recombinant cd8a  (Sino Biological)

Journal: International Journal of Molecular Sciences

Article Title: Microbubble-Protected Oncolytic Virotherapy Targeted by Sonoporation Induces Tumor Necrosis and T-Lymphocyte Infiltration in Humanized Mice Bearing Triple-Negative Breast Cancer

doi: 10.3390/ijms252413697

Figure Lengend Snippet: Immunohistochemical detection of CD8 + T-cells in tumors. CD8 + T cell tumor infiltrates (TILs) are few in the control (saline) group, and sporadic in the Pembrolizumab treated group; however, their expression significantly increases in both OV groups, and their number is dramatically increased in both MB groups. The magnification of the upper panels is 100×, and the magnification of the lower panels is 200×.

Article Snippet: Primary antibodies for the detection of immune cells included a rabbit recombinant multiclonal anti-CD4 (RM1013, 1:1000 dilution, Abcam), a rabbit recombinant monoclonal anti-CD8a (Clone EPR21769, 1:2000 dilution, Abcam), and a rabbit monoclonal anti-CD25/IL-2Ra (Clone SP176, 1:100 dilution, Abcam).

Techniques: Immunohistochemical staining, Control, Saline, Expressing

Journal: Cell Genomics

Article Title: CXCR4 orchestrates the TOX-programmed exhausted phenotype of CD8 + T cells via JAK2/STAT3 pathway

doi: 10.1016/j.xgen.2024.100659

Figure Lengend Snippet: scRNA-seq analysis identified that CXCR4 inhibition enhanced anti-PD-1 immunotherapy by attenuating the T ex phenotype (A) Schematic representation of experimental design and treatment. (B) UMAP plot displaying the 11 major cell types of U14-derived xenograft tumors based on scRNA-seq. (C) UMAP plot displaying the 11 major cell types of U14-derived xenograft tumors in different treatment groups. (D) The proportions and cell number of cell types in each sample. (E) UMAP plot displaying the CD8 + T cells of U14-derived xenograft tumors in different treatment groups. (F) Volcano plot showing the DEGs of CD8 + T cells between combination-treated and control groups. (G) UMAP plot and RNA velocity displaying the four major cell types of CD8 + T cells based on scRNA-seq data. (H) Pseudotime heatmap displaying the genes involved in the developmental process of CD8 + T cells.

Article Snippet: Recombinant Anti-CD8 alpha antibody, human , Abcam , Cat# ab237709; RRID: AB_2892677.

Techniques: Inhibition, Derivative Assay, Control

Journal: Cell Genomics

Article Title: CXCR4 orchestrates the TOX-programmed exhausted phenotype of CD8 + T cells via JAK2/STAT3 pathway

doi: 10.1016/j.xgen.2024.100659

Figure Lengend Snippet: CXCR4 inhibition promoted CD8 + T cell activation by mediating abundant CD8 + TCR clonotypes and increasing MHC-I molecule expression (A) UMAP plot displaying the distribution of TCR types in cell types based on scTCR-seq. (B) The number of CDR3 (AA), TCRα, and TCRβ in different treatment groups. (C) The number of clonotypes shows differences among different treatment groups. (D) UMAP plot displaying the distribution of TCR types in CD8 + T cells based on scTCR-seq. (E) Column chart showing the number of clone types in different treatment groups. (F) Network plot showing the TCR specificity groups with at least one cell from the different treatments. (G) Violin plots of MHC-I expression levels in dendritic cells, monocytes/macrophages, and B cells within different treatment groups. p values were calculated by the Mann-Whitney test in GraphPad Prism. Each group was compared with the control group, and a significant p value is indicated by ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Recombinant Anti-CD8 alpha antibody, human , Abcam , Cat# ab237709; RRID: AB_2892677.

Techniques: Inhibition, Activation Assay, Expressing, MANN-WHITNEY, Control

Journal: Cell Genomics

Article Title: CXCR4 orchestrates the TOX-programmed exhausted phenotype of CD8 + T cells via JAK2/STAT3 pathway

doi: 10.1016/j.xgen.2024.100659

Figure Lengend Snippet: CXCR4 inhibition mitigated the TOX -mediated T ex phenotype by JAK2-STAT3 pathway regulation to reduce p-STAT3 and TOX interactions (A) Western blots showing the results of the JAK-STAT pathway and the exhausted markers after plerixafor or NUCC-390 treatment in Jurkat T cells. The JAK-STAT pathway and the exhausted markers were then detected by western blot after use of the CXCR4 antagonist (plerixafor) and JAK2-STAT3 inhibitor (SD-1029) in Jurkat T cells. (B) Western blots showing the results of the T ex phenotype in the JAK2-STAT3 pathway treated by motixafortide in Jurkat and mouse T cells. (C) Co-immunoprecipitation assay showing the interaction between p-STAT3 and TOX in human and mouse T cells. (D) Co-immunoprecipitation assay and western blot results of the spleens in Cxcr4 flox/flox Lck Cre and Cxcr4 flox/flox mice show the interactions between p-STAT3 and TOX, and the changes of T ex phenotype and the phosphorylation of the JAK2-STAT3 pathway. (E) Mean enrichment profiles along gene bodies (±3 kb) of peaks and TSS profile plots for p-STAT3 and TOX ChIP-seq treated by plerixafor in Jurkat T cells. TSS, transcription start site; TES, transcription end site. (F) Mean enrichment profiles along gene bodies and TSS profile plots of peaks for p-STAT3 and TOX ChIP-seq treated by plerixafor in mouse spleen. (G) TSS profile plots of ICGs for p-STAT3 and TOX ChIP-seq treated by plerixafor in Jurkat T cells. (H) Schematic representation of the experimental design. (I) Pictures of the anatomy of intestinal tissues with different treatments. (J) Kaplan-Meier analysis of mice bearing tumors transplanted with CD8 + T cells pretreated with vehicle control, SD-1029, or STAT3-IN-9. p values were determined using a log-rank test in GraphPad Prism.

Article Snippet: Recombinant Anti-CD8 alpha antibody, human , Abcam , Cat# ab237709; RRID: AB_2892677.

Techniques: Inhibition, Western Blot, Co-Immunoprecipitation Assay, ChIP-sequencing, Control

Journal: Cell Genomics

Article Title: CXCR4 orchestrates the TOX-programmed exhausted phenotype of CD8 + T cells via JAK2/STAT3 pathway

doi: 10.1016/j.xgen.2024.100659

Figure Lengend Snippet: Single-cell chromatin accessibility profiles revealed that CXCR4 deficiency epigenetically orchestrated the exhausted and functional phenotype of CD8 + T cells (A) Schematic representation of the experimental design for scATAC-seq and scRNA-seq. (B) UMAP plot displaying the 14 major cell types of spleen based on scATAC-seq and scRNA-seq. (C) Violin plots showing the expression levels of Cxcr4 in the CD8 + T cell cluster. (D) mIHC staining of spleen tissues validated the expression levels of CXCR4 and exhausted markers in CD8 + T cells. (E) The UMAP plot displays eight subtypes of CD8 + T cells based on scATAC-seq and scRNA-seq. The violin plot shows the marker genes associated with eight subtypes of CD8 + T cells. (F) The proportions of eight subtypes of CD8 + T cells in each sample. (G) Peaks plots in exhausted markers show the peaks in CD8 + T cells and other cell clusters excluded T cells in each sample. (H) Peaks plots in functional markers show the peaks in CD8 + T cells and other cell clusters excluded T cells in each sample. (I) Assessment of chromatin accessibility levels in the CD8 + T cell cluster via Cicero. (J) Statistical analysis of IHC staining with different treatments. p values were determined using an unpaired, parametric t test in GraphPad Prism. ∗∗∗∗ p < 0.0001.

Article Snippet: Recombinant Anti-CD8 alpha antibody, human , Abcam , Cat# ab237709; RRID: AB_2892677.

Techniques: Functional Assay, Expressing, Staining, Marker, Immunohistochemistry

Journal: Cell Genomics

Article Title: CXCR4 orchestrates the TOX-programmed exhausted phenotype of CD8 + T cells via JAK2/STAT3 pathway

doi: 10.1016/j.xgen.2024.100659

Figure Lengend Snippet: Clinical applications demonstrated that CXCR4 expression was associated with ICB response status in cancers (A–D) mIHC staining in surgical samples from cervical cancer undergoing NACT with cisplatin plus paclitaxel and combined with anti-PD-1 treatment (A). Statistical analysis of the expression of p63 (B), percentage of CXCR4 + CD8 + and GZMB + CD8 + T cells (C), and number of CD8 + T cells (D) in samples. p values were determined using the Mann-Whitney test in GraphPad Prism. (E) The expression levels of CXCR4 in responder (R) and non-responder (NR) groups from ICBatlas. Patients who achieved complete response or partial response were categorized as R, while those experiencing stable disease or progressive disease were classified as NR. (F) CXCR4 expression in melanoma before (pre-) and after (post-) the anti-CTLA4 therapy (ipilimumab). The DEGs in RNA-seq were calculated by DESeq2 with parameters (|log 2 fold change| > 1 and p < 0.05). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Recombinant Anti-CD8 alpha antibody, human , Abcam , Cat# ab237709; RRID: AB_2892677.

Techniques: Expressing, Staining, MANN-WHITNEY, RNA Sequencing Assay

Journal: Cell Genomics

Article Title: CXCR4 orchestrates the TOX-programmed exhausted phenotype of CD8 + T cells via JAK2/STAT3 pathway

doi: 10.1016/j.xgen.2024.100659

Figure Lengend Snippet:

Article Snippet: Recombinant Anti-CD8 alpha antibody, human , Abcam , Cat# ab237709; RRID: AB_2892677.

Techniques: Recombinant, Lysis, Activation Assay, Staining, Magnetic Beads, Control, Amplification, Immunohistochemistry, Cell Isolation, RNA Extraction, Sonication, Chromatin Immunoprecipitation, Software