anti cd8  (Boster Bio)

Journal: Cell Transplantation

Article Title: Leflunomide Inhibits rat-to-Mouse Cardiac Xenograft Rejection by Suppressing Adaptive Immune Cell Response and NF-κB Signaling Activation

doi: 10.1177/09636897211054503

Figure Lengend Snippet: Phenotypic characteristics of grafts. Xenografts and allografts were recovered at POD 6. Relative mRNA expression of IL-2, IFN-γ, IL-4, and TNF-α in (A) xenograft and (B) allografts measured by qRT-PCR ( n = 3 mice/group). (C) Immunohistochemistry staining of CD20 + (brown) in xenograft and allograft; bar indicates 20 µm and quantified. (D) IgM in grafts were examined by immunohistochemistry staining; bar indicates 20 µm and quantified. Immunohistochemistry staining of CD4 + (brown), and CD8 + (brown) in (E) xenograft and (H) allografts. (Original magnification: ×400). Data are presented as mean ± SEM from three independent experiments. * P

Article Snippet: Immunohistochemical staining was performed as described previously , the sections were then stained with primary antibody anti-CD4 Ab (Boster, Wuhan, China), anti-CD8 Ab (Boster), anti-CD20 Ab (Boster), anti-IgM (Boster), and secondary antibody goat anti-rabbit IgG (GB23303; Boster, Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Mouse Assay, Immunohistochemistry, Staining

Journal: Cell Transplantation

Article Title: Leflunomide Inhibits rat-to-Mouse Cardiac Xenograft Rejection by Suppressing Adaptive Immune Cell Response and NF-κB Signaling Activation

doi: 10.1177/09636897211054503

Figure Lengend Snippet: The effect of LEF treatment on CMR- and AVR- mediated immune responses. (A) MLR responses. Recipient splenocytes were isolated at POD 6 (responders) and irradiated naïve SD or BALB/c splenocytes (stimulators) were co-cultured for three days. Data are representative of three independent experiments. (B) Absolute numbers of splenocytes in LEF -treated and normal-saline treated mice recipients. Naïve C57BL/6 mice are shown for comparison ( n = 3 mice/group). (C) Representative proportion of CD4 + and CD8 + T cells in recipient splenocytes. A total of 1 × 10 6 splenocytes were isolated at POD 6, and the percentage of CD4 + and CD8 + T cells was determined by flow cytometry ( n = 3 mice/group). (D) Representative proportion of CD19 + B cells in recipient splenocytes. A total of 1 × 10 6 splenocytes were isolated at POD 6, and the percentage of CD19 + B cells was determined by flow cytometry ( n = 3 mice/group). (E) The number of CD3 + T cells in recipient spleen were determined by flow cytometry ( n = 3 mice/group). (F) The number of CD19 + B cells in recipient spleen were determined by flow cytometry ( n = 3 mice/group). (G) Representative proportion of donor-specific antibodies in recipient serum. Serum was collected from xenograft and allograft recipients at POD 6, and the percent of IgG1, IgG2a, and IgM was determined by flow cytometry ( n = 3 mice/group). (H) Serum levels of proinflammatory cytokines. Peripheral blood was collected at POD 6 and IFN-γ serum levels were measured by ELISA ( n = 3 mice/group). (I) CD3 + T cells were isolated from naïve C57BL/6 mice, and co-cultured with anti-CD3 and anti-CD28 monoclonal antibody in the absence and presence of LEF for 3 days, the supernatant was collected and measured by ELISA ( n = 3 separate experiments). Data are presented as the mean ± SEM of three independent experiments. * P

Article Snippet: Immunohistochemical staining was performed as described previously , the sections were then stained with primary antibody anti-CD4 Ab (Boster, Wuhan, China), anti-CD8 Ab (Boster), anti-CD20 Ab (Boster), anti-IgM (Boster), and secondary antibody goat anti-rabbit IgG (GB23303; Boster, Wuhan, China).

Techniques: Isolation, Irradiation, Cell Culture, Mouse Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: World Journal of Gastrointestinal Oncology

Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

doi: 10.4251/wjgo.v14.i6.1124

Figure Lengend Snippet: Detection of mitochondrial damage and apoptosis in CD8 + T cells. Activated CD8 + T cells, co-cultured with Huh-7 cells for 3 d, were detected by flow cytometry. A: JC-1; B: Reactive oxygen species; C: Apoptosis. The data represent the mean ± SE; b P

Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

Techniques: Cell Culture, Flow Cytometry

Journal: World Journal of Gastrointestinal Oncology

Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

doi: 10.4251/wjgo.v14.i6.1124

Figure Lengend Snippet: Regulation of CD8 + T cell function by carbonyl cyanide-3-chlorophenylhydrazone. A-C: Activated CD8 + T cells were cultured in RPMI 1640 supplemented with 5 μmol/L or 10 μmol/L carbonyl cyanide-3-chlorophenylhydrazone for 3 d, respectively. The levels of indicated molecules in CD8 + T cells were detected by flow cytometry. A: PD-1; B: PRF1; C: Granzyme B. The data represent the mean ± SE; d P

Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

Techniques: Cell Function Assay, Cell Culture, Flow Cytometry

Journal: World Journal of Gastrointestinal Oncology

Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

doi: 10.4251/wjgo.v14.i6.1124

Figure Lengend Snippet: Regulation of CD8 + T cell function by Mdivi-1. A-C: Activated CD8 + T cells were co-cultured with Huh-7 cells in RPMI 1640 containing 5 μmol/L Mdivi-1 for 3 d. The levels of indicated molecules in CD8 + T cells were detected by flow cytometry. A: PD-1; B: PRF1; C: Granzyme B. The data represent the mean ± SE; c P

Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

Techniques: Cell Function Assay, Cell Culture, Flow Cytometry

Journal: World Journal of Gastrointestinal Oncology

Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

doi: 10.4251/wjgo.v14.i6.1124

Figure Lengend Snippet: Effect of glutamine deprivation on CD8 + T cells co-cultured with hepatoma cells. Activated CD8 + T cells were co-cultured with Huh-7 cells in RPMI 1640 containing no glutamine (0Gln) for 3 d. The levels of indicated molecules in CD8 + T cells were detected by flow cytometry. A: PD-1; B: PRF1; C: Granzyme B; D: JC-1; E: Reactive oxygen species; F: Apoptosis. The data represent the mean ± SE; a P

Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

Techniques: Cell Culture, Flow Cytometry

Journal: World Journal of Gastrointestinal Oncology

Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

doi: 10.4251/wjgo.v14.i6.1124

Figure Lengend Snippet: PD-1 expression by infiltrating CD8 + T cells in liver tissues. Tissues were subjected to fluorescent double staining with anti-CD8 and anti-PD-1 antibodies. A: Hepatocellular carcinoma tissues; B: Paracancerous tissues. Scale bar: 400 μm.

Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

Techniques: Expressing, Double Staining

Journal: World Journal of Gastrointestinal Oncology

Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

doi: 10.4251/wjgo.v14.i6.1124

Figure Lengend Snippet: Evaluations of CD8 + T cell function. Activated CD8 + T cells were co-cultured with Huh-7 cells for 3 d. Flow cytometry was used to detect the expression of indicated molecules in CD8 + T cells. A: PD-1; B: PRF1; C: Granzyme B (GZMB). D and E: Quantitative real-time polymerase chain reaction was used to detect the mRNA levels in CD8 + T cells (D: PRF1 ; E: GZMB ). The data represent the mean ± SE; a P

Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

Techniques: Cell Function Assay, Cell Culture, Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction

Journal: World Journal of Gastrointestinal Oncology

Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

doi: 10.4251/wjgo.v14.i6.1124

Figure Lengend Snippet: Differential gene expression in CD8 + T cells. Activated CD8 + T cells were co-cultured with Huh-7 cells for 3 d. Total RNA was extracted from CD8 + T cells for transcriptome sequencing. A: GO enrichment plot of upregulated genes; B: GO enrichment plot of downregulated genes.

Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

Techniques: Expressing, Cell Culture, Sequencing

Journal: World Journal of Gastrointestinal Oncology

Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

doi: 10.4251/wjgo.v14.i6.1124

Figure Lengend Snippet: Transmission electron microscopy images of mitochondria in CD8 + T cells. Activated CD8 + T cells were co-cultured with Huh-7 cells for 3 d. The morphology of mitochondria in CD8 + T cells was detected by transmission electron microscopy. A: Control; B: Co-culture. Scale bar: 2 μm.

Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

Techniques: Transmission Assay, Electron Microscopy, Cell Culture, Co-Culture Assay

Journal: World Journal of Gastrointestinal Oncology

Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

doi: 10.4251/wjgo.v14.i6.1124

Figure Lengend Snippet: CD8 + T cell infiltration in liver tissues. A: Hepatocellular carcinoma tissues; B: Paracancerous tissues. Scale bar: 200 μm.

Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

Techniques:

Journal: World Journal of Gastrointestinal Oncology

Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

doi: 10.4251/wjgo.v14.i6.1124

Figure Lengend Snippet: Altered CD8 + T cell function by glutamine deficiency. Activated CD8 + T cells were cultured in RPMI 1640 containing no glutamine (0Gln) for 3 d. The levels of indicated molecules in CD8 + T cells were detected by flow cytometry. A: PD-1; B: PRF1; C: Granzyme B; D: JC-1; E: Reactive oxygen species; F: Apoptosis. The data represent the mean ± SE; b P

Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

Techniques: Cell Function Assay, Cell Culture, Flow Cytometry

Journal: Theranostics

Article Title: USP7 targeting modulates anti-tumor immune response by reprogramming Tumor-associated Macrophages in Lung Cancer

doi: 10.7150/thno.47137

Figure Lengend Snippet: Targeting USP7 can activate the p38 MAPK pathway to reprogram TAMs. ( A ) The strategy for sorting TAMs in TME by flow cytometry. ( B ) Heat maps illustrating the differentially expressed M1- and M2-related genes in TAMs between the P5091 group and the Control group based on the results of RNA sequencing. ( C ) RT-PCR further verifying the differentially expressed genes of sorted TAMs in each group. Data are presented as the mean ± SEM (n = 3). ( D ) KEGG analysis identifying 20 most obviously enriched pathways based on the differentially expressed genes of the two groups. ( E ) Western blotting detection of the expression of JNK, p-JNK, ERK1/2, p-ERK1/2, p38, p-p38, and β-actin in IL-4/13-BMDM M2 cells treated with P5091 (10 µM) at the indicated time points. ( F ) Flow cytometry analysis of CFSE expression on the surface of CD8 + T cells in the presence of conditioned medium of IL-4/13-induced BMDM M2 cells from various indicated treatments. Treatments indicated: DMSO stimulation, P5091 (10 µM) stimulation, P5091 (10 µM) stimulation in the presence of inhibitors of p38 (SB203580, 10 µM), JNK (SP600125, 10 µM), Erk1/2 (U0126-EtOH, 10 µM). Data are presented as the mean ± SEM (n = 4).

Article Snippet: CD8 antibody was purchased from BOSTER Biological Technology Co., Ltd (Wuhan, China).

Techniques: Flow Cytometry, RNA Sequencing Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing

Journal: Theranostics

Article Title: USP7 targeting modulates anti-tumor immune response by reprogramming Tumor-associated Macrophages in Lung Cancer

doi: 10.7150/thno.47137

Figure Lengend Snippet: Targeting USP7 inhibits tumor growth and induces local anti-tumor immunity in vivo . ( A ) Harvested and photographed tumors in the P5091 and the Control group. ( B ) Lewis tumor growth following P5091 or Vehicle treatment in vivo. Data are presented as the mean ± SEM (n = 6 per group). ( C ) Spider diagram of the tumor volume growth in each mouse from the P5091 group and the Control group. ( D ) Gating strategy for detection of the TAMs by flow cytometry. ( E-G ) Proportions of M1 ( E ) and M2 ( F ), and M1/M2 ratio ( G ) in the TME of the P5091 group and the Control group. ( H-M ) Percentages of MDSC ( H ), Treg ( I ), CD4 + T ( J ), CD8 + T ( K ), Th1 ( L ), and CTLs ( M ) within the TME in each group. Data are presented as the mean ± SEM (n = 7) for ( E-M ).

Article Snippet: CD8 antibody was purchased from BOSTER Biological Technology Co., Ltd (Wuhan, China).

Techniques: In Vivo, Flow Cytometry

Journal: Theranostics

Article Title: USP7 targeting modulates anti-tumor immune response by reprogramming Tumor-associated Macrophages in Lung Cancer

doi: 10.7150/thno.47137

Figure Lengend Snippet: Targeting USP7 activates local and systemic anti-tumor immunity. ( A ) Multicolor immunofluorescence detection of TAMs and CTLs in the TME of the P5091 group and the Control group. ( B-F ) Cytokines IFN-γ ( B ), TNF-α ( C ), IL-2 ( D ), IL-5 ( E ), and IL-6 ( F ) in the TME of each group were detected by Mul-Analyte Flow Assay Kit. ( G-K ) Proportion of Treg ( G ), CD4 + T ( H ), CD8 + T ( I ), Th1 ( J ), and CTLs ( K ) in the spleen of each group. Data are presented as the mean ± SEM (n = 7) for ( B-K ).

Article Snippet: CD8 antibody was purchased from BOSTER Biological Technology Co., Ltd (Wuhan, China).

Techniques: Immunofluorescence

Journal: Theranostics

Article Title: USP7 targeting modulates anti-tumor immune response by reprogramming Tumor-associated Macrophages in Lung Cancer

doi: 10.7150/thno.47137

Figure Lengend Snippet: Targeting USP7 inhibits murine M2 phenotype and function in vitro . (A) The mRNA expression of common genes related to DUBs was detected by RT-PCR in M0, M1 (LPS/IFN-γ M1), and M2 (IL-4/13 M2 and IL-10 M2) induced from ANA-1. ( B ) The mRNA expression of USP7 in MΦs, M1, and M2 induced from BMDMs was detected by RT-PCR. ( C ) Western blotting showing the expression of USP7 in MΦ, M1, and M2 induced from BMDMs. ( D-E ) Flow cytometry analyses of the expression of CD206 in IL-4/13-induced BMDM M2 cells, which had been treated with P5091 (5 μM or 10 μM) for 24 h. Data are presented as the mean ± SEM (n = 3). ( F ) Detection of the expression of USP7 in BMDMs, which were transfected with either NC- or USP7-siRNA by western blotting. ( G ) The expression of CD206 in BMDMs of NC-siRNA and USP7-siRNA group was detected by flow cytometry. ( H-I ) Flow cytometry analyses of CFSE expression on the surface of CD8 + T cells in the presence of conditioned medium from either DMSO- or P5091- (5 μM or 10 μM) treated IL-4/13-induced BMDM M2 cells. Data are presented as the mean ± SEM (n = 3).

Article Snippet: CD8 antibody was purchased from BOSTER Biological Technology Co., Ltd (Wuhan, China).

Techniques: In Vitro, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Flow Cytometry, Transfection

Journal: Translational Lung Cancer Research

Article Title: Radiotherapy programs neutrophils to an antitumor phenotype by inducing mesenchymal-epithelial transition

doi: 10.21037/tlcr-21-152

Figure Lengend Snippet: RT-recruited neutrophils exhibit potent antitumor activity. Representative LLC tumor samples showing the expression levels of CD8 (A). TAN counts were positively correlated with CD8 levels (all P

Article Snippet: CD8, CD11b, vimentin, and E-cadherin staining was performed with anti-CD8 (BOSTER, Wuhan, China; 1:400), anti-CD11b (Abcam; 1:200), anti-vimentin (Cell Signaling Technology, CST, Boston, MA, USA; 1:300), and anti-E-cadherin (CST; 1:300), respectively.

Techniques: Activity Assay, Expressing