Journal: Nature Communications

Article Title: Hypoxia induces HIF1α-dependent epigenetic vulnerability in triple negative breast cancer to confer immune effector dysfunction and resistance to anti-PD-1 immunotherapy

doi: 10.1038/s41467-022-31764-9

Figure Lengend Snippet: a Schematic diagram showing the establishment of humanized mice (humice) with human immune system reconstituted in NIKO mice. The presence of human CD45 + cells, NK cells, CD4 + and CD8 + T cells in the mice’s peripheral system was validated by flow cytometry. b Primary LM2 tumor size in humice (control, n = 14; Keytruda, n = 14; ENT, n = 12; PX478, n = 14; ENT + Keytruda, n = 16; PX478 + Keytruda, n = 16) and NIKO mice (control, n = 10; ENT + Keytruda, n = 10; PX478 + Keytruda, n = 10), at Day 21 of treatments. c Lung metastasis of humice (control, n = 6; Keytruda, n = 6; ENT, n = 6; PX478, n = 6; ENT + Keytruda, n = 7; PX478 + Keytruda, n = 7) and NIKO mice (control, n = 5; ENT + Keytruda, n = 5; PX478 + Keytruda, n = 5) bearing LM2 tumors at Day 35 assessed by bioluminescence (BLI) measurement. d Representative bioluminescence (BLI) images showing the lung metastasis of humice and NIKO mice. e Flow cytometric analysis of LM2 tumors harvested from humanized mice. IFNγ, TNFα, and granzyme B expression was examined in tumor-infiltrating human CD8 + T cells and NK cells. N = 5 for each group. f Flow cytometry analysis of LM2 tumors harvested from humanized mice. Expressions of human PD-L1 and PD-L2 were examined in total living cells dissociated from LM2 tumors. N = 5 for each group. Quantification data of flow cytometry ( e , f ) are presented as a box and whiskers, with median values and whiskers of minimum and maximum values. Data for b and c were presented as mean ± SD . P values were determined by one-way ( e , f ) or two-way ( b , c ) ANOVA with Turkey’s test. Source data are provided as a source data file.

Article Snippet: The following antibodies were used for staining, anti-activated pimonidazole FITC antibody (Hypoxyprobe, CAT# HP2-200kit, dilution 1:200), anti-mouse HIF1α APC antibody (R&D Systems, CAT# IC1935A, dilution 1:50), anti-mouse CD3 BV421 antibody (BD Biosciences, CAT# 564008, dilution 1:100), anti-mouse CD45 Percp-Vio700 antibody (Miltenyi Biotec, CAT# 130-110-663, dilution 1:100) anti-mouse CD8 APC-Vio770 antibody (Miltenyi Biotec, CAT# 130-120-737, dilution 1:100), anti-mouse Nkp46 APC antibody (Miltenyi Biotec, CAT# 130-112-202, dilution 1:100), anti-mouse CD4 BV650 antibody (Biolegend, CAT# 563747, dilution 1:100), anti-mouse TIM-3 BV711 antibody (Biolegend, CAT# 119727, dilution 1:100), anti-mouse PD-1 PE-Vio770 (Miltenyi Biotec, CAT# 130-120-391, dilution 1:100), anti-mouse IFNγ PE (Miltenyi Biotec, CAT# 130-117-352, dilution 1:100), anti-mouse TNFα BV711 (BD Biosciences, CAT# 563944, dilution 1:100), anti-mouse/human granzyme B FITC (Miltenyi Biotec, Cat#130-118-430, dilution 1:100), anti-mouse PD-L1 BV786 antibody (BD Biosciences, CAT# 741014, dilution 1:100), anti-mouse PD-L2 FITC antibody (Miltenyi Biotec, Cat# 130-102-222, dilution 1:100), anti-human CD45 FITC antibody (BD Biosciences, CAT# 304006, dilution 1:100), anti-human CD3 PE antibody (Biolegend, CAT# 300308, dilution 1:100) anti-human CD8 APC-Cy7 antibody (BD Biosciences, CAT# 557834, dilution 1:100), anti-human CD56 BV711 antibody (Biolegend, CAT# 318336, dilution 1:100), anti-human CD4 APC antibody (Biolegend, CAT# 300514, dilution 1:100), anti-human IFNγ BV785 (Biolegend, CAT# 502542, dilution 1:100), anti-human TNFα BV650 (Biolegend, CAT# 502398, dilution 1:100), anti-human Granzyme B BV421 (BD Biosciences, Cat# 563389, dilution 1:100), anti-human PD-L1 PE-Cy7 antibody (Biolegend, CAT# 374506, dilution 1:100), anti-human PD-L2 PE antibody (Miltenyi Biotec, CAT# 130-098-530, dilution 1:100).

Techniques: Flow Cytometry, Control, Expressing

Journal: Ophthalmic plastic and reconstructive surgery

Article Title: PD-L1 and PD-L2 Expression Levels Are Low in Primary and Secondary Adenoid Cystic Carcinomas of the Orbit: Therapeutic Implications

doi: 10.1097/IOP.0000000000001585

Figure Lengend Snippet: Expression of PD-L1 and PD-L2, and CD8 T-lymphocyte infiltration

Article Snippet: The following antibodies were used: PD-L1 (rabbit monoclonal antibody clone E1L3N, 1:30 dilution, Cell Signaling Technology, Danvers, MA), PD-L2 (rabbit monoclonal antibody clone D7U8C, 1:100 dilution, Cell Signaling Technology, Danvers, MA), and CD8 (rabbit polyclonal antibody, #ab4055, 1:200 dilution, Abcam, Cambridge, MA).

Techniques: Expressing, Standard Deviation

Journal: Ophthalmic plastic and reconstructive surgery

Article Title: PD-L1 and PD-L2 Expression Levels Are Low in Primary and Secondary Adenoid Cystic Carcinomas of the Orbit: Therapeutic Implications

doi: 10.1097/IOP.0000000000001585

Figure Lengend Snippet: Immunohistochemical staining for PD-L1, PD-L2, and CD8 in adenoid cystic carcinoma. A, PD-L1 positively immunnonstains the apical membranes of some tumor cells (arrow). B, PD-L1 positively immunostains the stromal cells (arrow) surrounding some tumors. In this case, the tumor cells do not express PD-L1. C, This anti-PD-L1 antibody also prominently highlights nerve fascicles in a number of the tumor samples examined (arrow). D, PD-L2 positively immunostains the cytoplasm and membranes of some tumor cells (arrow). E, PD-L2 positively immunostains clusters of S. In this case, the T do not express PD-L2. F, Rare CD8-positive T-lymphocytes are present infiltrating the tumor, but greater numbers are appreciated in the surrounding stroma (A–F, immunoperoxidase reaction, diaminobenzidine chromogen, hematoxylin counterstain, original magnification ×40, ×40, ×10, ×40, ×40, ×20, with additional optical magnification of ×3 for A–E and ×2 for F). PD-L, programmed cell death ligand; S, stromal cells; T, tumor cells.

Article Snippet: The following antibodies were used: PD-L1 (rabbit monoclonal antibody clone E1L3N, 1:30 dilution, Cell Signaling Technology, Danvers, MA), PD-L2 (rabbit monoclonal antibody clone D7U8C, 1:100 dilution, Cell Signaling Technology, Danvers, MA), and CD8 (rabbit polyclonal antibody, #ab4055, 1:200 dilution, Abcam, Cambridge, MA).

Techniques: Immunohistochemical staining, Staining

Journal: Ophthalmic plastic and reconstructive surgery

Article Title: PD-L1 and PD-L2 Expression Levels Are Low in Primary and Secondary Adenoid Cystic Carcinomas of the Orbit: Therapeutic Implications

doi: 10.1097/IOP.0000000000001585

Figure Lengend Snippet: Immunohistochemical staining for PD-L1, PD-L2, and CD8 in normal lacrimal gland. A, PD-L1 positively immunostains occasional lacrimal gland acini in a membranous pattern (arrow). B, PD-L2 positively immunostains the cytoplasm and membranes of small clusters of stromal cells (arrow) distributed among the lacrimal gland acini. C, Low numbers of CD8-positive T-lymphocytes are scattered throughout the lacrimal gland (A–C, immunoperoxidase reaction, diaminobenzidine chromogen, hematoxylin counterstain, original magnification ×20, ×20, ×20, with additional ×3 optical magnification for A–C). PD-L, programmed cell death ligand.

Article Snippet: The following antibodies were used: PD-L1 (rabbit monoclonal antibody clone E1L3N, 1:30 dilution, Cell Signaling Technology, Danvers, MA), PD-L2 (rabbit monoclonal antibody clone D7U8C, 1:100 dilution, Cell Signaling Technology, Danvers, MA), and CD8 (rabbit polyclonal antibody, #ab4055, 1:200 dilution, Abcam, Cambridge, MA).

Techniques: Immunohistochemical staining, Staining

Journal: Ophthalmic plastic and reconstructive surgery

Article Title: PD-L1 and PD-L2 Expression Levels Are Low in Primary and Secondary Adenoid Cystic Carcinomas of the Orbit: Therapeutic Implications

doi: 10.1097/IOP.0000000000001585

Figure Lengend Snippet: Review of previous studies of PD-L1 or PD-L2 expression in adenoid cystic carcinoma

Article Snippet: The following antibodies were used: PD-L1 (rabbit monoclonal antibody clone E1L3N, 1:30 dilution, Cell Signaling Technology, Danvers, MA), PD-L2 (rabbit monoclonal antibody clone D7U8C, 1:100 dilution, Cell Signaling Technology, Danvers, MA), and CD8 (rabbit polyclonal antibody, #ab4055, 1:200 dilution, Abcam, Cambridge, MA).

Techniques: Expressing, In Situ Hybridization

Journal: Journal of Clinical Medicine

Article Title: A New Histology-Based Prognostic Index for Aggressive T-Cell lymphoma: Preliminary Results of the “TCL Urayasu Classification”

doi: 10.3390/jcm13133870

Figure Lengend Snippet: Summary of the tumor immunohistochemical findings in the patients with TCL ( n = 16).

Article Snippet: The primary antibodies against the major proteins involved in anticancer drug metabolism included (1) GRP94: Proteintech (Rosemont, IL 60018, USA), clone 1H10B7 (this monoclonal antibody was generated against the N-terminal region of full-length HSP90b1); (2) CYP3A4: Sigma-Aldrich (St. Louis, MO 63103, USA), SAB1400064 (this polyclonal antibody was generated against CYP3A4); (3) AKR1C3: Proteintech, 11194-1-AP (this polyclonal antibody was generated against AKRC3); (4) MDR1 (P-glycoprotein): Proteintech, 22336-1-AP (this polyclonal antibody was generated against MDR1); (5) MRP1 (CD9): Proteintech, 60232-1-IG (this monoclonal antibody was generated against the N-terminal region of full-length MRP1); (6) TGF beta1: Proteintech, 21898-1-AP (this polyclonal antibody was generated against TGF-beta); (7) GRP78: Proteintech, 66574-1-IG (this monoclonal antibody was generated against the N-terminal region of full-length GRP78); (8) glutathione S-transferase kappa1 (GST): Proteintech, 14535-1-AP (this polyclonal antibody was generated against GST1); (9) thymidine phosphorylase: Abcam (Cambridge, UK), ab226917 (this polyclonal antibody was generated against thymidine phosphorylase); (10) MRP4 (ABCC4): SANTA CRUZ BIOTECHNOLOGY (Dallas, TX 75220, USA), SC-376262 (this monoclonal antibody was generated against the N-terminal region of full-length MRP4 (amino acid 1-280)); (11) CYP2B6: LifeSpan BioSciences, Inc. (Seattle, WA 98121, USA), LS-C352084 (this polyclonal antibody was generated against CYP2B6); (12) TNF1 alpha: Sigma-Aldrich, SAB4502982 (this polyclonal antibody was generated against TNF1 alpha); (13) PD-1; (14) PD-L1: Proteintech (Rosemont, IL, USA), 66248-1-IG, mouse IgG1 monoclonal antibody, clone 2B11D11; (15) PD-L2: Proteintech (Rosemont, IL, USA), 18251-1-AP 16, rabbit IgG polyclonal antibody; (16) P53: Cell Signaling Technology, Inc. (3 Trask Lane Danvers, MA 01923, USA), DO-7 mouse monoclonal antibody #48818; (17) c-MYC: Abcam (Kendall Sq Cambridge, MA 02139, USA), Y69 clone ab32072; (18) ENT-1 (equilibrative nucleoside transporter 1): Proteintech (Rosemont, IL, USA), 1337-1-AP rabbit IgG polyclonal antibody; (19) AKR1B1: Sigma-Aldrich (3050 Spruce Street Saint Louis, MO 63103, USA), rabbit polyclonal antibody HPA052751; (20) AKR1B10: Sigma-Aldrich (3050 Spruce Street Saint Louis, MO 63103, USA), rabbit monoclonal antibody HPA020280.

Techniques: Immunohistochemical staining, Silver Staining