Journal: Cancer Immunology Research

Article Title: Programmed Cell Death Ligand 1 Expression in Osteosarcoma

doi: 10.1158/2326-6066.cir-13-0224

Figure Lengend Snippet: Figure 1. PDL1 expression in cancer and noncancer cell lines. A, relative expression of PDL1 from total RNA isolated from cell lines. B, representative PDL1 protein expression levels in corresponding high- and low- expressing cell lines by IHC. C, representative PDL1 protein expression levels in corresponding high- and low- expressing cell lines by flow cytometry.

Article Snippet: Unfortunately, the ProSci rabbit polyclonal failed to detect PDL1 on the positive control cell line SKOV3, and the XW mouse mAb exhibited unspecific binding on the negative control cell line MCF7 (Supplementary Fig. S1).

Techniques: Expressing, Isolation, Cytometry

Journal: Cancer Immunology Research

Article Title: Programmed Cell Death Ligand 1 Expression in Osteosarcoma

doi: 10.1158/2326-6066.cir-13-0224

Figure Lengend Snippet: Figure 2. Relative expression of PDL1 in 38 osteosarcoma specimens. PDL1 expression was evaluated from total RNA by quantitative real-time RT-PCR and showed that the expression levels range over 4 log (5,000-fold).

Article Snippet: Unfortunately, the ProSci rabbit polyclonal failed to detect PDL1 on the positive control cell line SKOV3, and the XW mouse mAb exhibited unspecific binding on the negative control cell line MCF7 (Supplementary Fig. S1).

Techniques: Expressing, Quantitative RT-PCR

Journal: Cancer Immunology Research

Article Title: Programmed Cell Death Ligand 1 Expression in Osteosarcoma

doi: 10.1158/2326-6066.cir-13-0224

Figure Lengend Snippet: Figure 3. Overall survival of 37 patients with osteosarcoma in relation to PDL1 gene expression. The median overall survival for PDL1-low patients was 89 months compared with 28 months for PDL1-high patients, which showed a trend but was not statistically significant (P ¼ 0.0544).

Article Snippet: Unfortunately, the ProSci rabbit polyclonal failed to detect PDL1 on the positive control cell line SKOV3, and the XW mouse mAb exhibited unspecific binding on the negative control cell line MCF7 (Supplementary Fig. S1).

Techniques: Gene Expression

Journal: Cancer Immunology Research

Article Title: Programmed Cell Death Ligand 1 Expression in Osteosarcoma

doi: 10.1158/2326-6066.cir-13-0224

Figure Lengend Snippet: Figure 4. Correlation between PDL1 gene expression and TILs by IHC. A, representative TILs in osteosarcoma tissues (400); score 0, no TILs; 1, rare/few TILs; 2, brisk/prominent TILs. B, significant positive correlation was shown between PDL1 gene expression and TILs in patients with osteosarcoma (P ¼ 0.0117).

Article Snippet: Unfortunately, the ProSci rabbit polyclonal failed to detect PDL1 on the positive control cell line SKOV3, and the XW mouse mAb exhibited unspecific binding on the negative control cell line MCF7 (Supplementary Fig. S1).

Techniques: Gene Expression

Journal: Cancer Immunology Research

Article Title: Programmed Cell Death Ligand 1 Expression in Osteosarcoma

doi: 10.1158/2326-6066.cir-13-0224

Figure Lengend Snippet: Figure 5. Characterization of the origins of metastases. A, PDL1 expression is significantly higher in metastatic osteosarcoma tumors that originate from lung than from other locations (P ¼ 0.0024). B, TILs also exhibit a positive correlation with pulmonary osteosarcoma metastasis compared with nonpulmonary metastasis (P ¼ 0.0443).

Article Snippet: Unfortunately, the ProSci rabbit polyclonal failed to detect PDL1 on the positive control cell line SKOV3, and the XW mouse mAb exhibited unspecific binding on the negative control cell line MCF7 (Supplementary Fig. S1).

Techniques: Expressing

Journal: Materials Today Bio

Article Title: Developing immune-regulatory materials using immobilized monosaccharides with immune-instructive properties

doi: 10.1016/j.mtbio.2020.100080

Figure Lengend Snippet: Flow cytometry data are shown as a heat map summarizing the expression profile of three surface markers (CD86, CD40, and CD274) on DCs after incubation on immobilized monosaccharide libraries in the presence of a TLR4 ligand (LPS). Unstimulated (immature) DCs were used as a negative control. All changes are expressed as a percentage of expression levels in mature (LPS-stimulated) DCs. A reduction in CD40 expression was observed in most conditions with the highest reduction in fucose-contained libraries compared to LPS alone. CD86 expression was higher in a number of conditions (particularly mixers of Gal1 and Gal2); however, these did not reach statistical significance. CD274 (PD-L1) expression showed a significant increase in Man1 and Gal2 combinations. Data are shown as mean ± SD of three independent donors where ∗ p <0.05, ∗∗ p <0.01, ∗∗∗ p <0.001, and ∗∗∗∗ p <0.0001.

Article Snippet: DCs were then incubated with labeled antibodies CD274 (APC clone REA1197), CD40 PE (clone HB14), and CD86 FITC (clone FM95) and isotype-matched mouse antibody controls (all from Miltenyi Biotec) for 20 min in the dark at 4 °C.

Techniques: Flow Cytometry, Expressing, Incubation, Negative Control

Journal: Scientific Reports

Article Title: Combining DNMT and HDAC6 inhibitors increases anti-tumor immune signaling and decreases tumor burden in ovarian cancer

doi: 10.1038/s41598-020-60409-4

Figure Lengend Snippet: DNMT1 protein levels are decreased by combination treatment of DNMTi and HDAC6i. ( A ) Ovarian cancer cell lines were treated as in Fig. and protein was extracted at Day 7 after treatment with IFN-gamma (IFN-γ+) (to assess MHC I and PD-L1 expression, in later figures) or control (IFN-γ -). Protein was isolated and immunoblots were run for the DNMT1 protein and α-tubulin as a loading control. Immunoblot membranes were cut and probed separately for DNMT1 (about 188 kDa) and α-tubulin (50 kDa). Cropped blots are shown here, and black lines indicate where one part of the blot ends and another begins. Figure shows the entire blot images. ( B ) The TykNu cell line was treated as in ( A ) and the protein synthesis cycloheximide added to cells on Day 7 for 0, 4, and 8 hours at 10 μM as indicated on the blot. Protein was isolated and immunoblots were run for the DNMT1 protein and α-tubulin as a loading control. Immunoblot membranes were cut and probed separately for DNMT1 (about 188 kDa) and α-tubulin (50 kDa). Cropped blots are shown here, and black lines indicate where one part of the blot ends and another begins. Figure shows the entire blot images. ( C ) Stable knockdowns of the HDAC6 protein were generated in the ID8 Trp53+/+ and Trp53−/− cell lines . Protein was extracted and immunoblots were run for the DNMT1 protein with B-actin as a loading control. Immunoblot membranes were probed for DNMT1 (about 188 kDa) and α-tubulin (50 kDa). Cropped blots are shown here and black lines indicate where one part of the blot ends and another begins. Figure shows the entire blot images. ( D ) Immunoblot showing knockdown of HDAC6 protein with a-Tubulin as a loading control. Protein was extracted and immunoblots were run for the HDAC6 protein with B-actin as a loading control. Immunoblots were probed for HDAC6 (131 kDa) and tubulin (50 kDa). Cropped blots are shown here and black lines indicate where one part of the blot ends and another begins. Figure shows the entire blot images. ( E ) Ovarian cancer cell lines were treated as in Fig. and RNA was extracted at Day 7. qRT-PCR was run for DNMT1, DNMT3a, and DNMT3b and TBP was used as a reference gene. *p < 0.05 compared to Mock.

Article Snippet: The antibodies used for immunoblotting included: DNMT1 (Sigma, D4692), PD-L1 (ProSci, 4059), HDAC1 (Cell Signaling, 2062), HDAC2 (Cell Signaling, 2540), HDAC6 (Assay Biotech, C0026), acetyl-alpha Tubulin (Cell Signaling, 3971), alpha-Tubulin (Cell Signaling, 3873).

Techniques: Expressing, Control, Isolation, Western Blot, Generated, Knockdown, Quantitative RT-PCR

Journal: OncoImmunology

Article Title: PD-L2 expression is correlated with the molecular and clinical features of glioma, and acts as an unfavorable prognostic factor

doi: 10.1080/2162402x.2018.1541535

Figure Lengend Snippet: Figure 1. Comparison of PD-L2 expression level in CGGA and TCGA cohorts with different WHO grades (a, c) and IDH status (b, d). PD-L2 was significantly increased in GRADE IV and IDH-wildtype gliomas in CGGA and TCGA data set. Photographs of immunohistochemical staining of PD-L2 in different grades of gliomas. Positive cells are stained brown. (e) Diffuse astrocytoma (WHO grade II). (f) Anaplastic astrocytoma (WHO grade III). (g) Glioblastoma multiforme (WHO grade IV). Magnification, x200. *, **, ***, and **** indicate p < 0.05, p < 0.01, p < 0.001 and p < 0.0001, respectively.

Article Snippet: Anti-PD-L2 antibody (18251–1-AP, dilution:1:200, Proteintech) was used to detect PD-L2 protein expression.

Techniques: Comparison, Expressing, Immunohistochemical staining, Staining

Journal: OncoImmunology

Article Title: PD-L2 expression is correlated with the molecular and clinical features of glioma, and acts as an unfavorable prognostic factor

doi: 10.1080/2162402x.2018.1541535

Figure Lengend Snippet: Figure 2. Comparison of PD-L2 expression level in CGGA and TCGA cohorts with different TCGA molecular subtypes (a, c), PD-L2 was significantly increased in mesenchymal subtype (p < 0.0001). ROC curves exhibited highly sensitivity in predicting mesenchymal subtype (b, d). Receiver operating characteristic (ROC) curve for mesenchymal subtype prediction in CGGA and TCGA datasets. ROC curve analysis showed that PD-L2 had highly sensitivity and specificity to predict mesenchymal subtype in CGGA and TCGA database. Area under curve(AUC) was 0.903 and 0.816, respectively.

Article Snippet: Anti-PD-L2 antibody (18251–1-AP, dilution:1:200, Proteintech) was used to detect PD-L2 protein expression.

Techniques: Comparison, Expressing

Journal: OncoImmunology

Article Title: PD-L2 expression is correlated with the molecular and clinical features of glioma, and acts as an unfavorable prognostic factor

doi: 10.1080/2162402x.2018.1541535

Figure Lengend Snippet: Figure 4. PD-L2 related inflammation activities in CGGA and TCGA cohorts. In pie charts, positive correlations are displayed in green and negative correlations in red. Color intensity and the size of the circle are proportional to the correlation coefficients.

Article Snippet: Anti-PD-L2 antibody (18251–1-AP, dilution:1:200, Proteintech) was used to detect PD-L2 protein expression.

Techniques:

Journal: OncoImmunology

Article Title: PD-L2 expression is correlated with the molecular and clinical features of glioma, and acts as an unfavorable prognostic factor

doi: 10.1080/2162402x.2018.1541535

Figure Lengend Snippet: Figure 3. The biology function of PD-L2 positively related genes in CGGA and TCGA cohorts. The biological functions related with immune response, T cell proliferation, defense response to virus, cytokine secretion, NF-kappaB signaling and cellular response to mechanical stimulus were significantly positively correlated with PD-L2 expression (p < 0.05).

Article Snippet: Anti-PD-L2 antibody (18251–1-AP, dilution:1:200, Proteintech) was used to detect PD-L2 protein expression.

Techniques: Virus, Expressing

Journal: OncoImmunology

Article Title: PD-L2 expression is correlated with the molecular and clinical features of glioma, and acts as an unfavorable prognostic factor

doi: 10.1080/2162402x.2018.1541535

Figure Lengend Snippet: Figure 6. Forest plot of hazard ratios for overall survival assessed by PD-L2 expression and clinicopathological factors. PD-L2 expression was an independent prognostic factor after adjusting age, grade, IDH status and 1p19q status in TCGA and CGGA datasets.

Article Snippet: Anti-PD-L2 antibody (18251–1-AP, dilution:1:200, Proteintech) was used to detect PD-L2 protein expression.

Techniques: Expressing

Journal: OncoImmunology

Article Title: PD-L2 expression is correlated with the molecular and clinical features of glioma, and acts as an unfavorable prognostic factor

doi: 10.1080/2162402x.2018.1541535

Figure Lengend Snippet: Figure 5. The PD-L2 survival curves of glioma, LGG and GBM in CGGA and TCGA cohorts. Kaplan–Meier survival analysis showed that high expression of PD-L2 conferred a significantly worse prognosis in glioma, LGG and GBM patients, respectively.

Article Snippet: Anti-PD-L2 antibody (18251–1-AP, dilution:1:200, Proteintech) was used to detect PD-L2 protein expression.

Techniques: Expressing

Journal: Oncotarget

Article Title: Circulating programmed death ligand-1 (cPD-L1) in non-small-cell lung cancer (NSCLC)

doi: 10.18632/oncotarget.24785

Figure Lengend Snippet: Baseline PD-L1 plasma levels according to the number of metastatic sites

Article Snippet: The level of PD-L1 was measured in plasma samples using the human (PD-L1/CD274) ELISA kit (CUSABIO, MD, USA).

Techniques:

Journal: Oncotarget

Article Title: Circulating programmed death ligand-1 (cPD-L1) in non-small-cell lung cancer (NSCLC)

doi: 10.18632/oncotarget.24785

Figure Lengend Snippet: PD-L1 plasma levels at baseline ( A ) and after 3 months of first-line treatment ( B ) in subgroup of patients receiving chemotherapy (red) or no chemotherapy (blue) agents.

Article Snippet: The level of PD-L1 was measured in plasma samples using the human (PD-L1/CD274) ELISA kit (CUSABIO, MD, USA).

Techniques:

Journal: Clinical and Experimental Immunology

Article Title: CMTM6: increased circulating level and up-regulated expression in labial salivary glands in patients with primary Sjogren’s syndrome

doi: 10.1093/cei/uxab003

Figure Lengend Snippet: Expression level of serum CMTM6, PD-L1, and PD-1 in pSS patients, non-pSS and HC. ( A ) Serum CMTM6 levels were significantly higher in pSS patients (257.04 [123.22, 704.41] pg/ml) compared to those in HC (65.02 [29.59, 121.35] pg/ml). ( B ) Serum PD-L1 levels in HC (86.51 ± 29.26 pg/ml) were also decreased than those in pSS patients (248.94 ± 114.73 pg/ml). ( C ) Serum PD-1 levels in HC (64.35 ± 26.33 pg/ml) were lower than those in pSS patients (93.72 [58.83, 153.62] pg/ml). ( D–E ) Correlation between CMTM6 and PD-L1 level ( r = 0.402, P = 0.004) and correlation between PD-1 and PD-L1 level ( r = 0.337, P = 0.017). Results are presented as the mean (SD) and median (IQR). Mann–Whitney U -test and Student t -test; Spearman Correlation. ∗∗ P < 0.01, ∗∗∗ P < 0.001, ns: not statistical significant.

Article Snippet: CMTM6 antibody (NBP1-31183) and CMTM6 human ELISA kit (NBP2-75298) were purchased from NOVUS; PD-1 antibody (66220-1-Ig), PD-L1 antibody (66248-1-Ig), CD8 antibody (66868-1-AP), CD20 antibody (24828-1-AP), and PD-L1 human ELISA kit (KE00074) were commercially from Proteintech; CD4 antibodies were purchased from Immunolway (YT0762); PD-1 human ELISA kit was bought from Invitrogen (BMS2214).

Techniques: Expressing, MANN-WHITNEY

Journal: Clinical and Experimental Immunology

Article Title: CMTM6: increased circulating level and up-regulated expression in labial salivary glands in patients with primary Sjogren’s syndrome

doi: 10.1093/cei/uxab003

Figure Lengend Snippet: Immunohistochemical staining(brown) of B, T Cells marker, CD20 ( A ), CD4 ( B ), and CD8 ( C ) in infiltrating cells and CMTM6 ( D ), PD-L1 ( E ), PD-1 ( F ) expression in minor salivary gland pSS patients (right) and non-pSS patients (left). Boxed areas represented the sites of the zoomed -in images in the right. All sections are shown at 200× (left) and 400× (right) magnifications. (A–C) The expressions of CD20, CD4, and CD8 in the minor salivary glands of pSS patients and non-pSS controls. (D) The expressions of CMTM6 tended to highly expressed on dual cells both in pSS and non-pSS and CMTM6 showed much stronger expression in ductal epithelial cells around periphery of lymphocytic foci of pSS patients. (E) The PD-L1 preotein expressed both in epithelia cells and periphery of lymphocytic foci as well as had higher expression in the glands of pSS patients. (F) The PD-1 strongly expressed in lymphocytic foci and showed higher expression in the glands of pSS. The PD-1-positve staining cells also located in the sites with CD20, CD4, and CD8-positive staining sites.

Article Snippet: CMTM6 antibody (NBP1-31183) and CMTM6 human ELISA kit (NBP2-75298) were purchased from NOVUS; PD-1 antibody (66220-1-Ig), PD-L1 antibody (66248-1-Ig), CD8 antibody (66868-1-AP), CD20 antibody (24828-1-AP), and PD-L1 human ELISA kit (KE00074) were commercially from Proteintech; CD4 antibodies were purchased from Immunolway (YT0762); PD-1 human ELISA kit was bought from Invitrogen (BMS2214).

Techniques: Immunohistochemical staining, Staining, Marker, Expressing

Journal: Clinical and Experimental Immunology

Article Title: CMTM6: increased circulating level and up-regulated expression in labial salivary glands in patients with primary Sjogren’s syndrome

doi: 10.1093/cei/uxab003

Figure Lengend Snippet: CMTM6, PD-L1, and PD-1 expression in labial gland of pSS patients with different CM grades and ESSDAI scores. ( A ) CMTM6 expression in CM = 4 ( n = 29) group was significantly higher than those in CM<2 ( n = 9) and CM = 3 ( n = 12) groups. ( B ) PD-L1 expression in CM = 4 group was higher than in those in CM<2 and CM = 3 groups but there was no expressional difference between GM<2 and GM = 3 groups. ( C ) PD-1 expression show significance between CM = 4 group and CM = 3 group but there was no difference between CM<2 and CM = 3 groups. ( D ) CMTM6 expression with ESSDAI scores>14 showed significance compared to those with ESSDAI scores>14 or 514 and those with 514 showed higher PD-1 expression than those patients with 5

Article Snippet: CMTM6 antibody (NBP1-31183) and CMTM6 human ELISA kit (NBP2-75298) were purchased from NOVUS; PD-1 antibody (66220-1-Ig), PD-L1 antibody (66248-1-Ig), CD8 antibody (66868-1-AP), CD20 antibody (24828-1-AP), and PD-L1 human ELISA kit (KE00074) were commercially from Proteintech; CD4 antibodies were purchased from Immunolway (YT0762); PD-1 human ELISA kit was bought from Invitrogen (BMS2214).

Techniques: Expressing, MANN-WHITNEY