b6h12 2 hybridoma  (ATCC)

Journal: Experimental and molecular pathology

Article Title: Facilitating cholangiocarcinoma inhibition by targeting CD47.

doi: 10.1016/j.yexmp.2024.104935

Figure Lengend Snippet: Fig. 2. CD47 and CD68 expressions in iCCA. Representative immunohistochemistry staining of (A) CD47 and (B) CD68-expressing cells. CD47 is categorized into negative, 1+ (weak), 2+ (moderate), and 3+ (intense) staining, and CD68-expressing cells are classified as light, moderate, and dense infiltrations. Bar = 100 μm, 200× magnification.

Article Snippet: Anti-human CD47 (B6H12.2) hybridomas were obtained from the ATCC (MD, VA, USA).

Techniques: Immunohistochemistry, Staining, Expressing

Journal: Experimental and molecular pathology

Article Title: Facilitating cholangiocarcinoma inhibition by targeting CD47.

doi: 10.1016/j.yexmp.2024.104935

Figure Lengend Snippet: Fig. 3. Kaplan–Meier survival analysis of patients with iCCA with different CD47 and CD68 expressions and distinct histological subtypes. Kaplan–Meier survival analysis of patients with iCCA with different CD47 (A, C, E) and CD68 (B, D, F) expressions. Total iCCA (A, B), papillary iCCA (C, D), and non-papillary subtype (E, F) were subdivided. Dense CD68-expressing cells in total iCCA and non-papillary iCCA were related to longer survival times. *P < 0.05, **P < 0.01.

Article Snippet: Anti-human CD47 (B6H12.2) hybridomas were obtained from the ATCC (MD, VA, USA).

Techniques: Expressing

Journal: Experimental and molecular pathology

Article Title: Facilitating cholangiocarcinoma inhibition by targeting CD47.

doi: 10.1016/j.yexmp.2024.104935

Figure Lengend Snippet: Fig. 4. CD47 alterations in CD47-deficient clones. (A) CD47 mutations in CD47-deficient clones. Parts of CD47 exon 1 DNA sequences were compared. Underline represents the CD47 gRNA-targeting sequences. Arrowheads indicate the predicted Cas9 cleavage site. Green characters represent protospacer adjacent motif (PAM) sequences. Red characters represent DNA mutation observed on CD47 deficient clones. A dash indicates deletion. The numbers in parentheses represent numbers of inserted (+) nucleotides. (B) CD47 and GFP expressions of KKU-213A and CD47-deficient clones. CD47 expressions on the surface of CCA cells were detected by flow cytometry using PerCP/Cy5.5 conjugated anti-human CD47 antibody. CD47 was not detected on clones #19, #23, and #28. GFP-positive cells represent CD47- CRISPR/Cas9-transduced cells. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Anti-human CD47 (B6H12.2) hybridomas were obtained from the ATCC (MD, VA, USA).

Techniques: Clone Assay, Mutagenesis, Flow Cytometry, CRISPR

Journal: Experimental and molecular pathology

Article Title: Facilitating cholangiocarcinoma inhibition by targeting CD47.

doi: 10.1016/j.yexmp.2024.104935

Figure Lengend Snippet: Fig. 5. The effects of CD47-deficiency in vitro and in vivo. (A) Cell numbers of KKU-213A and CD47 deficient clones #19, #23, and #28 were determined by formazan-based MTS assay. Cell proliferation was presented as fold change calculated by dividing OD492 at indicated times with OD492 at 8 h. The data are presented as mean ± SD from three independent experiments. (B) Phagocytic indices of CD47-deficient clones #19, #23, and #28 compared to KKU-213A. CSFE-labeled KKU- 213A and CD47-deficient clones were incubated with MDMs, and phagocytosed cells were CSFE+/CD11b+ cells. The phagocytic index was calculated by deter mining phagocytic cells per 100 MDMs. (C) Photograph of transplanted tumor on day 14 after injection. The black circles represented missing tumors. (D) Tumor weight was obtained from KKU-213A, CD47 deficient clones #19 and #28 subcutaneously injected into mice. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Anti-human CD47 (B6H12.2) hybridomas were obtained from the ATCC (MD, VA, USA).

Techniques: In Vitro, In Vivo, Clone Assay, MTS Assay, Labeling, Incubation, Injection

Journal: Translational Oncology

Article Title: Attenuation of CD47-SIRPα Signal in Cholangiocarcinoma Potentiates Tumor-Associated Macrophage-Mediated Phagocytosis and Suppresses Intrahepatic Metastasis 1 2

doi: 10.1016/j.tranon.2018.10.007

Figure Lengend Snippet: High CD47 expressions protected CCA from macrophage phagocytosis, and these effects were abolished by disrupting CD47-SIRPα interaction. (A) Expressions of CD47 in HCC (▲) and CCA (●) tissues were expressed by H-score. Each spot represents an individual sample. Representative pictures of HCC and CCA are shown. Bar = 100 μm. (B) CD47 expressions in HCC (HepG2 and Huh7) and CCA (KKU-055, KKU-100, KKU-213, and HuCCT1) cell lines were determined by flow cytometry. White histogram represents anti-CD47–stained cells, while gray histogram represents isotype control staining. Numbers on upper-right corners are the mean fluorescent intensities (MFI) of anti-CD47–stained cells normalized by those of isotype control. (C) Phagocytic indices of anti-CD47 blocking antibody (B6H12.2) treatment compared to nonblocking antibody (2D3). (D) Comparison of anti-CD47 (B6H12.2) and anti-SIRPα (SE5A5) treatments. Representative pictures of phagocytic cells are presented with phagocytosed cells indicated by the arrow heads. IgG was isotype control IgG. ** P < .01, *** P < .001.

Article Snippet: Anti-human CD47 (B6H12.2) hybridoma was obtained from ATCC (MD, USA).

Techniques: Flow Cytometry, Staining, Control, Blocking Assay, Comparison

Journal: Translational Oncology

Article Title: Attenuation of CD47-SIRPα Signal in Cholangiocarcinoma Potentiates Tumor-Associated Macrophage-Mediated Phagocytosis and Suppresses Intrahepatic Metastasis 1 2

doi: 10.1016/j.tranon.2018.10.007

Figure Lengend Snippet: Anti-CD47 treatment reduced cancer colonization in spleen and liver but increased macrophage infiltration. Splenic (A) and liver (B) weights of mice in control (white box) and anti-CD47–treated group (black box) are presented. Red fluorescent intensities per time (total counts/second) detected from spleens (C) and livers (D) are shown. Representative images are presented in small insets. 1 = control, 2 = anti-CD47 treated, 3 = non–cancer-injected (NRJ) organ. Hematoxylin and eosin and immunohistochemistry staining of CK19 (bile duct marker) and F4/80 (mouse macrophage marker) is demonstrated in spleen (E) and liver (F). NRJ = corresponding area of spleen and liver from NRJ mouse not injected with CCA cells. Bar = 100 μm. Percentages of macrophage per cancer area in spleens (G) and livers (H) of control and anti-CD47–treated group are presented ( n = 4/group). (I) % CK19-positive area in anterior and posterior right lobes of livers from both groups ( n = 4/group). Small insets show representative images of livers. Bars show minimum to maximum numbers. * P < .05.

Article Snippet: Anti-human CD47 (B6H12.2) hybridoma was obtained from ATCC (MD, USA).

Techniques: Control, Injection, Immunohistochemistry, Staining, Marker

Journal: Translational Oncology

Article Title: Attenuation of CD47-SIRPα Signal in Cholangiocarcinoma Potentiates Tumor-Associated Macrophage-Mediated Phagocytosis and Suppresses Intrahepatic Metastasis 1 2

doi: 10.1016/j.tranon.2018.10.007

Figure Lengend Snippet: Treatment with anti-CD47 antibody potentiated the phagocytic activities of all macrophage subgroups. (A) Representative images of CCA-phagocytosed resting, M1, M2, and TAM-like subgroups. Arrowheads indicated phagocytosed cells in isotype control IgG- and anti-CD47–treated groups. Bar = 100 μm. (B) Phagocytic indices of IgG- (○) and anti-CD47–treated (●) resting, M1, M2, and TAM-like MDMs. Individual spots each represent the mean phagocytic index from an individual donor. Line compared the phagocytic indices from the same donor. * P < .05.

Article Snippet: Anti-human CD47 (B6H12.2) hybridoma was obtained from ATCC (MD, USA).

Techniques: Control