anti il27 p28  (Bio X Cell)

Journal: bioRxiv

Article Title: Oncolytic HSV-IL27 expression improves CD8 T cell function and therapeutic activity in syngeneic glioma models

doi: 10.1101/2025.05.12.653429

Figure Lengend Snippet: (A) Schematic illustration of C134 and C027. C134 is a second-generation oHSV with deletion of the γ134.5 genes and insertion of the human cytomegalovirus (HCMV) IRS1 gene in the UL3/UL4 intergenic region. C027 was generated from C134 by insertion of bicistronic copies of murine IL-27 (mIL27) in the deleted γ134.5 regions under the MND promoter. (B) mIL27 (p28) secretion from C027 or C134 infected (MOI=1) CT-2A cell supernatants at 24- and 48-hours post-infection measured by ELISA. (C) Viral replication in CT-2A cells infected with C027, C134, or WT HSV (MOI=1) demonstrating viral replication kinetics. Experimental design schematics and representative Kaplan-Meier survival curves in (D) CT-2A (n=11 per cohort), (F) KR158 (n=5- 10 per cohort), and (H) SB28 (n=10 per cohort) immunocompetent syngeneic murine glioma models (Log- Rank test of significance) after treatment with Saline or equivalent PFU (1×10 7 ) of C027 or C134 (parental control). In vivo viral recovery from murine brain tumors (E) CT-2A, (G) KR158, and (I) SB28 at two days post- treatment with C134 (n=3) or C027 (n=3). For B, C, E, G, and I, data are mean ± SEM with each shape representing one replicate or animal. Statistical analyses were performed using two-way analysis of variance with Holm-Šídák’s correction for multiple comparisons (B), unpaired two-tailed Student’s t-test (E, G, and I), or Log-Rank tests of significance (D, F, and H). IC, intracranial; oHSV, oncolytic herpes simplex virus; PFU, plaque forming unit; MOI, multiplicity of infection; IL, interleukin; Tx, treatment.

Article Snippet: Antibodies used for in vivo experiments include: anti-CD8a (2.43, Leinco), anti-IL27 p28 (MM27.7B1, BioXCell), rat IgG2b isotype (LTF-2, BioXCell), mouse IgG2a isotype (N123, Leinco) and were administered by intraperitoneal injection on indicated days.

Techniques: Generated, Infection, Enzyme-linked Immunosorbent Assay, Saline, Control, In Vivo, Two Tailed Test, Virus

Journal: iScience

Article Title: Interferon-γ and IL-27 positively regulate type 1 regulatory T cell development during adaptive tolerance

doi: 10.1016/j.isci.2025.112308

Figure Lengend Snippet:

Article Snippet: Mouse Anti-Mouse IL-27 p28 (clone MM27.7B1) , BioXCell , Cat #BE0326; RRID:AB_2819053.

Techniques: Recombinant, Saline, Staining, Red Blood Cell Lysis, Sequencing, Software

Journal: bioRxiv

Article Title: Interferon-γ and IL-27 positively regulate type 1 regulatory T-cell development during adaptive tolerance

doi: 10.1101/2024.06.21.598825

Figure Lengend Snippet: Tg4 Nr4a3 -Tocky Il10 -eGFP Ifng -YFP mice were administered 4 mg/kg [4Y]-MBP in PBS s.c. and 1 mg αIFNγ or αIgG1 isotype in 200 μL PBS i.p. and spleens were harvested at 24 hr. Representative flow plots showing Il10 -eGFP against Ifng -YFP above and Ifng -YFP + -derived TCRvβ8.1/8.2 against NK1.1 below (A) , and TCRvβ8.1/8.2 against NK1.1 above and NK1.1 + -derived Il10 -eGFP against Ifng -YFP below (B) . Summary of Ifng -YFP + frequency (C) from (A) and Ifng -YFP + frequency from NK1.1 + (D) from (B) . Linear regression (pearson’s correlation) of CD4 + Nr4a3 -Timer + Il10 -eGFP + against NK1.1 + TCRvβ8.1/8.2 − (E) . Tg4 Nr4a3 -Tocky Il10 -eGFP mice were given 200 μg αNK1.1 or αIgG2a isotype in 200 μL PBS i.p and 48 hrs later were immunised with 4 mg/kg [4Y]-MBP in PBS s.c. and. and spleens were harvested 24 hr later. Representative spectral cytometry plots showing NK1.1 against FSC-A (F) . Summary of NK1.1 + (G) from (F) . Representative spectral cytometry plots at 24 hr showing Il10 -eGFP against CD4 (H) . Summary of Il10 -eGFP + frequency (I) and ICOS (J) and OX40 (K) MFI of CD4 + Nr4a3 -Timer + T-cells. Tg4 Nr4a3 -Tocky Il10 -eGFP mice were administered 4 mg/kg [4Y]-MBP in PBS s.c. and 0.5 mg αIFNγ, αIL-27, a combination of both or a mixed IgG1 and αIgG2a isotype in 200 μL PBS i.p. and spleens were harvested 24 hours later. Representative spectral cytometry plots at 24 hr showing Il10 -eGFP against CD4 in Nr4a3 -Timer + (L) . Summary of Il10 -eGFP + frequency within the CD4 + Nr4a3 -Timer + T-cells. (M). Analysis for additive effect of treatment by factoring αIFNγ and αIL-27 as two vairables (N) . (C, D, G, I-K, M, N) bars represent median with interquartile range. Statistical analysis by Mann-Whitney U Test ( C,D,I-K ), Kruskal-Wallis test with Dunn’s multiple comparisons (M). N = 4 (A-E) , N = 8 (F-K) , N = 15 (Combo), N = 16 (Isotype) N = 17 (αIFNγ) and N = 17 (αIL-27) (L-N) per treatment.

Article Snippet: For in vivo blockade experiments, in vivo grade anti-IL-27p28 (clone MM27.7B1, mouse IgG2a, BioXcell), or anti-IFNγ (clone XMG1.2, kind gift from Prof Anne Cooke, University of Cambridge, rat IgG1) were administered through intraperitoneal injection as indicated in figure legends.

Techniques: Derivative Assay, Cytometry, MANN-WHITNEY

Journal: EMBO Molecular Medicine

Article Title: IL ‐27 produced during acute malaria infection regulates Plasmodium ‐specific memory CD4 + T cells

doi: 10.15252/emmm.202317713

Figure Lengend Snippet: A–D B6, Ebi3 −/− , and Il27 −/− mice were transferred with PbT‐II cells a day before Pcc infection. Levels of parasitemia, proportion of PbT‐II cells in CD4 + T cells in peripheral blood (PB), as well as spleen (SP) profiles were monitored. Gating strategy for CD4 + T and PbT‐II cell analysis is described in Fig . (A) Experimental scheme for adoptive transfer of PbT‐II to B6, Ebi3 −/− , and Il27 −/− mice. (B, C) Parasitemia levels and proportions of CD4 + T and PbT‐II cells in PB of B6 (WT) vs. Ebi3 −/− mice (B; n = 4 mice/group) and B6 (WT) vs. Il27 −/− mice (C; n = 4 mice/ group). Representative data of 2 independent experiments are shown. (D) CD4 + T and PbT‐II cell profile in the spleen of B6 (WT) vs. Il27 −/− mice ( n = 6, 8, 9, 9 for WT and n = 6, 7, 10, 10 for Il27 −/− mice at day 7, 14, 21, 28 post infection (pi), respectively). Data are pooled from 2, 2, 3 and 3 independent experiments for day 7, 14, 21, and 28 timepoint, respectively. E, F Adoptive transfer of PbT‐II cells to B6 mice later infected with Pcc. Anti‐IL‐27 mAb was administered for regular, early, and late αIL‐27 groups at −1 to 19, −1 to 7, and 11 to 19 dpi, respectively, and IgG control group received IgG between −1 and 19 dpi. (E) Experimental scheme of IL‐27 neutralization with anti‐IL‐27 mAb. (F) Parasitemia levels and proportions of CD4 + T and PbT‐II cells in PB during weekly monitoring ( n = 3 mice/group). Dose effect results were replicated in Fig . G Adoptive transfer of PbT‐II to Pcc‐infected B6 mice administered with either IgG or anti‐IL‐27 mAb during the early phase (between −1 and 7 days after infection). Total number of cells, proportion, and number of CD4 + T and PbT‐II cells in the spleen were monitored ( n = 9, 6, 11, 15 for IgG‐treated and n = 9, 7, 11, 15 mice for anti‐IL‐27 mAb‐treated mice at day 7, 14, 21, 28 days pi, respectively). Data are pooled from 2, 2, 3 and 4 independent experiments for day 7, 14, 21, and 28 timepoint, respectively. Data information: Statistical significance in (B), (C), (D), and (G) was assessed by unpaired two‐tailed Student's t ‐tests ( P values (< 0.05) shown in black) or Mann–Whitney U test ( P values (< 0.05) shown in brown) for comparing WT to EBI3 −/− /Il27 −/− mice or IgG‐ to anti‐IL‐27 mAb‐treated mice per timepoint, depending on normality assessment. Error bars represent SD in all graphs. Source data are available online for this figure.

Article Snippet: InVivoMAb anti‐mouse IL‐27 p28 (250 ug per mouse) , Bio X Cell , Clone: MM27.7B1; BE0326; RRID: AB_2819053.

Techniques: Infection, Cell Analysis, Adoptive Transfer Assay, Control, Neutralization, Two Tailed Test, MANN-WHITNEY

Journal: EMBO Molecular Medicine

Article Title: IL ‐27 produced during acute malaria infection regulates Plasmodium ‐specific memory CD4 + T cells

doi: 10.15252/emmm.202317713

Figure Lengend Snippet: Gating strategy for flow cytometry analysis of PbT‐II cells. Samples were stained for CD4, TCRβ, CD45.1, and CD45.2 to identify PbT‐II cells. APCCy7 anti‐CD45.1 and Bv605 anti‐CD45.2 mAbs for the congenic markers and Bv510 anti‐TCRβ mAb were maintained in all panels. For CD4 staining, Bv711 anti‐CD4 mAb was used in Figs , , and , and ; Pacific Blue‐anti‐CD4 mAb for Figs , and , and , and , and FITC anti‐CD4 mAb for Figs and and . B6 mice transferred with PbT‐II cells were administered with anti‐IL27 mAb in 4 different conditions or IgG control at timepoints indicated and were infected with Pcc ( n = 4 mice/group). Parasitemia levels, proportion of CD4 + T cells, and proportions of PbT‐II cells in CD4 + T cells in PB were monitored. Representative data of two independent experiments are shown. Data information: Statistical significance was assessed by one‐way ANOVA followed by Tukey's multiple comparison test in (B) and by Student's t test in (C). P values (< 0.05) are shown. Small letters in (B) indicate significant differences compared to IgG control (a = 4‐dose, b = 3‐dose, c = 2‐dose, d = 1‐dose treatment), and P ‐values are: day 7, b = 0.010; day 21, a = < 0.001, b = 0.015, c = 0.010, d = 0.022; day 28, a = 0.035 (middle graph); day 7, a = 0.031, c = 0.022, d = 0.009; day 14, a = 0.023, b = 0.029; day 21, a = < 0.001, b = 0.002, c = < 0.001, d = 0.018; day 28, a = 0.001, c = 0.044, d = 0.036 (right graph). Error bars represent SD.

Article Snippet: InVivoMAb anti‐mouse IL‐27 p28 (250 ug per mouse) , Bio X Cell , Clone: MM27.7B1; BE0326; RRID: AB_2819053.

Techniques: Flow Cytometry, Staining, Control, Infection, Comparison

Journal: EMBO Molecular Medicine

Article Title: IL ‐27 produced during acute malaria infection regulates Plasmodium ‐specific memory CD4 + T cells

doi: 10.15252/emmm.202317713

Figure Lengend Snippet: B6 mice were transferred with PbT‐II cells, infected with Pcc, and were treated with either IgG or anti‐IL‐27 mAb between −1 and 7 days after infection ( n = 1 biological replicate per timepoint). PbT‐II cells were prepared from spleen at day 28 pi, stained for CD4/TCR/CD45.1 and for CD127, KLRG1, and CD49d with TotalSeq antibodies, sort purified, and processed for scRNA‐seq and CITE‐Seq analysis. Details of the experiment are found in Fig . A–G Comparative analysis of scRNA‐seq data from IgG and anti‐IL‐27 mAb‐treated PbT‐II cells. (A) UMAP plot colored of day 28 PbT‐II cells from IgG control ( n = 7,491) and anti‐IL‐27 mAb‐treated mice ( n = 4,944) after unsupervised clustering of pooled single cell data from the two groups, with clusters colored by gene expression profiles. Cluster labels were harmonized to reflect similar gene expression patterns in the clusters at day 7 pi (Fig ) and anti‐IL27 mAb day 7–28 PbT‐II analysis (Fig ). (B) UMAP clustering of PbT‐II cells colored by cell cycle profiles. (C) CITE‐seq analysis of PbT‐II cells for IgG2a (isotype control), CD127, KLRG1, and CD49d, shown in the same UMAP clustering as (A). (D) Proportions (%) of each cluster within PbT‐II cells, with bar graph sizes shown relative to the total number of PbT‐II cells in IgG (36.8 × 10 ) and anti‐IL‐27 mAb treated (265.7 × 10 ) mice. (E) Ridge plots of PbT‐II cells showing the expression of published CD4 + T cell signature genes (Ciucci et al , ). (F) Violin plots comparing the expression of the CD4 + T cell signature genes. (G) Dot plots showing the expression of Th1‐, Tfh‐, Tcmp‐, and proliferation‐associated genes in each cluster. Dot colors represent the intensity of expression, while dot size represents the proportion of cells with the corresponding expression. H Volcano plot of differentially expressed genes between major clusters 1* and 1** within PbT‐II cells from anti‐IL‐27‐treated mice and corresponding Gene Ontology enrichment analysis for the upregulated genes in each group using Metascape. Source data are available online for this figure.

Article Snippet: InVivoMAb anti‐mouse IL‐27 p28 (250 ug per mouse) , Bio X Cell , Clone: MM27.7B1; BE0326; RRID: AB_2819053.

Techniques: Infection, Staining, Purification, Control, Gene Expression, Expressing

Journal: EMBO Molecular Medicine

Article Title: IL ‐27 produced during acute malaria infection regulates Plasmodium ‐specific memory CD4 + T cells

doi: 10.15252/emmm.202317713

Figure Lengend Snippet:

Article Snippet: InVivoMAb anti‐mouse IL‐27 p28 (250 ug per mouse) , Bio X Cell , Clone: MM27.7B1; BE0326; RRID: AB_2819053.

Techniques: Marker, Staining, FACS, Purification, Software, Cell Isolation

Journal: EMBO Molecular Medicine

Article Title: IL ‐27 produced during acute malaria infection regulates Plasmodium ‐specific memory CD4 + T cells

doi: 10.15252/emmm.202317713

Figure Lengend Snippet:

Article Snippet: For IL‐27 neutralization, anti‐IL‐27p28 mAb (250 ug/mouse; Clone: MM27.7B1; Bio X Cell) was injected i.p. 1 day before and 2, 5, 7 days after Pcc infection, unless otherwise specified in the experimental schemes.

Techniques: Marker, Staining, FACS, Purification, Software, Cell Isolation