Cadherin-17-specific antibodies and cytotoxic cells for cancer therapy
A cadherin, cytotoxicity technology applied in the field of cancer immunotherapy, which can solve the problems of limited expression and no observed partial or complete response
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example 1
[0136] Example 1 Research Design and Methods
[0137] An orthotopic tumor xenograft model was used to test the effect of Lic5 alone or in combination with cisplatin or epirubicin on tumor growth inhibition and animal survival. Human HCC cell lines MHCC97-L (97L) and PLC / PRF / 5 (PLC) were used. Lic5, cisplatin, and epirubicin treatment alone inhibited the growth of cultured HCC cells using a cell proliferation assay, while a more significant reduction was observed when either cisplatin or epirubicin was used together with Lic5. Similar growth inhibition trends were observed when the same experimental group was applied to orthotopic tumor-bearing nude mice.
[0138] Treatment with Lic5 enhanced the survival of orthotopic tumor-bearing nude mice when compared to controls. The combined Lic5 and epirubicin group produced the best survival among all experimental groups. For the next stage of antibody humanization, the complementarity determining regions (CDRs) on the variable re...
example 3
[0141] Example 3 In Vivo Cell Proliferation Assay
[0142] A predetermined number of 97L and PLC cells were seeded onto 96-well culture plates for 1 day. Mouse IgG (Sigma-Aldrich, St. Louis, MO, USA) at 150 μg / ml was used as a control for Lic5. After treatment, proliferation of treated cells was measured using Cell Proliferation Kit I (Roche, Indianapolis, IN, USA) as described above. Briefly, treated cells were used to incubate MTT reagent for 4 hours. The colorimetric signal was then measured after addition of 10% SDS. To confirm the results an independent cell proliferation MTT assay was performed at least twice.
example 4
[0143] Example 4 Orthotopic Xenograft Model
[0144] Male nude mice were obtained from the Laboratory Animal Unit of the University of Hong Kong, Hong Kong. Animals were housed in individually ventilated cages on a 12 h light / 12 h dark cycle and had free access to autoclaved water and food. The animal experiments performed in this study were approved by the committee that permits the use of live animals in our Faculty for Teaching and Research (CULATR). To allow in vivo visualization of growing orthotopic liver tumors, a pre-established luciferase-labeled 97L cell line was used. 5x106 luciferase-labeled 97L cells were injected subcutaneously into nude mice to allow subcutaneous tumor growth. These subcutaneous tumors were harvested when they reached 200-300 mm3 in size and used as tumor inoculations to establish orthotopic liver tumors in isolated groups of nude mice following standard procedures. Five days after orthotopic tumor implantation, mice received the following ...
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