Journal: Molecular imaging

Article Title: Comparison of Tumor Non-specific and PD-L1 Specific Imaging by Near-Infrared Fluorescence/Cerenkov Luminescence Dual-Modality In-situ Imaging.

doi: 10.1177/15353508241261473

Figure Lengend Snippet: Figure 2. 131I-Atezolizumab and 131I-IgG treatment effect evaluation. (a) Tumor volume change. (b) Mouse body weight changes. (c) CLI of mice 11 days after administration. (d) HE staining and Ki-67 immunohistochemical chemistry.

Article Snippet: Briefly, 100 μl of phosphate buffer (PB, 0.25 mol/L, pH7.4) and 100 μg Atezolizumab (HY-P9904, MCE, USA) or human nonspecific IgG1 (IgG, BE0297, BioXcell, USA) were added to the coating tube containing 50 μg Iodogen and mixed, and about 185 to 330 MBq Na131I (Chengdu Gaotong Isotope Co., Ltd, China) was added and reacted 15 min after mixing at room temperature.

Techniques: Staining, Immunohistochemical staining

Journal: Molecular imaging

Article Title: Comparison of Tumor Non-specific and PD-L1 Specific Imaging by Near-Infrared Fluorescence/Cerenkov Luminescence Dual-Modality In-situ Imaging.

doi: 10.1177/15353508241261473

Figure Lengend Snippet: Figure 1. In-vitro cell binding assay. 131I-IgG and131I-Atezolizumab cell binding experiments in different cell lines (Hela, A375, H460, RKO, and HCT8). RKO cells and 131I-Atezolizumab had the highest binding capacity.

Article Snippet: Briefly, 100 μl of phosphate buffer (PB, 0.25 mol/L, pH7.4) and 100 μg Atezolizumab (HY-P9904, MCE, USA) or human nonspecific IgG1 (IgG, BE0297, BioXcell, USA) were added to the coating tube containing 50 μg Iodogen and mixed, and about 185 to 330 MBq Na131I (Chengdu Gaotong Isotope Co., Ltd, China) was added and reacted 15 min after mixing at room temperature.

Techniques: In Vitro, Cell Binding Assay, Binding Assay

Journal: Molecular imaging

Article Title: Comparison of Tumor Non-specific and PD-L1 Specific Imaging by Near-Infrared Fluorescence/Cerenkov Luminescence Dual-Modality In-situ Imaging.

doi: 10.1177/15353508241261473

Figure Lengend Snippet: Figure 3. IR-Atezolizumab and IR-IgG NIRF imaging in RKO model. IR-Atezolizumab was injected into mouse 1 and 2, IR-IgG was injected into mouse 3 and 4. Mouse 1 and 2 had no tracer uptake, while mouse 3 and 4 had significant tracer uptake in the pharyngeal lymphatic ring (dotted line).

Article Snippet: Briefly, 100 μl of phosphate buffer (PB, 0.25 mol/L, pH7.4) and 100 μg Atezolizumab (HY-P9904, MCE, USA) or human nonspecific IgG1 (IgG, BE0297, BioXcell, USA) were added to the coating tube containing 50 μg Iodogen and mixed, and about 185 to 330 MBq Na131I (Chengdu Gaotong Isotope Co., Ltd, China) was added and reacted 15 min after mixing at room temperature.

Techniques: Imaging, Injection

Journal: Molecular imaging

Article Title: Comparison of Tumor Non-specific and PD-L1 Specific Imaging by Near-Infrared Fluorescence/Cerenkov Luminescence Dual-Modality In-situ Imaging.

doi: 10.1177/15353508241261473

Figure Lengend Snippet: Figure 4. CLI in different mouse models. (a)131I-IgG CLI in different mouse models (Hela, A375, H460, RKO, and HCT8 models);131I-Atezolizumab CLI in HCT8* model. (b) The relative fluorescence intensity (tumor-to-muscle uptake ratio) of 131I-IgG or 131I-Atezolizumab in different mouse models within 1 to 5 days (n = 3/group). (c) Growth curves of tumors of different models (n = 3/ group). Hela, A375, H460, RKO,HCT8 model injected with 131I-IgG, HCT8* model injected with 131I-Atezolizumab, and the dotted line represents the time of drug injection.

Article Snippet: Briefly, 100 μl of phosphate buffer (PB, 0.25 mol/L, pH7.4) and 100 μg Atezolizumab (HY-P9904, MCE, USA) or human nonspecific IgG1 (IgG, BE0297, BioXcell, USA) were added to the coating tube containing 50 μg Iodogen and mixed, and about 185 to 330 MBq Na131I (Chengdu Gaotong Isotope Co., Ltd, China) was added and reacted 15 min after mixing at room temperature.

Techniques: Injection

Journal: Molecular imaging

Article Title: Comparison of Tumor Non-specific and PD-L1 Specific Imaging by Near-Infrared Fluorescence/Cerenkov Luminescence Dual-Modality In-situ Imaging.

doi: 10.1177/15353508241261473

Figure Lengend Snippet: Figure 6. NIRF/CL in-situ distribution imaging at 168 h after administration. (a and b) IR-Atezolizumab NIRF imaging in RKO model. (c and d) 131I-IgG CLI in RKO model. (e and f) IR-Atezolizumab NIRF imaging in HCT8 model. (g and h) 131I-IgG CLI in HCT8 model. The dotted line is the mean fluorescence intensity of the muscle.

Article Snippet: Briefly, 100 μl of phosphate buffer (PB, 0.25 mol/L, pH7.4) and 100 μg Atezolizumab (HY-P9904, MCE, USA) or human nonspecific IgG1 (IgG, BE0297, BioXcell, USA) were added to the coating tube containing 50 μg Iodogen and mixed, and about 185 to 330 MBq Na131I (Chengdu Gaotong Isotope Co., Ltd, China) was added and reacted 15 min after mixing at room temperature.

Techniques: In Situ, Imaging

Journal: bioRxiv

Article Title: B cell receptor silencing reveals the origin of high-grade B cell lymphomas with MYC and BCL2 rearrangements

doi: 10.1101/2024.07.13.603066

Figure Lengend Snippet: (A) Representative (n=6) sIGK/IGL FACS analyses of COH-DHL1 cells. Cell frequencies within each quadrant are indicated. (B) PCR amplification of IGKC (upper panel) in COH-DHL1 cells, 293T cells and PBMC. Amplification of RPLP0 controlled for DNA input. Genomic PCR for the KDE rearrangement was performed on the same sample set (lower panel). Data represent n=3 experiments. The asterisk indicates nonspecific bands. (C) IGM and IG light chain IHC and RNA analysis of the primary HGBCL-DH- BCL2 - BCL6 case and the COH-THL1 cell line derivative, analyzed at diagnosis (left panel) and after relapse (right panel). (D) Representative (n=6) FACS analysis of COH-THL1 cells before (left plot) and after (mid and right plots) gating on the indicated subsets. Numbers indicate cell frequencies (E). Representative (n=5) FACS analysis for sIGK/L of COH-THL1 bulk cultures (left panel), purified IGH UND variants (mid panel) and derivatives analyzed 2 weeks later (right panel). Numbers indicate cell frequencies. (F) Normalized RAG1/2 transcripts in IGH UND COH-DHL1 and COH-THL1 bulk cultures relative to 697 leukemic cells, measured by qRT-PCR. 293T were included as negative controls. Data represent n=3 experiments. (G) Normalized RAG1/2 transcripts in FACS-purified IGH + and IGH UND COH-THL1 cells, relative to 697 leukemic cells, quantified by RT-PCR. Data represent n=3 experiments. (H) Co-cultures of sCD79B + (IGH + ) and sCD79B lo (IGH UND ) COH-THL1 cells were treated for 72 h with Polatuzumab (Pola)-Vedotin, unconjugated Polatuzumab (Pola), or left untreated (-), and assessed by FACS for frequency of sCD79B + cells. Data represent n=3 experiments. Numbers indicate cell frequencies in the corresponding quadrants. (I) Mean fluorescence intensity (MFI) values representing sCD79B levels quantified by flow cytometry for the indicated lymphoma cell lines. IGH + COH-THL1 cells were separated into three subsets according to sIG light chain usage. (J) Cell viability measured 72 h post Polatuzumab-Vedotin/Polatuzumab treatment for the indicated DLBCL, BL and HGBCL-DH- BCL2(-BCL6) cell lines, measured by flow cytometry. Data are relative to untreated cells. Surface BCR status is indicated. Data represent n=3 experiments.

Article Snippet: HT (IG UND ), RAMOS (IGM + ), DoGKIT (IG UND ), COH-THL1 (IGM + ), COH-THL1 (IGH UND ), and COH-DHL1 (IGH UND ) human lymphoma lines were treated with Polatuzumab (P)-Vedotin (V) or Polatuzumab (P) (MedChemExpress).

Techniques: Amplification, Biomarker Discovery, Purification, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Fluorescence, Flow Cytometry

Journal: Acta pharmaceutica Sinica. B

Article Title: Anti-PD-L1 antibody enhances curative effect of cryoablation via antibody-dependent cell-mediated cytotoxicity mediating PD-L1 high CD11b + cells elimination in hepatocellular carcinoma.

doi: 10.1016/j.apsb.2022.08.006

Figure Lengend Snippet: Figure 5 Anti-PD-L1 antibody triggered NK cell-mediated ADCC effect to eliminating CD11bþ myeloid cells. (A, B) Representative flow cytometry plots of CD49bþ NK cells gated on CD45þ cells and granzyme Bþ cells gated on NK cells in all groups. (C) Flow cytometric quantification of CD49bþ (NK) population, and expression of granzyme Bþ populations in all groups. n Z 6 for each group, *P < 0.05, **P < 0.01, ***P < 0.001. ns, no significant difference, t-tests. All data are mean SEM. (D) H22 cells were inoculated subcutaneously into BALB/c mice. Twelve days later, cryoablation or microwave-ablation was proceeded, with corresponding antibody being infused via tail injection every 3 days. ADCC blocking assay employed by anti-mouse CD16 neutralizing antibody. Anti-mouse IgG was used as negative control. Tumor volume was shown. n Z 6 for each groups, *P < 0.05, **P < 0.01, ***P < 0.001. ns, no significant difference, t-tests. All data are mean SEM. (E) Immunofluorescence staining of tumor tissue sections. CD11b were stained with Alexa Fluor 647, CD49b was stained with Alexa Fluor 488, Tunel was stained by Alexa Fluor 594. Scale bar Z 20 mm.

Article Snippet: This mix was then added to anti-IFN-g pre-coated plates from the human IFN-g ELISpot assay kit (DKW22-1000-096s; Dakewei, China), along with the following control groups: no antibody, isotype IgG1 control (MCE, HY-P9900, Newark, NJ, USA), and positive control (phytohemagglutinin stimulation).

Techniques: Cytometry, Expressing, Injection, Blocking Assay, Negative Control, Staining, TUNEL Assay

Journal: Acta pharmaceutica Sinica. B

Article Title: Anti-PD-L1 antibody enhances curative effect of cryoablation via antibody-dependent cell-mediated cytotoxicity mediating PD-L1 high CD11b + cells elimination in hepatocellular carcinoma.

doi: 10.1016/j.apsb.2022.08.006

Figure Lengend Snippet: Figure 8 Bavencio induced enhanced ADCC effect than mutant Fc antibodies in eliminating human HCC infiltrating PD-L1þ CD33bþ myeloid cells in vitro. (A) Surgically resected human HCC tumor and autologous PBMCs were collected from three individuals. Three different parts of each tumor was collected considering intratumoral heterogeneity. CD33þ myeloid cells and CD45- tumor cells from tumor were sorted as target cells, and CD56þ NK cells from PBMCs for effector cells. NK cells were co-cultured with target cells as previously described. (B) Quantification of flow cytometry revealed PD-L1 expressed on tumor cell, T cell and myeloid cells in tumor microenvironment for three patients. (C) Representative images for IFN-g Elispot assay revealed the different antibodies induced ADCC effect. Target cells derived from one of the three part of tumor in HCC-1 candidate. PHA was regarded as positive control, no antibodies added and Isotype IgG1 was control group. (D) The number of IFN-g spots/1 105 cells in each group. n Z 3 for each patient, *P < 0.05, **P < 0.01, ***P < 0.001. ns, no significant difference, t- tests. All data are mean SEM.

Article Snippet: This mix was then added to anti-IFN-g pre-coated plates from the human IFN-g ELISpot assay kit (DKW22-1000-096s; Dakewei, China), along with the following control groups: no antibody, isotype IgG1 control (MCE, HY-P9900, Newark, NJ, USA), and positive control (phytohemagglutinin stimulation).

Techniques: Mutagenesis, In Vitro, Cell Culture, Cytometry, Enzyme-linked Immunospot, Derivative Assay, Positive Control, Control

Journal: Nature Communications

Article Title: CD200R1-CD200 checkpoint inhibits phagocytosis differently from SIRPα-CD47 to suppress tumor growth

doi: 10.1038/s41467-025-60456-3

Figure Lengend Snippet: a Flow cytometry analysis of CD200 (top) or tumor antigens (CD20, CD38, CD123, DLL3 and SLAMF7; bottom) on various human tumor cell lines. Red curves, CD200 mAb or tumor antigen-specific mAbs. Filled curves, Ctrl mAbs. b As per Fig. , except using IL-4-primed human blood monocyte-derived macrophages and human tumor cells in the presence of samalizumab (human IgG1, Fc-silent) or Ctrl mAb MOPC21 (human IgG1, Fc-silent). IgG opsonization was performed using CD20 mAb rituximab (SLVL, 721.221), CD38 mAb daratumumab (NCI-H929), CD123 mAb talacotuzumab (KG-1a), DLL3 mAb rovalpituzumab (NCI-H209), or SLAMF7 mAb elotuzumab (SK-MEL-28) ( n = 3 or 4). c As per Fig. 5b, except using the indicated cell lines and samalizumab (human IgG1, Fc-silent), CD47 mAb B6H12 (human IgG1, Fc-silent), or Ctrl mAb MOPC21 (human IgG1, Fc-silent). Tumor cells were opsonized as detailed for Fig. 5b, ( n = 3). d –f Subcutaneous injection of SLVL cells in NSG mice, followed by intraperitoneal injection of rituximab (mouse IgG2a version) combined with samalizumab or Ctrl mAb every 2 days, ( n = 8). d Schematic representation of the experimental workflow. e Tumor volume over time. f Tumor weight. g –i Subcutaneous injection of 721.221 cells in NSG mice, followed by intraperitoneal injection of rituximab (mouse IgG2a version) combined with samalizumab or Ctrl mAbs every 2 days, ( n = 10). g Schematic representation of the experimental workflow. h Tumor volume over time. i Tumor weight. Data are from three to four ( a, b ), three ( c ), or two ( d-i ) independent experiments. Each symbol represents one human sample or one mouse. Data are presented as mean ± s.e.m. Statistical analysis: two-tailed t-test ( f , i ), with multiple comparisons ( b, c , e , h ). ns, not significant. See also Supplementary Fig. .

Article Snippet: For cell opsonization, the following mAbs were used: Tac mAb 7G7 (BioXCell); human CD20 mAb rituximab biosimilar (BioXCell); human CD123 mAb talacotuzumab (MedChemExpress, Princeton, NJ); human CD38 mAb daratumumab (MedChemExpress); human DLL3 mAb rovalpituzumab (MedChemExpress).

Techniques: Flow Cytometry, Derivative Assay, Injection, Two Tailed Test