Journal: medRxiv

Article Title: Anomalous epithelial variations and ectopic inflammatory response in chronic obstructive pulmonary disease

doi: 10.1101/2020.12.03.20242412

Figure Lengend Snippet: A. Representative images immunostained with CCL2, TNC, and CXCL8 with AT2 cell markers (SFTPC or ABCA3) in COPD and never-smoker lung sections. B . Violin plots for COL4A1, COL4A2, and COL4A3 in AT1 clusters. C. Violin plots for FKBP5 in selected cell types. a) present dataset; b) combined publicly-available datasets.

Article Snippet: Antibodies and reagents were as follows: anti-PD-L1 (rabbit, 1:1000, #ab205921, Abcam, Cambridge, UK); anti-TNC (rabbit, 1:100, #HPA004823, Sigma-Aldrich); anti-CCL2 (rabbit, 1:100, #HPA019163, Sigma-Aldrich); anti-RAGE (rabbit, 1:1000, #ab216329, Abcam); anti-SFTPC (rabbit, 1:1000, #HPA010928, Sigma-Aldrich); anti-ABCA3 (mouse, 1:1000, #WMAB-ABCA3-17, Seven Hills Bioreagents, OH, USA); anti- macrophage inflammatory protein 3 alpha (rabbit, 1:1000, #ab224188, Abcam); anti- CXCL1 (mouse, 1:100, #MAB275, R&D Systems, MN, USA); anti-CXCL8 (mouse, 1:100, #MAB208, R&D Systems); and Hoechst 33242 (1:1000, #H342, Sigma-Aldrich).

Techniques:

Journal: medRxiv

Article Title: Anomalous epithelial variations and ectopic inflammatory response in chronic obstructive pulmonary disease

doi: 10.1101/2020.12.03.20242412

Figure Lengend Snippet: A. 100% stacked bar charts of the percentage of cell populations for AT1 subtype clusters, AT2 subtype clusters, and basal lineage clusters. B. Bar charts displaying the percentages of subpopulations such as AT1-B, AT2-A, AT2-B, club, goblet, and basal clusters across patient states. C. a UMAP plot of epithelial cells focusing on the AT2-C cluster (left). The cell population of the AT2-C cluster (right). The y-axis represents the ratio of cells in the AT2-C cluster across patient states. Brackets () represent the percentage of cells of the AT2-C cluster based on epithelial cells in each of the patient states. D. Violin plots of representative inflammatory-related genes displaying the AT2-C (iAT2) cluster. E. Violin plots for CXCL1 and CXCL8 in AT2 cells in combined publicly-available datasets. F. Representative images immunostained with CD274 (PD-L1), CXCL1 and CXCL20 with AT2 cell markers (SFTPC or ABCA3) in COPD and never-smoker lung sections. Scale bars, 25 μm (left), 12.5 μm (right).

Article Snippet: Antibodies and reagents were as follows: anti-PD-L1 (rabbit, 1:1000, #ab205921, Abcam, Cambridge, UK); anti-TNC (rabbit, 1:100, #HPA004823, Sigma-Aldrich); anti-CCL2 (rabbit, 1:100, #HPA019163, Sigma-Aldrich); anti-RAGE (rabbit, 1:1000, #ab216329, Abcam); anti-SFTPC (rabbit, 1:1000, #HPA010928, Sigma-Aldrich); anti-ABCA3 (mouse, 1:1000, #WMAB-ABCA3-17, Seven Hills Bioreagents, OH, USA); anti- macrophage inflammatory protein 3 alpha (rabbit, 1:1000, #ab224188, Abcam); anti- CXCL1 (mouse, 1:100, #MAB275, R&D Systems, MN, USA); anti-CXCL8 (mouse, 1:100, #MAB208, R&D Systems); and Hoechst 33242 (1:1000, #H342, Sigma-Aldrich).

Techniques:

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Concomitant NAFLD Facilitates Liver Metastases and PD-1-Refractory by Recruiting MDSCs via CXCL5/CXCR2 in Colorectal Cancer

doi: 10.1016/j.jcmgh.2024.04.008

Figure Lengend Snippet: NAFLD promotes CXCL5 and recruits CXCR2 + MDSCs accumulation in liver metastases. ( A ) Relative CXCL5 level in peripheral blood of each tumor-bearing C57BL/6 mice models, n = 4. Student t test. ( B ) Quantification of CXCL5 concentration in peripheral blood of mice in each group, n = 5. One-way ANOVA. ( C ) Quantification of CXCL5 concentration in liver homogenate of mice in each group, n = 4. One-way ANOVA. ( D ) Quantification of CXCL5 concentration in tumor tissues of each tumor-bearing C57BL/6 mice models, n = 3. Student t test. ( E ) UMAP visualization of liver metastatic tissues, paratumoral tissues, and liver tissues profiled in all groups ( F ) colored by sample or subpopulation ( red rectangles represent granulocytes), n = 6. ( G ) Graph of cellular identification results, UMAP cluster map revealing 8 specific clusters representing the major tissue types. ( H ) UMAP cluster map showing CXCR2 expression of all samples’ cell types. Violet dots give the fields of the main clusters of interest. ( I ) Representative histograms of CXCR2 expression in MDSCs (CD45 + CD11b + Gr-1 + ) and percentages (%) of CXCR2 expression in each group were analyzed by FC, n = 6, one-way ANOVA. All data are presented as mean ± SD. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001, and ns, not significant.

Article Snippet: Rabbit anti-human CXCR2 , N/A , Proteintech; 20634-1-AP.

Techniques: Concentration Assay, Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Concomitant NAFLD Facilitates Liver Metastases and PD-1-Refractory by Recruiting MDSCs via CXCL5/CXCR2 in Colorectal Cancer

doi: 10.1016/j.jcmgh.2024.04.008

Figure Lengend Snippet: NAFLD promotes CXCL5 and recruits CXCR2 + MDSCs accumulation in liver metastases. ( A ) Mouse XL Cytokine Array detects multiple cytokines, chemokines, growth factors, and other soluble proteins in serum. Data shown are from a 5-minute exposure to Bio-Rad, top: CRLM group, bottom : NAFLD with CRLM group. ( B ) Relative cytokine levels in serum of each tumor-bearing C57BL/6 mice models. ( C ) Localization of CXCL5 and CXCR2 reactivity in tissue samples of ob/ob and WT group, acquired by confocal imaging. Scale bars represent 100 μm, upper panel ; 20 μm, lower panel .

Article Snippet: Rabbit anti-human CXCR2 , N/A , Proteintech; 20634-1-AP.

Techniques: Imaging

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Concomitant NAFLD Facilitates Liver Metastases and PD-1-Refractory by Recruiting MDSCs via CXCL5/CXCR2 in Colorectal Cancer

doi: 10.1016/j.jcmgh.2024.04.008

Figure Lengend Snippet: CXCR2 + MDSCs are recruited to liver via CXCL5 secreted by Kupffer cells. ( A ) Cell clusters of CXCL5 source. Representative images of mIF for CXCL5 and candidate clusters (CD146/SMA/CD31/F4/80) expression in the liver. Scale bars represent 50 μm. ( B ) Representative images of immunohistochemical staining for CXCL5 and CXCR2 expression in the liver. Scale bars represent 50 μm. Mean intensity of CXCL5 and CXCR2 expression in the livers was analyzed by TG tissue imaging cytometry system, n = 4, one-way ANOVA. ( C ) Correlation between the mean intensity of CXCL5 and CXCR2 in each group. Pearson test. ( D ) Localization of CXCL5 and CXCR2 reactivity in tissue samples of each group, acquired by confocal imaging. Scale bars represent 100 μm, upper panel ; 20 μm, lower panel . ( E ) Macroscopic views of representative livers are shown of CXCL5 KO+NAFLD+CRLM and CXCL5 KO+CRLM groups. Yellow arrow indicates tumors. ( F ) Liver metastasis foci number of CXCL5 KO+NAFLD+CRLM and CXCL5 KO+CRLM groups, n = 4–5, Student t test. ( G ) Representative data show that CXCL5 KO+NAFLD+CRLM and CXCL5 KO+CRLM groups of CD45 + /CD11b + /Gr-1 + MDSCs expression on day 12 in the liver, which percentages (%) of MDSCs (CD45 + C11b + Gr-1 + ) in CD45 + cells were analyzed by FC, n = 4–5, Student t test. ( H ) Percentages (%) of CXCR2 expression in MDSCs (CD45 + CD11b + Gr-1 + ) in each group were analyzed by flow cytometry, n = 4–5, Student t test. All data are presented as mean ± SD. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001, and ns, not significant.

Article Snippet: Rabbit anti-human CXCR2 , N/A , Proteintech; 20634-1-AP.

Techniques: Expressing, Immunohistochemical staining, Staining, Imaging, Cytometry, Flow Cytometry

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Concomitant NAFLD Facilitates Liver Metastases and PD-1-Refractory by Recruiting MDSCs via CXCL5/CXCR2 in Colorectal Cancer

doi: 10.1016/j.jcmgh.2024.04.008

Figure Lengend Snippet: CXCR2 + MDSCs are recruited to liver via CXCL5 secreted by Kupffer cells. ( A ) Cell clusters of CXCL5 source were analyzed by Akoya system. One-way ANOVA. ( B ) Liver weights of CXCL5 KO+NAFLD+CRLM and CXCL5 KO+CRLM groups, n = 4–5, Student test. ( C ) Liver/body weight ratio of CXCL5 KO+NAFLD+CRLM and CXCL5 KO+CRLM groups, n = 4–5, Student t test. ( D ) Liver weights of CXCL5 KO+NAFLD+CRLM and WT+NAFLD+CRLM groups, n = 4–5, Student test. ( E ) Liver/body weight ratio of CXCL5 KO+NAFLD+CRLM and WT+NAFLD+CRLM groups, n = 4–5, Student t test. ( F ) Liver metastasis foci number of CXCL5 KO+NAFLD+CRLM and WT+NAFLD+CRLM groups, n = 4–5, Student t test. ( G ) Representative data show that CXCL5 KO+NAFLD+CRLM and WT+NAFLD+CRLM groups of CD45 + /CD11b + /Gr-1 + MDSCs expression on day 12 in the liver, which percentages (%) of MDSCs (CD45 + C11b + Gr-1 + ) in CD45 + cells were analyzed by FC, n = 4–5, Student t test. ( H ) Percentages (%) of CXCR2 expression in MDSCs (CD45 + CD11b + Gr-1 + ) in each group were analyzed by FC, n = 4–5, Student t test. ( I ) Localization of CXCL5 and CXCR2 reactivity in tissue samples of CXCL5 KO+NAFLD+CRLM and CXCL5 KO+CRLM groups, acquired by confocal imaging. Scale bars represent 100 μm, upper panel ; 20 μm, lower panel . All data are presented as mean ± SD. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001, and ns, not significant.

Article Snippet: Rabbit anti-human CXCR2 , N/A , Proteintech; 20634-1-AP.

Techniques: Expressing, Imaging

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Concomitant NAFLD Facilitates Liver Metastases and PD-1-Refractory by Recruiting MDSCs via CXCL5/CXCR2 in Colorectal Cancer

doi: 10.1016/j.jcmgh.2024.04.008

Figure Lengend Snippet: Reparixin inhibits CXCR2 + MDSC infiltration and CRLM growth. ( A ) Treatment experimental scheme. After Luci-MC38 cell inoculation, PBS or Reparixin was injected at days 3, 5, 7, 9, and 11, Mice were euthanized on day 12 for analyses. ( B ) In vivo bioluminescence imaging of the 4 groups (CRLM, CRLM+Reparixin, NAFLD+CRLM, and NAFLD+CRLM+Reparixin) at indicated time points of day 6. ( C ) Macroscopic views of representative livers are shown of 4 groups (CRLM, CRLM+Reparixin, NAFLD+CRLM, and NAFLD+CRLM+Reparixin). Yellow arrow indicates tumors. ( D ) Liver weights of each group. Samples were collected on day 12 after Luciferase-MC38 cell inoculation and treatment; n = 5–6. One-way ANOVA. ( E ) Liver/body weight ratio of each group; n = 5–6. One-way ANOVA. ( F ) Representative data show that each group of CD45 + /CD11b + /Gr-1 + MDSCs expression on day 12 in the liver, which percentages (%) of MDSCs (CD45 + C11b + Gr-1 + ) in CD45 + cells were analyzed by FC. One-way ANOVA. ( G ) Representative histograms of CXCR2 expression in MDSCs (CD45 + CD11b + Gr-1 + ) and percentages (%) of CXCR2 expression in each group were analyzed by FC. One-way ANOVA. ( H ) Co-localization of CXCL5 and CXCR2 reactivity in tissue samples of liver metastases, NAFLD with liver metastasis, and their separately treated liver tissues, acquired by confocal imaging. Scale bars represent 100 μm, upper panel ; 20 μm, lower panel , n = 6. ( I ) Therapeutic effects of Reparixin-21d on NAFLD. Macroscopic views of representative liver tissues, Oil Red O staining, and immunohistochemistry staining of liver sections for each group. Scale bars represent 100 μm. Mean density of immunohistochemical staining in the Reparixin-21d on NAFLD, n = 4, Student t test. All data are presented as mean ± SD. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001, and ns, not significant.

Article Snippet: Rabbit anti-human CXCR2 , N/A , Proteintech; 20634-1-AP.

Techniques: Injection, In Vivo, Imaging, Luciferase, Expressing, Staining, Immunohistochemistry, Immunohistochemical staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Concomitant NAFLD Facilitates Liver Metastases and PD-1-Refractory by Recruiting MDSCs via CXCL5/CXCR2 in Colorectal Cancer

doi: 10.1016/j.jcmgh.2024.04.008

Figure Lengend Snippet: Reparixin inhibits CXCR2 + MDSC infiltration and CRLM growth. ( A ) Differential therapeutic effects of Reparixin-12d or 15d on NAFLD. Macroscopic views of representative liver tissues and Oil Red O staining of liver sections for each group. Scale bars represent 50 μm. Oil Red O staining positive area rate of NAFLD and the 12d ( B ) or 15d ( C ) treatment group, n = 4, Student t test. All data are presented as mean ± SD. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001, and ns, not significant.

Article Snippet: Rabbit anti-human CXCR2 , N/A , Proteintech; 20634-1-AP.

Techniques: Staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Concomitant NAFLD Facilitates Liver Metastases and PD-1-Refractory by Recruiting MDSCs via CXCL5/CXCR2 in Colorectal Cancer

doi: 10.1016/j.jcmgh.2024.04.008

Figure Lengend Snippet: Reparixin overcomes anti-PD-1 treatment refractory and achieves long-term survival over half mice with CRLM. ( A ) Scheme of combination treatment with or without antibody to PD-1. ∗Treatment intraperitoneally, after Luci-MC38 cell inoculation, isotype-IgG, anti-PD-1 antibodies, Reparixin were injected every other day starting on day 3, with the combination treatment group also received at these time points. ( B ) Body weight trends of mice in the combination treatment group. n = 9. ( C ) Representative liver tissues stained with 5 markers (CD8, programmed death-ligand 1 [PD-L1], CXCL5, CXCR2, and CD11b). The image acquisition of all markers occurs simultaneously. Individual markers (or select combinations of markers) can then be displayed. Five-color overlay ( top left ) and 2/3-color insets (border) of a representative sampling of the simultaneously acquired markers. Scale bars represent 20 μm. Top : PD-1 treatment group, bottom : Reparixin treatment group. ( D ) Quantification of CD8 + PD-L1 + lymphocytes by AKOYA, n = 3/group, 5 views, one-way ANOVA. ( E ) Quantification of CXCR2 + CD11b + cluster by AKOYA, n = 3/group, 3 views, one-way ANOVA. All data are presented as mean ± SD. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001, and ns, not significant.

Article Snippet: Rabbit anti-human CXCR2 , N/A , Proteintech; 20634-1-AP.

Techniques: Injection, Staining, Sampling

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Concomitant NAFLD Facilitates Liver Metastases and PD-1-Refractory by Recruiting MDSCs via CXCL5/CXCR2 in Colorectal Cancer

doi: 10.1016/j.jcmgh.2024.04.008

Figure Lengend Snippet: Reparixin overcomes anti-PD-1 treatment refractory and achieves long-term survival over half mice with CRLM. ( A ) In vivo bioluminescence imaging of each treatment group at indicated time points, n = 6–9. ( B ) In vivo bioluminescence imaging of the combination treatment group at day 12 and day 58, n = 6. ( C ) Body weight trends of NAFLD with CRLM mice treated with different protocols, n = 6–9. ( D ) Survival curves of NAFLD+CRLM mice, n = 6–9. Log-rank (Mantel-Cox) test compared with the group of NAFLD+CRLM+isotype-IgG. ( E ) Survival curves of CRLM mice, n = 7–8. Log-rank (Mantel-Cox) test compared with the group of CRLM+isotype-IgG. ( F and G ) Liver weights of each group, n = 6–9, one-way ANOVA. ( H ) Macroscopic views of representative livers are shown of 4 groups (NAFLD+CRLM+isotype-IgG, NAFLD+CRLM+PD-1, NAFLD+CRLM+Reparixin, and NAFLD+CRLM+PD-1+Reparixin), n = 6–9. ( I ) Representative images of H&E staining of liver sections are shown of each group. Black box indicates lymphocytes. Scale bars represent 1 mm, upper panel ; 100 μm, lower panel. ( J ) Representative liver tissues stained with 5 markers (CD8, PD-L1, CXCL5, CXCR2, and CD11b). The image acquisition of all markers occurs simultaneously. Individual markers (or select combinations of markers) can then be displayed. Six-color overlay ( top left ) and 2/3-color insets (border) of a representative sampling of the simultaneously acquired markers. Left: control group, right: combination treatment group. Scale bars represent 20 μm. Quantification of CD8 + lymphocytes in liver metastases of mice in each treatment group, one-way ANOVA. All data are presented as mean ± SD. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001, and ns, not significant.

Article Snippet: Rabbit anti-human CXCR2 , N/A , Proteintech; 20634-1-AP.

Techniques: In Vivo, Imaging, Staining, Sampling, Control

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Concomitant NAFLD Facilitates Liver Metastases and PD-1-Refractory by Recruiting MDSCs via CXCL5/CXCR2 in Colorectal Cancer

doi: 10.1016/j.jcmgh.2024.04.008

Figure Lengend Snippet: NAFLD increases risk of liver metastases in CRC patients. ( A–C ) Representative immunohistochemistry-stained liver sections of CXCL5 ( A ), CXCR2 ( B ), and CD11b + ( C ) expression in each group. Scale bars represent 100 μm: upper panel ; 20 μm: lower panel . ( D ) Representative H&E-stained sections of liver tissues and adipocyte infiltration were assessed between liver metastasis with fatty liver (n = 6) or without fatty liver (n = 5). Scale bars represent 100 μm: upper panel ; 20 μm: lower panel . Mean ± SD, Student t test. ( E ) Comparison of BMI in synCRLM + and synCRLM - cohorts. Mean ± standard error of the mean, Student t test. ( F ) Multivariate logistic regression analysis of the significant predictors for synCRLM. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001, and ns, not significant.

Article Snippet: Rabbit anti-human CXCR2 , N/A , Proteintech; 20634-1-AP.

Techniques: Immunohistochemistry, Staining, Expressing, Comparison

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Concomitant NAFLD Facilitates Liver Metastases and PD-1-Refractory by Recruiting MDSCs via CXCL5/CXCR2 in Colorectal Cancer

doi: 10.1016/j.jcmgh.2024.04.008

Figure Lengend Snippet: NAFLD increases the risk of liver metastases in CRC patients. ( A–C ) Representative immunohistochemistry-stained liver sections of CXCL5 ( A ), CXCR2 ( B ), and CD11b + ( C ) expression between liver metastasis with fatty liver (n = 9) or without fatty liver (n = 6). Scale bars represent 100 μm: upper panel ; 20 μm: lower panel . ( D ) Prevalence of synCRLM in normal weight and obesity cohort. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001, and ns, not significant.

Article Snippet: Rabbit anti-human CXCR2 , N/A , Proteintech; 20634-1-AP.

Techniques: Immunohistochemistry, Staining, Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Concomitant NAFLD Facilitates Liver Metastases and PD-1-Refractory by Recruiting MDSCs via CXCL5/CXCR2 in Colorectal Cancer

doi: 10.1016/j.jcmgh.2024.04.008

Figure Lengend Snippet: Immunohistochemical Staining

Article Snippet: Rabbit anti-human CXCR2 , N/A , Proteintech; 20634-1-AP.

Techniques: Immunohistochemical staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Concomitant NAFLD Facilitates Liver Metastases and PD-1-Refractory by Recruiting MDSCs via CXCL5/CXCR2 in Colorectal Cancer

doi: 10.1016/j.jcmgh.2024.04.008

Figure Lengend Snippet: Fluorescence-Activated Cell Sorting

Article Snippet: Rabbit anti-human CXCR2 , N/A , Proteintech; 20634-1-AP.

Techniques: Fluorescence

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Concomitant NAFLD Facilitates Liver Metastases and PD-1-Refractory by Recruiting MDSCs via CXCL5/CXCR2 in Colorectal Cancer

doi: 10.1016/j.jcmgh.2024.04.008

Figure Lengend Snippet: mIF Staining

Article Snippet: Rabbit anti-human CXCR2 , N/A , Proteintech; 20634-1-AP.

Techniques: Concentration Assay