ip0065 critical  (Bio X Cell)

Journal: JACC: Basic to Translational Science

Article Title: Interleukin-18 Inhibition Aggravates Atherosclerosis in Jak2 V617F Clonal Hematopoiesis

doi: 10.1016/j.jacbts.2025.101463

Figure Lengend Snippet: Experimental Design Ldlr −/− mice were transplanted with a mixture of bone marrow (BM) from Mx1-Cre-Jak2 VF (20%) and wild-type mice (80%) to generate a Jak2 VF clonal hematopoiesis (CH) model or with BM from Mx1-Cre (20%) and wild-type mice as controls. At 4 weeks (wks) after bone marrow transplantation (BMT), mice received 2 pIpC (100 μg per dose) injections to induce Cre . Two wks later, mice were fed a Western-type diet (WTD) for a period of 8 and 10 wks or 16 wks. At 4 wks of WTD feeding, mice were injected with IgG or anti–IL-18 for a period of 4, 6, or 12 wks, respectively. The graphical illustrations were created with BioRender.com.

Article Snippet: At 4 weeks after WTD feeding, mice were injected intraperitoneally with IL-18 antibodies (Bio X Cell, BE0237); 500 μg dissolved in 100 μL of in vivo dilution buffer (Bio X Cell, IP0070) or IgG isotype control (Bio X Cell, BE0089); 500 μg dissolved in 100 μL of in vivo dilution buffer (Bio X Cell, IP0070) 3 times per week with a 55- to 60-hour interval between injections.

Techniques: Transplantation Assay, Western Blot, Injection

Journal: JACC: Basic to Translational Science

Article Title: Interleukin-18 Inhibition Aggravates Atherosclerosis in Jak2 V617F Clonal Hematopoiesis

doi: 10.1016/j.jacbts.2025.101463

Figure Lengend Snippet: The Impact of IL-18 Antibody Treatment on Lesion Area and Necrotic Core Formation in Jak2 VF CH Mice (A) Representative hematoxylin and eosin (H&E) images of aortic root lesions. Necrotic cores are marked by dashed lines. Scale bar, 200 μm. (B) Quantification of lesion area and (C) necrotic core area. (D) Number of foam cell rich lesions, collagen rich lesions, and advanced lesions with necrotic cores per group based on H and E staining and expressed per number of total observations. For B to D, n = 12-15 mice for 8-wk, n = 14 for 10-wk, and n = 10-14 for the 16-wk cohort. For B and C, statistical analysis was performed using 2-way analysis of variance with Tukey’s multiple comparison test and for D using the chi-square test. Statistical differences are indicated on the graphs, with P values >0.05 omitted. Statistically significant differences by treatment and genotype factors are presented below the graphs. Abbreviations as in .

Article Snippet: At 4 weeks after WTD feeding, mice were injected intraperitoneally with IL-18 antibodies (Bio X Cell, BE0237); 500 μg dissolved in 100 μL of in vivo dilution buffer (Bio X Cell, IP0070) or IgG isotype control (Bio X Cell, BE0089); 500 μg dissolved in 100 μL of in vivo dilution buffer (Bio X Cell, IP0070) 3 times per week with a 55- to 60-hour interval between injections.

Techniques: Staining, Comparison

Journal: JACC: Basic to Translational Science

Article Title: Interleukin-18 Inhibition Aggravates Atherosclerosis in Jak2 V617F Clonal Hematopoiesis

doi: 10.1016/j.jacbts.2025.101463

Figure Lengend Snippet: IL-18 Antibody Treatment Enhances Fibrous Cap Thickness and Collagen Content in Atherosclerotic Lesions (A) Representative images of Masson’s Trichrome staining showing collagen staining in lesions. Black bars indicate cap thickness. Scale bar, 200 μm. (B) Total collagen staining (blue) in lesions and (C) Quantification of fibrous cap thickness (n = 12-15 mice for 8-wk, n = 14 for 10-wk, and n = 10-14 for 16-wk cohort) at 8, 10, and 16 wks WTD feeding. For B and C statistical analysis was performed using 2-way analysis of variance with Tukey’s multiple comparison test. Statistical differences are indicated on the graphs, with P values >0.05 omitted. Statistically significant differences by treatment and genotype factors are presented below the graphs. Abbreviations as in .

Article Snippet: At 4 weeks after WTD feeding, mice were injected intraperitoneally with IL-18 antibodies (Bio X Cell, BE0237); 500 μg dissolved in 100 μL of in vivo dilution buffer (Bio X Cell, IP0070) or IgG isotype control (Bio X Cell, BE0089); 500 μg dissolved in 100 μL of in vivo dilution buffer (Bio X Cell, IP0070) 3 times per week with a 55- to 60-hour interval between injections.

Techniques: Staining, Comparison

Journal: JACC: Basic to Translational Science

Article Title: Interleukin-18 Inhibition Aggravates Atherosclerosis in Jak2 V617F Clonal Hematopoiesis

doi: 10.1016/j.jacbts.2025.101463

Figure Lengend Snippet: IL-18 Antibody Treatment Decreased Cl-GSDMD levels, IFN-γ, and AIM2 expression in Atherosclerotic Lesions of Jak2 VF CH Mice (A and B) IFN-γ immunostaining. (A) Representative images. Interferon (IFN)-γ is shown in green. (B) Quantification was performed as mean fluorescence intensity (MFI) (n = 12-15 mice for 8-wk, n = 12 for 10-wk, and n = 10 for 16-wk cohort). (C-D) AIM2 inflammasome expression in macrophages in lesions. (C) Representative IF images of aortic root sections stained for AIM2 (magenta) and Mac2 (green). Arrows indicate macrophages positive for AIM2. Quantification was performed as MFI of AIM2 signal within Mac2 + macrophages using ImageJ (D) quantification, Mac2 + areas were thresholded to create a binary mask, which was applied to the AIM2 channel, and the mean gray value of AIM2 signal within Mac2 + regions was measured (n = 12 mice for 10-wk and n = 8-14 for 16-wk cohort). (E and F) Cleaved Gasdermin D (Cl-GSDMD) expression in macrophages in the lesions. (E) Representative images. Cl-GSDMD is shown in magenta. Cl-GSDMD–positive macrophages (white spots) highlighted by yellow arrows in lesions (F) Quantification (n = 15 mice for 10-wk and n = 10 for 16-wk cohort). Scale bar for all images, 100 μm. For B, D, and F, statistical analysis was performed using 2-way analysis of variance with Tukey’s multiple comparison test. Abbreviations as in .

Article Snippet: At 4 weeks after WTD feeding, mice were injected intraperitoneally with IL-18 antibodies (Bio X Cell, BE0237); 500 μg dissolved in 100 μL of in vivo dilution buffer (Bio X Cell, IP0070) or IgG isotype control (Bio X Cell, BE0089); 500 μg dissolved in 100 μL of in vivo dilution buffer (Bio X Cell, IP0070) 3 times per week with a 55- to 60-hour interval between injections.

Techniques: Expressing, Immunostaining, Fluorescence, Staining, Comparison

Journal: JACC: Basic to Translational Science

Article Title: Interleukin-18 Inhibition Aggravates Atherosclerosis in Jak2 V617F Clonal Hematopoiesis

doi: 10.1016/j.jacbts.2025.101463

Figure Lengend Snippet: IL-18 Antibody Treatment Increases Apoptotic Macrophages and Impairs Efferocytosis (A and B) Apoptotic cleaved (Cl)-Caspase-3 + macrophages. (A) Representative images. Cl-caspase-3 is shown in magenta. Cl-caspase-3 positive macrophages (white spots) highlighted by yellow arrows in lesions. (B) Quantification (n = 13-15 mice for 10-wk and n = 7-10 for 16-wk cohort). (C and D) TUNEL assay. (C) TUNEL + staining visualized in red spots; dead macrophages (yellow spots) indicated by yellow arrows. (D) Quantification (n = 15 mice per group for 10-wk and n = 10 for 16-wk cohort). (E-F) GSDME, a pyroptotic marker, in macrophages. (E) Representative images. GSDME is shown in magenta. GSDME-positive macrophages (white spots) highlighted by yellow arrows in lesions. (F) Quantification (n = 15 mice per group for 10-wk and n = 10 for 16-wk cohort). (G) Representative images and in situ efferocytosis assessment (n = 15 mice per group for 10-wk and n = 10 for 16-wk cohort). Scale bar for all images, 100 μm. For B, D, and F, statistical analysis was performed using 2-way analysis of variance with Tukey’s multiple comparison test. Abbreviations as in .

Article Snippet: At 4 weeks after WTD feeding, mice were injected intraperitoneally with IL-18 antibodies (Bio X Cell, BE0237); 500 μg dissolved in 100 μL of in vivo dilution buffer (Bio X Cell, IP0070) or IgG isotype control (Bio X Cell, BE0089); 500 μg dissolved in 100 μL of in vivo dilution buffer (Bio X Cell, IP0070) 3 times per week with a 55- to 60-hour interval between injections.

Techniques: TUNEL Assay, Staining, Marker, In Situ, Comparison

Journal: JACC: Basic to Translational Science

Article Title: Interleukin-18 Inhibition Aggravates Atherosclerosis in Jak2 V617F Clonal Hematopoiesis

doi: 10.1016/j.jacbts.2025.101463

Figure Lengend Snippet: Singe Cell RNA-Seq Analysis of Atherosclerotic Plaques Shows Reduced Expression of Efferocytosis Genes in Resident-Like Macrophages (A) UMAP of integrated scRNA-sequencing of CD45 + plaque cells isolated from aortic arches of control mice, Jak2 VF mice, and Jak2 VF mice with IL-18 antibodies. (B) Percentage of lesional CD45 + cells within each cluster. (C) Violin plots depicting gene expression levels of efferocytosis markers Axl , Mertk , and Cd36 in control, Jak2 VF , and Jak2 VF mice treated with IL-18 antibody, derived from scRNA-seq data. (D) Representative aortic images with DAPI, MAC2, and AXL staining. DAPI is shown in blue, MAC2 is shown in green, and AXL is shown in magenta. AXL-positive macrophages (white spots) are highlighted by yellow arrows in the lesions. (E) Quantification of lesional AXL positive macrophages (n = 15/group) after 10 weeks of WTD feeding. (F) Violin plots depicting gene expression levels of MHC II genes in Jak2 VF and Jak2 VF mice treated with IL-18 antibody, derived from scRNA-seq data. Scale bar, 100 μm. Statistical analyses were performed using the Wilcoxon rank sum test with Bonferroni correction in C and F. For E, statistical analysis was performed using 2-way analysis of variance with Tukey’s multiple comparison test. Statistical differences are indicated on the graphs, with P values >0.05 omitted. Statistically significant differences by treatment and genotype factors are presented below the graphs. Abbreviations as in .

Article Snippet: At 4 weeks after WTD feeding, mice were injected intraperitoneally with IL-18 antibodies (Bio X Cell, BE0237); 500 μg dissolved in 100 μL of in vivo dilution buffer (Bio X Cell, IP0070) or IgG isotype control (Bio X Cell, BE0089); 500 μg dissolved in 100 μL of in vivo dilution buffer (Bio X Cell, IP0070) 3 times per week with a 55- to 60-hour interval between injections.

Techniques: RNA Sequencing, Expressing, Sequencing, Isolation, Control, Gene Expression, Derivative Assay, Staining, Comparison