human il-11 antibody  (Bio-Techne corporation)

Journal: Nature

Article Title: Glioblastoma-instructed astrocytes suppress tumour-specific T cell immunity

doi: 10.1038/s41586-025-08997-x

Figure Lengend Snippet: a , Pearson correlation of TNFSF10 in the TCGA glioblastoma dataset. b , IL11 expression in a glioblastoma scRNA-seq dataset 4 . OPC, oligodendrocyte progenitor cell; NPC, neural progenitor cell; AC, astrocyte; MES, mesenchymal. c , EGFRvIII immunohistochemistry and IL-11 + glioblastoma cell immunofluorescence. d , Peritumoural IL11RA + TRAIL + astrocyte immunofluorescence in glioblastoma. e , Glioblastoma survival stratified by the first and fourth quartile IL11 expression (data from TCGA). f , IL-11 signalling score (WP2332) in GL261 Aldh1l1 cre-ERT2/TdTomato reporter cell scRNA-seq ( Fig. 2b ). g , IL-11 immunofluorescence in GL261 at 15 days. h , IL-11 concentration in GL261-implanted brain lysates at 15 days (n = 3 each). i , Astrocyte STAT3 phosphorylation after treatment with recombinant mouse IL-11 (rmIL-11) ( n = 3 control, n = 5 TCM). j – l , TNFSF10 expression by quantitative PCR (qPCR) ( j , k ) and flow cytometry ( l ) of primary mouse astrocytes treated with rmIL-11 ( j : n = 23 control, n = 14 rmIL-11, l : n = 3 control, n = 5 rmIL-11) and human astrocytes treated with human IL-11 (rhIL-11) ( k ; n = 5 control, n = 11 rhIL-11). m – o , TNFSF10 expression by qPCR ( m , n ) and flow cytometry ( o ) in IL-11-treated mouse ( m , n = 4 each) and human ( n , n = 5 each) astrocytes with STAT3 inhibition. p , q , TNFSF10 expression in GL261 TCM-treated mouse ( p ) and U87 TCM-treated human ( q ) astrocytes and IL-11 neutralization ( n = 5 each). r , GL261 survival after astrocyte Il11ra1 inactivation ( n = 18 NT sgRNA, n = 28 sgIl11ra1). Weight fold change in survivors, sgIl11ra1 1.064 ± 0.028 g, NT sgRNA 1.035 ± 0 g (sgIl11ra1 versus NT sgRNA, P = 0.7015 two-tailed t -test). s , t , TRAIL + ( s ) and pSTAT3 + TRAIL + ( t ) TdTomato + reporter cells in GL261 at 15 days ( n = 3 NT sgRNA, n = 5 sgIl11ra1). u , Apoptotic CD4 + and CD8 + tumour-infiltrating T cells ( n = 5 each). v , Survival in IL11 -overexpressing GL261 mice ( n = 10 each). w–y , TRAIL + ( w ) and pSTAT3 + TRAIL + TdTomato + reporter cells ( x ), and apoptotic T cells ( y ) in IL11 -overexpressing GL261 mice at 12 days ( n = 5 each). Data are the mean ± s.e.m. Unpaired two-tailed t -test. log-rank (Mantel–Cox) test ( e , r , v ). n indicates biologically independent samples. Scale bars, 25 μm ( d , right) 50 μm ( c , right; g , bottom; d , left bottom), 100 μm ( c , left; d , left top), 500 μm ( g , top).

Article Snippet: We used the following monoclonal antibodies: TRAIL blocking antibody (Thermo Fisher Scientific, 16–5951-85) and its respective rat anti-IgG2aκ isotype control (Thermo Fisher Scientific, 16–4321-82) were used at 5 μg ml −1 ; human/mouse cross-reactive anti-IL11 blocking antibody (R&D Systems, MAB218) and its respective mouse anti-IgG2aκ Isotype control (Thermo Fisher Scientific, 14472485) were used at 10 μg ml −1 .

Techniques: Derivative Assay, Expressing, Immunohistochemistry, Immunofluorescence, Concentration Assay, Phospho-proteomics, Recombinant, Control, Real-time Polymerase Chain Reaction, Flow Cytometry, Inhibition, Neutralization, Two Tailed Test

Journal: Nature

Article Title: Glioblastoma-instructed astrocytes suppress tumour-specific T cell immunity

doi: 10.1038/s41586-025-08997-x

Figure Lengend Snippet: ( a,b ) IL11 expression per cell type (a) and GBM subtype (b) in TCGA after cell-type deconvolution. ( c ) IL11 (left) and IL11RA expression by cell type in scRNA-seq 22 . ( d ) Spatially annotated bulk RNA-seq from Ivy Glioblastoma Atlas showing expression correlation of STAT3 signalling program (M5897) and TRAIL-driven apoptosis signalling (M23448) split by anatomical structure. ( e ) Spatial transcriptomics analysis of human GBM performed with SPATA2 86 showing spatially-weighted correlation of IL11 expression with GBM subtype metaprograms 32 . ( f ) IL11RA expression by time to recurrence in GBM-associated astrocytes analysed by scRNA-seq in Fig. 1b . ( g ) IL11RA expression in GBM-associated astrocytes determined by scRNA-seq. ( h ) Il11ra1 expression in GL261-associated astrocytes determined by scRNA-seq. ( i-k ) IL-11 quantified by ELISA in conditioned media from primary human GBM lines (i, n = 4 each), primary mouse astrocytes or GL261 cultures (j, n = 3 astrocytes, n = 6 GL261), or genetically-engineered glioma models (k, n = 4 each). ( l ) pSTAT3 (Y705) in primary murine astrocytes stimulated with rmIL-11 for 15 mins. ( m ) TNFSF10 promoter luciferase reporter activity in HepG2 cells treated with rhIL-11 for 24 or 48 h. ( n ) Tnfsf10 expression measured by qPCR in purified cultures of either microglia, oligodendrocytes, or neurons overnight with recombinant IL-11. ( o ) IL15 and TGFB1 expression in GBM cells analysed by scRNA-seq 4 . ( p ) Tnfsf10 expression measured by qPCR in primary mouse astrocyte cultures stimulated with the indicated cytokines. Data shown as mean ± SEM. Unpaired two-tailed t-test and two-way ANOVA used. n indicates biologically independent samples.

Article Snippet: We used the following monoclonal antibodies: TRAIL blocking antibody (Thermo Fisher Scientific, 16–5951-85) and its respective rat anti-IgG2aκ isotype control (Thermo Fisher Scientific, 16–4321-82) were used at 5 μg ml −1 ; human/mouse cross-reactive anti-IL11 blocking antibody (R&D Systems, MAB218) and its respective mouse anti-IgG2aκ Isotype control (Thermo Fisher Scientific, 14472485) were used at 10 μg ml −1 .

Techniques: Expressing, RNA Sequencing, Enzyme-linked Immunosorbent Assay, Luciferase, Activity Assay, Purification, Recombinant, Two Tailed Test

Journal: Nature

Article Title: Glioblastoma-instructed astrocytes suppress tumour-specific T cell immunity

doi: 10.1038/s41586-025-08997-x

Figure Lengend Snippet: ( a ) Cleaved caspase-3/7 + and cleaved caspase-8 + CD4/8 + T cells in co-cultures with GL261 TCM pre-treated astrocytes with IL-11 neutralization (n = 6 CD4 + groups; n = 3 CD8 + anti-Isotype; n = 4 CD8 + anti-IL-11). ( b ) Cleaved caspase-3/7 + and cleaved caspase-8 + CD4/8 + T cells co-cultured with rmIL-11-treated astrocytes (n = 11 CD4 + control; n = 6 CD4 + rmIL-11; n = 7 CD8 + control; n = 4 CD8 + rmIL-11). ( c ) Percentage of cleaved caspase-3/7 + (left) and cleaved caspase-8 + (right) CD4 + or CD8 + T cells co-cultured with rmIL-11 pre-treated astrocytes and TRAIL-blocking antibodies (n = 6 CD4 + ; n = 3 CD8 + anti-Isotype; n = 4 CD8 + anti-TRAIL). ( d ) Percentage of cleaved caspase-3/7 + CD4 + T cells co-cultured with astrocytes pre-treated with rmIL-11 in combination with STAT3 inhibitor or vehicle. TRAIL-blocking or control antibodies, or recombinant TRAIL were added at start of co-culture. (n = 6 each). ( e ) Validation of astrocyte-specific Il11ra1 knockdown by immunofluorescence. ( f ) Absolute cell count of the indicated compartments in GL261-bearing mice following astrocyte-specific Il11ra1 or non-targeting control inactivation. ( g,h ) IL-11 concentration in TCM (g) (n = 4 empty vector; n = 2 IL-11 OE) and growth dynamics (h) (n = 6 empty vector; n = 2 IL-11 OE) of empty vector transfected or IL-11 overexpressing (OE) GL261 cells (n = 3 each). ( i ) Relative increase in bioluminescence tumour size between days 6 and 11 in control or IL-11-overexpressing tumours. ( j ) Analysis of the Il11 promoter showing multiple AHR consensus binding sites. ( k,l ) Expression of Il11 (left) and Cyp1b1 (right) in GL261 cells harboring control- or AHR-deletion (k) or a constitutively activated form of AHR (l). ( m,n ) Mouse GL261 (m) or human U87 (n) GBM cells stimulated with L-Kynurenine (Kyn) alone or in combination with AHR inhibitor CH223191 (AHRi). Data shown as mean ± SEM. Unpaired two-tailed t-test and two-way ANOVA used. n indicates biologically independent samples.

Article Snippet: We used the following monoclonal antibodies: TRAIL blocking antibody (Thermo Fisher Scientific, 16–5951-85) and its respective rat anti-IgG2aκ isotype control (Thermo Fisher Scientific, 16–4321-82) were used at 5 μg ml −1 ; human/mouse cross-reactive anti-IL11 blocking antibody (R&D Systems, MAB218) and its respective mouse anti-IgG2aκ Isotype control (Thermo Fisher Scientific, 14472485) were used at 10 μg ml −1 .

Techniques: Expressing, Neutralization, Cell Culture, Control, Blocking Assay, Recombinant, Co-Culture Assay, Biomarker Discovery, Knockdown, Immunofluorescence, Cell Counting, Concentration Assay, Plasmid Preparation, Transfection, Binding Assay, Two Tailed Test