mouse bladder cancer cell line mb49  (Millipore)

Journal: Cancer Research Communications

Article Title: Development of an Anti-canine PD-L1 Antibody and Caninized PD-L1 Mouse Model as Translational Research Tools for the Study of Immunotherapy in Humans

doi: 10.1158/2767-9764.CRC-22-0468

Figure Lengend Snippet: cPD-L1 antibodies enhance antitumor immunity in the caninized PD-L1 syngeneic mouse model. A, Knock-in strategy of the caninized PD-L1 mice (c57BL/c background). B, Validation of cPD-L1 protein expression in the MB49 cPD-L1 cells. Flow cytometric analysis of membrane located mPD-L1 and cPD-L1 protein in MB49 cells expressing cPD-L1 (MB49 cPD-L1 ) or MB49 parental cells. C, Immunofluorescence staining and protein expression pattern of mPD-L1 and cPD-L1 in MB49 or MB49 cPD-L1 tumor masses from the caninized PD-L1 mice. DAPI, nuclear counterstaining. Scale bar, 100 μm. D, Interaction of cPD-1 or mPD-1 protein with cPD-L1 or mPD-L1 protein with or without cPD-L1 antibody, 12C. His-tagged canine or mPD-L1 protein was immobilized on the Ni-NTA 96-well plate, and HRP-conjugated anti-human IgG Fc-specific secondary with mPD-1-hFc or cPD-1-hFc protein was added. OD 450 was measured to quantify the amount of bound PD-1 protein. E, Binding of cPD-L1 antibodies, 12C and 3C, with human PD-L1 (hPD-L1), mPD-L1, and cPD-L1 proteins. His-tagged human PD-L1, mPD-L1, or cPD-L1 protein was immobilized on the Ni-NTA 96-well plate, and anti-cPD-L1 antibodies, 12C or 3C, with HRP-conjugated anti-canine IgG-specific secondary was added. OD 450 was measured to quantify the amount of bound PD-L1 antibodies. Ab, antibody. F, Tumor growth of MB49 cPD-L1 in the caninized PD-L1 mice treated with cPD-L1 antibody, 12C or 3C. The IgG isotype of 12C and 3C antibodies is mouse IgG1 which is equivalent to human IgG4. Tumors were measured at the indicated timepoints ( n = 8 per group). At the endpoint, the tumors were dissected. G–I, Immunofluorescence staining, and protein expression pattern of CD8 and granzyme B in MB49 tumor masses from IgG-, 12C-, or 3C-treated mice. DAPI, nuclear counterstaining. Scale bar, 100 μm. Representative images of immunostaining of CD8 and granzyme B in the MB49 tumor mass (G). CD8 (H) and granzyme B (I) were quantified using Gen5 software (BioTek). n = 10. Treatment with the PD-L1 antibody did not affect kidney function (serum creatinine; J ) or liver enzyme activity (ALT; K ), measured in blood collected at the end of the experiment. ALT, alanine aminotransferase.

Article Snippet: The BT549 human breast cancer and MB49 mouse bladder cancer cell lines were obtained from ATCC and Millipore Sigma, respectively.

Techniques: Knock-In, Biomarker Discovery, Expressing, Membrane, Immunofluorescence, Staining, Binding Assay, Immunostaining, Software, Activity Assay

Journal: Frontiers in Endocrinology

Article Title: Enhanced Mortality to Metastatic Bladder Cancer Cell Line MB49 in Vasoactive Intestinal Peptide Gene Knockout Mice

doi: 10.3389/fendo.2017.00162

Figure Lengend Snippet: Kaplan–Meier plot comparing death rates between vasoactive intestinal peptide knockout (VIP KO) controls, VIP KO mice with MB49, C57BL/6 controls, and C57BL/6 mice with MB49 ( p = 0.002).

Article Snippet: Using a mouse bladder cancer cell line, MB49, obtained from Timothy Ratliff (Purdue University College of Veterinary Medicine), we created a model using leg injections of the cancer cells to test whether loss of the VIP gene leads to increased mortality and/or morbidity from bladder cancer metastases, compared to control C57BL/6 mice.

Techniques: Knock-Out