Journal: Frontiers in Immunology

Article Title: High expression ITGA2 affects the expression of MET, PD-L1, CD4 and CD8 with the immune microenvironment in pancreatic cancer patients

doi: 10.3389/fimmu.2023.1209367

Figure Lengend Snippet: The study found that the expression of ITGA2, CD4 and CD8 in pancreatic cancer tissues and adjacent tissues were analyzed using HIC. The left scar bar represents 100× and the right scar bar represents 400×. (A) The results indicated that ITGA2 exhibited low, medium, and high expression levels in pancreatic cancer, and the H-score between the adjacent and cancerous tissues showed significant differences (P<0.05).In pancreatic cancer, the expression levels of CD4 (B) and CD8 (C) can be classified as low, medium, or high. The H-score indicates a significant difference (P < 0.05) between adjacent and cancer tissues for both CD4 and CD8.

Article Snippet: Immunohistochemical methods were used, with serial sections made at a thickness of 4μm.After dehydration and hydration, the sections were repaired with sodium citrate antigen, blocked with goat serum, blocked with peroxide, added with the first antibody ITGA2 monoclonal antibody (1:400, bioss, Beijing, China, No : BSM-52613R); CD4 monoclonal antibody (1:50, bioss, Beijing, China, No:bsm-52469R); CD8 monoclonal antibody (1:1500, bioss, Beijing, China, No:bs-23447R) and the second antibody IGg polymer (1:200, bioss, Beijing, China, No: bs-0295G) under high temperature and high pressure environment, and finally stained with DAB, In this study, hematoxylin was used for blue reverse staining, followed by dehydration and fixation.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: High expression ITGA2 affects the expression of MET, PD-L1, CD4 and CD8 with the immune microenvironment in pancreatic cancer patients

doi: 10.3389/fimmu.2023.1209367

Figure Lengend Snippet: Correlation and Prognostic Signifificance between the expression levels of ITGA2, CD4,and CD8 in Pancreatic Cancer. (A) A scatter plot graph illustrates that the percentage of ITGA2 positive cells directly correlates with the number of CD4 positive cells (r= -0.344, P<0.05). (B) A scatter plot graph illustrates that the percentage of ITGA2 positive cells directly correlates with the number of CD8 positive cells (r= -0.398, P<0.05). (C) Patients with ITGA2+++ exhibited a shorter survival compared to patients with ITGA2++ and ITGA2+ (median survival: 9 vs. 9 vs. 9.5 months, P=0.033). (D) Patients with CD4+++ had a longer survival compared to patients with CD4++ and CD4+ (median survival: 16 vs. 11 vs. 9 months, P=0.001 ). (E) Patients with CD8+++ had a longer survival compared to patients with CD8++ and CD8+ (median survival: 10 vs. 13 vs. 9 months, P=0.002 ). + repersent weakly positive, ++ represent positive, +++ represent strong positive.

Article Snippet: Immunohistochemical methods were used, with serial sections made at a thickness of 4μm.After dehydration and hydration, the sections were repaired with sodium citrate antigen, blocked with goat serum, blocked with peroxide, added with the first antibody ITGA2 monoclonal antibody (1:400, bioss, Beijing, China, No : BSM-52613R); CD4 monoclonal antibody (1:50, bioss, Beijing, China, No:bsm-52469R); CD8 monoclonal antibody (1:1500, bioss, Beijing, China, No:bs-23447R) and the second antibody IGg polymer (1:200, bioss, Beijing, China, No: bs-0295G) under high temperature and high pressure environment, and finally stained with DAB, In this study, hematoxylin was used for blue reverse staining, followed by dehydration and fixation.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: High expression ITGA2 affects the expression of MET, PD-L1, CD4 and CD8 with the immune microenvironment in pancreatic cancer patients

doi: 10.3389/fimmu.2023.1209367

Figure Lengend Snippet: The relationship between the expression of ITGA2, CD4 and CD8 in pancreatic cancer tissues and the clinicopathological features.

Article Snippet: Immunohistochemical methods were used, with serial sections made at a thickness of 4μm.After dehydration and hydration, the sections were repaired with sodium citrate antigen, blocked with goat serum, blocked with peroxide, added with the first antibody ITGA2 monoclonal antibody (1:400, bioss, Beijing, China, No : BSM-52613R); CD4 monoclonal antibody (1:50, bioss, Beijing, China, No:bsm-52469R); CD8 monoclonal antibody (1:1500, bioss, Beijing, China, No:bs-23447R) and the second antibody IGg polymer (1:200, bioss, Beijing, China, No: bs-0295G) under high temperature and high pressure environment, and finally stained with DAB, In this study, hematoxylin was used for blue reverse staining, followed by dehydration and fixation.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: High expression ITGA2 affects the expression of MET, PD-L1, CD4 and CD8 with the immune microenvironment in pancreatic cancer patients

doi: 10.3389/fimmu.2023.1209367

Figure Lengend Snippet: Univariate and multivariate analysis of Cox model for clinical prognostic factors of pancreatic cancer (n = 62).

Article Snippet: Immunohistochemical methods were used, with serial sections made at a thickness of 4μm.After dehydration and hydration, the sections were repaired with sodium citrate antigen, blocked with goat serum, blocked with peroxide, added with the first antibody ITGA2 monoclonal antibody (1:400, bioss, Beijing, China, No : BSM-52613R); CD4 monoclonal antibody (1:50, bioss, Beijing, China, No:bsm-52469R); CD8 monoclonal antibody (1:1500, bioss, Beijing, China, No:bs-23447R) and the second antibody IGg polymer (1:200, bioss, Beijing, China, No: bs-0295G) under high temperature and high pressure environment, and finally stained with DAB, In this study, hematoxylin was used for blue reverse staining, followed by dehydration and fixation.

Techniques: Expressing

Journal:

Article Title: KDEL and KKXX Retrieval Signals Appended to the Same Reporter Protein Determine Different Trafficking between Endoplasmic Reticulum, Intermediate Compartment, and Golgi Complex

doi: 10.1091/mbc.E02-08-0468

Figure Lengend Snippet: Different presence of CD8-K and CD8-E19 at ER exit sites in transiently transfected HuH-7 cells. Conventional thin section electron microscopy (a and b) and cryoimmunoelectron microscopy (c–k) of parental (a and b) and transiently transfected HuH-7 cells expressing CD8-K (c–f) and CD8-E19 (g–k) proteins. The ultrastructural analysis revealed the presence of long ER cisternae and outer nuclear membranes mostly studded with ribosomes (arrowheads in a and b) and occasionally showing smooth areas with emerging protrusions (arrows in a and b) facing IC units (asterisk in a). Immunogold labeling with anti-CD8 polyclonal antibody and protein A-colloidal gold conjugates were dense and unevenly distributed over ER cisternae (c), whereas the gold particles frequently were localized on the smooth ribosome-free areas of the membranes and clustered over protrusions extending from them (e and f, arrows). Similar clustering of gold particles was observed in protrusions extending from the outer nuclear membranes (d, arrow). Immunolabeling was also present on IC units (d, asterisk). In contrast, immunogold labeling of CD8-E19–expressing cells was homogeneously distributed over either outer nuclear membranes (g) or ER cisternae (k) and the protrusions extending from the ER (arrows in h–j) were often unlabeled but sometimes labeled (j). M, mitochondria; er, endoplasmic reticulum; NM, nuclear membrane; Nu, nucleus. Bars, 0.1 μm.

Article Snippet: Ultrathin cryosections were collected with sucrose and methylcellulose and incubated with polyclonal anti-CD8 antibody followed by 10-nm-diameter protein A-colloidal gold conjugates (British BioCell International, Cardiff, United Kingdom).

Techniques: Transfection, Electron Microscopy, Microscopy, Expressing, Labeling, Immunolabeling

Journal: Frontiers in Immunology

Article Title: Gastrointestinal stromal tumors regulate macrophage M2 polarization through the MIF/CXCR4 axis to immune escape

doi: 10.3389/fimmu.2024.1431535

Figure Lengend Snippet: Cell counts of every cluster in two samples.

Article Snippet: Immunohistochemistry and immunofluorescence were performed on the tissue microarray with the following antibodies: rabbit polyclonal antibodies against human CD8 (ZEN Bio, Lot No:10011860), CD206 (Protein tech, Lot No:00080496), MIF (ZEN Bio, Lot No:20200101), and CXCR4 (ZEN Bio, Lot No: BJ10221968), and monoclonal mouse antibody against human CD68 (Abcam, Lot No:00098190).

Techniques:

Journal: Frontiers in Immunology

Article Title: Gastrointestinal stromal tumors regulate macrophage M2 polarization through the MIF/CXCR4 axis to immune escape

doi: 10.3389/fimmu.2024.1431535

Figure Lengend Snippet: Immunohistochemical characterization of CD8, CD68, MIF, CD206 and CXCR4 in GIST samples.

Article Snippet: Immunohistochemistry and immunofluorescence were performed on the tissue microarray with the following antibodies: rabbit polyclonal antibodies against human CD8 (ZEN Bio, Lot No:10011860), CD206 (Protein tech, Lot No:00080496), MIF (ZEN Bio, Lot No:20200101), and CXCR4 (ZEN Bio, Lot No: BJ10221968), and monoclonal mouse antibody against human CD68 (Abcam, Lot No:00098190).

Techniques: Immunohistochemical staining

Journal: Frontiers in Immunology

Article Title: Gastrointestinal stromal tumors regulate macrophage M2 polarization through the MIF/CXCR4 axis to immune escape

doi: 10.3389/fimmu.2024.1431535

Figure Lengend Snippet: (A) (A) There were differences in CD8+T cell infiltration among different groups (0.35 ± 0.15 vs 0.27 ± 0.10 vs 0.09 ± 0.02, P =0.003); (B) There were differences in the infiltration of CD68+macrophage cells among different groups (0.38 ± 0.09 vs 0.64 ± 0.17 vs 0.98 ± 0.19, P =0.03). (B) (A) As the risk increases, the expression of CXCR4 gradually increases, and the difference is statistically significant (0.43 ± 0.24 vs 0.90 ± 0.68 vs 1.64 ± 0.53, P <0.001); (B) As the risk increases, the expression of MIF gradually increases, and the difference is statistically significant (0.009 ± 0.003 vs 0.70 ± 0.02 vs 0.12 ± 0.03, P <0.001); (C) As the risk increases, the expression of CD206 gradually increases, and the difference is statistically significant (0.04 ± 0.01 vs 0.07 ± 0.0.02 vs 0.15 ± 0.03, P<0.001); The statistical method is non-parametric test, n=80, * P <0.05; ** P <0.01.

Article Snippet: Immunohistochemistry and immunofluorescence were performed on the tissue microarray with the following antibodies: rabbit polyclonal antibodies against human CD8 (ZEN Bio, Lot No:10011860), CD206 (Protein tech, Lot No:00080496), MIF (ZEN Bio, Lot No:20200101), and CXCR4 (ZEN Bio, Lot No: BJ10221968), and monoclonal mouse antibody against human CD68 (Abcam, Lot No:00098190).

Techniques: Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Synergistic immunomodulatory effect of synbiotics pre- and postoperative resection of pancreatic ductal adenocarcinoma: a randomized controlled study

doi: 10.1007/s00262-024-03686-6

Figure Lengend Snippet: Tissue expression of CD8 + cells and INF-γ in subjected groups

Article Snippet: The other two slides were underwent immunohistochemical procedure as incubated with anti-CD8 polyclonal antibody (YPA2235, Biospes, Chongqing, China), and anti- IFNγ polyclonal antibody (YPA2285, Biospes, Chongqing, China), at dilution of 1:100.

Techniques: Expressing, Probiotics

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Synergistic immunomodulatory effect of synbiotics pre- and postoperative resection of pancreatic ductal adenocarcinoma: a randomized controlled study

doi: 10.1007/s00262-024-03686-6

Figure Lengend Snippet: The clinicopathological parameters on the expression of INFγ and CD8 + T cells in the tumor tissues of all groups

Article Snippet: The other two slides were underwent immunohistochemical procedure as incubated with anti-CD8 polyclonal antibody (YPA2235, Biospes, Chongqing, China), and anti- IFNγ polyclonal antibody (YPA2285, Biospes, Chongqing, China), at dilution of 1:100.

Techniques: Expressing