Journal: Cancer Management and Research

Article Title: Tumor-Associated CD163 + M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer

doi: 10.2147/CMAR.S257692

Figure Lengend Snippet: Immunohistochemistry staining for PD-L1, CD68, and CD163 expression in cervical squamous cell carcinoma tissues. The left panel shows negative staining for PD-L1 and low macrophage density. The right panel shows positive staining for PD-L1 and high macrophage density.

Article Snippet: After placing in a microwave oven for pretreatment (PD-L1: EDTA antigen repair solution for 30 min, pH 9; CD68 and CD163: sodium citrate buffer for 15 min, pH 6), endogenous peroxidase was inhibited by 3% H 2 O 2 for 30 min, then the sections were incubated with 10% normal goat serum for 40 min. Primary antibodies composed of rabbit anti-PD-L1 antibody 28–8 (diluted 1:100, ab205921, Abcam, Cambridge, UK), rabbi anti-CD68 antibody (diluted 1:50, BA3638, Boster, California, UK), or rabbit anti-CD163 antibody (diluted 1:200, bs-2527R, bioss, Beijing, China), were applied overnight in a moist chamber at 4°C.

Techniques: Immunohistochemistry, Staining, Expressing, Negative Staining

Journal: Cancer Management and Research

Article Title: Tumor-Associated CD163 + M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer

doi: 10.2147/CMAR.S257692

Figure Lengend Snippet: Density of CD68- and CD163-Positive Cells in the Tumor Field and Clinical Features in 120 Samples of Cervical Squamous Cell Carcinoma

Article Snippet: After placing in a microwave oven for pretreatment (PD-L1: EDTA antigen repair solution for 30 min, pH 9; CD68 and CD163: sodium citrate buffer for 15 min, pH 6), endogenous peroxidase was inhibited by 3% H 2 O 2 for 30 min, then the sections were incubated with 10% normal goat serum for 40 min. Primary antibodies composed of rabbit anti-PD-L1 antibody 28–8 (diluted 1:100, ab205921, Abcam, Cambridge, UK), rabbi anti-CD68 antibody (diluted 1:50, BA3638, Boster, California, UK), or rabbit anti-CD163 antibody (diluted 1:200, bs-2527R, bioss, Beijing, China), were applied overnight in a moist chamber at 4°C.

Techniques: Biomarker Discovery

Journal: Cancer Management and Research

Article Title: Tumor-Associated CD163 + M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer

doi: 10.2147/CMAR.S257692

Figure Lengend Snippet: Correlation of PD-L1 expression and density of CD68-and CD163-positive cells in cervical cancer. Notes: ( A ) PD-L1 expression in tumor cells and density ofCD68. ( B ) PD-L1 expression in tumor cells and density of CD163. ( C ) PD-L1 expression in the stroma and density of CD68. ( D ) PD-L1 expression in the stroma and density of CD163.

Article Snippet: After placing in a microwave oven for pretreatment (PD-L1: EDTA antigen repair solution for 30 min, pH 9; CD68 and CD163: sodium citrate buffer for 15 min, pH 6), endogenous peroxidase was inhibited by 3% H 2 O 2 for 30 min, then the sections were incubated with 10% normal goat serum for 40 min. Primary antibodies composed of rabbit anti-PD-L1 antibody 28–8 (diluted 1:100, ab205921, Abcam, Cambridge, UK), rabbi anti-CD68 antibody (diluted 1:50, BA3638, Boster, California, UK), or rabbit anti-CD163 antibody (diluted 1:200, bs-2527R, bioss, Beijing, China), were applied overnight in a moist chamber at 4°C.

Techniques: Expressing

Journal: Cancer Management and Research

Article Title: Tumor-Associated CD163 + M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer

doi: 10.2147/CMAR.S257692

Figure Lengend Snippet: Relationship between the density of CD68- and CD163-positive cells and the expression of PD-L1 in cervical cancer. ( A ) Density of CD68 in PD-L1 positive and negative expression groups in TC. ( B ) Density of CD163 in PD-L1 positive and negative expression groups in TC. ( C ) Density of CD68 in PD-L1 positive and negative expression groups in stroma. ( D ) Density of CD163 in PD-L1 positive and negative expression groups in stroma.

Article Snippet: After placing in a microwave oven for pretreatment (PD-L1: EDTA antigen repair solution for 30 min, pH 9; CD68 and CD163: sodium citrate buffer for 15 min, pH 6), endogenous peroxidase was inhibited by 3% H 2 O 2 for 30 min, then the sections were incubated with 10% normal goat serum for 40 min. Primary antibodies composed of rabbit anti-PD-L1 antibody 28–8 (diluted 1:100, ab205921, Abcam, Cambridge, UK), rabbi anti-CD68 antibody (diluted 1:50, BA3638, Boster, California, UK), or rabbit anti-CD163 antibody (diluted 1:200, bs-2527R, bioss, Beijing, China), were applied overnight in a moist chamber at 4°C.

Techniques: Expressing

Journal: Cancer Management and Research

Article Title: Tumor-Associated CD163 + M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer

doi: 10.2147/CMAR.S257692

Figure Lengend Snippet: Univariate and Multivariate Analysis of Factors Associated with PD-L1 Expression in Tumor Cells

Article Snippet: After placing in a microwave oven for pretreatment (PD-L1: EDTA antigen repair solution for 30 min, pH 9; CD68 and CD163: sodium citrate buffer for 15 min, pH 6), endogenous peroxidase was inhibited by 3% H 2 O 2 for 30 min, then the sections were incubated with 10% normal goat serum for 40 min. Primary antibodies composed of rabbit anti-PD-L1 antibody 28–8 (diluted 1:100, ab205921, Abcam, Cambridge, UK), rabbi anti-CD68 antibody (diluted 1:50, BA3638, Boster, California, UK), or rabbit anti-CD163 antibody (diluted 1:200, bs-2527R, bioss, Beijing, China), were applied overnight in a moist chamber at 4°C.

Techniques: Expressing, Biomarker Discovery

Journal: Cancer Management and Research

Article Title: Tumor-Associated CD163 + M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer

doi: 10.2147/CMAR.S257692

Figure Lengend Snippet: Recurrence-free and overall survival curves in CC patients. ( A ) Kaplan–Meier OS curves according to PD-L1 expression in tumor cells ( B ) Kaplan–Meier OS curves according to PD-L1 expression in stroma ( C ) Kaplan–Meier OS curves according to the density of CD68-positive cells ( D ) Kaplan–Meier OS curves according to the density of CD163-positive cells ( E ) Kaplan–Meier RFS curves according to PD-L1 expression in tumor cells ( F ) Kaplan–Meier RFS curves according to PD-L1 expression in stroma ( G ) Kaplan–Meier RFS curves according to the density of CD68-positive cells ( H ) Kaplan–Meier RFS curves according to the density of CD163-positive cells. (The density of CD68/CD163 above median = high CD68/CD163; The density of CD68/CD163 below median = low CD68/CD163; p -values obtained from Log-rank tests.).

Article Snippet: After placing in a microwave oven for pretreatment (PD-L1: EDTA antigen repair solution for 30 min, pH 9; CD68 and CD163: sodium citrate buffer for 15 min, pH 6), endogenous peroxidase was inhibited by 3% H 2 O 2 for 30 min, then the sections were incubated with 10% normal goat serum for 40 min. Primary antibodies composed of rabbit anti-PD-L1 antibody 28–8 (diluted 1:100, ab205921, Abcam, Cambridge, UK), rabbi anti-CD68 antibody (diluted 1:50, BA3638, Boster, California, UK), or rabbit anti-CD163 antibody (diluted 1:200, bs-2527R, bioss, Beijing, China), were applied overnight in a moist chamber at 4°C.

Techniques: Expressing

Journal: Cancer Management and Research

Article Title: Tumor-Associated CD163 + M2 Macrophage Infiltration is Highly Associated with PD-L1 Expression in Cervical Cancer

doi: 10.2147/CMAR.S257692

Figure Lengend Snippet: Univariate Analysis of Overall and Recurrence-Free Survival

Article Snippet: After placing in a microwave oven for pretreatment (PD-L1: EDTA antigen repair solution for 30 min, pH 9; CD68 and CD163: sodium citrate buffer for 15 min, pH 6), endogenous peroxidase was inhibited by 3% H 2 O 2 for 30 min, then the sections were incubated with 10% normal goat serum for 40 min. Primary antibodies composed of rabbit anti-PD-L1 antibody 28–8 (diluted 1:100, ab205921, Abcam, Cambridge, UK), rabbi anti-CD68 antibody (diluted 1:50, BA3638, Boster, California, UK), or rabbit anti-CD163 antibody (diluted 1:200, bs-2527R, bioss, Beijing, China), were applied overnight in a moist chamber at 4°C.

Techniques: Expressing

Journal: Journal of biomedical materials research. Part A

Article Title: Evaluation of the tissue response to alginate encapsulated islets in an omentum pouch model

doi: 10.1002/jbm.a.35769

Figure Lengend Snippet: Immunohistochemical staining around alginate microbeads. Spleen was used as a control for markers CD68 (A), CD163 (D), CCR7 (G). CD68+ stain is observed (B, C)as well as CCR7+ (H,I). CD163+ staining was not observed (E, F). (C, F, I) higher resolution images of the inset area.

Article Snippet: The secondary antibody was added Biotinylated anti-mouse IgG (Vector, BA-2001, for CD68 and CD163) and Bio-tinylated anti-rabbit IgG (Vector, BA-1000, for CCR7) at a dilution of 1:200 for all antibodies.

Techniques: Immunohistochemical staining, Staining

Journal: Journal of biomedical materials research. Part A

Article Title: Evaluation of the tissue response to alginate encapsulated islets in an omentum pouch model

doi: 10.1002/jbm.a.35769

Figure Lengend Snippet: Masson’s Trichrome (A, B) indicates the presence of a chronic inflammatory response (red color) around both stable and failed alginate microbeads with islets and the presence of collagen for a small subset of alginate microbeads without islets. CCR7 stain indicates the presence of inflammatory macrophages (M1) in stable and failed alginate microbeads (C,D) while CD163 pro-healing macrophage phenotype (M2) was not observed in the groups with islets (E), the presence of pro-healing macrophage (CD163+) is observed in stable and failed alginate microbeads without islets (F). Arrows indicate macrophage presence.

Article Snippet: The secondary antibody was added Biotinylated anti-mouse IgG (Vector, BA-2001, for CD68 and CD163) and Bio-tinylated anti-rabbit IgG (Vector, BA-1000, for CCR7) at a dilution of 1:200 for all antibodies.

Techniques: Staining

Journal: Scientific Reports

Article Title: Interleukin-33 produced by M2 macrophages and other immune cells contributes to Th2 immune reaction of IgG4-related disease

doi: 10.1038/srep42413

Figure Lengend Snippet: ( A ) mRNA expression levels of innate immune cells in SGs from controls (n = 10), and patients with SS (n = 10) and IgG4-RD (n = 7). Expression levels of innate immune cells markers for macrophages (CD68), M2 macrophages (CD163), mDC (CD11c), and pDC (CD123) were estimated quantitatively as described in the Methods section. Statistically significant differences between groups were determined by Mann–Whitney U tests (* p < 0.05, ** p < 0.01). ( B ) Distribution of innate immune cells in SGs from representative controls and patients with SS and IgG4-RD. Counterstaining was performed with Mayer’s hematoxylin (blue). Scale bars, 100 μm.

Article Snippet: Anti-IL-33, -CD68, -CD163, and -CD123 mouse monoclonal antibodies were used to analyze IL-33 (clone: Nessy-1, Enzo, Japan), CD123 (clone: NCL-CD123, Leica Biosystems, Germany), CD68 (clone: ab955; Abcam, USA), and CD163 (clone: NCL-CD163, Leica Biosystems, Germany) molecules, respectively.

Techniques: Expressing, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Interleukin-33 produced by M2 macrophages and other immune cells contributes to Th2 immune reaction of IgG4-related disease

doi: 10.1038/srep42413

Figure Lengend Snippet: Double immunofluorescence staining performed with IL-33 (green), innate immune cells markers for macrophages (CD68), M2 macrophages (CD163), mDC (CD11c), and pDC (CD123) (red), and DAPI for nuclei staining (blue), as described in the Methods section. Scale bars, 50 μm.

Article Snippet: Anti-IL-33, -CD68, -CD163, and -CD123 mouse monoclonal antibodies were used to analyze IL-33 (clone: Nessy-1, Enzo, Japan), CD123 (clone: NCL-CD123, Leica Biosystems, Germany), CD68 (clone: ab955; Abcam, USA), and CD163 (clone: NCL-CD163, Leica Biosystems, Germany) molecules, respectively.

Techniques: Double Immunofluorescence Staining, Staining