Journal: Frontiers in Immunology

Article Title: In Vivo Priming of Peritoneal Tumor-Reactive Lymphocytes With a Potent Oncolytic Virus for Adoptive Cell Therapy

doi: 10.3389/fimmu.2021.610042

Figure Lengend Snippet: Vaccinia virus expressing IL15/IL15Rα (vvDD-IL15/Rα) treatment leads to enrichment of memory CD8 + T cells with an activated phenotype. Immunocompetent B6 mice were inoculated IP with 5.0x10 5 MC38-luc cells (day −9), randomized and treated with intraperitoneal (IP) injection of PBS, vvDD or vvDD-IL15/Rα at a dose of 5.0x107 pfu/mouse (day 0). Ten days following i.p. oncolytic virotherapy, peritoneal CD8 + T cells were isolated via peritoneal lavage and characterized via flow cytometry (in addition to day 3, 6, 8, and 14 for figure A). (A) Regional delivery of vvDD-IL15/Rα greatly enhanced and prolonged peritoneal CD8 + T cell infiltration on day 6 to 14 following virotherapy. (B) The amount of peritoneal central memory (Tcm, CD44 + CD62L + ) and effector memory (Tem, CD44 + CD62L - ) T cells is increased 10 days following vvDD-IL15/Rα injection. (C) Representative flow cytometry charts of memory CD8 + T cell subsets. vvDD-IL15Rα treatment resulted in increased Ki67 (D) , CXCR3 (E) , CD122 (F) , Tim3 - PD1 int (G) , CD103 (H) , and CD44 + PD-1 + (I) staining compared to vvDD or PBS 10 days following injection. PD-1 high (J) , TIGIT + (K) , and Tim-3 + (L) staining were not increased after vvDD-IL15/Rα treatment compared to vvDD or PBS treatment. Representative flow cytometry charts of PD-1 expression by treatment group are depicted in . N=8–12. All values presented as mean ± SEM. *p < 0.05. **p < 0.01. ***p < 0.001. ****p < 0.0001. Data are combined from two independent experiments.

Article Snippet: FITC conjugated anti-human CD122 (clone REA167) was purchased from Miltenyi Biotec.

Techniques: Virus, Expressing, Injection, Isolation, Flow Cytometry, Staining

Journal: Frontiers in Immunology

Article Title: In Vivo Priming of Peritoneal Tumor-Reactive Lymphocytes With a Potent Oncolytic Virus for Adoptive Cell Therapy

doi: 10.3389/fimmu.2021.610042

Figure Lengend Snippet: Characterization of human peritoneal CD8 + and CD4 + T cells derived from peritoneal metastasized tumors. Peritoneal fluid was collected from 14 patients undergoing peritoneal surgery, and peritoneal lymphocytes were extracted and characterized. (A) Flowchart of flow cytometry gating to identify peritoneal CD4 and CD8 T cells. Single cells gating, dead cells exclusion via Zombie aqua staining. Subsequently CD3 + cells were separated into CD4 + or CD8 + cells. (B) Distribution of naïve T cells (Tn, CD45RO - CCR7 + ), central memory T cells (Tcm, CD45RO + CCR7 + ), effector memory T cells (Tem, CD45RO + CCR7 - ) and highly differentiated effector T cells (Temra, CD45RO - CCR7 - ) gated from CD8 + T cells. (C) Representative flow cytometry chart of peritoneal memory CD8 + T cell subsets. (D) Levels of 4-1BB and Ox40 on CD8 + T cells were analyzed. (E–H) Analysis of PD-1 and Tim-3 demonstrated the majority of CD8 + cells were PD-1 high and an average of 20% expressed Tim-3. Representative flow charts of PD-1 and Tim-3 expression are shown in (F, H) respectively. (I, J) (representative flow chart): The shared beta-subunit of the IL-2- and IL15-receptor (CD122) showed a mean expression in peritoneal CD8 + T cells of 29.1%. (K–M) Peritoneal CD8 + T cells from 7 patients were tested for secretion of cytotoxic IFN-γ by ELISA (K) and 4-1BB surface expression by flow (L, M) (representative images) after overnight coculture with correlating tumor digest. Monocytes were used as negative controls, CD3/CD28 dynabeads as a positive control. All values presented as mean ± SEM. *p < 0.05. **p < 0.01. ***p < 0.001. ****p < 0.0001.

Article Snippet: FITC conjugated anti-human CD122 (clone REA167) was purchased from Miltenyi Biotec.

Techniques: Derivative Assay, Flow Cytometry, Staining, Expressing, Enzyme-linked Immunosorbent Assay, Positive Control

Journal: Molecular Medicine Reports

Article Title: Expression, immunogenicity and clinical significance analysis of thyroid-stimulating hormone receptor fusion proteins

doi: 10.3892/mmr.2025.13639

Figure Lengend Snippet: Changes of specific regulatory T cell subsets in mice after immunization. (A) Flow cytometry analysis of changes in CD4 + CD25 + T cells in the hTSHR289, hTSHR290 and hTSHR410 groups after immunization. (B) Flow cytometry analysis of changes in CD8 + CD122 + T cells in the hTSHR289, hTSHR290 and hTSHR410 groups after immunization. *P<0.05 (One-way ANOVA with Tukey's post hoc test. hTSHR, human thyroid-stimulating hormone receptor.

Article Snippet: The analyte detectors were as follows: CD3 + antibody (cat. no. 565643; Becton, Dickinson and Company), CD4 + antibody (cat. no. F21004A02; Multi Sciences Biotech), CD8 + antibody (cat. no. F2100801; Multi Sciences Biotech), CD25 antibody (cat. no. E-AB-F1102C; Wuhan Elabscience Biotechnology Co., Ltd.) and CD122 antibody (cat. no. E-AB-F1029D; Wuhan Elabscience Biotechnology Co., Ltd.).

Techniques: Flow Cytometry

Journal: NAR Genomics and Bioinformatics

Article Title: Evolutionary genetic algorithm identifies IL2RB as a potential predictive biomarker for immune-checkpoint therapy in colorectal cancer

doi: 10.1093/nargab/lqab016

Figure Lengend Snippet: Evolutionary genetic algorithm based tool (ACE) highlights IL2RB as the most common intersection between analyses of nine immune checkpoint markers in CRC, across both TCGA mRNA microarray and RNA-seq datasets. ( A and B ) UpSet plot showing the number of intersections produced by ACE gene lists for each biomarker in both matched microarray data and RNA-seq data, respectively.

Article Snippet: Immunohistochemistry was performed for IL2RB (Polyclonal Anti-IL2RB Antibody; Atlas Antibodies, Voltavägen, Sweden; catalogue number: HPA062657; 1:1000 dilution; 15 min incubation at room temperature) on the Leica BOND-MAX automated immunostainer (Antigen retrieval: ER2 for 20 min; Detection chemistry: Bond Polymer Refine Detection and Enhancer).

Techniques: Microarray, RNA Sequencing, Produced, Biomarker Discovery

Journal: NAR Genomics and Bioinformatics

Article Title: Evolutionary genetic algorithm identifies IL2RB as a potential predictive biomarker for immune-checkpoint therapy in colorectal cancer

doi: 10.1093/nargab/lqab016

Figure Lengend Snippet: Transcriptional quantification of microenvironment and prognostic value of IL2RB mRNA expression in CRC. ( A ) Correlation of IL2RB expression with microenvironment cell population (MCP) scores for individual cell types in TCGA CRC microarray data. ( B ) Representative CD3 and CD8 IHC images generated from an in-house TMA split by median IL2RB mRNA expression using matched transcriptional profiles from GSE103479 (×5 magnification). ( C ) Kaplan–Meier curve showing improved DFS for patients with higher expression of IL2RB in CRC TCGA RNA-seq dataset ( n = 322, log rank P = 0.011). ( D ) Kaplan–Meier curve showing improved DFS for patients with higher expression of IL2RB in GSE39582 ( n = 519, log rank P value = 0.0057).

Article Snippet: Immunohistochemistry was performed for IL2RB (Polyclonal Anti-IL2RB Antibody; Atlas Antibodies, Voltavägen, Sweden; catalogue number: HPA062657; 1:1000 dilution; 15 min incubation at room temperature) on the Leica BOND-MAX automated immunostainer (Antigen retrieval: ER2 for 20 min; Detection chemistry: Bond Polymer Refine Detection and Enhancer).

Techniques: Expressing, Microarray, Generated, RNA Sequencing

Journal: NAR Genomics and Bioinformatics

Article Title: Evolutionary genetic algorithm identifies IL2RB as a potential predictive biomarker for immune-checkpoint therapy in colorectal cancer

doi: 10.1093/nargab/lqab016

Figure Lengend Snippet: Comprehensive comparative assessment of IL2RB IHC and immune markers in stage II/III MSI-CRC ( n = 115) from an in-house TMA. ( A ) Dot plot showing mean IL2RB positive cells/mm2. ( B ) Representative image of IL2RB IHC (×20 magnification). ( C ) Boxplot and representative images (×5 magnification) comparing PDL1+ tumor cells by IHC in IL2RB high and low patients ( P = 0.058). ( D ) Boxplot and representative images (×5 magnification) comparing density of ICOS+ cells by IHC in IL2RB high and low patients ( P = 0.1956). ( E ) Boxplot and representative images (×5 magnification) comparing density of CD3+ positive cells by IHC in IL2RB high and low patients ( P = 0.0103). ( F ) Boxplot and representative images (×5 magnification) comparing density of CD4+ positive cells by IHC in IL2RB high and low patients ( P = 0.006). ( G ) Boxplot and representative images (×5 magnification) comparing density of FOXP3+ positive cells by IHC in IL2RB high and low patients ( P <0.0001). ( H ) Boxplot and representative images (×5 magnification) comparing density of CD8+ positive cells by IHC in IL2RB high and low patients ( P = 0.2012). Significance determined using Mann–Whitney test. IL2RB status determined using 43rd percentile from transcriptional analysis

Article Snippet: Immunohistochemistry was performed for IL2RB (Polyclonal Anti-IL2RB Antibody; Atlas Antibodies, Voltavägen, Sweden; catalogue number: HPA062657; 1:1000 dilution; 15 min incubation at room temperature) on the Leica BOND-MAX automated immunostainer (Antigen retrieval: ER2 for 20 min; Detection chemistry: Bond Polymer Refine Detection and Enhancer).

Techniques: MANN-WHITNEY

Journal: NAR Genomics and Bioinformatics

Article Title: Evolutionary genetic algorithm identifies IL2RB as a potential predictive biomarker for immune-checkpoint therapy in colorectal cancer

doi: 10.1093/nargab/lqab016

Figure Lengend Snippet: Single-Cell RNA-seq characterisation of IL2RB in CRC. ( A ) Dot plot showing expression of IL2RB (log2 FPKM +1) in publically available (GSE81861) single cell RNA-seq profiles from 11 CRC patients compared across cell type (ANOVA, P < 0.0001 (****)). ( B ) Heatmap comparing log2 FPKM+1 expression of PDCD1, PDL1, LAG3, CTLA4, ICOS and TIGIT in IL2RB+ CD8+ T cells ( n = 1192) derived from single cell MC38 Colon cancer anti-PD1 and anti-GITR treated mouse models RNA-seq dataset (GSE120909).

Article Snippet: Immunohistochemistry was performed for IL2RB (Polyclonal Anti-IL2RB Antibody; Atlas Antibodies, Voltavägen, Sweden; catalogue number: HPA062657; 1:1000 dilution; 15 min incubation at room temperature) on the Leica BOND-MAX automated immunostainer (Antigen retrieval: ER2 for 20 min; Detection chemistry: Bond Polymer Refine Detection and Enhancer).

Techniques: RNA Sequencing, Expressing, Derivative Assay

Journal: Journal of Clinical Laboratory Analysis

Article Title: High‐dimensional single‐cell proteomics analysis reveals the landscape of immune cells and stem‐like cells in renal tumors

doi: 10.1002/jcla.23155

Figure Lengend Snippet: Purified antibodies about the immune cell‐centric panel

Article Snippet: CD25/IL‐2R , 149Sm , 2A3 , Fluidigm , 3149010B.

Techniques: Purification