serum il  (Cusabio)

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 impairs early B-cell development in the bone marrow. (A) Experimental scheme for AAV-mediated IL-27 delivery and analysis of splenic and BM B-cell compartments. (B–D) Splenic B-cell analysis 10 weeks after AAV-Ctrl or AAV-IL-27 treatment (n = 2 per group). (B) Frequency of total splenic B cells. (C) CIBERSORTx-based deconvolution of splenic B-cell subsets, including clustering and pseudotime analysis (left) and relative subset distribution (right). (D) Cd93 expression (TPM) in splenic B cells. (E–G) Analysis of splenic and BM B-cell compartments 3 weeks after treatment (n = 4 per group). (E) Representative flow cytometry plots. (F) Frequencies of B220 + CD19 − and B220 − CD19 + populations in the BM and spleen. (G) Relative distribution of B-cell developmental subsets in the BM. (H–I) Kinetics of B-cell populations following IL-27 treatment (n = 2 per group at each time point). (H) Frequency and absolute number of BM B220 + B cells. (I) Proportions of B-cell subsets within the B220 + compartment at the indicated time points. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Cell Analysis, Expressing, Flow Cytometry

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: Exogenous IL-27 inhibits CLP formation in the bone marrow. (A) qPCR analysis of B-lineage- and myeloid-associated transcription factors in BM cells 5 days after AAV-Ctrl or AAV-IL-27 administration (n = 3 per group). (B) qPCR analysis of CEBPA expression in Reh cells treated with hIL-27 and DAC for 3 days (n = 3 per group). (C–D) Frequencies and absolute numbers of CLPs in WT and CD19 Cre ;IL-27R fl/fl mice (n = 3 per group). (E–F) BM B-cell frequency (E) and proportion of B220 + CD43 hi IgM − B cells (F) in WT and CD19 Cre ;IL-27R fl/fl mice 2 weeks after AAV treatment (n = 3 for WT Ctrl group; n = 3 for WT IL-27 group; n = 3 for CD19 Cre Ctrl group; n = 4 for CD19 Cre IL-27 group). (G–H) BM B-cell frequency (G) and proportion of B220 + CD43 hi IgM − B cells (H) in CD4 Cre ;IL-27R fl/fl and Lyz2 Cre ;IL-27R fl/fl mice 2 weeks after AAV treatment (n = 3 for CD4 Cre Ctrl group; n = 3 for CD4 Cre IL-27 group; n = 5 for Lyz2 Cre Ctrl group; n = 6 for Lyz2 Cre IL-27 group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Expressing

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: Exogenous IL-27 reshapes immune reconstitution after bone marrow transplantation. (A) Experimental scheme. WT mice underwent BM transplantation followed by AAV-Ctrl or AAV-IL-27 treatment. Splenic and BM immune reconstitution was analysed at 10 weeks after treatment (n = 4 mice for the Ctrl group; n = 5 mice for the IL-27 group). (B) Percentages and absolute numbers of CD45 + splenocytes. (C–F) Frequencies of major immune cell populations within CD45 + splenocytes: myeloid cells (C), macrophages (D), T and B cells (E), and NK cells (F). (G) Frequency of CD4 + T cells among splenic T cells. (H) Proportion of Tregs among CD4 + T cells. (I–J) BM B-cell reconstitution. Frequency of BM B220 + B cells (I) and distribution of B220 + B-cell subsets (J). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Transplantation Assay

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 inhibits B-cell development through both direct and indirect mechanisms. (A) Schematic of the BM chimera model. WT (WTR) or IL-27R −/− (KOR) recipients were treated with AAV-Ctrl or AAV-IL-27. Donor BM cells were a 1:1 mix of WT (CD45.1 + ) and IL-27R −/− (CD45.1 − ) cells. Analyses were performed at 5 weeks (B–E; n = 3 per group) or 10 weeks (F–I; n = 5 per group) post-transplantation. (B) Percentage of CD19 + B cells from CD45.1 + or CD45.1 − (IL-27R −/− ) donors in spleen and BM of WTR and KOR mice. (C) Absolute numbers of total CD19 + B cells in spleen and BM of WTR and KOR mice. (D) Absolute numbers of CD19 + B cells from CD45.1 + or CD45.1 − (IL-27R −/− ) donors in spleen and BM of WTR and KOR mice. (E) Percentage of B220 + CD19 − B cells from CD45.1 + or CD45.1 − (IL-27R −/− ) donors in BM of WTR and KOR mice. (F) Absolute numbers of B220 + CD19 - B cells from CD45.1 + or CD45.1 - (IL-27R −/− ) donors in BM of WTR and KOR mice. (G) Frequencies and absolute numbers of BM B220 + B cells derived from CD45.1 + or IL-27R −/− donors in WTR mice. (H) Proportion of CD43 hi IgM − subsets from CD45.1 + or IL-27R −/− donors in BM B220 + B cells in WTR mice. (I–J) Frequencies of CLPs (I) and their donor-specific contributions (J) in BM chimeras. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Transplantation Assay, Derivative Assay

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: Exogenous IL-27 indirectly inhibits BM B-ALL cells. (A) Experimental design. GFP + B-ALL cells were transferred into WT mice, followed by AAV-Ctrl or AAV-IL-27 administration. BM and splenic B-ALL cells were analysed 9–10 days later (n = 3 per group). (B–C) Representative flow cytometry plots and quantification of GFP + B-ALL and GFP − B cells. (D) GSVA analysis of transcriptional changes in BM B-ALL cells isolated from mice in (A). (E) Gene Ontology (GO) enrichment analysis of DEGs between Ctrl and IL-27 groups in (A) (adjusted P < 0.05, |log 2 fold change| > 1). The outer ring indicates enriched GO terms, the middle ring represents relative gene expression changes in the IL-27 group, and the inner ring shows background gene counts, with colour intensity reflecting enrichment significance. Enriched pathways include interferon-γ-mediated signalling, interferon-β responses, adhesion-related processes, protozoan defence responses, calmodulin-dependent kinase signalling, and negative regulation of STAT tyrosine phosphorylation. (F–G) Expression of apoptosis- and cell cycle-related genes in BM B-ALL cells from (A). (H) Experimental design. WT and IL-27R −/− mice transplanted with GFP + B-ALL cells were treated with AAV-Ctrl or AAV-IL-27 (n = 4 for WT Ctrl group; n = 4 for WT IL-27 group; n = 5 for IL-27R −/− Ctrl group; n = 5 for IL-27R −/− IL-27 group). (I–J) Representative flow plots and quantification of BM B-ALL cells. (K–L) Flow cytometry plots and quantification of BM B-ALL cells in CD4 Cre ;IL-27R fl/fl , CD19 Cre ;IL-27R fl/fl , and Lyz2 Cre ;IL-27R fl/fl mice (n = 2 for CD4 Cre Ctrl group; n = 2 for CD4 Cre IL-27 group; n = 3 for CD19 Cre Ctrl group; n = 3 for CD19 Cre IL-27 group; n = 3 for Lyz2 Cre Ctrl group; n = 3 for Lyz2 Cre IL-27 group). (M) Flow cytometry plots and quantification of BM B-ALL cells in Rag1 −/− mice (n = 3 per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Flow Cytometry, Isolation, Gene Expression, Phospho-proteomics, Expressing

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 reshapes the BM niche, reducing B-cell support and leukaemogenesis. (A) Pseudotime analysis of BM B-cell subsets (C0–C10). (B) Expression of representative marker genes across B-cell clusters in (A). (C–G) BM cells were harvested 8 days after AAV-Ctrl or AAV-IL-27 administration for RNA-seq analysis (n = 3 per group). (C) CIBERSORTx analysis showing increased pre-pro-B and cycling pro-B subsets after IL-27 treatment. (D) KEGG pathway enrichment analysis highlighting alterations in adhesion-related pathways. (E–F) Heatmaps of transcription factors (E) and adhesion molecules (F). (G) RNA-seq analysis of genes shown in the figure in BM cells. (H) RNA-seq analysis of genes shown in the figure in human MSCs with or without hIL-27 treatment (n = 2 per group). (I) qPCR analysis of genes shown in the figure in murine MSCs with or without IL-27 (n = 3 per group). (J) RNA-seq analysis of genes shown in the figure in BM Gr-1 + cells 3 weeks after AAV-Ctrl or AAV-IL-27 administration (n = 2 per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Expressing, Marker, RNA Sequencing

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 exhibits therapeutic potential while modulating B-cell compartments. (A–B) Experimental design of combination therapy in WT (A) and Rag1 −/− (B) mice, with corresponding Kaplan–Meier survival analysis. (C–D) Experimental design of IL-27R −/− CAR-T cell therapy combined with AAV-Ctrl or AAV-IL-27 in B-ALL-bearing mice, with Kaplan–Meier survival curves. B-ALL cells were transplanted either prior to chemotherapy (C) or after chemotherapy (D). (E–F) NSG mice transplanted with human G-PBSCs were injected intramuscularly with AAV-Ctrl or hIL-27. 10 days later, frequencies and absolute numbers of B cells in the BM and spleen were analysed (n = 5 per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; log-rank test (A–D) and unpaired Student's t-test (E–F).

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Injection

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 impairs early B-cell development in the bone marrow. (A) Experimental scheme for AAV-mediated IL-27 delivery and analysis of splenic and BM B-cell compartments. (B–D) Splenic B-cell analysis 10 weeks after AAV-Ctrl or AAV-IL-27 treatment (n = 2 per group). (B) Frequency of total splenic B cells. (C) CIBERSORTx-based deconvolution of splenic B-cell subsets, including clustering and pseudotime analysis (left) and relative subset distribution (right). (D) Cd93 expression (TPM) in splenic B cells. (E–G) Analysis of splenic and BM B-cell compartments 3 weeks after treatment (n = 4 per group). (E) Representative flow cytometry plots. (F) Frequencies of B220 + CD19 − and B220 − CD19 + populations in the BM and spleen. (G) Relative distribution of B-cell developmental subsets in the BM. (H–I) Kinetics of B-cell populations following IL-27 treatment (n = 2 per group at each time point). (H) Frequency and absolute number of BM B220 + B cells. (I) Proportions of B-cell subsets within the B220 + compartment at the indicated time points. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Cell Analysis, Expressing, Flow Cytometry

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: Exogenous IL-27 inhibits CLP formation in the bone marrow. (A) qPCR analysis of B-lineage- and myeloid-associated transcription factors in BM cells 5 days after AAV-Ctrl or AAV-IL-27 administration (n = 3 per group). (B) qPCR analysis of CEBPA expression in Reh cells treated with hIL-27 and DAC for 3 days (n = 3 per group). (C–D) Frequencies and absolute numbers of CLPs in WT and CD19 Cre ;IL-27R fl/fl mice (n = 3 per group). (E–F) BM B-cell frequency (E) and proportion of B220 + CD43 hi IgM − B cells (F) in WT and CD19 Cre ;IL-27R fl/fl mice 2 weeks after AAV treatment (n = 3 for WT Ctrl group; n = 3 for WT IL-27 group; n = 3 for CD19 Cre Ctrl group; n = 4 for CD19 Cre IL-27 group). (G–H) BM B-cell frequency (G) and proportion of B220 + CD43 hi IgM − B cells (H) in CD4 Cre ;IL-27R fl/fl and Lyz2 Cre ;IL-27R fl/fl mice 2 weeks after AAV treatment (n = 3 for CD4 Cre Ctrl group; n = 3 for CD4 Cre IL-27 group; n = 5 for Lyz2 Cre Ctrl group; n = 6 for Lyz2 Cre IL-27 group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Expressing

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: Exogenous IL-27 reshapes immune reconstitution after bone marrow transplantation. (A) Experimental scheme. WT mice underwent BM transplantation followed by AAV-Ctrl or AAV-IL-27 treatment. Splenic and BM immune reconstitution was analysed at 10 weeks after treatment (n = 4 mice for the Ctrl group; n = 5 mice for the IL-27 group). (B) Percentages and absolute numbers of CD45 + splenocytes. (C–F) Frequencies of major immune cell populations within CD45 + splenocytes: myeloid cells (C), macrophages (D), T and B cells (E), and NK cells (F). (G) Frequency of CD4 + T cells among splenic T cells. (H) Proportion of Tregs among CD4 + T cells. (I–J) BM B-cell reconstitution. Frequency of BM B220 + B cells (I) and distribution of B220 + B-cell subsets (J). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Transplantation Assay

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 inhibits B-cell development through both direct and indirect mechanisms. (A) Schematic of the BM chimera model. WT (WTR) or IL-27R −/− (KOR) recipients were treated with AAV-Ctrl or AAV-IL-27. Donor BM cells were a 1:1 mix of WT (CD45.1 + ) and IL-27R −/− (CD45.1 − ) cells. Analyses were performed at 5 weeks (B–E; n = 3 per group) or 10 weeks (F–I; n = 5 per group) post-transplantation. (B) Percentage of CD19 + B cells from CD45.1 + or CD45.1 − (IL-27R −/− ) donors in spleen and BM of WTR and KOR mice. (C) Absolute numbers of total CD19 + B cells in spleen and BM of WTR and KOR mice. (D) Absolute numbers of CD19 + B cells from CD45.1 + or CD45.1 − (IL-27R −/− ) donors in spleen and BM of WTR and KOR mice. (E) Percentage of B220 + CD19 − B cells from CD45.1 + or CD45.1 − (IL-27R −/− ) donors in BM of WTR and KOR mice. (F) Absolute numbers of B220 + CD19 - B cells from CD45.1 + or CD45.1 - (IL-27R −/− ) donors in BM of WTR and KOR mice. (G) Frequencies and absolute numbers of BM B220 + B cells derived from CD45.1 + or IL-27R −/− donors in WTR mice. (H) Proportion of CD43 hi IgM − subsets from CD45.1 + or IL-27R −/− donors in BM B220 + B cells in WTR mice. (I–J) Frequencies of CLPs (I) and their donor-specific contributions (J) in BM chimeras. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Transplantation Assay, Derivative Assay

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: Exogenous IL-27 indirectly inhibits BM B-ALL cells. (A) Experimental design. GFP + B-ALL cells were transferred into WT mice, followed by AAV-Ctrl or AAV-IL-27 administration. BM and splenic B-ALL cells were analysed 9–10 days later (n = 3 per group). (B–C) Representative flow cytometry plots and quantification of GFP + B-ALL and GFP − B cells. (D) GSVA analysis of transcriptional changes in BM B-ALL cells isolated from mice in (A). (E) Gene Ontology (GO) enrichment analysis of DEGs between Ctrl and IL-27 groups in (A) (adjusted P < 0.05, |log 2 fold change| > 1). The outer ring indicates enriched GO terms, the middle ring represents relative gene expression changes in the IL-27 group, and the inner ring shows background gene counts, with colour intensity reflecting enrichment significance. Enriched pathways include interferon-γ-mediated signalling, interferon-β responses, adhesion-related processes, protozoan defence responses, calmodulin-dependent kinase signalling, and negative regulation of STAT tyrosine phosphorylation. (F–G) Expression of apoptosis- and cell cycle-related genes in BM B-ALL cells from (A). (H) Experimental design. WT and IL-27R −/− mice transplanted with GFP + B-ALL cells were treated with AAV-Ctrl or AAV-IL-27 (n = 4 for WT Ctrl group; n = 4 for WT IL-27 group; n = 5 for IL-27R −/− Ctrl group; n = 5 for IL-27R −/− IL-27 group). (I–J) Representative flow plots and quantification of BM B-ALL cells. (K–L) Flow cytometry plots and quantification of BM B-ALL cells in CD4 Cre ;IL-27R fl/fl , CD19 Cre ;IL-27R fl/fl , and Lyz2 Cre ;IL-27R fl/fl mice (n = 2 for CD4 Cre Ctrl group; n = 2 for CD4 Cre IL-27 group; n = 3 for CD19 Cre Ctrl group; n = 3 for CD19 Cre IL-27 group; n = 3 for Lyz2 Cre Ctrl group; n = 3 for Lyz2 Cre IL-27 group). (M) Flow cytometry plots and quantification of BM B-ALL cells in Rag1 −/− mice (n = 3 per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Flow Cytometry, Isolation, Gene Expression, Phospho-proteomics, Expressing

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 reshapes the BM niche, reducing B-cell support and leukaemogenesis. (A) Pseudotime analysis of BM B-cell subsets (C0–C10). (B) Expression of representative marker genes across B-cell clusters in (A). (C–G) BM cells were harvested 8 days after AAV-Ctrl or AAV-IL-27 administration for RNA-seq analysis (n = 3 per group). (C) CIBERSORTx analysis showing increased pre-pro-B and cycling pro-B subsets after IL-27 treatment. (D) KEGG pathway enrichment analysis highlighting alterations in adhesion-related pathways. (E–F) Heatmaps of transcription factors (E) and adhesion molecules (F). (G) RNA-seq analysis of genes shown in the figure in BM cells. (H) RNA-seq analysis of genes shown in the figure in human MSCs with or without hIL-27 treatment (n = 2 per group). (I) qPCR analysis of genes shown in the figure in murine MSCs with or without IL-27 (n = 3 per group). (J) RNA-seq analysis of genes shown in the figure in BM Gr-1 + cells 3 weeks after AAV-Ctrl or AAV-IL-27 administration (n = 2 per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Expressing, Marker, RNA Sequencing

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 exhibits therapeutic potential while modulating B-cell compartments. (A–B) Experimental design of combination therapy in WT (A) and Rag1 −/− (B) mice, with corresponding Kaplan–Meier survival analysis. (C–D) Experimental design of IL-27R −/− CAR-T cell therapy combined with AAV-Ctrl or AAV-IL-27 in B-ALL-bearing mice, with Kaplan–Meier survival curves. B-ALL cells were transplanted either prior to chemotherapy (C) or after chemotherapy (D). (E–F) NSG mice transplanted with human G-PBSCs were injected intramuscularly with AAV-Ctrl or hIL-27. 10 days later, frequencies and absolute numbers of B cells in the BM and spleen were analysed (n = 5 per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; log-rank test (A–D) and unpaired Student's t-test (E–F).

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Injection

Journal: eLife

Article Title: IL-27 limits HSPC differentiation during infection and protects from stem cell exhaustion

doi: 10.7554/eLife.105876

Figure Lengend Snippet: ( A ) Schematic of infection and TAM dosing strategy for Procr-Ai6 mice. ( B ) Procr-Ai6 mice were infected and representative flow plots of long-term hematopoietic stem cells (LTHSCs) (Lin (CD3, NK1.1, B220, Ly6G) − , Sca-1 + , CD117 (cKit) + , CD135 − , CD48 − , CD150 + ) from naive (left) and infected (right) mice are shown. Proportions of zsGreen + LTHSCs were then quantified in both groups. ( C ) Proportions of zsGreen + monocytes (CD3 − , B220 − , CD11b + , Ly6C + , Ly6G − ) in the spleen and peritoneal exudate cells (PECs) of naive and infected mice. ( D ) Schematic of infection of WT and Il27 –/– mice for E and F. ( E ) Numbers of LTHSCs and granulocyte progenitors (GPs) (CD3 − , NK1.1 − , B220 − , CD117 + , CD34 + , CD16/32 hi , Ly6C + , CD135 − , CD115 − ) in the bone marrow of WT and Il27 –/– infected mice throughout infection. ( F ) Monocyte progenitors (MPs) (CD3 − , NK1.1 − , B220 − , CD117 + , CD34 + , CD16/32 hi , Ly6C + , CD135 − , CD115 + ; orange box) in the bone marrow of infected WT and Il27 −/− mice throughout infection. Representative flow plots are shown (left) and quantified (right). Statistical significance was tested by one-way ANOVA with Sidak’s correction. *, **, and *** correspond to p-values ≤0.05, 0.01, and 0.001, respectively. N = 3–5 mice/group and data shown are representative of two to three repeated experiments. Error bars indcate standard error of the mean (SEM).

Article Snippet: Il27ra –/– (Jackson labs; strain # 018078; RRID: IMSR_JAX:018078 ) were bred in house, as were Il27 –/– bio mice, originally generated by Lexicon Pharmaceuticals, Inc and previously described ( ).

Techniques: Infection

Journal: eLife

Article Title: IL-27 limits HSPC differentiation during infection and protects from stem cell exhaustion

doi: 10.7554/eLife.105876

Figure Lengend Snippet: ( A ) The proportion of zsGreen + monocyte dendritic cell progenitors (MDPs) (CD3 − , NK1.1 − , B220 − , CD117 + , CD34 + , CD16/32 lo , CD115 + ), monocytes (CD3 − , B220 − , Ly6G − , CD115 + , CD135 − , CD117 − , CD11b + , Ly6C + ), and neutrophils (CD3 − , B220 − , CD11b + , CD115 − , Ly6G + ) in the BM of naive and infected Procr-Ai6 mice. ( B ) The proportion of zsGreen + CCR2 hi/lo CX3CR1 hi/lo monocytes in the spleen and peritoneal exudate cells (PECs) in naive and infected Procr-Ai6 mice. ( C ) Number of MDPs in WT and Il27 –/– mice in the bone marrow during infection. ( D ) Numbers of monocytes in the liver in WT and Il27 −/− throughout infection. ( E ) WT and Il27 –/– mice were injected I.V. with fluorescent anti-CD45 to label immune cells in the vasculature. Numbers of I.V. label - and CCR2 hi CX3CR1 lo monocytes in the liver were then quantified throughout infection. ( F ) IFNγ-Thy1.1 reporter mice were infected and treated with an anti-p28 blocking antibody. Numbers of IFNγ + cells in the bone marrow were then assessed by flow cytometry. Representative flow plots are shown (left) and quantified (right). Statistical significance was tested by Welch’s t -test ( A, F ) or one-way ANOVA with Sidak’s correction ( B–E ). *, **, ***, and **** correspond to p-values ≤0.05, 0.01, 0.001, and 0.0001, respectively. N = 3–5 mice/group and data shown are representative of two to three repeated experiments. Error bars indcate SEM.

Article Snippet: Il27ra –/– (Jackson labs; strain # 018078; RRID: IMSR_JAX:018078 ) were bred in house, as were Il27 –/– bio mice, originally generated by Lexicon Pharmaceuticals, Inc and previously described ( ).

Techniques: Infection, Injection, Blocking Assay, Flow Cytometry

Journal: eLife

Article Title: IL-27 limits HSPC differentiation during infection and protects from stem cell exhaustion

doi: 10.7554/eLife.105876

Figure Lengend Snippet: ( A ) WT mice were infected, and the bone marrow harvested throughout infection. BM was either washed with media or cultured for 24 hr, and supernatant collected. Both the wash and supernatant were analyzed by ELISA for IL-27p28. ( B ) Il27 GFP reporter mice were infected and their bone marrow analyzed by flow cytometry throughout infection. A representative flow plot at 5 dpi of bulk marrow is shown (left) and proportion of immune cell contribution to the GFP + population measured (right). N=3 biological replicates, with two technical replicates performed. ( C ) Numbers of GFP + cells in the spleen, blood, and BM of infected mice were quantified throughout infection (left). Proportions of GFP + CD45 + cells were analyzed by cell type in the blood, spleen, and peritoneal exudate cell (PEC) during infection (right). ( D ) Femurs from infected Procr tdTomato+ Il27 GFP+ mice were harvested, sectioned, and imaged at ×40 magnification. The total imaged femur is shown (top), with zoomed-in sections shown below for increased detail of cellular localization (bottom). Arrows indicate areas of localization with Procr tdTomato+ and Il27 GFP+ cells. Numbers of Procr tdTomato+ and Il27 GFP+ cells were then counted in eight randomly selected regions throughout the marrow (see ). GFP + cells were normalized to tdTomato + cells and the eight counted regions classified as areas of hyper- or hypo-vascularization according to proximity of the region to the labeled vasculature. This allowed quantification of the localization of cells (bottom right). N = 3–5 mice/group, and data shown are representative of two repeated experiments. Error bars indcate SEM.

Article Snippet: Il27ra –/– (Jackson labs; strain # 018078; RRID: IMSR_JAX:018078 ) were bred in house, as were Il27 –/– bio mice, originally generated by Lexicon Pharmaceuticals, Inc and previously described ( ).

Techniques: Infection, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Labeling

Journal: eLife

Article Title: IL-27 limits HSPC differentiation during infection and protects from stem cell exhaustion

doi: 10.7554/eLife.105876

Figure Lengend Snippet: ( A ) Il27 GFP mice were infected, and the numbers of each CD45 + , immune cell type that expressed GFP in the BM or periphery were quantified throughout infection. ( B ) Levels of GFP expression were measured in bulk monocytes (red) and dendritic cells (blue) in the bone marrow (left) and spleen (right). ( C ) Femurs from naive Il27 GFP mice were stained for Sca-1 and imaged at ×40 magnification. Zoomed-in images are shown (right) of the indicated areas, with the yellow arrows indicating positively stained, Sca-1 + cells. ( D ) The merged image shown in is separated by channel (GFP, tdTomato, and AF647, from top to bottom). Eight randomly selected regions that were used for quantification of either GFP + or tdTomato + cells are shown (top), and the quantification of cells in each region is shown next to the corresponding fluorescence channel (right). N = 3–5 mice/group, and data shown are representative of two repeated experiments. Error bars indcate SEM.

Article Snippet: Il27ra –/– (Jackson labs; strain # 018078; RRID: IMSR_JAX:018078 ) were bred in house, as were Il27 –/– bio mice, originally generated by Lexicon Pharmaceuticals, Inc and previously described ( ).

Techniques: Infection, Expressing, Staining, Fluorescence