mouse il 38  (R&D Systems)

Journal: mAbs

Article Title: IL-38 blockade induces anti-tumor immunity by abrogating tumor-mediated suppression of early immune activation

doi: 10.1080/19420862.2023.2212673

Figure Lengend Snippet: IL-38 is targeted by B cells in cancer patients and is highly expressed in multiple tumor types. (a) Identification of IL-38 as the specific target of IMM20130 assessed by protein microarray. Binding signal and S-Scores shown for the top 200 proteins based on binding signal. (b) Assessment of IL-38 transcript expression in tumor and normal tissue using TCGA RNAseq data. Samples were normalized for IL-38 expression and subdivided based on high (brown) and low (green) expression using thresholds set according to a value of 10–15% of normal tissues. Numbers in each plot represent frequency of IL-38 positive samples in all tumor samples using this threshold. (c) Immunohistochemical analysis of IL-38 expression in three cancer types. Head and neck squamous carcinoma (HNSC), lung tumors comprising multiple subtypes including adenocarcinoma and squamous cell carcinoma, and cervical squamous carcinoma (CESC) were stained with IMM20324 or isotype control antibodies to detect IL-38 expression. Pathological scoring was performed according to staining density. Representative examples of specimens with a range of expression intensity within each indication are shown. (d) Average intensity of IL-38 expression across cancer indications tested. Intensity scoring analysis was performed using a 7-point scale, with 0 corresponding to no expression and 6 being strong staining. Scoring was performed in both cytoplasmic and nuclear compartments and averaged to calculate final scores.

Article Snippet: After washing, the MSD plate was incubated with biotinylated mouse anti-human IL-38 (clone H127C, Thermo Fisher Scientific, #14-7385-82) at final concentration of 100 ng/mL and Sulfo-Tag-Streptavidin (MesoScale Discovery, #L15×B) at final concentration of 100 ng/mL for 1 h at RT.

Techniques: Microarray, Binding Assay, Expressing, Immunohistochemical staining, Staining

Journal: mAbs

Article Title: IL-38 blockade induces anti-tumor immunity by abrogating tumor-mediated suppression of early immune activation

doi: 10.1080/19420862.2023.2212673

Figure Lengend Snippet: IL-38 expression is associated with reduced immune infiltration and HPV status in head and neck squamous carcinoma. (a) Signature analysis of immune lineages in IL-38 high versus IL-38 low head and neck tumors. Negative values represent reduced expression of lineage-associated genes and lower infiltration. Positive values represent increased infiltration of indicated lineages. HNSC: head and neck squamous cell carcinoma; LUSC: lung squamous cell carcinoma; ESCA: esophageal carcinoma: LUAD: Lung adenocarcinoma; CESC: cervical squamous cell carcinoma and endocervical adenocarcinoma: SKCM: skin cutaneous melanoma. (b) IL-38 expression in HPV-negative and HPV-positive populations of head and neck cancers.

Article Snippet: After washing, the MSD plate was incubated with biotinylated mouse anti-human IL-38 (clone H127C, Thermo Fisher Scientific, #14-7385-82) at final concentration of 100 ng/mL and Sulfo-Tag-Streptavidin (MesoScale Discovery, #L15×B) at final concentration of 100 ng/mL for 1 h at RT.

Techniques: Expressing

Journal: mAbs

Article Title: IL-38 blockade induces anti-tumor immunity by abrogating tumor-mediated suppression of early immune activation

doi: 10.1080/19420862.2023.2212673

Figure Lengend Snippet: IMM20324 is specific for IL-38, binds human and mouse protein with different affinities and neutralizes binding to putative human IL-38 receptors. (a) Identification of IL-38 as the specific target of IMM20324H assessed by protein microarray. Binding signal and S-scores shown for the top 200 proteins based on binding signal. (b) (Top) Binding of recombinantly produced IMM20324 mIgG2a antibody to recombinant human and mouse IL-38 using BLI. Two-fold titrations (1.52–0.095 µM) were performed to generate K D values for binding to species-specific proteins. (Bottom) Values shown are average of two experiments. (c) ELISA-based binding analysis of IMM20324 to recombinant full-length human and mouse IL-38 using direct ELISA. (d) Flow cytometry-based binding to human and mouse IL-38 overexpressing CHO cells. Cells were fixed and permeabilized to assess binding of intracellular IL-38. Mock-transfected controls, in addition to isotype staining controls for human (gray) and mouse (black) cells also shown on graph. (e) Inhibition of IL-38 receptor binding to its receptors. Inhibition of binding to IL-36R is shown in the circles, IL1RAPL1 is shown in squares. Isotype control-treated samples are in blue, while samples treated with IMM20324 are shown in red.

Article Snippet: After washing, the MSD plate was incubated with biotinylated mouse anti-human IL-38 (clone H127C, Thermo Fisher Scientific, #14-7385-82) at final concentration of 100 ng/mL and Sulfo-Tag-Streptavidin (MesoScale Discovery, #L15×B) at final concentration of 100 ng/mL for 1 h at RT.

Techniques: Binding Assay, Microarray, Produced, Recombinant, Enzyme-linked Immunosorbent Assay, Direct ELISA, Flow Cytometry, Transfection, Staining, Inhibition

Journal: mAbs

Article Title: IL-38 blockade induces anti-tumor immunity by abrogating tumor-mediated suppression of early immune activation

doi: 10.1080/19420862.2023.2212673

Figure Lengend Snippet: Targeting of IL-38 inhibits tumor growth and elicits immunological memory against EMT6 breast tumors in vivo . (a–e) Mice were randomized at D1 based on tumor volume and treated with three different doses of IMM20324H or a PBS vehicle control twice per week for 6 total doses. (a) Mean tumor volume up to D17 (b) Tumor volume on D17, the final day all mice were on study. (c) Individual growth curves for each mouse in the indicated group up to D45. (d–e) EMT6 cells were implanted into the opposite flank of IMM20324H-treated mice following full regression of the primary tumor. No additional antibody was administrated during secondary challenge. (d) Secondary EMT6 tumor growth curves compared to untreated age-matched mice. (e) Survival curves of rechallenged mice showing % survival per group. * p <0.05.

Article Snippet: After washing, the MSD plate was incubated with biotinylated mouse anti-human IL-38 (clone H127C, Thermo Fisher Scientific, #14-7385-82) at final concentration of 100 ng/mL and Sulfo-Tag-Streptavidin (MesoScale Discovery, #L15×B) at final concentration of 100 ng/mL for 1 h at RT.

Techniques: In Vivo

Journal: Virology Journal

Article Title: Neutralization of interleukin-38 exacerbates coxsackievirus B3-induced acute myocarditis in mice

doi: 10.1186/s12985-021-01687-w

Figure Lengend Snippet: Elevated expression of IL-38 and IL-36R in AVMC. A Representative images of IL-38 immunohistochemistry of heart tissue from the control (n = 10) and AVMC (n = 13) groups (Brown granules, magnification × 400). B Morphometric quantitation of cardiac IL-38 protein expression. C ELISA analysis of serum IL-38 levels. D Relative cardiac mRNA expression of IL-38 and IL-36R detected by RT-qPCR. E–F Negative association of cardiac IL-38 mRNA expression with bodyweight loss and cardiac pathological scores on day 7 post-infection. Each point represents an individual mouse. G–H Negative association of serum IL-38 levels with bodyweight loss and cardiac pathological scores on day 7 post-infection. Each point represents an individual mouse. ** P < 0.01, compared to control group. Data are expressed as mean ± SD

Article Snippet: Mouse Anti-IL-38 Abs and IgG 2A isotype control were obtained from R&D Systems, Minneapolis, MN, USA.

Techniques: Expressing, Immunohistochemistry, Quantitation Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Infection

Journal: Virology Journal

Article Title: Neutralization of interleukin-38 exacerbates coxsackievirus B3-induced acute myocarditis in mice

doi: 10.1186/s12985-021-01687-w

Figure Lengend Snippet: Neutralization of IL-38 reduced survival rate and impaired cardiac function. Male BALB/c mice were infected with CVB3 on day 0 and then treated with i.p. injection of PBS, Anti-IL-38 Abs, or IgG 2A isotype control on day 0 and 4 after CVB3 infection. A Survival analysis in the Sham (n = 10), PBS (n = 15), IL-38N (n = 15), and IgG (n = 15) groups after CVB3 infection. B The bodyweight changes were monitored daily until day 7. C Representative M-mode echocardiography images of the left ventricle in different groups on day 7 post-infection. D–G LVESD, LVEDD, FS, and EF were measured on day 7 post-infection. * P < 0.05, ** P < 0.01, compared to Sham group; # P < 0.05, ## P < 0.01, compared to IgG group. Data are expressed as mean ± SD

Article Snippet: Mouse Anti-IL-38 Abs and IgG 2A isotype control were obtained from R&D Systems, Minneapolis, MN, USA.

Techniques: Neutralization, Infection, Injection

Journal: Virology Journal

Article Title: Neutralization of interleukin-38 exacerbates coxsackievirus B3-induced acute myocarditis in mice

doi: 10.1186/s12985-021-01687-w

Figure Lengend Snippet: Neutralization of IL-38 exacerbated CVB3-induced myocarditis. A Values of HW/BW in the Sham (n = 10), PBS (n = 11), IL-38N (n = 5), and IgG (n = 11) groups. B Cardiac pathological scores in different groups. Each point represents an individual mouse. C Representative images of myocardial histopathology H&E staining in different groups (magnification × 200 and × 400). * P < 0.05, ** P < 0.01, compared to Sham group; # P < 0.05, ## P < 0.01, compared to IgG group

Article Snippet: Mouse Anti-IL-38 Abs and IgG 2A isotype control were obtained from R&D Systems, Minneapolis, MN, USA.

Techniques: Neutralization, Histopathology, Staining

Journal: Virology Journal

Article Title: Neutralization of interleukin-38 exacerbates coxsackievirus B3-induced acute myocarditis in mice

doi: 10.1186/s12985-021-01687-w

Figure Lengend Snippet: Neutralization of IL-38 regulated Th1 and Th17 cells differentiation in vivo. A Representative flow cytometry dot plots of splenic CD4 + IFN-γ + Th1 cells and CD4 + IL-17 + Th17 cells in the Sham, PBS, IL-38N, and IgG groups. Numbers in upper left quadrants and lower right quadrants indicate the separate percentages of CD4 + Th1 cells and CD4 + Th17 cells. B Abundance of Th1 cells in different groups. C Cardiac mRNA expression levels of T-bet in different groups. D Abundance of Th17 cells in different groups. E Cardiac mRNA expression levels of RORγt in different groups. ** P < 0.01, compared to Sham group; # P < 0.05, ## P < 0.01, compared to IgG group. Data are expressed as mean ± SD

Article Snippet: Mouse Anti-IL-38 Abs and IgG 2A isotype control were obtained from R&D Systems, Minneapolis, MN, USA.

Techniques: Neutralization, In Vivo, Flow Cytometry, Expressing

Journal: Virology Journal

Article Title: Neutralization of interleukin-38 exacerbates coxsackievirus B3-induced acute myocarditis in mice

doi: 10.1186/s12985-021-01687-w

Figure Lengend Snippet: Neutralization of IL-38 reduced IFN-γ production, increased IL-17, TNF-α, and IL-6 levels, and promoted CVB3 replication. Cardiac mRNA expression levels of IFN-γ, IL-17, TNF-α, and IL-6 were detected by RT-qPCR, and serum protein levels of these cytokines were measured by ELISA in the Sham, PBS, IL-38N, and IgG groups. A–D IFN-γ, IL-17, TNF-α, and IL-6, respectively. E Levels of cardiac CVB3 RNA in different groups. F Levels of cardiac CVB3 titers in different groups. ** P < 0.01, compared to Sham group; # P < 0.05, ## P < 0.01, compared to IgG group. Data are expressed as mean ± SD

Article Snippet: Mouse Anti-IL-38 Abs and IgG 2A isotype control were obtained from R&D Systems, Minneapolis, MN, USA.

Techniques: Neutralization, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay