Journal: PLOS Neglected Tropical Diseases

Article Title: Immune checkpoint TIM-3 defines hyperactivated NK cells and predicts fatal outcome in severe fever with thrombocytopenia syndrome

doi: 10.1371/journal.pntd.0013928

Figure Lengend Snippet: (A) Representative flow cytometry plots showing TIM-3 expression on NK cells from healthy donors and SFTS patients. (B) Frequency of NK cells (CD3 ⁻ CD56⁺) among peripheral blood mononuclear cells (PBMCs) in healthy donors (N = 14), recovered SFTS patients (N = 17), and deceased SFTS patients (N = 4). (C) Frequency of TIM-3 ⁺ NK cells among total NK cells in the same cohorts. (D) Paired longitudinal analysis of peripheral TIM-3 + NK cell frequencies among NK cells in SFTS patients (N = 4) during the acute phase and approximately two weeks into recovery. Paired longitudinal analysis of soluble TIM-3 (E) , Galectin-9 (F) , TNF-α (G) , IFN-γ (H) in plasma of SFTS patients (N = 9) during the acute and recovery phase. Each dot represents an individual donor. Box plots show the full range (min to max). Statistical analysis was performed using Kruskal–Wallis H test for (B); one-way ANOVA with Tukey’s multiple comparisons test for (C) and paired t-test for (D-H). IFN-γ, interferon gamma; NK, natural killer; sGalectin-9, soluble Galectin-9; SFTS, severe fever with thrombocytopenia syndrome; TNF-α, tumor necrosis factor alpha.

Article Snippet: This study aimed to characterize the expression profile of TIM-3 expression on peripheral NK cells in SFTS, explore the role of TIM-3 + NK cells in disease progression, and evaluate soluble TIM-3 (sTIM-3) and Galectin-9 (sGalectin-9) as prognostic biomarkers.

Techniques: Flow Cytometry, Expressing, Clinical Proteomics

Journal: PLOS Neglected Tropical Diseases

Article Title: Immune checkpoint TIM-3 defines hyperactivated NK cells and predicts fatal outcome in severe fever with thrombocytopenia syndrome

doi: 10.1371/journal.pntd.0013928

Figure Lengend Snippet: (A) Proportions and expression levels of immune checkpoint molecules in circulating NK cells. (B) Volcano plot showing differentially expressed genes between TIM-3 + and TIM-3 - NK cells. (C) Enriched pathways identified by Metascape comparing TIM-3 + and TIM-3 - NK cells. (D) Expression levels of HAVCR2 (encoding TIM-3) and its ligand genes ( CEACAM1 , HMGB1 , LGALS9 ) across NK cell subsets. (E) Feature plots displaying the expression of HAVCR2 and LGALS9 in NK cells. (F) Cell-cell communication analysis by CellChat showing enhanced interactions between NK cells and other immune populations in fatal SFTS compared to recovered cases. (G) Chord diagram illustrating LGALS9 -associated signaling pathways enriched in NK cells from fatal cases, including the LGALS9 – HAVCR2 axis. Box plots show the median and interquartile range. Statistical analysis was performed using the Wilcoxon rank-sum test. DC, dendritic cell; NK, natural killer cell; SFTS, severe fever with thrombocytopenia syndrome.

Article Snippet: This study aimed to characterize the expression profile of TIM-3 expression on peripheral NK cells in SFTS, explore the role of TIM-3 + NK cells in disease progression, and evaluate soluble TIM-3 (sTIM-3) and Galectin-9 (sGalectin-9) as prognostic biomarkers.

Techniques: Expressing, Protein-Protein interactions

Journal: PLOS Neglected Tropical Diseases

Article Title: Immune checkpoint TIM-3 defines hyperactivated NK cells and predicts fatal outcome in severe fever with thrombocytopenia syndrome

doi: 10.1371/journal.pntd.0013928

Figure Lengend Snippet: (A) Representative flow cytometry plots showing granzyme B expression in TIM-3+ and TIM-3- NK cells isolated from peripheral blood of SFTS patients at disease onset and two weeks into recovery. (B-D) Quantification of granzyme B (B), perforin (C) and CD107a (D) expression in TIM-3+ and TIM-3- NK cells at disease onset (N = 3, 3, 6, respectively); median values are indicated. For CD107a detection, PBMCs of SFTS patients at disease onset were stimulated with PMA and ionomycin for 6 hours. (E) Paired longitudinal analysis of granzyme B expression in TIM-3+ and TIM-3- NK cells from matched samples at disease onset and recovery phase (N = 3). Statistical analysis was performed using paired t-tests. NK, natural killer; PMA, phorbol 12-myristate 13-acetate; SFTS, severe fever with thrombocytopenia syndrome.

Article Snippet: This study aimed to characterize the expression profile of TIM-3 expression on peripheral NK cells in SFTS, explore the role of TIM-3 + NK cells in disease progression, and evaluate soluble TIM-3 (sTIM-3) and Galectin-9 (sGalectin-9) as prognostic biomarkers.

Techniques: Flow Cytometry, Expressing, Isolation

Journal: PLOS Neglected Tropical Diseases

Article Title: Immune checkpoint TIM-3 defines hyperactivated NK cells and predicts fatal outcome in severe fever with thrombocytopenia syndrome

doi: 10.1371/journal.pntd.0013928

Figure Lengend Snippet: (A-C) Quantification of granzyme B (A) , perforin (B) and CD107a (C) expression in NK cells with and without αTIM-3 antibody treatment after PMA/Ionomycin stimulation (N = 6); median values are indicated. (D) Flow cytometric analysis of IFN-γ expression in NK cells following αTIM-3 antibody blockade after PMA/Ionomycin stimulation. (E) Quantification of IFN-γ expression in NK cells with or without αTIM-3 antibody treatment (N = 6); box plots show minimum to maximum values. (F) Quantification of IFN-γ concentration in NK cells supernatant with and without αTIM-3 antibody treatment after stimulation (N = 6). (G) Flow cytometric analysis of TNF-α expression in NK cells following TIM-3 blockade after PMA/Ionomycin stimulation. (H) Quantification of TNF-α expression in NK cells with or without αTIM-3 antibody treatment (N = 6); box plots show minimum to maximum values. (I) Cytokine concentration in NK cells supernatant with and without αTIM-3 antibody treatment after stimulation using a CBA kit (N = 6); data are presented as mean ± SD. Statistical analysis was performed using paired t-test. αTIM-3 Ab, anti-TIM-3 antibody; CBA, cytometric bead array; IFN-γ, interferon gamma; MFI, mean fluorescence intensity; SD, standard deviation; TNF-α, tumor necrosis factor alpha.

Article Snippet: This study aimed to characterize the expression profile of TIM-3 expression on peripheral NK cells in SFTS, explore the role of TIM-3 + NK cells in disease progression, and evaluate soluble TIM-3 (sTIM-3) and Galectin-9 (sGalectin-9) as prognostic biomarkers.

Techniques: Expressing, Concentration Assay, Fluorescence, Standard Deviation

Journal: PLOS Neglected Tropical Diseases

Article Title: Immune checkpoint TIM-3 defines hyperactivated NK cells and predicts fatal outcome in severe fever with thrombocytopenia syndrome

doi: 10.1371/journal.pntd.0013928

Figure Lengend Snippet: (A) Serum levels of soluble TIM-3 (sTIM-3) and Galectin-9 in healthy individuals (N = 10), recovered SFTS patients (N = 17), and deceased SFTS patients (N = 4). (B) Pearson correlation analysis between sTIM-3 levels, Galectin-9 levels, and the frequency of TIM-3 + NK cells among NK cells with clinical characteristics. (C) ROC curves of serum Galectin-9, sTIM-3, and the frequency of TIM-3 ⁺ NK cells in predicting fatal outcome in the same cohort. (D) ROC curves of serum Galectin-9 and sTIM-3 in the validation cohort (N = 104), with AUC, sensitivity, and specificity calculated using the Youden index. (E) Kaplan–Meier survival analysis stratified by high vs. low serum Galectin-9 levels in the validation cohort (N = 104), using the ROC-derived optimal cutoff. Statistical significance was assessed using the Mann-Whitney test for (A), and Pearson correlation analysis with the Benjamini-Hochberg correction for (B), DeLong’s test for (C-D) and the log-rank test for (E). sTIM-3, soluble T cell immunoglobulin and mucin domain-3; Gal-9, galectin-9; CCI, Charlson comorbidity index; PLT, platelet count; AST, aspartate aminotransferase; Cr, creatinine; ROC, receiver operating characteristic curve; AUC, area under the ROC curve.

Article Snippet: This study aimed to characterize the expression profile of TIM-3 expression on peripheral NK cells in SFTS, explore the role of TIM-3 + NK cells in disease progression, and evaluate soluble TIM-3 (sTIM-3) and Galectin-9 (sGalectin-9) as prognostic biomarkers.

Techniques: Biomarker Discovery, Derivative Assay, MANN-WHITNEY