two-tailed Student’s t -test in g and unpaired two-way ANOVA with Bonferroni for the posttest in h and i were used to measure significance. The error bars indicate mean ± s.e.m. ns, not significant. * P < 0.05, *** P < 0.001.

Article Snippet: For IL-12, IL-15, IL-18 and CCL2 neutralizing, mice were i.p. injected with 200 μg per mouse of anti-IL-12 antibody (clone R1-5D9), anti-CD122 (IL-15Rβ) antibody (clone TM-Beta1), anti-IL-18 antibody (clone YIGIF74-1G7) and anti-CCL2 antibody (clone 2H5) (all from BioXCell).

Techniques: Expressing, Staining, Two Tailed Test