Chronic pathogen-expressing cell lines
View Patent ↗This application provides a method to establish and construct cell lines expressing pathogens without destruction of the host cells. The invention allows for the formation of cell lines for the purpose of continuous expression, release, and harvesting of the pathogen and maintain the consistency of the final biological pro duct. Although the invention is intended for pathogen antigen expression, the invention allows for the production of any antigen by the described methods. The establishment of a chronically infected celline can be used for reagent, diagnostic, quantification, or vaccine purposes. We have used the procedure to select for a host cell line that naturally adapts to HIV-1 replication without affecting the host cell's ability to survive. This allowed for the establishment of a chronic HIV-1 expressing cell line that continuously expresses HIV-1 particles.
1. A biologically constructed cell line modified to chronically produce budding particles, said cell line comprising
components for production of said budding particles, said components comprising an integrated exogenous gene which expresses an antigen of an infectious pathogen;
wherein said cell line chronically produces non-infectious budding particles comprising said antigen.
2. The biologically constructed cell line of claim 1 wherein said cell line tolerates the continuous expression and assembly of intact budding particles.
3. The biologically constructed cell line of claim 2 wherein said intact budding particles are released into the cell line culture supernatant.
4. The cell line of claim 1 wherein said exogenous gene expresses a protein, a polypeptide, an antibody, a glycolipid, a lipid, a pathogen, a tumor, or an infectious agent.
5. The cell line of claim 1 wherein said components are introduced by infection into said cell line.
6. The cell line of claim 3 wherein said components are recombinantly introduced by an expression vector into the host cell line.
7. The cell line of claim 1 wherein said components are recombinantly introduced by an expression vector into the host cell line.
8. The cell line of claim 4 , wherein said pathogen is the entire pathogen.
9. The cell line of claim 4 , wherein said infectious agent is the entire agent.
10. A method for establishing a cell line according to claim 1 , said method comprising integrating an exogenous gene which expresses an antigen of a pathogen and
recombinantly introducing components into said cell line to produce non-infectious budding particles that comprise said antigen.
11. The cell line of claim 1 wherein said particles are human immunodeficiency virus type 1 particles.
12. The cell line of claim 1 wherein said budding particles further comprise at least one antigen fragment associated with a primary surface molecule and at least one co-stimulatory molecule.
13. The cell line of claim 12 modified to express a molecule(s) that can inhibit a human pathogen.
14. The cell line of claim 13 wherein said pathogen is biological.
15. The cell line of claim 1 wherein said antigen is an HIV-1 antigen.
16. The cell line of claim 1 wherein said antigen is presented on said particles and capable of initiating an immune response.
17. The cell line of claim 1 wherein said exogenous gene expresses a hybrid chimera of antigens from more than one infectious pathogen.