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CLL alters gut bacterial diversity and generates a dysbiotic microbiome compared with WT. A, Study schematic illustrating transgenic <t>Eμ-TCL1</t> and WT B6 mice were monitored with serial blood and fecal pellet sample collection over 12 months. DNA isolated from fecal pellets was subject to 16S rRNA sequencing at the indicated time points. At study end, mice were sacrificed, and tissues were harvested ( n = 7–13 mice/genotype). B, CLL disease burden was measured by the percentage of CD45 + /CD19 + /CD5 + cells in the peripheral blood via flow cytometric analysis. Asterisks denote the significance between Eμ-TCL1 mice over disease course (*, P < 0.05; **, P < 0.01). Hashtags denote the significance between WT B6 and Eμ-TCL1 mice at each time point ( ## , P < 0.01; ### , P < 0.001). Two-way ANOVA with Dunnett multiple comparisons was applied for testing. C, Alpha diversity measured by the Shannon diversity index in transgenic Eμ-TCL1 mice compared with age-matched WT B6 mice at 7, 10, and 12 months (*, P < 0.05). Data are presented as box-and-whisker plots, with box edges representing the 25th and 75th percentiles, the center line showing the median value, and whiskers extending to minimum and maximum values. Asterisks denote the significance between WT B6 and Eμ-TCL1 mice at each time point. Unpaired Welch t test was applied for testing. D, PCoA plot depicting Bray–Curtis distance as a measure of beta diversity between transgenic Eμ-TCL1 mice (blue) and age-matched WT B6 mice (red) at all time points (4 – 12 months). E, Alpha diversity measured by the Shannon diversity index in transgenic Eμ-TCL1 mice over time (4–12 months). Data are presented as box-and-whisker plots, with box edges representing the 25th and 75th percentiles, the center line showing the median value, and whiskers extending to minimum and maximum values. F, PCoA plot depicting Bray–Curtis distance as a measure of beta diversity in transgenic Eμ-TCL1 mice over time (4–12 months). G, Relative abundance bar plots of bacterial taxa present in the gut microbiome of Eμ-TCL1 and WT B6 mice over 12 months. Sequences with a quality score of Q28 and higher were assembled for paired-end sequencing using default parameters of Phredd33 encoding and included in the analysis. Per these parameters, a single 4-month WT B6 sample met this criterion and was retained during this step in the analysis. Additional WT B6 samples in the three subsequent time points (7, 10, and 12 months) passed this quality control check and as such were included. Remainder includes all remaining taxa present in the microbiome at decreased abundance.

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1) Product Images from "Gut Microbiome Profiling in Eμ-TCL1 Mice Reveals Intestinal Changes and a Dysbiotic Signature Specific to Chronic Lymphocytic Leukemia"

Article Title: Gut Microbiome Profiling in Eμ-TCL1 Mice Reveals Intestinal Changes and a Dysbiotic Signature Specific to Chronic Lymphocytic Leukemia

Journal: Cancer Research Communications

doi: 10.1158/2767-9764.CRC-25-0022

CLL alters gut bacterial diversity and generates a dysbiotic microbiome compared with WT. A, Study schematic illustrating transgenic Eμ-TCL1 and WT B6 mice were monitored with serial blood and fecal pellet sample collection over 12 months. DNA isolated from fecal pellets was subject to 16S rRNA sequencing at the indicated time points. At study end, mice were sacrificed, and tissues were harvested ( n = 7–13 mice/genotype). B, CLL disease burden was measured by the percentage of CD45 + /CD19 + /CD5 + cells in the peripheral blood via flow cytometric analysis. Asterisks denote the significance between Eμ-TCL1 mice over disease course (*, P < 0.05; **, P < 0.01). Hashtags denote the significance between WT B6 and Eμ-TCL1 mice at each time point ( ## , P < 0.01; ### , P < 0.001). Two-way ANOVA with Dunnett multiple comparisons was applied for testing. C, Alpha diversity measured by the Shannon diversity index in transgenic Eμ-TCL1 mice compared with age-matched WT B6 mice at 7, 10, and 12 months (*, P < 0.05). Data are presented as box-and-whisker plots, with box edges representing the 25th and 75th percentiles, the center line showing the median value, and whiskers extending to minimum and maximum values. Asterisks denote the significance between WT B6 and Eμ-TCL1 mice at each time point. Unpaired Welch t test was applied for testing. D, PCoA plot depicting Bray–Curtis distance as a measure of beta diversity between transgenic Eμ-TCL1 mice (blue) and age-matched WT B6 mice (red) at all time points (4 – 12 months). E, Alpha diversity measured by the Shannon diversity index in transgenic Eμ-TCL1 mice over time (4–12 months). Data are presented as box-and-whisker plots, with box edges representing the 25th and 75th percentiles, the center line showing the median value, and whiskers extending to minimum and maximum values. F, PCoA plot depicting Bray–Curtis distance as a measure of beta diversity in transgenic Eμ-TCL1 mice over time (4–12 months). G, Relative abundance bar plots of bacterial taxa present in the gut microbiome of Eμ-TCL1 and WT B6 mice over 12 months. Sequences with a quality score of Q28 and higher were assembled for paired-end sequencing using default parameters of Phredd33 encoding and included in the analysis. Per these parameters, a single 4-month WT B6 sample met this criterion and was retained during this step in the analysis. Additional WT B6 samples in the three subsequent time points (7, 10, and 12 months) passed this quality control check and as such were included. Remainder includes all remaining taxa present in the microbiome at decreased abundance.

Figure Legend Snippet: CLL alters gut bacterial diversity and generates a dysbiotic microbiome compared with WT. A, Study schematic illustrating transgenic Eμ-TCL1 and WT B6 mice were monitored with serial blood and fecal pellet sample collection over 12 months. DNA isolated from fecal pellets was subject to 16S rRNA sequencing at the indicated time points. At study end, mice were sacrificed, and tissues were harvested ( n = 7–13 mice/genotype). B, CLL disease burden was measured by the percentage of CD45 + /CD19 + /CD5 + cells in the peripheral blood via flow cytometric analysis. Asterisks denote the significance between Eμ-TCL1 mice over disease course (*, P < 0.05; **, P < 0.01). Hashtags denote the significance between WT B6 and Eμ-TCL1 mice at each time point ( ## , P < 0.01; ### , P < 0.001). Two-way ANOVA with Dunnett multiple comparisons was applied for testing. C, Alpha diversity measured by the Shannon diversity index in transgenic Eμ-TCL1 mice compared with age-matched WT B6 mice at 7, 10, and 12 months (*, P < 0.05). Data are presented as box-and-whisker plots, with box edges representing the 25th and 75th percentiles, the center line showing the median value, and whiskers extending to minimum and maximum values. Asterisks denote the significance between WT B6 and Eμ-TCL1 mice at each time point. Unpaired Welch t test was applied for testing. D, PCoA plot depicting Bray–Curtis distance as a measure of beta diversity between transgenic Eμ-TCL1 mice (blue) and age-matched WT B6 mice (red) at all time points (4 – 12 months). E, Alpha diversity measured by the Shannon diversity index in transgenic Eμ-TCL1 mice over time (4–12 months). Data are presented as box-and-whisker plots, with box edges representing the 25th and 75th percentiles, the center line showing the median value, and whiskers extending to minimum and maximum values. F, PCoA plot depicting Bray–Curtis distance as a measure of beta diversity in transgenic Eμ-TCL1 mice over time (4–12 months). G, Relative abundance bar plots of bacterial taxa present in the gut microbiome of Eμ-TCL1 and WT B6 mice over 12 months. Sequences with a quality score of Q28 and higher were assembled for paired-end sequencing using default parameters of Phredd33 encoding and included in the analysis. Per these parameters, a single 4-month WT B6 sample met this criterion and was retained during this step in the analysis. Additional WT B6 samples in the three subsequent time points (7, 10, and 12 months) passed this quality control check and as such were included. Remainder includes all remaining taxa present in the microbiome at decreased abundance.

Techniques Used: Transgenic Assay, Isolation, Sequencing, Whisker Assay, Control

Compositional analysis of microbial communities as CLL disease progresses. A, LEfSe analysis demonstrating enrichment of specific bacterial taxa at the phylum level in transgenic Eμ-TCL1 mice ( n = 12) and WT B6 mice ( n = 10) at 12 months of age. LDA, linear discriminant analysis. B, Predicted microbial function pathways at the phylum level in transgenic Eμ-TCL1 mice and WT B6 mice at 12 months based on 16S rRNA sequencing data analyzed using ggPICRUSt2. ECM, extracellular matrix.

Figure Legend Snippet: Compositional analysis of microbial communities as CLL disease progresses. A, LEfSe analysis demonstrating enrichment of specific bacterial taxa at the phylum level in transgenic Eμ-TCL1 mice ( n = 12) and WT B6 mice ( n = 10) at 12 months of age. LDA, linear discriminant analysis. B, Predicted microbial function pathways at the phylum level in transgenic Eμ-TCL1 mice and WT B6 mice at 12 months based on 16S rRNA sequencing data analyzed using ggPICRUSt2. ECM, extracellular matrix.

Techniques Used: Transgenic Assay, Sequencing

Disturbances in the intestinal tract of Eμ-TCL1 mice with advanced CLL disease. A, Representative hematoxylin and eosin images of small intestine tissue in WT B6 and Eμ-TCL1 mice at 12 months ( n = 7–13 mice/genotype). Hematoxylin and eosin stains: scale bar, 100 μm (left and middle) and 200 pixels (right). B and C, Representative images of small intestine (serial sections) from 12-month-old WT B6 and Eμ-TCL1 mice ( n = 4–5 mice/genotype) stained with antibody against human TCL1 ( B ) and murine CD19 ( C ). Histoscore data are presented as violin plots illustrating the empirical distribution of data. The black, dashed line represents the median. Asterisks denote the significance of the histoscores between WT B6 and Eμ-TCL1 mice (***, P < 0.001). Unpaired Welch t test was applied for testing.

Figure Legend Snippet: Disturbances in the intestinal tract of Eμ-TCL1 mice with advanced CLL disease. A, Representative hematoxylin and eosin images of small intestine tissue in WT B6 and Eμ-TCL1 mice at 12 months ( n = 7–13 mice/genotype). Hematoxylin and eosin stains: scale bar, 100 μm (left and middle) and 200 pixels (right). B and C, Representative images of small intestine (serial sections) from 12-month-old WT B6 and Eμ-TCL1 mice ( n = 4–5 mice/genotype) stained with antibody against human TCL1 ( B ) and murine CD19 ( C ). Histoscore data are presented as violin plots illustrating the empirical distribution of data. The black, dashed line represents the median. Asterisks denote the significance of the histoscores between WT B6 and Eμ-TCL1 mice (***, P < 0.001). Unpaired Welch t test was applied for testing.

Techniques Used: Staining

Differential expression of markers pertaining to intestinal integrity and inflammation in WT B6 and Eμ-TCL1 mice. ELISA was used to assess the concentration of tight junction markers in plasma of 12-month-old WT B6 and Eμ-TCL1 mice: CLDN2 ( A ), CLDN3 ( B ), zonulin ( C ), and sCD14 ( D ). Asterisks denote significance between WT B6 and Eμ-TCL1 mice (**, P < 0.01; ***, P < 0.001). Unpaired Welch t test was applied for testing.

Figure Legend Snippet: Differential expression of markers pertaining to intestinal integrity and inflammation in WT B6 and Eμ-TCL1 mice. ELISA was used to assess the concentration of tight junction markers in plasma of 12-month-old WT B6 and Eμ-TCL1 mice: CLDN2 ( A ), CLDN3 ( B ), zonulin ( C ), and sCD14 ( D ). Asterisks denote significance between WT B6 and Eμ-TCL1 mice (**, P < 0.01; ***, P < 0.001). Unpaired Welch t test was applied for testing.

Techniques Used: Quantitative Proteomics, Enzyme-linked Immunosorbent Assay, Concentration Assay, Clinical Proteomics

Dysbiotic microbiome is observed in the aggressive syngeneic model of CLL. A, Study schematic illustrating WT B6 mice were engrafted intravenously via the tail vein with 1e 7 spleen-derived lymphocytes from a moribund Eμ-TCL1 mouse (CLL engraftment). Engrafted mice ( n = 10) were monitored with serial blood and fecal pellet sample collections over the study course of 7 weeks. B, CLL disease burden was measured by the percentage of CD45 + /CD19 + /CD5 + cells in the peripheral blood via flow cytometric analysis starting 1 week after engraftment. Asterisks denote the significance between leukemic mice over time (***, P < 0.001). One-way ANOVA with Dunnett multiple comparisons was applied for testing. C, Alpha diversity measured by the Shannon index in leukemic mice over 7 weeks. Data are presented as box-and-whisker plots, with box edges representing the 25th and 75th percentiles, the center line showing the median value, and whiskers extending to minimum and maximum values. One-way ANOVA with Dunnett multiple comparisons was applied for testing. D, PCoA plot depicting Bray–Curis distance as a measure of beta diversity in leukemic mice over 7 weeks. E, Relative abundance bar plots of bacterial taxa present in the gut microbiome of diseased mice over 7 weeks following engraftment. Unassigned includes all remaining taxa present in the microbiome at decreased abundance. F, LEfSe analysis demonstrating enrichment of specific bacterial taxa at the phylum level in leukemic mice at the time of engraftment and 1 and 3 weeks after engraftment. Eng./engraft., engraftment. LDA, linear discriminant analysis.

Figure Legend Snippet: Dysbiotic microbiome is observed in the aggressive syngeneic model of CLL. A, Study schematic illustrating WT B6 mice were engrafted intravenously via the tail vein with 1e 7 spleen-derived lymphocytes from a moribund Eμ-TCL1 mouse (CLL engraftment). Engrafted mice ( n = 10) were monitored with serial blood and fecal pellet sample collections over the study course of 7 weeks. B, CLL disease burden was measured by the percentage of CD45 + /CD19 + /CD5 + cells in the peripheral blood via flow cytometric analysis starting 1 week after engraftment. Asterisks denote the significance between leukemic mice over time (***, P < 0.001). One-way ANOVA with Dunnett multiple comparisons was applied for testing. C, Alpha diversity measured by the Shannon index in leukemic mice over 7 weeks. Data are presented as box-and-whisker plots, with box edges representing the 25th and 75th percentiles, the center line showing the median value, and whiskers extending to minimum and maximum values. One-way ANOVA with Dunnett multiple comparisons was applied for testing. D, PCoA plot depicting Bray–Curis distance as a measure of beta diversity in leukemic mice over 7 weeks. E, Relative abundance bar plots of bacterial taxa present in the gut microbiome of diseased mice over 7 weeks following engraftment. Unassigned includes all remaining taxa present in the microbiome at decreased abundance. F, LEfSe analysis demonstrating enrichment of specific bacterial taxa at the phylum level in leukemic mice at the time of engraftment and 1 and 3 weeks after engraftment. Eng./engraft., engraftment. LDA, linear discriminant analysis.

Techniques Used: Derivative Assay, Whisker Assay

Antibiotic-mediated microflora ablation delays CLL development, fostering a similar dysbiotic microbiome. A, WT B6 mice were randomly assigned to either receive an antibiotic cocktail (Abx) or normal drinking water (water). Five days before CLL engraftment, mice receiving antibiotics were gavaged with a high-dose Abx. In parallel, mice in the antibiotic arm received antibiotics in drinking water until study end (left). Control mice were gavaged with similar volumes of water and received normal drinking water until study end (left). Following the initial 5-day oral gavage, a baseline fecal pellet sample was obtained, and CLL was initiated via adoptive transfer of Eμ-TCL1 spleen-derived lymphocytes (right; CLL engraftment). Study arm mice were housed in separate cages and monitored over 7 weeks with serial sampling of blood to monitor CLL burden and fecal pellet sample collection for microbiome profiling ( n = 10 mice/cohort). Spleens were harvest at 9 weeks after engraftment for further immunophenotyping. B, CLL disease burden was measured by the percentage of CD45 + /CD19 + /CD5 + cells in the peripheral blood via flow cytometric analysis beginning at 1 week after engraftment ( n = 10 mice/cohort). Asterisks denote the significance between antibiotic-receiving or water-receiving mice over time (*, P < 0.05). Hashtags denote the significance between antibiotic-receiving and water-receiving mice at each time point (#, P < 0.05; ##, P < 0.01). Two-way ANOVA with Dunnett multiple comparisons was applied for testing. C–F, Abundances of cell types found in the blood of antibiotic-receiving and water-receiving mice: non-malignant B cells (CD45 + /CD19 + /CD5 − ; C ) T cells (CD45 + /CD3 + ; D ) monocytic-like MDSCs (M-MDSCs; CD45 + /B220 − /CD3 − /CD11b + /Ly6C + /Ly6G − ; E ), and granulocytic-like MDSCs (G-MDSCs; CD45 + /B220−/CD3 − /CD11b + /Ly6C lo /Ly6G + ; F ). Asterisks denote the significance between antibiotic-receiving and water-receiving mice over time (*, P < 0.05). Unpaired Welch t test was applied for testing. G, PCoA plot depicting Bray–Curtis distance as a measure of beta diversity between Abx (blue) and water (red) arms. H, Alpha diversity measured by the Shannon index in antibiotic and water cohorts at time of engraftment and 1, 3, 5, and 7 weeks after engraftment. Data are presented as box-and-whisker plots, with box edges representing the 25th and 75th percentiles, the center line showing the median value, and whiskers extending to minimum and maximum values. Asterisks denote the significance between antibiotic-receiving and water-receiving mice at each time point (***, P < 0.001). Unpaired Welch t test was applied for testing. I, Relative abundance bar plots of bacterial taxa present at the phylum level in the gut microbiome of antibiotic-receiving leukemic mice and water-receiving leukemic mice. Remainder includes all remaining taxa present in the microbiome at decreased abundance. Engraft., engraftment; MDSCs, myeloid-derived suppressor cells; post-eng., after engraftment.

Figure Legend Snippet: Antibiotic-mediated microflora ablation delays CLL development, fostering a similar dysbiotic microbiome. A, WT B6 mice were randomly assigned to either receive an antibiotic cocktail (Abx) or normal drinking water (water). Five days before CLL engraftment, mice receiving antibiotics were gavaged with a high-dose Abx. In parallel, mice in the antibiotic arm received antibiotics in drinking water until study end (left). Control mice were gavaged with similar volumes of water and received normal drinking water until study end (left). Following the initial 5-day oral gavage, a baseline fecal pellet sample was obtained, and CLL was initiated via adoptive transfer of Eμ-TCL1 spleen-derived lymphocytes (right; CLL engraftment). Study arm mice were housed in separate cages and monitored over 7 weeks with serial sampling of blood to monitor CLL burden and fecal pellet sample collection for microbiome profiling ( n = 10 mice/cohort). Spleens were harvest at 9 weeks after engraftment for further immunophenotyping. B, CLL disease burden was measured by the percentage of CD45 + /CD19 + /CD5 + cells in the peripheral blood via flow cytometric analysis beginning at 1 week after engraftment ( n = 10 mice/cohort). Asterisks denote the significance between antibiotic-receiving or water-receiving mice over time (*, P < 0.05). Hashtags denote the significance between antibiotic-receiving and water-receiving mice at each time point (#, P < 0.05; ##, P < 0.01). Two-way ANOVA with Dunnett multiple comparisons was applied for testing. C–F, Abundances of cell types found in the blood of antibiotic-receiving and water-receiving mice: non-malignant B cells (CD45 + /CD19 + /CD5 − ; C ) T cells (CD45 + /CD3 + ; D ) monocytic-like MDSCs (M-MDSCs; CD45 + /B220 − /CD3 − /CD11b + /Ly6C + /Ly6G − ; E ), and granulocytic-like MDSCs (G-MDSCs; CD45 + /B220−/CD3 − /CD11b + /Ly6C lo /Ly6G + ; F ). Asterisks denote the significance between antibiotic-receiving and water-receiving mice over time (*, P < 0.05). Unpaired Welch t test was applied for testing. G, PCoA plot depicting Bray–Curtis distance as a measure of beta diversity between Abx (blue) and water (red) arms. H, Alpha diversity measured by the Shannon index in antibiotic and water cohorts at time of engraftment and 1, 3, 5, and 7 weeks after engraftment. Data are presented as box-and-whisker plots, with box edges representing the 25th and 75th percentiles, the center line showing the median value, and whiskers extending to minimum and maximum values. Asterisks denote the significance between antibiotic-receiving and water-receiving mice at each time point (***, P < 0.001). Unpaired Welch t test was applied for testing. I, Relative abundance bar plots of bacterial taxa present at the phylum level in the gut microbiome of antibiotic-receiving leukemic mice and water-receiving leukemic mice. Remainder includes all remaining taxa present in the microbiome at decreased abundance. Engraft., engraftment; MDSCs, myeloid-derived suppressor cells; post-eng., after engraftment.

Techniques Used: Control, Adoptive Transfer Assay, Derivative Assay, Sampling, Whisker Assay

Image Search Results

Journal: Cancer Research Communications

Article Title: Gut Microbiome Profiling in Eμ-TCL1 Mice Reveals Intestinal Changes and a Dysbiotic Signature Specific to Chronic Lymphocytic Leukemia

doi: 10.1158/2767-9764.CRC-25-0022

Figure Lengend Snippet: CLL alters gut bacterial diversity and generates a dysbiotic microbiome compared with WT. A, Study schematic illustrating transgenic Eμ-TCL1 and WT B6 mice were monitored with serial blood and fecal pellet sample collection over 12 months. DNA isolated from fecal pellets was subject to 16S rRNA sequencing at the indicated time points. At study end, mice were sacrificed, and tissues were harvested ( n = 7–13 mice/genotype). B, CLL disease burden was measured by the percentage of CD45 + /CD19 + /CD5 + cells in the peripheral blood via flow cytometric analysis. Asterisks denote the significance between Eμ-TCL1 mice over disease course (*, P < 0.05; **, P < 0.01). Hashtags denote the significance between WT B6 and Eμ-TCL1 mice at each time point ( ## , P < 0.01; ### , P < 0.001). Two-way ANOVA with Dunnett multiple comparisons was applied for testing. C, Alpha diversity measured by the Shannon diversity index in transgenic Eμ-TCL1 mice compared with age-matched WT B6 mice at 7, 10, and 12 months (*, P < 0.05). Data are presented as box-and-whisker plots, with box edges representing the 25th and 75th percentiles, the center line showing the median value, and whiskers extending to minimum and maximum values. Asterisks denote the significance between WT B6 and Eμ-TCL1 mice at each time point. Unpaired Welch t test was applied for testing. D, PCoA plot depicting Bray–Curtis distance as a measure of beta diversity between transgenic Eμ-TCL1 mice (blue) and age-matched WT B6 mice (red) at all time points (4 – 12 months). E, Alpha diversity measured by the Shannon diversity index in transgenic Eμ-TCL1 mice over time (4–12 months). Data are presented as box-and-whisker plots, with box edges representing the 25th and 75th percentiles, the center line showing the median value, and whiskers extending to minimum and maximum values. F, PCoA plot depicting Bray–Curtis distance as a measure of beta diversity in transgenic Eμ-TCL1 mice over time (4–12 months). G, Relative abundance bar plots of bacterial taxa present in the gut microbiome of Eμ-TCL1 and WT B6 mice over 12 months. Sequences with a quality score of Q28 and higher were assembled for paired-end sequencing using default parameters of Phredd33 encoding and included in the analysis. Per these parameters, a single 4-month WT B6 sample met this criterion and was retained during this step in the analysis. Additional WT B6 samples in the three subsequent time points (7, 10, and 12 months) passed this quality control check and as such were included. Remainder includes all remaining taxa present in the microbiome at decreased abundance.

Article Snippet: Next, the tissue sections were incubated overnight with primary antibody [anti-human TCL1 (CST4042S) 1:400 or anti-mouse CD19 ( CST90176 ) 1:100; Cell Signaling Technologies] prepared in 2.5% normal horse serum.

Techniques: Transgenic Assay, Isolation, Sequencing, Whisker Assay, Control

Journal: Cancer Research Communications

Article Title: Gut Microbiome Profiling in Eμ-TCL1 Mice Reveals Intestinal Changes and a Dysbiotic Signature Specific to Chronic Lymphocytic Leukemia

doi: 10.1158/2767-9764.CRC-25-0022

Figure Lengend Snippet: Compositional analysis of microbial communities as CLL disease progresses. A, LEfSe analysis demonstrating enrichment of specific bacterial taxa at the phylum level in transgenic Eμ-TCL1 mice ( n = 12) and WT B6 mice ( n = 10) at 12 months of age. LDA, linear discriminant analysis. B, Predicted microbial function pathways at the phylum level in transgenic Eμ-TCL1 mice and WT B6 mice at 12 months based on 16S rRNA sequencing data analyzed using ggPICRUSt2. ECM, extracellular matrix.

Techniques: Transgenic Assay, Sequencing

Journal: Cancer Research Communications

Article Title: Gut Microbiome Profiling in Eμ-TCL1 Mice Reveals Intestinal Changes and a Dysbiotic Signature Specific to Chronic Lymphocytic Leukemia

doi: 10.1158/2767-9764.CRC-25-0022

Figure Lengend Snippet: Disturbances in the intestinal tract of Eμ-TCL1 mice with advanced CLL disease. A, Representative hematoxylin and eosin images of small intestine tissue in WT B6 and Eμ-TCL1 mice at 12 months ( n = 7–13 mice/genotype). Hematoxylin and eosin stains: scale bar, 100 μm (left and middle) and 200 pixels (right). B and C, Representative images of small intestine (serial sections) from 12-month-old WT B6 and Eμ-TCL1 mice ( n = 4–5 mice/genotype) stained with antibody against human TCL1 ( B ) and murine CD19 ( C ). Histoscore data are presented as violin plots illustrating the empirical distribution of data. The black, dashed line represents the median. Asterisks denote the significance of the histoscores between WT B6 and Eμ-TCL1 mice (***, P < 0.001). Unpaired Welch t test was applied for testing.

Techniques: Staining

Journal: Cancer Research Communications

Article Title: Gut Microbiome Profiling in Eμ-TCL1 Mice Reveals Intestinal Changes and a Dysbiotic Signature Specific to Chronic Lymphocytic Leukemia

doi: 10.1158/2767-9764.CRC-25-0022

Figure Lengend Snippet: Differential expression of markers pertaining to intestinal integrity and inflammation in WT B6 and Eμ-TCL1 mice. ELISA was used to assess the concentration of tight junction markers in plasma of 12-month-old WT B6 and Eμ-TCL1 mice: CLDN2 ( A ), CLDN3 ( B ), zonulin ( C ), and sCD14 ( D ). Asterisks denote significance between WT B6 and Eμ-TCL1 mice (**, P < 0.01; ***, P < 0.001). Unpaired Welch t test was applied for testing.

Techniques: Quantitative Proteomics, Enzyme-linked Immunosorbent Assay, Concentration Assay, Clinical Proteomics

Journal: Cancer Research Communications

Article Title: Gut Microbiome Profiling in Eμ-TCL1 Mice Reveals Intestinal Changes and a Dysbiotic Signature Specific to Chronic Lymphocytic Leukemia

doi: 10.1158/2767-9764.CRC-25-0022

Figure Lengend Snippet: Dysbiotic microbiome is observed in the aggressive syngeneic model of CLL. A, Study schematic illustrating WT B6 mice were engrafted intravenously via the tail vein with 1e 7 spleen-derived lymphocytes from a moribund Eμ-TCL1 mouse (CLL engraftment). Engrafted mice ( n = 10) were monitored with serial blood and fecal pellet sample collections over the study course of 7 weeks. B, CLL disease burden was measured by the percentage of CD45 + /CD19 + /CD5 + cells in the peripheral blood via flow cytometric analysis starting 1 week after engraftment. Asterisks denote the significance between leukemic mice over time (***, P < 0.001). One-way ANOVA with Dunnett multiple comparisons was applied for testing. C, Alpha diversity measured by the Shannon index in leukemic mice over 7 weeks. Data are presented as box-and-whisker plots, with box edges representing the 25th and 75th percentiles, the center line showing the median value, and whiskers extending to minimum and maximum values. One-way ANOVA with Dunnett multiple comparisons was applied for testing. D, PCoA plot depicting Bray–Curis distance as a measure of beta diversity in leukemic mice over 7 weeks. E, Relative abundance bar plots of bacterial taxa present in the gut microbiome of diseased mice over 7 weeks following engraftment. Unassigned includes all remaining taxa present in the microbiome at decreased abundance. F, LEfSe analysis demonstrating enrichment of specific bacterial taxa at the phylum level in leukemic mice at the time of engraftment and 1 and 3 weeks after engraftment. Eng./engraft., engraftment. LDA, linear discriminant analysis.

Techniques: Derivative Assay, Whisker Assay

Journal: Cancer Research Communications

Article Title: Gut Microbiome Profiling in Eμ-TCL1 Mice Reveals Intestinal Changes and a Dysbiotic Signature Specific to Chronic Lymphocytic Leukemia

doi: 10.1158/2767-9764.CRC-25-0022

Figure Lengend Snippet: Antibiotic-mediated microflora ablation delays CLL development, fostering a similar dysbiotic microbiome. A, WT B6 mice were randomly assigned to either receive an antibiotic cocktail (Abx) or normal drinking water (water). Five days before CLL engraftment, mice receiving antibiotics were gavaged with a high-dose Abx. In parallel, mice in the antibiotic arm received antibiotics in drinking water until study end (left). Control mice were gavaged with similar volumes of water and received normal drinking water until study end (left). Following the initial 5-day oral gavage, a baseline fecal pellet sample was obtained, and CLL was initiated via adoptive transfer of Eμ-TCL1 spleen-derived lymphocytes (right; CLL engraftment). Study arm mice were housed in separate cages and monitored over 7 weeks with serial sampling of blood to monitor CLL burden and fecal pellet sample collection for microbiome profiling ( n = 10 mice/cohort). Spleens were harvest at 9 weeks after engraftment for further immunophenotyping. B, CLL disease burden was measured by the percentage of CD45 + /CD19 + /CD5 + cells in the peripheral blood via flow cytometric analysis beginning at 1 week after engraftment ( n = 10 mice/cohort). Asterisks denote the significance between antibiotic-receiving or water-receiving mice over time (*, P < 0.05). Hashtags denote the significance between antibiotic-receiving and water-receiving mice at each time point (#, P < 0.05; ##, P < 0.01). Two-way ANOVA with Dunnett multiple comparisons was applied for testing. C–F, Abundances of cell types found in the blood of antibiotic-receiving and water-receiving mice: non-malignant B cells (CD45 + /CD19 + /CD5 − ; C ) T cells (CD45 + /CD3 + ; D ) monocytic-like MDSCs (M-MDSCs; CD45 + /B220 − /CD3 − /CD11b + /Ly6C + /Ly6G − ; E ), and granulocytic-like MDSCs (G-MDSCs; CD45 + /B220−/CD3 − /CD11b + /Ly6C lo /Ly6G + ; F ). Asterisks denote the significance between antibiotic-receiving and water-receiving mice over time (*, P < 0.05). Unpaired Welch t test was applied for testing. G, PCoA plot depicting Bray–Curtis distance as a measure of beta diversity between Abx (blue) and water (red) arms. H, Alpha diversity measured by the Shannon index in antibiotic and water cohorts at time of engraftment and 1, 3, 5, and 7 weeks after engraftment. Data are presented as box-and-whisker plots, with box edges representing the 25th and 75th percentiles, the center line showing the median value, and whiskers extending to minimum and maximum values. Asterisks denote the significance between antibiotic-receiving and water-receiving mice at each time point (***, P < 0.001). Unpaired Welch t test was applied for testing. I, Relative abundance bar plots of bacterial taxa present at the phylum level in the gut microbiome of antibiotic-receiving leukemic mice and water-receiving leukemic mice. Remainder includes all remaining taxa present in the microbiome at decreased abundance. Engraft., engraftment; MDSCs, myeloid-derived suppressor cells; post-eng., after engraftment.

Techniques: Control, Adoptive Transfer Assay, Derivative Assay, Sampling, Whisker Assay

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