Journal: The EMBO Journal

Article Title: Poxvirus dsDNA genomes differentially activate AIM2 or NLRP3 inflammasomes in human primary cells

doi: 10.1038/s44318-025-00690-z

Figure Lengend Snippet: ( A–D ) PMA-differentiated THP-1 macrophages were either pretreated for 3 h with LPS (200 ng/mL), or overnight with 500 U/mL IFN-γ or IFN-α and infected with VACV WR WT at a multiplicity of infection (MOI) of 5 for 6 h. IL-1β in the supernatant was measured by homogeneous time-resolved fluorescence (HTRF) ( A ). Cell death was measured by LDH release, normalized to cells lysed in 1% Triton X-100 ( B ). Membrane permeabilization was quantified by the uptake of DNA dye DRAQ7 over 6 h in an Incucyte Live-Cell Imaging system. Representative images after 6 h ( C ) and graphs of normalized DRAQ7 uptake over 6 h are displayed ( D ). ( E ) Scheme of inflammasome reporter caspase-1 CARD -EGFP (C1C-EGFP), and detection of C1C-EGFP specks with flow cytometry, by plotting height (EGFP-H) against width (EGFP-W) of the EGFP signal. Data were from a representative sample of PMA-differentiated THP-1 C1C-EGFP cells pretreated with IFN-γ and infected with VACV WR WT. ( F ) PMA-differentiated THP-1 macrophages expressing doxycycline (dox)-inducible C1C-EGFP were treated with LPS, IFN-γ, or IFN-α as mentioned before and infected with VACV at an MOI 5 for 6 h in the presence of 40 μM VX-765 (VX). Infected cells were harvested, fixed in 4% PFA, and speck assembly was quantified by flow cytometry. ( G ) Genome structure of recombinant VACV strains expressing C1C-EGFP from the J2R early promoter (pE). The transgene was inserted into the VACV TK locus by homologous recombination. ( H – L ) PMA-differentiated THP-1 macrophages were pretreated as mentioned before and infected with the indicated recombinant VACV at an MOI of 5 in the presence ( H ) or absence ( I – L ) of VX for 6 h. Infection (EGFP + cells, left) and C1C-EGFP speck assembly in infected or mock-treated cells was quantified by flow cytometry ( H ). IL-1β in the supernatant was measured by HTRF ( I ), and cell death by LDH release ( J ) as described before. Representative images of cells infected in the presence of DRAQ7 for 6 h ( K ), and graphs of normalized infection (EGFP + , left) and DRAQ7 uptake (right) over 6 h ( L ) are displayed. Scale bar = 100 µm. Data represents average values (with individual data points) from N = 3 independent experiments ± SEM. .

Article Snippet: The following small compound inhibitors and reagents were used: CRID3 (MCC950) (Tocris), cycloheximide (Sigma-Aldrich), Cytarabine (AraC) (Abcam), doxycycline (Biomol), G140 (Invivogen), H-151 (Biozol), human IFN-α (PBL Assay Science), human IFN-γ (Immunotools), LPS-EK Ultrapure (Invivogen), PMA (phorbol 12-myristate 13-acetate) (Sigma-Aldrich), and VX-765/belnacasan (Selleckchem).

Techniques: Infection, Fluorescence, Membrane, Live Cell Imaging, Flow Cytometry, Expressing, Recombinant, Homologous Recombination

Journal: The EMBO Journal

Article Title: Poxvirus dsDNA genomes differentially activate AIM2 or NLRP3 inflammasomes in human primary cells

doi: 10.1038/s44318-025-00690-z

Figure Lengend Snippet: ( A ) PMA-differentiated THP-1 cells were either left untreated or pretreated overnight with 500 U/mL IFN-γ and infected with the indicated viruses at an MOI of 5. Six hours post infection, IL-1β in the supernatant was measured by homogeneous time-resolved fluorescence (HTRF), and cell death was measured by LDH release, normalized to cells lysed in 1% Triton X-100. Data represent values with individual data points from N = 3 independent experiments ± SEM.

Article Snippet: The following small compound inhibitors and reagents were used: CRID3 (MCC950) (Tocris), cycloheximide (Sigma-Aldrich), Cytarabine (AraC) (Abcam), doxycycline (Biomol), G140 (Invivogen), H-151 (Biozol), human IFN-α (PBL Assay Science), human IFN-γ (Immunotools), LPS-EK Ultrapure (Invivogen), PMA (phorbol 12-myristate 13-acetate) (Sigma-Aldrich), and VX-765/belnacasan (Selleckchem).

Techniques: Infection, Fluorescence

Journal: The EMBO Journal

Article Title: Poxvirus dsDNA genomes differentially activate AIM2 or NLRP3 inflammasomes in human primary cells

doi: 10.1038/s44318-025-00690-z

Figure Lengend Snippet: ( A – C ) Human monocyte-derived macrophages (hMDMs) differentiated in GM-CSF, ( D – F ) normal human epidermal keratinocytes (NHEK), ( G – I ) CD14 + monocytes, and ( J ) PMA-differentiated THP-1 macrophages were left untreated or pretreated with IFN-γ overnight. Cells were infected with indicated recombinant VACV strains at an MOI of 5 ( A – F , J ) or MOI 10 ( G – I ), in the presence ( A , C , D , F , G , I , J ) or absence ( B , E , H ) of VX, and where indicated, in the presence of 2.5 µM CRID3. Cells were harvested and fixed 6 h post infection. Infection (EGFP + cells) and C1C-EGFP speck assembly in infected or mock-treated cells was quantified by flow cytometry ( A , D , G , J ); infection data (EGFP + cells) corresponding to the experiment shown in panel ( J ) is displayed in Fig. . Membrane permeabilization was quantified by the uptake of DNA dye DRAQ7 over 6 h in an Incucyte live-cell imaging system ( B , E , H ). Scale bar = 100 μm. Samples for confocal microscopy were fixed and stained with 5 μg/mL wheat germ agglutinin (WGA)-Alexa Fluor (AF) 647 and 4 μM Hoechst 33342 ( C , F , I ). Scale bar = 5 μm. ( K ) PMA-differentiated WT or indicated knockout THP-1 cells were left untreated or pretreated with IFN-γ overnight and infected with the indicated reporter VACV strains at an MOI of 5 in the presence of VX. C1C-EGFP speck assembly was quantified as before. The corresponding infection data (EGFP + cells) is shown in Fig. . ( L , M ) NHEK ( L ) or PMA-differentiated THP-1 cells ( M ) constitutively expressing C1C-EGFP were left untreated or pretreated with IFN-γ overnight and infected with VACV WT or monkeypox virus (MPXV) at an MOI of 5, and where indicated, in the presence of CRID3. Cells were harvested and fixed 6 h post infection. VACV- or MPXV-infected cells were stained with rabbit polyclonal anti-H5 serum and AF647-labeled goat anti-rabbit IgG antibodies, and measured by flow cytometry. Corresponding infection data (αH5-AF647 + cells) is shown in Fig . C1C-EGFP speck assembly was analyzed in infected or mock-treated cells. Data represent average values (with individual data points) from N = 3 independent donors (primary cells) ± SEM, or from N = 3 independent experiments ± SEM (THP-1 cells). .

Article Snippet: The following small compound inhibitors and reagents were used: CRID3 (MCC950) (Tocris), cycloheximide (Sigma-Aldrich), Cytarabine (AraC) (Abcam), doxycycline (Biomol), G140 (Invivogen), H-151 (Biozol), human IFN-α (PBL Assay Science), human IFN-γ (Immunotools), LPS-EK Ultrapure (Invivogen), PMA (phorbol 12-myristate 13-acetate) (Sigma-Aldrich), and VX-765/belnacasan (Selleckchem).

Techniques: Derivative Assay, Infection, Recombinant, Flow Cytometry, Membrane, Live Cell Imaging, Confocal Microscopy, Staining, Knock-Out, Expressing, Virus, Labeling

Journal: The EMBO Journal

Article Title: Poxvirus dsDNA genomes differentially activate AIM2 or NLRP3 inflammasomes in human primary cells

doi: 10.1038/s44318-025-00690-z

Figure Lengend Snippet: ( A – I ) Human monocyte-derived macrophages (hMDMs) differentiated in GM-CSF ( A , D , E ), normal human epidermal keratinocytes (NHEK) ( B , F , G ), or human primary CD14 + monocytes ( C , H , I ), were left untreated or pretreated with IFN-γ overnight. Cells were infected with VACV C1C-EGFP at an MOI of 5 ( A , B ) or MOI 10 ( C ), in the absence of VX, and where indicated in the presence of 2.5 µM CRID3. Membrane permeabilization was quantified by the uptake of DNA dye DRAQ7 over 6 h in an Incucyte Live-Cell Imaging system ( A – C ). DRAQ7 uptake normalized to cell confluency are shown. IL-1β in the supernatant was measured by homogeneous time resolved fluorescence (HTRF) ( D , F , H ). Cell death was measured by LDH release and normalized to cells lysed in 1% Triton X-100 ( E , G , I ). ( J , K ) PMA-differentiated WT ( J ) or indicated knockout THP-1 cells ( K ) were left untreated or pretreated with IFN-γ overnight and infected with indicated VACV strains in the presence of VX, and where indicated, CRID3. Infection levels are displayed here; specking data from the same experiments are shown in Fig. . ( L , M ) NHEK ( L ) and PMA-differentiated THP-1 cells ( M ) constitutively expressing C1C-EGFP were left untreated or pretreated with IFN-γ overnight and infected with VACV or monkeypox virus (MPXV) at an MOI of 5 in the presence of VX, and where indicated, CRID3. Cells were fixed 6 h post infection and stained with rabbit polyclonal anti-H5 serum and AF647-labeled goat anti-rabbit IgG antibodies. Infected cells (EGFP + ) were analyzed in infected or mock-treated cells by flow cytometry as described before. Infection levels are displayed here; specking data from the same experiments are displayed in Fig. . Data represent average values (with individual data points) from N = 3 independent donors (primary cells) ± SEM, or from N = 3 independent experiments ± SEM (THP-1 cells).

Article Snippet: The following small compound inhibitors and reagents were used: CRID3 (MCC950) (Tocris), cycloheximide (Sigma-Aldrich), Cytarabine (AraC) (Abcam), doxycycline (Biomol), G140 (Invivogen), H-151 (Biozol), human IFN-α (PBL Assay Science), human IFN-γ (Immunotools), LPS-EK Ultrapure (Invivogen), PMA (phorbol 12-myristate 13-acetate) (Sigma-Aldrich), and VX-765/belnacasan (Selleckchem).

Techniques: Derivative Assay, Infection, Membrane, Live Cell Imaging, Fluorescence, Knock-Out, Expressing, Virus, Staining, Labeling, Flow Cytometry

Journal: The EMBO Journal

Article Title: Poxvirus dsDNA genomes differentially activate AIM2 or NLRP3 inflammasomes in human primary cells

doi: 10.1038/s44318-025-00690-z

Figure Lengend Snippet: ( A ) Genome structure of recombinant VACV strains expressing C1C-EGFP from the J2R early promoter (pE) and bivalent nanobodies from a synthetic early/late promoter (pE/L). Transgenes were inserted into the VACV TK locus by homologous recombination. ( B ) Scheme of AIM2 inflammasome components indicating the VACV-encoded nanobodies used to perturb inflammasome activation. ( C – E ) Human MDMs differentiated with GM-CSF ( C ), NHEKs ( D ), and CD14 + monocytes ( E ) were left untreated or treated with IFN-γ overnight. Cells were infected with VACV C1C-EGFP expressing the indicated nanobodies at an MOI of 5 ( C , D ) or 10 ( E ), in the presence of VX and, where indicated, CRID3. Six hours post infection, cells were harvested, fixed, and infection and C1C-EGFP speck assembly was analyzed by flow cytometry. Average values (with individual data points) from N = 3 independent donors ± SEM are displayed. .

Article Snippet: The following small compound inhibitors and reagents were used: CRID3 (MCC950) (Tocris), cycloheximide (Sigma-Aldrich), Cytarabine (AraC) (Abcam), doxycycline (Biomol), G140 (Invivogen), H-151 (Biozol), human IFN-α (PBL Assay Science), human IFN-γ (Immunotools), LPS-EK Ultrapure (Invivogen), PMA (phorbol 12-myristate 13-acetate) (Sigma-Aldrich), and VX-765/belnacasan (Selleckchem).

Techniques: Recombinant, Expressing, Homologous Recombination, Activation Assay, Infection, Flow Cytometry

Journal: The EMBO Journal

Article Title: Poxvirus dsDNA genomes differentially activate AIM2 or NLRP3 inflammasomes in human primary cells

doi: 10.1038/s44318-025-00690-z

Figure Lengend Snippet: ( A – F ) PMA-differentiated THP-1 cells were either left untreated or treated with IFN-γ overnight and infected with VACV (MOI 5) expressing C1C-EGFP and the indicated nanobodies for 6 h in the presence ( A ) or absence ( B – F ) of VX (see Fig. for overview of virus strains). Cells were harvested, fixed, and infected, as well as ASC speck assembly were analyzed by flow cytometry as described before ( A ). IL-1β in the supernatant was measured by HTRF ( B ) and cell death was measured by LDH release, normalized to cells lysed in 1% Triton X-100 ( C ). ( D – F ) DRAQ7 uptake was monitored over 6 h in an Incucyte live-cell imaging system. Representative images after 6 h ( D ), and graphs showing infection (EGFP + cells) and DRAQ7 uptake normalized to confluency in untreated ( E ), and IFN-γ-treated ( F ) cells over 6 h are displayed. Scale bar = 100 µm. Data represent average values (with individual data points) from N = 3 independent experiments ± SEM ( A – C ). Graphs represent average values from N = 3 independent experiments ± SEM ( E , F ). ( G ) CD14 + monocytes were treated with IFN-γ overnight and proteins were immunoprecipitated with Sepharose beads covalently coupled to VHH AIM2-2 or VHH Enhancer , an EGFP-specific control nanobody. Enriched proteins are displayed in a volcano plot, demonstrating specific enrichment of AIM2 from IFN-γ-treated CD14 + monocytes with VHH AIM2-2 , but not VHH Enhancer .

Article Snippet: The following small compound inhibitors and reagents were used: CRID3 (MCC950) (Tocris), cycloheximide (Sigma-Aldrich), Cytarabine (AraC) (Abcam), doxycycline (Biomol), G140 (Invivogen), H-151 (Biozol), human IFN-α (PBL Assay Science), human IFN-γ (Immunotools), LPS-EK Ultrapure (Invivogen), PMA (phorbol 12-myristate 13-acetate) (Sigma-Aldrich), and VX-765/belnacasan (Selleckchem).

Techniques: Infection, Expressing, Virus, Flow Cytometry, Live Cell Imaging, Immunoprecipitation, Control

Journal: The EMBO Journal

Article Title: Poxvirus dsDNA genomes differentially activate AIM2 or NLRP3 inflammasomes in human primary cells

doi: 10.1038/s44318-025-00690-z

Figure Lengend Snippet: ( A – D ) The indicated cell types were treated with IFN-γ as before. ( A , B ) mRNA transcript levels of AIM2 were quantified by qPCR. The fold change of AIM2 expression upon IFN-γ treatment was normalized to HPRT transcript levels. ( C ) PMA-differentiated THP-1 cells or NHEKs were lysed and AIM2 protein levels were analyzed by immunoblot. ( D ) CD14 + monocytes and GM-CSF hMDMs were lysed and AIM2 was immunoprecipitated using VHH AIM2-2 covalently coupled to sepharose beads. AIM2 protein levels were analyzed by immunoblot. ( E ) Two monoclonal THP-1 ΔAIM2 cell lines were lentivirally transduced to dox-inducibly express AIM2. Cells were PMA-differentiated, treated with dox and IFN-γ as indicated, and infected with VACV C1C-EGFP at an MOI of 5 in the presence of VX, followed by analysis of infection and C1C-EGFP speck formation as before. ( F ) HEK293 cells expressing ASC-EGFP constitutively and either NLRP3 or AIM2 upon dox induction were either left untreated or treated with dox for 4 h. Cells were infected with VACV or MPXV (MOI 5) and harvested and fixed after 6 h. Infected cells were stained with anti-H5 serum as in Fig. . Infection and ASC-EGFP speck assembly was analyzed by flow cytometry. Data represent average values (with individual data points) from N = 3 independent donors (primary cells) ± SEM, or from N = 3 independent experiments ± SEM (Cell lines). .

Article Snippet: The following small compound inhibitors and reagents were used: CRID3 (MCC950) (Tocris), cycloheximide (Sigma-Aldrich), Cytarabine (AraC) (Abcam), doxycycline (Biomol), G140 (Invivogen), H-151 (Biozol), human IFN-α (PBL Assay Science), human IFN-γ (Immunotools), LPS-EK Ultrapure (Invivogen), PMA (phorbol 12-myristate 13-acetate) (Sigma-Aldrich), and VX-765/belnacasan (Selleckchem).

Techniques: Expressing, Western Blot, Immunoprecipitation, Infection, Staining, Flow Cytometry

Journal: The EMBO Journal

Article Title: Poxvirus dsDNA genomes differentially activate AIM2 or NLRP3 inflammasomes in human primary cells

doi: 10.1038/s44318-025-00690-z

Figure Lengend Snippet: ( A ) A549 cells were infected with an MOI 5 of recombinant VACV expressing EGFP under the control of the J2R early (VACV E EGFP), the G8R intermediate (VACV I EGFP) or the F17R late (VACV L EGFP) promoter, where indicated, in the presence of 50 µM CHX or 50 µM AraC. EGFP expression was quantified by flow cytometry. ( B ) PMA-differentiated THP-1 macrophages constitutively expressing C1C-TagBFP were pretreated with IFN-γ overnight and infected with VACV E EGFP in the presence of VX and the indicated inhibitors for 6 h. Infection (EGFP + ) and inflammasome assembly (C1C-TagBFP specks) in TagBFP + cells, or in infected (EGFP + ) cells, were quantified by flow cytometry as before. ( C , D ) PMA-differentiated THP-1 cells were infected with VACV expressing core protein A4 fused to EGFP (EGFP-A4), and C1C-mCherry at an MOI of 2 for 6 h, where indicated, in the presence of AraC. Where indicated, PMA-differentiated THP-1 ΔAIM2 were infected with VACV EGFP-A4 E C1C-mCherry to express C1C-mCherry and treated with 1 µg/mL PA and 0.1 µg/mL LFn-MxiH to activate NLRC4 as a control. Cells were fixed and stained for cellular and viral DNA with Hoechst 33342. Cells were recorded by confocal microscopy, and representative images of at least three independent experiments are displayed ( C ). Purple arrowheads indicate MVs or cores with genomes (positive for Hoechst and EGFP), while yellow arrowheads highlight released DNA genomes (positive for Hoechst, no EGFP). Scale bar = 5 µm. Hoechst-positive VACV genomes and C1C-mCherry specks were detected, and the fraction of C1C-mCherry specks in close proximity (<1 µm) to released viral genomes was quantified. A total of at least 100 specks per condition were analyzed in N = 3 independent repeats. ( E – G ) Primary CD14 + monocytes were infected with VACV C1C-EGFP at an MOI of 10 in the presence ( E , F ) or absence ( G ) of VX, as well as the indicated inhibitors. Six hours post infection, cells were harvested, fixed, and infection (EGFP + ) and C1C-EGFP speck assembly were analyzed by flow cytometry ( E , F ). Membrane permeabilization was quantified by the uptake of DNA dye DRAQ7 over 6 h ( G ). Data represent values with individual data points from N = 3 independent donors (primary cells), or from N = 3 independent experiments (cell lines) ± SEM. .

Article Snippet: The following small compound inhibitors and reagents were used: CRID3 (MCC950) (Tocris), cycloheximide (Sigma-Aldrich), Cytarabine (AraC) (Abcam), doxycycline (Biomol), G140 (Invivogen), H-151 (Biozol), human IFN-α (PBL Assay Science), human IFN-γ (Immunotools), LPS-EK Ultrapure (Invivogen), PMA (phorbol 12-myristate 13-acetate) (Sigma-Aldrich), and VX-765/belnacasan (Selleckchem).

Techniques: Infection, Recombinant, Expressing, Control, Flow Cytometry, Staining, Confocal Microscopy, Membrane

Journal: eBioMedicine

Article Title: Spatial transcriptomics uncovers lipid metabolic dysregulation driving immune-stroma crosstalk in Sjögren's Disease

doi: 10.1016/j.ebiom.2025.106074

Figure Lengend Snippet: MGLL -mediated glycerolipid downregulation was associated with weakened saliva secretion and enhanced antigen presentation in SjD-SGECs. (A) The cellular components of salivary gland. (B) The comparison of glycerolipid metabolic activity in SGECs between controls and patients with SjD revealed by scMetabolism analysis (HRA003613). (C) The glycerolipid metabolic score was positively correlated with saliva secretion score in SjD derived SGECs (SjD n = 11). (D) Ranks of glycerolipid metabolic genes by p value. (E) The expression of MGLL was downregulated in seromucous acini, mucous acini, and ductal cells in SjD. (F) Spatially mapped expression of MGLL in salivary glands (SjD-1). (G) Immunohistochemistry showing the MAGL signal was obviously lower in SjD salivary glands compared to controls (Con, n = 5; SjD, n = 7). Scale bar = 200 μm. (H) Bar plot showing SjD had higher proportion of MGLL low SGECs than controls. (I) The volcano plot for differentially expressed genes of MGLL high compared to MGLL low SGECs. |Log 2 (fold change)|>0.25, p value < 0.001. (J) Pathway enrichment of highly expressed genes in MGLL high SGECs and MGLL low SGECs. Colour scale represents adjust p value of each pathway, and circle size represents counts of genes. (K) Heatmap of saliva secretion associated genes and antigen presentation genes ordered by MGLL ( GSE173808 ). Colour scale represents Z-score normalised by row. (L) Correlation between log 2 (normalised intensity) of MGLL and focus score as well as proportion of CD45 + cells in labial salivary gland ( GSE173808 ) (control n = 39, SjD n = 75). (M) qPCR of several MHC-I/II genes relative to β-actin in salivary gland epithelial cells A253 treated with different doses of MAGL inhibitor ABX-1431 for 24 h (n = 3 samples per group). (N) Correlation between log 2 (normalised intensity) of IFNG and MGLL gene ( GSE173808 ). (O) Illustration of A253 treatment with IFN-γ and qPCR of MGLL relative to β-actin in A253 cells treated with different doses of IFN-γ for 6 h (n = 3 samples per group). (P) Representative Western blot image and quantification of MAGL relative to β-actin in A253 cells treated with different doses of IFN-γ for 24 h (n = 3 samples per group). Data are expressed as boxplot with minimum, maximum, and median values (B), and mean ± SD (G, M, O, and P), respectively. Statistical analyses were performed using Wilcoxon test (B), Pearson correlation (C, L, and N), Student's t test (G), and one-way ANOVA followed by Tukey's post-hoc test (M, O, and P), respectively. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. SGECs: salivary gland epithelial cells; SjD: Sjögren's Disease.

Article Snippet: Cells were then stimulated with 50–100 ng/ml recombinant human IFN-γ (HY–P7025, MCE, CHN) or 0–25 μM MAGL selective inhibitor ABX-1431 (HY-117632, MCE, CHN) for indicated time before cells were lysed.

Techniques: Immunopeptidomics, Comparison, Activity Assay, Derivative Assay, Expressing, Immunohistochemistry, Control, Western Blot

Journal: eBioMedicine

Article Title: Spatial transcriptomics uncovers lipid metabolic dysregulation driving immune-stroma crosstalk in Sjögren's Disease

doi: 10.1016/j.ebiom.2025.106074

Figure Lengend Snippet: Diagram illustrating alterations of the inflammatory niches in SjD SGs. Lymphoid foci organised around ducts within SGs in patients with SjD, comprising of T cells, B cells, plasma cells, macrophages, dendritic cells, and fibroblasts, etc. Chemokine gradients established by fibroblasts ( CXCL12/14 ), pericytes ( CCL2/19/21 ), and SGECs ( CCL28, CXCL1/2/17 ) are critical for immune recruitment, while ACKR1 high endothelial cells and CCL2 high fibroblasts act as important spatial organisers. In T/B cells, upregulated metabolism of phospholipids and their key metabolic genes including PIK3CD/CG , PIKFYVE , and PLCG2 were associated with enhanced T/B-cell activation and production of cytotoxic effectors (granzymes and perforin) as well as proinflammatory cytokines (including IFNs). IFN-γ induced suppression of monoacylglycerol lipase disrupts glycerolipid metabolism in SGECs, leading to enhanced antigen presentation and impaired saliva secretion. SG: salivary gland; SGECs: salivary gland epithelial cells; IFN: interferon.

Article Snippet: Cells were then stimulated with 50–100 ng/ml recombinant human IFN-γ (HY–P7025, MCE, CHN) or 0–25 μM MAGL selective inhibitor ABX-1431 (HY-117632, MCE, CHN) for indicated time before cells were lysed.

Techniques: Clinical Proteomics, Activation Assay, Immunopeptidomics