Journal: Developmental Cell

Article Title: Defining the identity and the niches of epithelial stem cells with highly pleiotropic multilineage potency in the human thymus

doi: 10.1016/j.devcel.2023.08.017

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Article Snippet: HumanKine® recombinant human IL-7 protein , Proteintech , cat# HZ-1281.

Techniques: Recombinant, Modification, Selection, RNA Extraction, Polymer, Cell Culture, Derivative Assay, Software

Journal: STAR Protocols

Article Title: Efficient generation of locus-specific human CAR-T cells with CRISPR/cCas12a

doi: 10.1016/j.xpro.2022.101321

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Article Snippet: IL-7 Protein, Human, Recombinant , SinoBiological , Cat# 11821-HNAE.

Techniques: Recombinant, Electroporation, Modification, SYBR Green Assay, Transfection, Enzyme-linked Immunosorbent Assay, Cytotoxicity Assay, DNA Extraction, Software, Real-time Polymerase Chain Reaction, Protein Purification, Plasmid Preparation, Synthesized, Flow Cytometry, Spectrophotometry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Interleukin-7-loaded oncolytic adenovirus improves CAR-T cell therapy for glioblastoma

doi: 10.1007/s00262-021-02856-0

Figure Lengend Snippet: oAD-IL7 successfully replicate in glioma. a Schematic representation of genome of the oncolytic viruses used in this study. b Monolayers of glioblastoma cell lines were infected by oAD-IL7 at the MOI of 10. Cells were imaged with a fluorescence microscope 24, 48, and 72 h after infection. Scale bar, 500 μm. c Cells were collected 72 h after infection for plaque assay. Each number of the virus tilter were calculated on average of three independent wells. d Oncolytic virus-induced GBM apoptosis was confirmed by western blot. Scale bar, 100 μm. e Cells were stained with IL7 antibody 72 h after infection and imaged with a fluorescence microscope. Green fluorescence represents the oAD-IL7 infection; red fluorescence represents the IL7 distribution; blue fluorescence represents the cell nuclei stained with DAPI. Scale bar, 50 nm. f Supernatants were collected for IL7 quantification. g The median tumor volume of the xenograft models treated by PBS, oAD, and oAD-IL7 was recorded. h Survival analysis of the three group of the xenograft models. *P < 0.05; **P < 0.01; ***P < 0.001 (two-tailed paired t test)

Article Snippet: A cDNA encoding human IL7 purchased from Sinobiological (Beijing, China) was cloned and then inserted the E1 region, and an E1A gene under the control of hTERT, followed by a E1B gene under the control of TATA box was synthesized by General Biosystems (Chuzhou, Anhui province, China) and inserted into the E3 region.

Techniques: Infection, Fluorescence, Microscopy, Plaque Assay, Virus, Western Blot, Staining, Two Tailed Test

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Interleukin-7-loaded oncolytic adenovirus improves CAR-T cell therapy for glioblastoma

doi: 10.1007/s00262-021-02856-0

Figure Lengend Snippet: oAD-IL7 enhanced the survival and efficacy of B7H3-CAR-T. a 1 × 10 B7H3-CAR-T or untransduced T cells underwent serial coculturing with 2 × 105 oncolytic virus-infected U87 cells for a total of 7 days. The GBM cells were refreshed on the 4th day of the serial coculture. b Cumulations of T cells expansion. c For proliferation analysis, T cells were stained with CytoTell Blue before serial coculture and collected for flow cytometry at the end of day 3 and day 7 to monitor the results of division. d For survival analysis, T cells were collected and stained with Annexin V and 7-AAD. e 51Cr-release assays were used to evaluate the lysis of GBM during the second round of coculture at the end of day 7. Data represent the mean ± SD of triplicate wells. *P < 0.05; **P < 0.01 (two-tailed paired t test)

Article Snippet: A cDNA encoding human IL7 purchased from Sinobiological (Beijing, China) was cloned and then inserted the E1 region, and an E1A gene under the control of hTERT, followed by a E1B gene under the control of TATA box was synthesized by General Biosystems (Chuzhou, Anhui province, China) and inserted into the E3 region.

Techniques: Virus, Infection, Staining, Flow Cytometry, Lysis, Two Tailed Test

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Interleukin-7-loaded oncolytic adenovirus improves CAR-T cell therapy for glioblastoma

doi: 10.1007/s00262-021-02856-0

Figure Lengend Snippet: oAD-IL7 prolonged the survival of tumor-infiltrating CAR-T cells in xenograft models. In a parallel experiment, the tumor-bearing mice underwent a delayed treatment since the 21st day after tumor inoculation. a The tumors of the mice treated with single B7H3-CAR-T therapy or combined therapy were imaged by MRI scanning at day 28. b The tumors were removed from the treated mice at day 30, and the immune infiltrate was evaluated by immunohistochemical staining according to the distribution of CD3+ cells. Scale bar, 200 μm. c Titration of oAD-IL7 in glioblastoma tissue by plaque assay. d The ratio of CD3 positive cells in tumor tissues removed from mice treated by combined therapy or single B7H3-CAR-T therapy according to the count of average CD3 + cells in each single field. e Representative flow cytometry analysis of the T cells in the tumor removed from mice underwent different therapies. f The expression of PD1 and LAG-3 of the CD3 positive cells in two groups. g The level of LAG-3 expression on the tumor-infiltrating T cells of two groups. h The level of PD1 expression on the tumor-infiltrating T cells of two groups. i Ki67 expression of the CD3 positive cells in two groups. j Expression of the B7H3 and CXAR of the tumors removed from tumor-bearing mice on day 21. Scale bar, 50 μm. k Expression of the B7H3 and CXAR of the tumors removed from tumor-bearing mice on day 30. Scale bar, 50 μm

Article Snippet: A cDNA encoding human IL7 purchased from Sinobiological (Beijing, China) was cloned and then inserted the E1 region, and an E1A gene under the control of hTERT, followed by a E1B gene under the control of TATA box was synthesized by General Biosystems (Chuzhou, Anhui province, China) and inserted into the E3 region.

Techniques: Immunohistochemical staining, Staining, Titration, Plaque Assay, Flow Cytometry, Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Interleukin-7-loaded oncolytic adenovirus improves CAR-T cell therapy for glioblastoma

doi: 10.1007/s00262-021-02856-0

Figure Lengend Snippet: Combination of oAD-IL7 and B7H3-CAR-T improved the anti-tumor effect in vivo. 2 × 105 GBM-LUCF cells were orthotopically injected into the NCG mice, and the bioluminescent imaging was performed twice a week since the 7th day after injection. The oncolytic virus was orthotopically injected on day 7, and the B7H3-CAR-T cells were systematically applied on day 9. a Experimental timeline for in vivo studies. b The representative bioluminescent images of the growth of glioblastoma over time were shown. c The expression of B7H3 and CXAR in the glioblastoma tissue of patient. Scale bar, 50 μm. d, e, and f Quantitated GBM-LUCF bioluminescence from each treatment group is displayed over time. g Average radiance of the three groups of the mice during the first 3 weeks. h Kaplan–Meier survival analysis of GBM-LUCF challenged mice after treatment with oncolytic virus and T cells. *P < 0.05; **P < 0.01; ***P < 0.001 (two-tailed paired t test). i H&E staining of the specimen removed from tumor-bearing mice that underwent a parallel experiment at day 30

Article Snippet: A cDNA encoding human IL7 purchased from Sinobiological (Beijing, China) was cloned and then inserted the E1 region, and an E1A gene under the control of hTERT, followed by a E1B gene under the control of TATA box was synthesized by General Biosystems (Chuzhou, Anhui province, China) and inserted into the E3 region.

Techniques: In Vivo, Injection, Imaging, Virus, Expressing, Two Tailed Test, Staining