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anti-human cd25-pe clone: bc96 (Thermo Fisher)

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Thermo Fisher anti-human cd25-pe clone: bc96

Anti Human Cd25 Pe Clone: Bc96, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 90 stars, based on 1 article reviews

anti-human cd25-pe clone: bc96 - by Bioz Stars, 2026-03

90/100 stars

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1) Product Images from "Protocol for mapping T cell activation using single-cell RNA-seq"

Article Title: Protocol for mapping T cell activation using single-cell RNA-seq

Journal: STAR Protocols

doi: 10.1016/j.xpro.2024.103409

Figure Legend Snippet:

Techniques Used: Recombinant, Cell Culture, Transferring, Software

Image Search Results

Journal: STAR Protocols

Article Title: Protocol for mapping T cell activation using single-cell RNA-seq

doi: 10.1016/j.xpro.2024.103409

Figure Lengend Snippet:

Article Snippet: Anti-human CD25-PE (clone: BC96) , eBioscience , Cat #12-0259-42; 1:200.

Techniques: Recombinant, Cell Culture, Transferring, Software

CD4 + CD25 + CD127 low UCB-Tregs suppress SLE-PBMC and shift their cell population distribution. (A) Functional analysis of ex vivo -expanded day 14 CD4 + CD25 + CD127 low UCB-Tregs on suppression of Tcon cells from healthy donor and SLE-PBMCs. Two-way ANOVA demonstrated that ratio ( p < 0.0001), group ( p = 0.0029), and interaction ( p = 0.0068) between UCB-Treg : Tcon and UCB-Treg : SLE-PBMC were statistically significant. Data are presented as mean ± SEM ( n = 3). p < 0.05 was considered statistically significant. ** p < 0.01; **** p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests. (B) Expression levels of surface and intracellular markers on tSNE map. Unstimulated or stimulated CD4 + CD25 + CD127 low UCB-Tregs, SLE-PBMCs, or CD4 + CD25 + CD127 low UCB-Treg plus SLE-PBMC co-cultures (1:1) with CD3/CD28 plus IL-2 were stained with Live/Dead dye, CD45, CD14, CD3, CD4, CD25, CD127, CD8, CD19, IgD, CD27, CD62L, Helios, FoxP3, CD56, and HLA-DR antibodies. Stained live CD45 + cells from all six treatments were gated, down-sampled to 10,000 cells per sample which were concatenated. tSNE was run on six samples and the resulting tSNE plots were displayed expression intensities of surface and intracellular markers for all treatments in the concatenated file. (C) Subset analysis on tSNE map. Events in the tSNE embeddings were overlaid with manually gated lymphocytes, CD3 + CD19 - T, CD4 + T, CD4 + CD25 + T, CD4 + CD25 + CD127 low Treg, CD4 + CD8 + T, CD8 + T, CD3 - CD19 + B, CD27 - IgD - DN B, CD27 + IgD - Memory B, CD27 - IgD + Naïve B, CD27 + IgD + Plasma, CD56 + NK cells, and CD14 + monocytes and displayed for all treatments in the concatenated file. Stained cells were acquired on a Cytek Aurora flow cytometer and analyzed using FlowJo software. (D) Quantification analysis of subsets. Unstimulated or stimulated CD4 + CD25 + CD127 low UCB-Tregs, SLE-PBMCs, or CD4 + CD25 + CD127 low UCB-Treg plus SLE-PBMC co-cultures (1:1) with CD3/CD28 plus IL-2 were stained with Live/Dead dye, CD45, CD14, CD3, CD4, CD25, CD127, CD8, CD19, IgD, CD27, CD62L, Helios, FoxP3, CD56, and HLA-DR antibodies. CD4 + T, CD8 + T, CD4 + CD8 + T, CD4 + CD25 + CD127 low Treg, CD3 - CD19 + B, CD56 + NK cells, and CD14 + monocytes were quantified. Data are presented as mean ± SEM ( n = 6). p < 0.05 was considered statistically significant. p < 0.0001 by one-way ANOVA test.

Journal: Frontiers in Immunology

Article Title: Allogeneic cord blood regulatory T cells decrease dsDNA antibody and improve albuminuria in systemic lupus erythematosus

doi: 10.3389/fimmu.2023.1217121

Figure Lengend Snippet: CD4 + CD25 + CD127 low UCB-Tregs suppress SLE-PBMC and shift their cell population distribution. (A) Functional analysis of ex vivo -expanded day 14 CD4 + CD25 + CD127 low UCB-Tregs on suppression of Tcon cells from healthy donor and SLE-PBMCs. Two-way ANOVA demonstrated that ratio ( p < 0.0001), group ( p = 0.0029), and interaction ( p = 0.0068) between UCB-Treg : Tcon and UCB-Treg : SLE-PBMC were statistically significant. Data are presented as mean ± SEM ( n = 3). p < 0.05 was considered statistically significant. ** p < 0.01; **** p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests. (B) Expression levels of surface and intracellular markers on tSNE map. Unstimulated or stimulated CD4 + CD25 + CD127 low UCB-Tregs, SLE-PBMCs, or CD4 + CD25 + CD127 low UCB-Treg plus SLE-PBMC co-cultures (1:1) with CD3/CD28 plus IL-2 were stained with Live/Dead dye, CD45, CD14, CD3, CD4, CD25, CD127, CD8, CD19, IgD, CD27, CD62L, Helios, FoxP3, CD56, and HLA-DR antibodies. Stained live CD45 + cells from all six treatments were gated, down-sampled to 10,000 cells per sample which were concatenated. tSNE was run on six samples and the resulting tSNE plots were displayed expression intensities of surface and intracellular markers for all treatments in the concatenated file. (C) Subset analysis on tSNE map. Events in the tSNE embeddings were overlaid with manually gated lymphocytes, CD3 + CD19 - T, CD4 + T, CD4 + CD25 + T, CD4 + CD25 + CD127 low Treg, CD4 + CD8 + T, CD8 + T, CD3 - CD19 + B, CD27 - IgD - DN B, CD27 + IgD - Memory B, CD27 - IgD + Naïve B, CD27 + IgD + Plasma, CD56 + NK cells, and CD14 + monocytes and displayed for all treatments in the concatenated file. Stained cells were acquired on a Cytek Aurora flow cytometer and analyzed using FlowJo software. (D) Quantification analysis of subsets. Unstimulated or stimulated CD4 + CD25 + CD127 low UCB-Tregs, SLE-PBMCs, or CD4 + CD25 + CD127 low UCB-Treg plus SLE-PBMC co-cultures (1:1) with CD3/CD28 plus IL-2 were stained with Live/Dead dye, CD45, CD14, CD3, CD4, CD25, CD127, CD8, CD19, IgD, CD27, CD62L, Helios, FoxP3, CD56, and HLA-DR antibodies. CD4 + T, CD8 + T, CD4 + CD8 + T, CD4 + CD25 + CD127 low Treg, CD3 - CD19 + B, CD56 + NK cells, and CD14 + monocytes were quantified. Data are presented as mean ± SEM ( n = 6). p < 0.05 was considered statistically significant. p < 0.0001 by one-way ANOVA test.

Article Snippet: APC-eFluor 780-conjugated mouse anti-human CD45 antibody (Ab) (HI30), Alexa Fluor-532-conjugated mouse anti-human CD3 Ab (UCHT1), FITC-conjugated mouse anti-human CD3 Ab (UCHT1), PerCP-Cyanine5.5-conjugated mouse anti-human CD8a Ab (RTA-T8), Super Bright 600-conjugated mouse anti-human CD19 Ab (SJ25C1), PE-conjugated mouse anti-human CD25 Ab (BC96), PE-Cy5-conjugated mouse anti-human CD127 Ab (eBioRDR5), APC-conjugated mouse anti-human CD56 Ab (CMSSB), FITC-conjugated mouse anti-human CD16 Ab (eBioCB16(CB16)), PerCP-eFluor 710-conjugated mouse anti-human CD14 Ab (61D3), PE-Cy7-conjugated mouse anti-human HLA-DR Ab (LN3), and LIVE/DEAD™ fixable Blue dye were purchased from Thermo Fisher Scientific.

Techniques: Functional Assay, Ex Vivo, Comparison, Expressing, Staining, Flow Cytometry, Software

CD4 + CD25 + CD127 low UCB-Tregs increase IL-10 secretion and decrease inflammatory cytokines in co-culture with SLE-PBMCs. CD4 + CD25 + CD127 low UCB-Tregs, HD-PBMCs, SLE-PBMCs, or UCB-Tregs plus SLE-PBMC co-cultures (1:1) were seeded at 1 × 10 6 cells per well in six-well plates and were stimulated with Human T-Activator CD3/CD28 in a 1 cell:1 bead ratio in X-VIVO 15 medium supplemented with 10% FBS, 2 mM L-glutamine, 1% penicillin–streptomycin, and 1,000 IU/ml IL-2. After 3 days or 7 days, cell culture supernatants were collected for cytokine analysis. Production levels of human IL-10 (A) , IFN-γ (B) , IP-10 (C) , TNF-α (D) , IL-6 (E) , and IL-17A (F) in the cell culture supernatants were assessed using Human Cytokine ELISA kits according to manufacturer’s instructions and production levels of sCD40L (G) , IL-1α (H) , and IL-21 (I) were measured using human cytokine/chemokine 71-plex discovery assay array. Data are presented as mean ± SEM ( n = 2–3). p < 0.05 was considered statistically significant. * p < 0.05, ** p < 0.01; *** p < 0.001; **** p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests.

Journal: Frontiers in Immunology

Article Title: Allogeneic cord blood regulatory T cells decrease dsDNA antibody and improve albuminuria in systemic lupus erythematosus

doi: 10.3389/fimmu.2023.1217121

Figure Lengend Snippet: CD4 + CD25 + CD127 low UCB-Tregs increase IL-10 secretion and decrease inflammatory cytokines in co-culture with SLE-PBMCs. CD4 + CD25 + CD127 low UCB-Tregs, HD-PBMCs, SLE-PBMCs, or UCB-Tregs plus SLE-PBMC co-cultures (1:1) were seeded at 1 × 10 6 cells per well in six-well plates and were stimulated with Human T-Activator CD3/CD28 in a 1 cell:1 bead ratio in X-VIVO 15 medium supplemented with 10% FBS, 2 mM L-glutamine, 1% penicillin–streptomycin, and 1,000 IU/ml IL-2. After 3 days or 7 days, cell culture supernatants were collected for cytokine analysis. Production levels of human IL-10 (A) , IFN-γ (B) , IP-10 (C) , TNF-α (D) , IL-6 (E) , and IL-17A (F) in the cell culture supernatants were assessed using Human Cytokine ELISA kits according to manufacturer’s instructions and production levels of sCD40L (G) , IL-1α (H) , and IL-21 (I) were measured using human cytokine/chemokine 71-plex discovery assay array. Data are presented as mean ± SEM ( n = 2–3). p < 0.05 was considered statistically significant. * p < 0.05, ** p < 0.01; *** p < 0.001; **** p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests.

Techniques: Co-Culture Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Comparison

Single injection of CD4 + CD25 + CD127 low UCB-Tregs halts disease progression in SLE xenografts. (A) Schematic summary of single injection of CD4 + CD25 + CD127 low UCB-Treg cell therapy for SLE in a SLE xenograft model. Female Rag2 -/- γc -/- mice were transplanted with 3 × 10 6 human SLE-PBMCs by intravenous injection on day 0 and divided into two groups (Control-single and Treatment-single, n = 3 per group). Single injection of 10 × 10 6 ex vivo expanded CD4 + CD25 + CD127 low UCB-Tregs was administered through t.v. on day 7 after SLE-PBMCs injection. (B) CD4 + CD25 + CD127 low UCB-Tregs improve body weight in SLE xenografts. Change of body weight was monitored twice per week until termination of experiments. Data are presented as mean ± SEM ( n = 3). p < 0.05 by Student t- test was considered statistically significant. (C) CD4 + CD25 + CD127 low UCB-Tregs improves median survival days. Kaplan–Meier analysis was performed to estimate median survival times. (D) CD4 + CD25 + CD127 low UCB-Tregs decrease human CD45 + cells. Mouse PBMC was procured from Control-single and Treatment-single recipients and analyzed for CD45 + cells as measured by flow cytometry at the indicated time points. p < 0.05 by two-tailed unpaired Student t- test was considered statistically significant. (E) CD4 + CD25 + CD127 low UCB-Tregs decrease human CD8 + cells. Mouse PBMC was procured from Control-single and Treatment-single recipients and analyzed for CD8 + cells as measured by flow cytometry at the indicated time points. Data are presented as mean ± SEM. p -values were obtained using two-tailed unpaired t- test with 95% confidence interval for evaluation of statistical significance compared with the untreated controls. p < 0.05 was considered statistically significant. (F) Comparison of phenotypes between Control-single and Treatment-single recipients at 8 weeks post engraftment of SLE-PBMCs. p < 0.05 was considered statistically significant. p < 0.0001 by one-way ANOVA test. (G) Photograph of representative mouse Control-single and Treatment-single arm exhibiting skin changes. Representative H&E and CD8 staining of mouse-affected skin tissue sections from Control-single and Treatment single arm. (H) Photograph of spleen and representative H&E, CD3, CD4, CD8, CD20, and Ki67 staining of spleen tissue sections from representative mouse Control-single and Treatment-single arm. (I) Quantification analysis of positive cells and H-score of spleen tissue sections. Immunochemistry images of human CD3 + , CD4 + , CD8 + , CD20 + , and Ki67 + cells were analyzed at ×40 magnification using HALO 3.3 software. Quantification analysis of human CD3 + , CD4 + , CD8 + , CD20 + , and Ki67 + cells and H-scores for human CD3, CD4, CD8, CD20, and Ki67 positivity were calculated using HALO 3.3 software. Data are presented as mean ± SEM ( n = 3). p < 0.05 was considered statistically significant. **** p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests. (J) Single injection of CD4 + CD25 + CD127 low UCB-Tregs decreased renal inflammation in vivo . Representative H&E, CD3, CD4, CD8, CD20, and Ki67 staining of kidney tissue sections. (K) Quantification analysis of positive cells and H-score of kidney tissue sections. Immunochemistry images of human CD3 + , CD4 + , CD8 + , CD20 + , and Ki67 + cells were analyzed at ×40 magnification using HALO 3.3 software. Quantification analysis of human CD3 + , CD4 + , CD8 + , CD20 + , and Ki67 + cells and H-scores for human CD3, CD4, CD8, CD20, and Ki67 positivity were calculated using HALO 3.3 software. Data are presented as mean ± SEM (n=5). p < 0.05 was considered statistically significant. (L) Single injection of CD4 + CD25 + CD127 low UCB-Tregs decreased albuminuria in SLE xenografts. Expression levels of urinary albumin, creatinine, and albumin/creatinine. Data are presented as mean ± SEM ( n = 6). p < 0.05 was considered statistically significant. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests.

Journal: Frontiers in Immunology

Article Title: Allogeneic cord blood regulatory T cells decrease dsDNA antibody and improve albuminuria in systemic lupus erythematosus

doi: 10.3389/fimmu.2023.1217121

Figure Lengend Snippet: Single injection of CD4 + CD25 + CD127 low UCB-Tregs halts disease progression in SLE xenografts. (A) Schematic summary of single injection of CD4 + CD25 + CD127 low UCB-Treg cell therapy for SLE in a SLE xenograft model. Female Rag2 -/- γc -/- mice were transplanted with 3 × 10 6 human SLE-PBMCs by intravenous injection on day 0 and divided into two groups (Control-single and Treatment-single, n = 3 per group). Single injection of 10 × 10 6 ex vivo expanded CD4 + CD25 + CD127 low UCB-Tregs was administered through t.v. on day 7 after SLE-PBMCs injection. (B) CD4 + CD25 + CD127 low UCB-Tregs improve body weight in SLE xenografts. Change of body weight was monitored twice per week until termination of experiments. Data are presented as mean ± SEM ( n = 3). p < 0.05 by Student t- test was considered statistically significant. (C) CD4 + CD25 + CD127 low UCB-Tregs improves median survival days. Kaplan–Meier analysis was performed to estimate median survival times. (D) CD4 + CD25 + CD127 low UCB-Tregs decrease human CD45 + cells. Mouse PBMC was procured from Control-single and Treatment-single recipients and analyzed for CD45 + cells as measured by flow cytometry at the indicated time points. p < 0.05 by two-tailed unpaired Student t- test was considered statistically significant. (E) CD4 + CD25 + CD127 low UCB-Tregs decrease human CD8 + cells. Mouse PBMC was procured from Control-single and Treatment-single recipients and analyzed for CD8 + cells as measured by flow cytometry at the indicated time points. Data are presented as mean ± SEM. p -values were obtained using two-tailed unpaired t- test with 95% confidence interval for evaluation of statistical significance compared with the untreated controls. p < 0.05 was considered statistically significant. (F) Comparison of phenotypes between Control-single and Treatment-single recipients at 8 weeks post engraftment of SLE-PBMCs. p < 0.05 was considered statistically significant. p < 0.0001 by one-way ANOVA test. (G) Photograph of representative mouse Control-single and Treatment-single arm exhibiting skin changes. Representative H&E and CD8 staining of mouse-affected skin tissue sections from Control-single and Treatment single arm. (H) Photograph of spleen and representative H&E, CD3, CD4, CD8, CD20, and Ki67 staining of spleen tissue sections from representative mouse Control-single and Treatment-single arm. (I) Quantification analysis of positive cells and H-score of spleen tissue sections. Immunochemistry images of human CD3 + , CD4 + , CD8 + , CD20 + , and Ki67 + cells were analyzed at ×40 magnification using HALO 3.3 software. Quantification analysis of human CD3 + , CD4 + , CD8 + , CD20 + , and Ki67 + cells and H-scores for human CD3, CD4, CD8, CD20, and Ki67 positivity were calculated using HALO 3.3 software. Data are presented as mean ± SEM ( n = 3). p < 0.05 was considered statistically significant. **** p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests. (J) Single injection of CD4 + CD25 + CD127 low UCB-Tregs decreased renal inflammation in vivo . Representative H&E, CD3, CD4, CD8, CD20, and Ki67 staining of kidney tissue sections. (K) Quantification analysis of positive cells and H-score of kidney tissue sections. Immunochemistry images of human CD3 + , CD4 + , CD8 + , CD20 + , and Ki67 + cells were analyzed at ×40 magnification using HALO 3.3 software. Quantification analysis of human CD3 + , CD4 + , CD8 + , CD20 + , and Ki67 + cells and H-scores for human CD3, CD4, CD8, CD20, and Ki67 positivity were calculated using HALO 3.3 software. Data are presented as mean ± SEM (n=5). p < 0.05 was considered statistically significant. (L) Single injection of CD4 + CD25 + CD127 low UCB-Tregs decreased albuminuria in SLE xenografts. Expression levels of urinary albumin, creatinine, and albumin/creatinine. Data are presented as mean ± SEM ( n = 6). p < 0.05 was considered statistically significant. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests.

Techniques: Injection, Ex Vivo, Flow Cytometry, Two Tailed Test, Comparison, Staining, Software, In Vivo, Expressing

Multiple injections of CD4 + CD25 + CD127 low UCB-Tregs increase Tregs, decrease CD8 + T cells, and improve skin inflammation in the SLE xenogeneic model. (A) Schematic summary of multiple CD4 + CD25 + CD127 low UCB-Treg cell therapy for SLE in a SLE xenograft model. Female Rag2 -/- γc -/- mice were transplanted with 3 × 10 6 human SLE-PBMCs by intravenous injection. After mice displayed human immune cells, they were divided into two groups (control and treatment, n = 7 mice/group), and 10 × 10 6 ex vivo expanded CD4 + CD25 + CD127 low UCB-Tregs were infused into SLE xenografts intravenously on day 30, day 35, day 45, and day 51 for treatment. (B) Sustained decrease in PB human CD45 + cells in CD4 + CD25 + CD127 low UCB-Treg recipients. Mouse PBMC was procured from Control-multiple and Treatment-multiple recipients and analyzed for CD45 + cells by flow cytometry at the indicated time points. Data are presented as mean ± SEM ( n = 7). p < 0.05 was considered statistically significant. ** p < 0.01 by two-way ANOVA with Tukey multiple comparison tests. (C) Sustained increase in PB human CD4 + CD25 high CD127 low Treg cells in CD4 + CD25 + CD127 low UCB-Treg recipients. Mouse PBMC was procured from Control-multiple and Treatment-multiple recipients and analyzed for CD4 + CD25 high CD127 low Treg cells by flow cytometry at the indicated time points. Data are presented as mean ± SEM ( n = 7). p -values were obtained using two-way analysis of variance (ANOVA) with Tukey multiple comparison test. p < 0.05 was considered statistically significant. **** p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests. (D) CD4 + CD25 + CD127 low UCB-Tregs decrease CD8 + T, CD45 + , and CD3 + T cells in multiple organs. At the time of euthanasia, PB and organs from Control-multiple and Treatment-multiple recipients were harvested and liquified and analyzed for human CD8 + T cells, CD45 + cells, and CD3 + T cells by flow cytometry at the indicated time points. Data are presented as mean ± SEM ( n = 5-7). p < 0.05 by Student t- test was considered statistically significant. (E) CD4 + CD25 + CD127 low UCB-Tregs decrease skin disease burden in SLE xenografts. Photographs of Control-multiple (upper panel) and Treatment-multiple (lower panel) were compared at 12 weeks. (F) Representative H&E staining of mouse skin tissue sections of Control-multiple (epidermal ulceration, subdermal lymphocyte infiltrate, disruption of hair follicles, and loss of subdermal adipose tissue) and Treatment-multiple (epidermal layer intact and clear visualization of the subdermal layers including hair follicles and adipose tissue).

Journal: Frontiers in Immunology

Article Title: Allogeneic cord blood regulatory T cells decrease dsDNA antibody and improve albuminuria in systemic lupus erythematosus

doi: 10.3389/fimmu.2023.1217121

Figure Lengend Snippet: Multiple injections of CD4 + CD25 + CD127 low UCB-Tregs increase Tregs, decrease CD8 + T cells, and improve skin inflammation in the SLE xenogeneic model. (A) Schematic summary of multiple CD4 + CD25 + CD127 low UCB-Treg cell therapy for SLE in a SLE xenograft model. Female Rag2 -/- γc -/- mice were transplanted with 3 × 10 6 human SLE-PBMCs by intravenous injection. After mice displayed human immune cells, they were divided into two groups (control and treatment, n = 7 mice/group), and 10 × 10 6 ex vivo expanded CD4 + CD25 + CD127 low UCB-Tregs were infused into SLE xenografts intravenously on day 30, day 35, day 45, and day 51 for treatment. (B) Sustained decrease in PB human CD45 + cells in CD4 + CD25 + CD127 low UCB-Treg recipients. Mouse PBMC was procured from Control-multiple and Treatment-multiple recipients and analyzed for CD45 + cells by flow cytometry at the indicated time points. Data are presented as mean ± SEM ( n = 7). p < 0.05 was considered statistically significant. ** p < 0.01 by two-way ANOVA with Tukey multiple comparison tests. (C) Sustained increase in PB human CD4 + CD25 high CD127 low Treg cells in CD4 + CD25 + CD127 low UCB-Treg recipients. Mouse PBMC was procured from Control-multiple and Treatment-multiple recipients and analyzed for CD4 + CD25 high CD127 low Treg cells by flow cytometry at the indicated time points. Data are presented as mean ± SEM ( n = 7). p -values were obtained using two-way analysis of variance (ANOVA) with Tukey multiple comparison test. p < 0.05 was considered statistically significant. **** p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests. (D) CD4 + CD25 + CD127 low UCB-Tregs decrease CD8 + T, CD45 + , and CD3 + T cells in multiple organs. At the time of euthanasia, PB and organs from Control-multiple and Treatment-multiple recipients were harvested and liquified and analyzed for human CD8 + T cells, CD45 + cells, and CD3 + T cells by flow cytometry at the indicated time points. Data are presented as mean ± SEM ( n = 5-7). p < 0.05 by Student t- test was considered statistically significant. (E) CD4 + CD25 + CD127 low UCB-Tregs decrease skin disease burden in SLE xenografts. Photographs of Control-multiple (upper panel) and Treatment-multiple (lower panel) were compared at 12 weeks. (F) Representative H&E staining of mouse skin tissue sections of Control-multiple (epidermal ulceration, subdermal lymphocyte infiltrate, disruption of hair follicles, and loss of subdermal adipose tissue) and Treatment-multiple (epidermal layer intact and clear visualization of the subdermal layers including hair follicles and adipose tissue).

Techniques: Injection, Ex Vivo, Flow Cytometry, Comparison, Staining, Disruption

CD4 + CD25 + CD127 low UCB-Tregs decrease tissue inflammation in SLE xenografts. To assess pathology and cellular infiltrates in the tissue, human CD3 + cells, CD4 + T cells, CD8 + T cells, CD20 + B cells, or Ki67 + cells in the spleen (A, B) and liver (C, D) of untreated (Control-multiple) and treated mice (Treatment-multiple) were detected by immunohistochemistry. Spleen (A) and liver (C) tissues were harvested, fixed with 10% buffered formalin, and embedded in paraffin for processing into 5-μm tissue sections. To assess pathology and cellular infiltrates in the tissues, de-paraffinized and rehydrated tissue sections were stained with Hematoxylin & Eosin. For immunohistochemistry, de-paraffinized and rehydrated tissue sections were subjected to heat-mediated antigen retrieval with sodium citrate buffer (pH 6), permeabilization, and blocking prior to staining with primary antibodies against human CD3, CD4, CD8, CD20, and Ki-67, and appropriate horseradish peroxidase-conjugated secondary antibodies were used to determine the subset of human cells in the mouse tissues. Immunochemistry images of human CD3 + , CD4 + , CD8 + , CD20 + , and Ki67 + cells were analyzed at ×40 magnification using HALO 3.3 software in spleen (B) and liver (D) . Quantification analysis of human CD3 + , CD4 + , CD8 + , CD20 + , and Ki67 + cells and H-scores for human CD3, CD4, CD8, CD20, and Ki67 positivity were calculated using HALO 3.3 software. Data are presented as mean ± SEM ( n = 6). p < 0.05 was considered statistically significant. p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests.

Journal: Frontiers in Immunology

Article Title: Allogeneic cord blood regulatory T cells decrease dsDNA antibody and improve albuminuria in systemic lupus erythematosus

doi: 10.3389/fimmu.2023.1217121

Figure Lengend Snippet: CD4 + CD25 + CD127 low UCB-Tregs decrease tissue inflammation in SLE xenografts. To assess pathology and cellular infiltrates in the tissue, human CD3 + cells, CD4 + T cells, CD8 + T cells, CD20 + B cells, or Ki67 + cells in the spleen (A, B) and liver (C, D) of untreated (Control-multiple) and treated mice (Treatment-multiple) were detected by immunohistochemistry. Spleen (A) and liver (C) tissues were harvested, fixed with 10% buffered formalin, and embedded in paraffin for processing into 5-μm tissue sections. To assess pathology and cellular infiltrates in the tissues, de-paraffinized and rehydrated tissue sections were stained with Hematoxylin & Eosin. For immunohistochemistry, de-paraffinized and rehydrated tissue sections were subjected to heat-mediated antigen retrieval with sodium citrate buffer (pH 6), permeabilization, and blocking prior to staining with primary antibodies against human CD3, CD4, CD8, CD20, and Ki-67, and appropriate horseradish peroxidase-conjugated secondary antibodies were used to determine the subset of human cells in the mouse tissues. Immunochemistry images of human CD3 + , CD4 + , CD8 + , CD20 + , and Ki67 + cells were analyzed at ×40 magnification using HALO 3.3 software in spleen (B) and liver (D) . Quantification analysis of human CD3 + , CD4 + , CD8 + , CD20 + , and Ki67 + cells and H-scores for human CD3, CD4, CD8, CD20, and Ki67 positivity were calculated using HALO 3.3 software. Data are presented as mean ± SEM ( n = 6). p < 0.05 was considered statistically significant. p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests.

Techniques: Immunohistochemistry, Staining, Blocking Assay, Software, Comparison

CD4 + CD25 + CD127 low UCB-Tregs decrease anti-dsDNA IgG Ab and improve renal function in SLE xenografts. (A) Multiple injections of CD4 + CD25 + CD127 low UCB-Tregs decrease anti-human dsDNA Ab. Plasma samples were collected from the EDTA-treated PB of untreated (Control-multiple) and CD4 + CD25 + CD127 low UCB-Tregs-injected mice (Treatment-multiple). Levels of anti-human dsDNA IgG Ab were measured using the Abnova assay kit. Data are presented as mean ± SEM ( n = 3). P -values were obtained using two-tailed unpaired t- test with 95% confidence interval for evaluation of statistical significance compared with the untreated controls. P < 0.05 was considered statistically significant. (B) Representative H&E, CD3, CD4, CD8, CD20, and Ki67 staining of kidney tissue sections of Control-multiple (upper panel) and Treatment-multiple (lower panel). (C) Quantification analysis of human CD3 + , CD4 + , CD8 + , CD20 + , and Ki67 + cells and H-scores for human CD3, CD4, CD8, CD20, and Ki67 positivity. Total positive cells and H-score were calculated using HALO 3.3 software. Data are presented as mean ± SEM ( n = 6). p < 0.05 was considered statistically significant. **** p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests. (D) CD4 + CD25 + CD127 low UCB-Tregs decrease albuminuria in SLE xenografts. At the time of euthanasia, at 12 weeks, urine samples were collected from Control-multiple and Treatment-multiple and levels of urinary albumin were assessed using the Exocell Albumin M assay kit. Data are presented as mean ± SEM ( n = 4). p -values were obtained using two-tailed unpaired t- test with 95% confidence interval for evaluation of statistical significance compared with the untreated controls. p < 0.05 was considered statistically significant.

Journal: Frontiers in Immunology

Article Title: Allogeneic cord blood regulatory T cells decrease dsDNA antibody and improve albuminuria in systemic lupus erythematosus

doi: 10.3389/fimmu.2023.1217121

Figure Lengend Snippet: CD4 + CD25 + CD127 low UCB-Tregs decrease anti-dsDNA IgG Ab and improve renal function in SLE xenografts. (A) Multiple injections of CD4 + CD25 + CD127 low UCB-Tregs decrease anti-human dsDNA Ab. Plasma samples were collected from the EDTA-treated PB of untreated (Control-multiple) and CD4 + CD25 + CD127 low UCB-Tregs-injected mice (Treatment-multiple). Levels of anti-human dsDNA IgG Ab were measured using the Abnova assay kit. Data are presented as mean ± SEM ( n = 3). P -values were obtained using two-tailed unpaired t- test with 95% confidence interval for evaluation of statistical significance compared with the untreated controls. P < 0.05 was considered statistically significant. (B) Representative H&E, CD3, CD4, CD8, CD20, and Ki67 staining of kidney tissue sections of Control-multiple (upper panel) and Treatment-multiple (lower panel). (C) Quantification analysis of human CD3 + , CD4 + , CD8 + , CD20 + , and Ki67 + cells and H-scores for human CD3, CD4, CD8, CD20, and Ki67 positivity. Total positive cells and H-score were calculated using HALO 3.3 software. Data are presented as mean ± SEM ( n = 6). p < 0.05 was considered statistically significant. **** p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests. (D) CD4 + CD25 + CD127 low UCB-Tregs decrease albuminuria in SLE xenografts. At the time of euthanasia, at 12 weeks, urine samples were collected from Control-multiple and Treatment-multiple and levels of urinary albumin were assessed using the Exocell Albumin M assay kit. Data are presented as mean ± SEM ( n = 4). p -values were obtained using two-tailed unpaired t- test with 95% confidence interval for evaluation of statistical significance compared with the untreated controls. p < 0.05 was considered statistically significant.

Techniques: Injection, Two Tailed Test, Staining, Software, Comparison

CD4 + CD25 + CD127 low UCB-Tregs decrease systemic inflammation in SLE xenografts. Plasma samples were collected from the EDTA-treated PB of untreated (Control-multiple) and CD4 + CD25 + CD127 low UCB-Tregs-infused mice (Treatment-multiple). Levels of human IFN-γ, IP-10, TNF-α, IL-17A, sCD40L, and IL-1α in the plasma of SLE xenografts were measured using the Human cytokine 42-plex Discovery assay kit. Data are presented as mean ± SEM ( n = 5). p < 0.05 by F -test was considered statistically significant. * p < 0.05, ** p < 0.01, **** p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests.

Journal: Frontiers in Immunology

Article Title: Allogeneic cord blood regulatory T cells decrease dsDNA antibody and improve albuminuria in systemic lupus erythematosus

doi: 10.3389/fimmu.2023.1217121

Figure Lengend Snippet: CD4 + CD25 + CD127 low UCB-Tregs decrease systemic inflammation in SLE xenografts. Plasma samples were collected from the EDTA-treated PB of untreated (Control-multiple) and CD4 + CD25 + CD127 low UCB-Tregs-infused mice (Treatment-multiple). Levels of human IFN-γ, IP-10, TNF-α, IL-17A, sCD40L, and IL-1α in the plasma of SLE xenografts were measured using the Human cytokine 42-plex Discovery assay kit. Data are presented as mean ± SEM ( n = 5). p < 0.05 by F -test was considered statistically significant. * p < 0.05, ** p < 0.01, **** p < 0.0001 by two-way ANOVA with Tukey multiple comparison tests.

Techniques: Comparison

GO‐Y030 controls metabolism of Tregs. A and B, Reactive oxygen species (ROS) expression in CD4 + CD25 + Tregs cultured with 1 μmol/L curcumin, 0.25 μmol/L GO‐Y030, or DMSO for 72 h. Data were pooled from four independent experiments. C, Oxygen consumption rate (OCR) was measured under basal conditions or following the addition of oligomycin, carbonylcyanide p‐(trifluoromethoxy) phenylhydrazone (FCCP), or rotenone and antimycin A. D‐G, Quantification of basal respiration (D), ATP production (E), maximal respiration (F), and proton leak (G) using the rate of OCR. Data were pooled for three independent experiments. One‐way ANOVA with post hoc Tukey's multiple comparison test was applied. Data shown are representative of three independent experiments. H and I, Western blotting of cultured Tregs with or without 0.25 µM GO‐Y030 for 72 h. Representative pictures at three independent experiments (H). Data were pooled at three independent experiments (n = 4, mean +standard deviation) (I). The relative expression of p‐S6 or S6 (/GAPDH) in DMSO Tregs was set as “1”

Journal: Cancer Science

Article Title: Curcumin analog GO‐Y030 boosts the efficacy of anti‐PD‐1 cancer immunotherapy

doi: 10.1111/cas.15136

Figure Lengend Snippet: GO‐Y030 controls metabolism of Tregs. A and B, Reactive oxygen species (ROS) expression in CD4 + CD25 + Tregs cultured with 1 μmol/L curcumin, 0.25 μmol/L GO‐Y030, or DMSO for 72 h. Data were pooled from four independent experiments. C, Oxygen consumption rate (OCR) was measured under basal conditions or following the addition of oligomycin, carbonylcyanide p‐(trifluoromethoxy) phenylhydrazone (FCCP), or rotenone and antimycin A. D‐G, Quantification of basal respiration (D), ATP production (E), maximal respiration (F), and proton leak (G) using the rate of OCR. Data were pooled for three independent experiments. One‐way ANOVA with post hoc Tukey's multiple comparison test was applied. Data shown are representative of three independent experiments. H and I, Western blotting of cultured Tregs with or without 0.25 µM GO‐Y030 for 72 h. Representative pictures at three independent experiments (H). Data were pooled at three independent experiments (n = 4, mean +standard deviation) (I). The relative expression of p‐S6 or S6 (/GAPDH) in DMSO Tregs was set as “1”

Article Snippet: PE‐conjugated, APC‐conjugated, or Pacific Blue–conjugated anti‐mouse Foxp3 antibody (FJK‐16S), FITC‐conjugated anti‐human TCR (IP26), PE‐conjugated anti‐human CD25 (BC96), PerCP‐Cyanine5.5–conjugated anti‐human CD4 (OKT4), eFluoro 660–conjugated anti‐human Foxp3 (PCH101), PE‐conjugated anti‐mouse PD‐1 (J43), and anti‐IFN‐γ (R4‐6A2) were purchased from eBioscience.

Techniques: Expressing, Cell Culture, Comparison, Western Blot, Standard Deviation

GO‐Y030 boosts levels of immune checkpoint inhibitors. A and B, Mean fluorescence intensity of PD‐1 in cultured Tregs (gated CD4 + Foxp3 + Zombie) in the presence of 10 ng/mL IL‐2 with or without 1.0 µmol/L curcumin or 0.25 μmol/L GO‐Y030 for 72 h. Data are representative of at least three independent experiments. C, Proliferation ratio of CFSE‐labeled CD8 + T cells isolated from CD45.1 mice and cultured with or without CD4 + CD25 + Tregs for 48 h (CD8 + T cells : CD4 + CD25 + Tregs = 1:0.5). Tregs were treated with 0.25 µmol/L GO‐Y030 or DMSO control for 3 d prior to the coculturing. The CD8 + CD45.1 + gated cell population is as shown. D, Relative suppressive ability of anti‐PD‐1 antibody. The percentage of suppression ability with control antibody in DMSO or GO‐Y030 was set as “1.” White: control antibody (10 μg/mL), black: anti‐PD‐1 (10 μg/mL). The horizontal bars represent the mean. Data were pooled from three independent experiments (n = 3, mean + standard deviation). E, Calculation of tumor volume (mm 3 ) in each day after 7 d of tumor injection (n = 7‐10, mean + standard deviation). Data were pooled from three independent experiments. F, Representative tumors in each group harvested at the end of experiments, as in (E). G, Immunohistochemistry of Foxp3 levels in tumor sites. Representative pictures from three independent experiments (n = 6). Scale bar, 50 µm. The original magnification is 40×. H, Absolute number of Foxp3 + Tregs in tumor‐infiltrating lymphocytes. One‐way ANOVA with post hoc Turkey's multiple comparison test was used (E, G, H). The graph shows mean and standard deviation

Journal: Cancer Science

Article Title: Curcumin analog GO‐Y030 boosts the efficacy of anti‐PD‐1 cancer immunotherapy

doi: 10.1111/cas.15136

Figure Lengend Snippet: GO‐Y030 boosts levels of immune checkpoint inhibitors. A and B, Mean fluorescence intensity of PD‐1 in cultured Tregs (gated CD4 + Foxp3 + Zombie) in the presence of 10 ng/mL IL‐2 with or without 1.0 µmol/L curcumin or 0.25 μmol/L GO‐Y030 for 72 h. Data are representative of at least three independent experiments. C, Proliferation ratio of CFSE‐labeled CD8 + T cells isolated from CD45.1 mice and cultured with or without CD4 + CD25 + Tregs for 48 h (CD8 + T cells : CD4 + CD25 + Tregs = 1:0.5). Tregs were treated with 0.25 µmol/L GO‐Y030 or DMSO control for 3 d prior to the coculturing. The CD8 + CD45.1 + gated cell population is as shown. D, Relative suppressive ability of anti‐PD‐1 antibody. The percentage of suppression ability with control antibody in DMSO or GO‐Y030 was set as “1.” White: control antibody (10 μg/mL), black: anti‐PD‐1 (10 μg/mL). The horizontal bars represent the mean. Data were pooled from three independent experiments (n = 3, mean + standard deviation). E, Calculation of tumor volume (mm 3 ) in each day after 7 d of tumor injection (n = 7‐10, mean + standard deviation). Data were pooled from three independent experiments. F, Representative tumors in each group harvested at the end of experiments, as in (E). G, Immunohistochemistry of Foxp3 levels in tumor sites. Representative pictures from three independent experiments (n = 6). Scale bar, 50 µm. The original magnification is 40×. H, Absolute number of Foxp3 + Tregs in tumor‐infiltrating lymphocytes. One‐way ANOVA with post hoc Turkey's multiple comparison test was used (E, G, H). The graph shows mean and standard deviation

Techniques: Fluorescence, Cell Culture, Labeling, Isolation, Control, Standard Deviation, Injection, Immunohistochemistry, Comparison

Journal: Immunity

Article Title: Longitudinal analysis reveals that delayed bystander CD8+ T cell activation and early immune pathology distinguish severe COVID-19 from mild disease

doi: 10.1016/j.immuni.2021.05.010

Figure Lengend Snippet:

Article Snippet: Anti-human CD25 PE (BC96) , Thermo , RRID: AB_1659682.

Techniques: Staining, Recombinant, Membrane, Enzyme-linked Immunosorbent Assay, Cell Counting, Flow Cytometry, Software, Enzyme-linked Immunospot, Cytometry, Luminex

Journal: Med (New York, N.y.)

Article Title: Mucosal-associated invariant T cell responses differ by sex in COVID-19

doi: 10.1016/j.medj.2021.04.008

Figure Lengend Snippet:

Article Snippet: PE anti-huamn CD25 (clone BC96) , ThermoFisher , Cat# 12-0259-42.

Techniques: Recombinant, Staining, Flow Cytometry, Software

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