Chronic pathogen-expressing cell lines

Abstract

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 maintaining the consistency of the final biological product. 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 cell line 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 - 19 . (canceled) 
   
   
       20 . A method for establishing a cell line modified to chronically produce non-infectious budding particles comprising an antigen of an infectious pathogen, said method comprising
 integrating an exogenous gene which expresses an antigen of an infectious pathogen and   recombinantly introducing components into said cell line to produce non-infectious budding particles that comprise said antigen.   
   
   
       21 - 33 . (canceled) 
   
   
       34 . A method for treating or preventing a disease in an animal, including humans, said method comprising:
 harvesting budding particles from a culture of a cell line modified to chronically produce non-infectious budding particles comprising an antigen of an infectious pathogen, and   administering, to the animal, a prophylactic or therapeutic effective amount of the harvested budding particles.   
   
   
       35 . The method of  claim 34  wherein said cell line tolerates the continuous expression and assembly of intact budding particles. 
   
   
       36 . The method of  claim 34  wherein said cell line releases intact budding particles are released into the cell line culture supernatant. 
   
   
       37 . The method of  claim 34  wherein said cell line comprises components introduced by infection into said cell line. 
   
   
       38 . The method of  claim 39  wherein said cell line comprises components recombinantly introduced by an expression vector into the cell line. 
   
   
       39 . The method of  claim 34  wherein said cell line comprises components recombinantly introduced by an expression vector into the cell line. 
   
   
       40 . The method of  claim 34  wherein said cell line produces human immunodeficiency virus type 1 particles. 
   
   
       41 . The method of  claim 34  wherein said budding particles further comprise at least one antigen fragment bound to a primary surface molecule and at least one co-stimulatory molecule. 
   
   
       42 . The method of  claim 34  wherein said cell line is modified to express a molecule(s) that can inhibit a human pathogen. 
   
   
       43 . The method of  claim 42  wherein said pathogen is biological. 
   
   
       44 . The method of  claim 34  wherein said cell line comprises an HIV-1 antigen. 
   
   
       45 . The method of  claim 34  wherein said cell line comprises an antigen presented on said particles and said antigen is capable of initiating an immune response. 
   
   
       46 . The method of  claim 34  wherein said cell line comprises an exogenous gene expressing a hybrid chimera of antigens from more than one infectious pathogen. 
   
   
       47 . A method for inducing an immune response in an animal, including humans, said method comprising
 harvesting budding particles from a culture of a cell line modified to chronically produce non-infectious budding particles comprising an antigen of an infectious pathogen, and   administering, to the animal, a prophylactic or therapeutic effective amount of the harvested budding particles.