Cancer immunotherapy by delivering class II MHC antigens using a VLP-replicon
A technology of replicon and composition, which is applied in the direction of cancer antigen components, antibody medical components, virus antigen components, etc., and can solve the problems of easy contamination, trouble, too long, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0123] Example 1 - Generation of an alphavirus-based gene expression system
[0124] Alphavirus gene expression systems are designed to allow VLP-mediated delivery of exogenous genes of interest (GOIs) or proteins of interest (POIs) to target cells with a low risk of causing cytopathic viral infection. Expression systems were designed using three vectors that can be expressed in packaging cell lines to generate transduced VLPs. One vector encodes an alphavirus-based expression construct, another vector encodes a retroviral gag protein to facilitate VLP formation, and a third vector encodes a fusion protein to mediate fusion of VLPs to host cells. Additionally, the system is constructed to work without the need for the presence of alphavirus structural proteins.
[0125] To accomplish this, an alphavirus-based DNA plasmid was generated with the following: cytomegalovirus promoter (CMV); followed by a retroviral packaging signal for the corresponding retroviral packaging protei...
Embodiment 2
[0131] Example 2 - Expression of various proteins in target cells transduced with VEE VLPs
[0132] The basic concept of RNA delivery using VLPs is shown in figure 2 . In this experiment, a breast cancer model was used. A replicon dual-promoter vector was developed for the expression of HLA-DR1 and CD80 ( image 3 ). 4T1-Luc2 breast cancer cells were infected with VLPs encoding these two antigens. Using anti-HLA-DR and CD80 antibodies, the results show that both proteins are indeed expressed in this cell. HLA-DR1 and CD80 have figure 2 The VLP-VEE replicon was expressed in VLP-transfected 4T1-Luc2 cells. Cells expressing HLA-DR1 were identified by anti-HLA-DR antibody labeled with FITC. Cells expressing CD80 were identified by anti-CD80 antibody labeled with PE.
[0133] To assess whether alphavirus replicons can express two separate proteins in the same cell, VEE replication with HLA-DR1 under the control of one subgenomic promoter and CD80 under the control of the ...
Embodiment 3
[0134] Example 3 - Expression of various proteins in target cells transduced with VEE VLPs
[0135] To test VLPs in a mouse animal system, 4T1-Luc2 cells (highly aggressive and metastatic breast cancer cells) were implanted into the mammary fat pads of 10 mice. Tumors were visible within a few days and became apparent and palpable (5-7 mm) within a week. 3 mice were injected intratumorally with VLPs expressing inert protein (GFP), 2 mice were injected with VLPs expressing human HLA-DR1 / CD80 (allogeneic), and 5 mice were left uninjected as controls.
[0136] After 1 week, tumors were surgically removed from all mice and followed for several weeks for tumor regrowth and metastasis. result( Figure 5) showed that 4 out of 5 uninjected mice (untreated controls) died within 4-6 weeks due to cancer regrowth and metastasis in lung, breast and liver. All 3 mice injected with VLP / GFP (non-specific protein control) also died within 4-6 weeks due to cancer regrowth and metastasis in l...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com