single chain variable fragment scfv  (Cold Spring Harbor Laboratory Meetings)

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Therapeutic potential of anti-ErbB3 chimeric antigen receptor natural killer cells against breast cancer

doi: 10.1007/s00262-024-03923-y

Figure Lengend Snippet: Antitumor effects of aErbB3 CAR-NK cells against SK-BR-3 in vivo. A Timetable of an in vivo experiment ( n = 5 per group). Female NSGA mice (6 weeks old) were subcutaneously injected with 1 × 10 7 cells of the breast cancer cell line SK-BR-3. Five days later, they received 5 × 10 6 cells of aCD19 and aErbB3 CAR-NK cells. B SK-BR-3 tumor growth curve as calculated from caliper measurements of tumor growth twice weekly after subcutaneous injection. C Bar graph of tumor weight. D Image of excised tumors in mice. Scale bar, 1 cm. E Representative flow cytometric dot plots showing hCD45 + and hCD56 + CAR-NK cells infiltrating the tumor tissue in each treatment group. F Bar graph showing the percentage of tumor-infiltrating lymphocytes (TILs; hCD45 + cells) in each group. G Immunofluorescence staining of tumor sections to visualize CAR-NK cell infiltration. Tumor sections were stained with DAPI (blue) for nuclei and hCD45-FITC (green) for human CAR-NK cells. Images represent 400× magnification (scale bar, 50 μm). H Quantification of CAR-NK cell infiltration based on FITC-positive (hCD45. + ) dots counted in 400× fields across three samples per group. Data are presented as the mean ± SD.* P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 based on the results of a two-way ANOVA for the ( B ) and one-way ANOVA ( C , F , H )

Article Snippet: The aErbB3 single-chain variable fragment (scFv) sequence was provided by ISU Abxis [ ] (Seongnam, Republic of Korea).

Techniques: In Vivo, Injection, Immunofluorescence, Staining

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Therapeutic potential of anti-ErbB3 chimeric antigen receptor natural killer cells against breast cancer

doi: 10.1007/s00262-024-03923-y

Figure Lengend Snippet: Manufacture and validation of aErbB3 CAR-NK cells using cord blood-derived NK cells. A Schematic diagram depicting the construction of the second-generation aCD19 CAR and aErbB3 CAR encoded in a pCDH lentiviral vector with an MSCV promoter (left). Bar graph of the titer of the lentivirus produced (right). B Schematic representation of CAR-NK production from cord blood-derived NK cells and subsequent evaluation. C Representative flow cytometric analysis of CAR expression in CD3 − CD56. + subsets 3 days post-transduction (left). Summary data is presented as a scatter plot of the percentage of CAR expression in the non-transduced (UTD) and transduced cells ( n = 9, mean ± SD) (right). D Determination of maintaining aCD19 CAR and aErbB3 CAR expression on transduced NK cells. E ELISA analysis of IL-15 secretion in aCD19 CAR-NK cells and aErbB3 CAR-NK cells. Control: Culture medium of non-transduced NK cells, P2A + GFP: culture medium of NK cells transduced with CAR lentivirus (P2A + GFP). P2A + IL-15: culture medium of NK cells transduced with CAR lentivirus (P2A + IL-15)

Article Snippet: The aErbB3 single-chain variable fragment (scFv) sequence was provided by ISU Abxis [ ] (Seongnam, Republic of Korea).

Techniques: Biomarker Discovery, Derivative Assay, Plasmid Preparation, Produced, Expressing, Transduction, Enzyme-linked Immunosorbent Assay, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Therapeutic potential of anti-ErbB3 chimeric antigen receptor natural killer cells against breast cancer

doi: 10.1007/s00262-024-03923-y

Figure Lengend Snippet: Antitumor activity of aErbB3 CAR-NK cells in vitro. A Flow cytometric analysis of ErbB3 expression in various cell lines. B Cytotoxicity of aErbB3 CAR-NK cells and aCD19 CAR-NK cells against each ErbB3-positive and -negative cell line. Flow cytometric analysis of CFSE + /FVD + -stained target cells after 4 h of co-culture at the indicated effector: target ( E : T ) ratio. C CD107a expression in aErbB3 CAR-NK cells and aCD19 CAR-NK cells when co-cultured with their respective cells. Target and effector cells were co-cultured at a 1:1 ratio for 4 h, and this was followed by flow cytometric analysis. D Quantification of IFN-γ present in supernatants collected after 24 h of co-culture with target cells through ELISA. E Antigen-specific proliferation of aErbB3 CAR-NK cells co-cultured with X-ray-irradiated MDA-MB-231 cells (ErbB3 − ) and X-ray-irradiated MDA-MB-453 cells (ErbB3 + ) for 5 days ( E : T ratio = 1:2). Data are presented as the mean ± SD of three independent experiments ( B, C, D, E ) using CBMC CAR-NK cells from three donors. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 based on the results of Student’s t-test

Article Snippet: The aErbB3 single-chain variable fragment (scFv) sequence was provided by ISU Abxis [ ] (Seongnam, Republic of Korea).

Techniques: Activity Assay, In Vitro, Expressing, Staining, Co-Culture Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Irradiation

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Therapeutic potential of anti-ErbB3 chimeric antigen receptor natural killer cells against breast cancer

doi: 10.1007/s00262-024-03923-y

Figure Lengend Snippet: Phenotyping of aCD19 and aErbB3 CAR-NK cells. A Comparative analysis of surface marker expression in aCD19 and aErbB3 CAR-NK cells. B Representative density plots illustrating the expression of these markers in aCD19 and aErbB3 CAR-NK cells. Data are presented as the mean ± SD of three independent experiments

Article Snippet: The aErbB3 single-chain variable fragment (scFv) sequence was provided by ISU Abxis [ ] (Seongnam, Republic of Korea).

Techniques: Marker, Expressing