Methods and compositions relating to viral latency

Abstract

Disclosed are compositions and methods that relate generally to viruses, and more particularly to the agents and their identification and use of anti-HIV agents which cause latently infected cells to reactivate.

1 - 13 . (canceled) 
     
     
         14 . A method of screening for a composition that activates a cell latently infected by a virus; the method comprising the steps of:
 a) creating a latently infected cell comprising the steps of: a) isolating uninfected primary CD4+, CD27+, CD45 RO− naïve T cells; b) priming the CD4+, CD27+, CD45RO− naïve T cells toward differentiation, wherein at least a portion of the primary CD4+, CD27+, CD45RO− naïve T cells differentiate into non-polarized CD4+, CD27+, CD45RO+ memory T cells; c) exposing the non-polarized cells of step b) to a lentivirus defective in env, wherein the lentivirus comprises one or more sequences of interest operatively inserted downstream of a lentiviral promoter, wherein the one or more sequences of interest are HIV genes that encode one or more HIV proteins, wherein the one or more HIV proteins comprise at least Tat and Rev, thereby creating a population of cells latently infected with the lentivirus; and d) stimulating the latently infected cells to reactivate the latent virus, wherein stimulating results in the expression of at least Tat and Rev;   b) exposing the cell to a test composition; and   c) determining if the latently infected cell becomes active.   
     
     
         15 . A composition identified by the method of  claim 14 . 
     
     
         16 . The method of  claim 14 , wherein the cell is exposed to CD3/CD28 antibodies during step b). 
     
     
         17 . The method of  claim 14 , wherein the cell is exposed to PHA during step b). 
     
     
         18 . The method of  claim 14 , wherein the virus is a retrovirus. 
     
     
         19 . The method of  claim 18 , wherein the retrovirus is selected from the group comprising HIV-1, HIV-2, SIV, XMRV, HTLV-1, HTLV-2, HTLV-3, or HTLV-4. 
     
     
         20 . The method of  claim 14 , wherein the virus is hepatitis B or hepatitis C. 
     
     
         21 . The method of  claim 14 , wherein the virus is a herpes virus. 
     
     
         22 . The method of  claim 21 , wherein the herpes virus is selected from the group consisting of HSV-1, HSV-2, VZV, EBV, CMV, HHV-6, HHV-7, and HHV-8. 
     
     
         23 . A cell line comprising non-polarized CD4+ cells that have been latently infected with a virus. 
     
     
         24 . The cell line of  claim 23 , wherein the virus is a retrovirus. 
     
     
         25 . The cell line of  claim 24 , wherein the retrovirus is selected from the group comprising HIV-1, HIV-2, SIV, XMRV, HTLV-1, HTLV-2, HTLV-3, or HTLV-4. 
     
     
         26 . The cell line of  claim 23 , wherein the virus is hepatitis B or hepatitis C. 
     
     
         27 . The cell line of  claim 23 , wherein the virus is a herpes virus. 
     
     
         28 . The cell line of  claim 27 , wherein the herpes virus is selected from the group consisting of HSV-1, HSV-2, VZV, EBV, CMV, HHV-6, HHV-7, and HHV-8. 
     
     
         29 . The cell line of  claim 23 , wherein the virus is env-. 
     
     
         30 . A method of reactivating a cell latently infected with virus, the method comprising activating NFAT in the absence of NF-κB or contacting the cell with IL-7 in the absence of NFAT. 
     
     
         31 - 47 . (canceled) 
     
     
         48 . A method of treating a subject with a latent viral infection, the method comprising:
 a) exposing the subject to a composition that reactivates cells latently infected with a virus; and   b) treating the subject with an antiviral agent identified by the method of  claim 14 .   
     
     
         49 - 52 . (canceled)