Journal: Acta Pharmaceutica Sinica. B

Article Title: Generation of α Gal-enhanced bifunctional tumor vaccine

doi: 10.1016/j.apsb.2022.03.002

Figure Lengend Snippet: Generation of END + -MHCC97H, Gal + -MHCC97H and END + /Gal + -MHCC97H cells. (A) Immunofluorescence was used to detect the expression of END, α 1,3 GT, and α 1,3 GT-END in MHCC97H cells. END + -MHCC97H cells were bound by FITC-conjugated anti-human END antibody; Gal + -MHCC97H cells were stained by Alexa Fluor 647-conjugated isolectin GS-IB4 lectin; END + /Gal + -MHCC97H cells were labeled by both fluorescent dye. MHCC97H cells were used as negative controls. Red: GS-IB4; Green: anti-human END antibody; Blue: nucleus. (Original magnification, 400 ×; Scale bar, 50 μm). (B) Expression of α 1,3 GT and END in MHCC97H, MHCC97H (pLVX-Puro), END + -MHCC97H, Gal + -MHCC97H and END + /Gal + -MHCC97H cells were assessed by flow cytometry. Quantitative analysis for immunofluorescence (C) and flow cytometry (D). ∗∗∗∗ P < 0.0001, ns stands for no difference. Data are representative of at least three individual experiments and are presented as mean ± SD.

Article Snippet: MHCC97H cells stably expressing α 1,3 GT or/and END and MHCC97H cells stably transduced with empty pLVX-Puro vector were generated using GenScript (Nanjing, China) and maintained in minimal essential medium (MEM; GIBCO) supplemented with 10% FBS, 100 U/mL penicillin/streptomycin, and 0.35 μg/mL puromycin (Sigma–Aldrich, St. Louis, MO, USA).

Techniques: Immunofluorescence, Expressing, Staining, Labeling, Flow Cytometry

Journal: Acta Pharmaceutica Sinica. B

Article Title: Generation of α Gal-enhanced bifunctional tumor vaccine

doi: 10.1016/j.apsb.2022.03.002

Figure Lengend Snippet: Detection of cell surface markers and production of IL-12p70 by five types of MHCC97H/DC fusion cells. Flow cytometry was used to assess the expression of cell surface markers, including CD83, CD86, HLA-ABC, HLA-DR, END and α Gal. Flow cytometry analysis of CD83, CD86, HLA-ABC, and HLA-DR (A) and quantitative analysis (B). (C) Flow cytometry analysis of END as well as α Gal. (D) Quantitative analysis. (E) ELISA was used to assay levels of IL-12p70 secreted after 48 h culture. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, ns stands for no difference. Data show mean ± SD and individual values from three independent experiments.

Article Snippet: MHCC97H cells stably expressing α 1,3 GT or/and END and MHCC97H cells stably transduced with empty pLVX-Puro vector were generated using GenScript (Nanjing, China) and maintained in minimal essential medium (MEM; GIBCO) supplemented with 10% FBS, 100 U/mL penicillin/streptomycin, and 0.35 μg/mL puromycin (Sigma–Aldrich, St. Louis, MO, USA).

Techniques: Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Generation of α Gal-enhanced bifunctional tumor vaccine

doi: 10.1016/j.apsb.2022.03.002

Figure Lengend Snippet: Cytotoxicity of T cells against 51 Cr-labeled target cells after induction by different types of MHCC97H/DC fusion cells, at different ratios of effector cells (T cells) to target cells (E:T). Target cells were (A) 293T, (B) A549, (C) HUVEC, (D) MHCC97H, (E) END + -MHCC97H, (F) Gal + -MHCC97H, and (G) END + /Gal + -MHCC97H/DC. (H) Cytotoxicity of T cells stimulated by END + /Gal + -MHCC97H/DC fusion cells was compared for other indicated target cells. In all cases, the ratio of T cells to target cells was 30:1. Measurement of IFN- γ -producing T cells following 7-day co-culture of with different types of MHCC97H/DC fusion cells. (I) Representative ELISPOT results. (J) Average numbers of IFN- γ -producing T cells per sample of 3 × 10 5 T cells. Spot numbers of Gal + -MHCC97H/DC and END + /Gal + -MHCC97H/DC fusion cells were significantly greater than that of other fusion cell types. ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, ns stands for no difference. Data are representative of at least three individual experiments and are presented as mean ± SD.

Article Snippet: MHCC97H cells stably expressing α 1,3 GT or/and END and MHCC97H cells stably transduced with empty pLVX-Puro vector were generated using GenScript (Nanjing, China) and maintained in minimal essential medium (MEM; GIBCO) supplemented with 10% FBS, 100 U/mL penicillin/streptomycin, and 0.35 μg/mL puromycin (Sigma–Aldrich, St. Louis, MO, USA).

Techniques: Labeling, Co-Culture Assay, Enzyme-linked Immunospot

Journal: Acta Pharmaceutica Sinica. B

Article Title: Generation of α Gal-enhanced bifunctional tumor vaccine

doi: 10.1016/j.apsb.2022.03.002

Figure Lengend Snippet: Antitumor effects of T cells induced by different types of MHCC97H/DC fusion cells. (A) Mice injected with MHCC97H cells (2 × 10 6 cells/mouse on Day 0) were treated with stimulated T cells once a week (from Day 5 to Day 33). (B) Tumor volume was measured every 5 days from Day 5 to Day 35. Tumors were significantly smaller in mice treated with T cells primed by END + /Gal + -MHCC97H/DC fusion cells than in mice treated with T cells primed by other types of fusion cells. ∗∗∗ P < 0.001 (by Day 35). (C) Kaplan–Meier survival curves of hepatoma-bearing nude mice ( n = 15 in each group). Curves were compared using the log-rank test. ∗∗∗∗ P < 0.0001.

Article Snippet: MHCC97H cells stably expressing α 1,3 GT or/and END and MHCC97H cells stably transduced with empty pLVX-Puro vector were generated using GenScript (Nanjing, China) and maintained in minimal essential medium (MEM; GIBCO) supplemented with 10% FBS, 100 U/mL penicillin/streptomycin, and 0.35 μg/mL puromycin (Sigma–Aldrich, St. Louis, MO, USA).

Techniques: Injection

Journal: Acta Pharmaceutica Sinica. B

Article Title: Generation of α Gal-enhanced bifunctional tumor vaccine

doi: 10.1016/j.apsb.2022.03.002

Figure Lengend Snippet: Mice had been injected with PBS or T cells stimulated by one of the following types of fusion cells: MHCC97H/DC, MHCC97H (pLVX-Puro)/DC, END + -MHCC97H/DC, Gal + -MHCC97H/DC, END + /Gal + -MHCC97H/DC. T cells stimulated by END + /Gal + -MHCC97H/DC fusion cells increased tumor cell apoptosis and inhibited their proliferation and microvessel formation in hepatoma-bearing nude mice. Cells apoptosis in tumor tissues of mice that received different treatments were detected by TUNEL. (A) TUNEL staining in the tumor tissues of mice that received different treatments (Original magnification, 200 ×; scale bar, 100 μm). (B) The mean and standard deviation of TUNEL staining in each group. (C) PCNA staining in tumor tissues of mice that received different treatments (Original magnification, 400 ×; scale bar, 50 μm). (D) PCNA-positive cells were counted in five randomly selected fields of tumor thin sections. Quantitative analysis for average numbers of positive cells. (E) Expression of CD31 in tumor tissues of mice that received different treatments (Original magnification, 200 ×; scale bar, 100 μm). (F) CD31-positive microvessels were counted in five randomly selected fields of tumor thin sections. Quantitative analysis for average numbers of microvessels. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. Data are representative of at least three individual experiments and are presented as mean ± SD.

Article Snippet: MHCC97H cells stably expressing α 1,3 GT or/and END and MHCC97H cells stably transduced with empty pLVX-Puro vector were generated using GenScript (Nanjing, China) and maintained in minimal essential medium (MEM; GIBCO) supplemented with 10% FBS, 100 U/mL penicillin/streptomycin, and 0.35 μg/mL puromycin (Sigma–Aldrich, St. Louis, MO, USA).

Techniques: Injection, TUNEL Assay, Staining, Standard Deviation, Expressing

Journal: PLoS ONE

Article Title: Suppression of AKT Anti-Apoptotic Signaling by a Novel Drug Candidate Results in Growth Arrest and Apoptosis of Hepatocellular Carcinoma Cells

doi: 10.1371/journal.pone.0054595

Figure Lengend Snippet: Cytotoxicity spectrum of HBF-0079 and the analogue IHVR-04042.

Article Snippet: MHCC97H were donated by Dr. Kunwar Shailubai (Synergy Pharmaceuticals, New York, NY).

Techniques: