Journal: Molecular cancer therapeutics
Article Title: Targeting TfR1 with the ch128.1/IgG1 antibody inhibits EBV driven lymphomagenesis in immunosuppressed mice bearing EBV + human primary B-cells
doi: 10.1158/1535-7163.MCT-21-0074
Figure Lengend Snippet: A, Representative flow cytometry plots of cells isolated from spleens of mice implanted with EBV− or EBV+ B-cell enriched preparations and treated with ch128.1/IgG1 or IgG1 control monoclonal antibody. Plots compare the percentage of murine CD45+ (mCD45+) and human CD45+ (hCD45+) cell populations (gated from live cells). Human CD3+ T-cell and CD19+ B-cell populations were gated from hCD45+ cells. B, Percentage of hCD45+ cells in peripheral blood, spleen, lymph node, and liver tissues for each EBV+ group treated with ch128.1/IgG1 or IgG1 control. The bars represent mean values of %hCD45+ B-cells; EBV+ group treated with ch128.1/IgG1: peripheral blood mean = 0.7%, spleen mean = 2.4%, lymph node mean = 2.6%, liver mean = 4.4%; EBV+ group treated with IgG1 isotype control: peripheral blood mean = 4.5%, spleen mean = 9.3%, lymph node mean = 10.5%, liver mean = 7.3%. C, Percentage of hCD19+ B-cells in peripheral blood, spleen, lymph node, and liver tissues. Percentages of hCD19+ B-cells were calculated from the total percentage of hCD45+ cells for each sample [(percentage of hCD19+) × (percentage of hCD45+ / 100%)]. The bars represent mean values of %hCD19+ B-cells; EBV+ group treated with ch128.1/IgG1: peripheral blood mean = 0.4%, spleen mean = 1.2%, lymph node mean = 0.8%, liver mean = 1.2%; EBV+ group treated with IgG1 isotype control: peripheral blood mean = 3.3%, spleen mean = 6.4%, lymph node mean = 7.6%, liver mean = 4.8%. Combined results shown in B and C are from 2 independent experiments. The results shown here for the EBV+ group treated with IgG1 control are from 12 of the 14 mice presented in Fig. 1B as two mice died before acquiring tissue at necropsy. Statistical comparisons were made for ch128.1/IgG1 versus IgG1 control treatments using nonparametric, unpaired Mann-Whitney tests, where *p = 0.01–0.05, **p = 0.001–0.01, and ns, not significant.
Article Snippet: IHC was performed using the following antibodies: polyclonal rabbit anti-human Kappa (κ) Light Chain (Agilent, catalog no. A019102–2); polyclonal rabbit anti-human Lambda (λ) Light Chain (Agilent, catalog no. A019302–2); monoclonal mouse anti-Epstein-Barr Virus, LMP Clone CS.1–4 (Agilent, catalog no. M0897); polyclonal rabbit anti-human CD3 (Agilent, catalog no. GA50361–2); and monoclonal mouse anti-human CD19 (Clone LE-CD19) (Agilent, catalog no. M729629–2).
Techniques: Flow Cytometry, Isolation, MANN-WHITNEY
Bio-Rad is a verified supplier 
![A, Representative flow cytometry plots of cells isolated from spleens of mice implanted with EBV− or EBV+ B-cell enriched preparations and treated with ch128.1/IgG1 or IgG1 control monoclonal antibody. Plots compare the percentage of murine CD45+ (mCD45+) and human CD45+ (hCD45+) cell populations (gated from live cells). Human CD3+ T-cell and <t>CD19+</t> B-cell populations were gated from hCD45+ cells. B, Percentage of hCD45+ cells in peripheral blood, spleen, lymph node, and liver tissues for each EBV+ group treated with ch128.1/IgG1 or IgG1 control. The bars represent mean values of %hCD45+ B-cells; EBV+ group treated with ch128.1/IgG1: peripheral blood mean = 0.7%, spleen mean = 2.4%, lymph node mean = 2.6%, liver mean = 4.4%; EBV+ group treated with IgG1 isotype control: peripheral blood mean = 4.5%, spleen mean = 9.3%, lymph node mean = 10.5%, liver mean = 7.3%. C, Percentage of <t>hCD19+</t> B-cells in peripheral blood, spleen, lymph node, and liver tissues. Percentages of hCD19+ B-cells were calculated from the total percentage of hCD45+ cells for each sample [(percentage of hCD19+) × (percentage of hCD45+ / 100%)]. The bars represent mean values of %hCD19+ B-cells; EBV+ group treated with ch128.1/IgG1: peripheral blood mean = 0.4%, spleen mean = 1.2%, lymph node mean = 0.8%, liver mean = 1.2%; EBV+ group treated with IgG1 isotype control: peripheral blood mean = 3.3%, spleen mean = 6.4%, lymph node mean = 7.6%, liver mean = 4.8%. Combined results shown in B and C are from 2 independent experiments. The results shown here for the EBV+ group treated with IgG1 control are from 12 of the 14 mice presented in Fig. 1B as two mice died before acquiring tissue at necropsy. Statistical comparisons were made for ch128.1/IgG1 versus IgG1 control treatments using nonparametric, unpaired Mann-Whitney tests, where *p = 0.01–0.05, **p = 0.001–0.01, and ns, not significant.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_9068/pmc08419068/pmc08419068__nihms-1712115-f0002.jpg)
