EIAV p26 deletion vaccine and diagnostic

Abstract

Disclosed herein is a vaccine which provides immunity to mammals from infection and/or disease caused by a lentivirus, such as equine infectious anemia virus, human immunodeficiency virus (HIV), feline immunodeficiency virus (FIV), bovine immunodeficiency virus (BIV) or simian immunodeficiency virus (SIV) said composition comprising a deletion in a gene that blocks replication of the virus in vivo. Said composition allows differentiation between vaccinated and non-vaccinated, but exposed, mammals and provides safety and immunity when administered as a vaccine to mammals. Preferably said composition encompasses at least one deletion in a lentivirus which allows mammals to be safely vaccinated and provides protection from exposure to wild-type lentiviruses. It also encompasses a marker vaccine in which a foreign gene is inserted into the gene-deleted region, said inserted gene providing a diagnostic tool for use in vaccinated mammals and, potentially, protection from infection from a foreign disease. The scope of the invention encompases an EIAV vaccine that allows equines to be safely vaccinated and protected from disease without converting to a seropositive status on the Coggin's Test or any other test which measures p26, said p26 antigen being expressed in disease-producing wild-type EIAVs. Additionally, said EIA vaccine virus cannot cause clinical disease in mammals or spread or shed to other mammals including equines. Finally, this invention encompasses a marker vaccine in which vaccinated equines can be distinguished from non-vaccinated equines by detection of a foreign gene in the vaccinated animals. A diagnostic test to detect this foreign gene or gene product is also described.

What is claimed:  
     
         1 . A vaccine that produces protection from disease and/or infection caused by a lentivirus comprising a lentivirus that lacks the ability to replicate in vivo.  
     
     
         2 . A vaccine that produces protection from disease and/or infection caused by a lentivirus comprising a lentivirus that lacks the ability to express a Capsid Antigen  
     
     
         3 . The vaccine according to  claim 2  wherein the lack of ability to express Capsid Antigen results from one or more deletions in a gene region selected from the group consisting of a portion of the gag gene, all of the gag gene and a gene regulating expression of the gag gene or an insertion of a stop codon in a gene affecting the expression of the gag gene.  
     
     
         4 . The vaccine according to  claim 1  wherein the lentivirus lacks the ability to replicate in vitro.  
     
     
         5 . The vaccine according to  claim 2  wherein the lentivirus lacks the ability to replicate in vitro.  
     
     
         6 . The vaccine according to  claim 4  wherein said gene-deleted lentivirus is produced in large quantities by a cell line transfected with the gene-deleted lentivirus.  
     
     
         7 . The vaccine according to  claim 5  wherein said gene-deleted lentivirus is produced in large quantities by a cell line transfected with the gene-deleted lentivirus.  
     
     
         8 . The vaccine according to  claim 1  which, safely and effectively immunizes mammals against disease and/or infection caused by a lentivirus and further allows for differentiation between vaccinated, non-vaccinated and wild-type exposed mammals.  
     
     
         9 . The vaccine according to  claim 2  which, safely and effectively immunizes mammals against disease and/or infection caused by a lentivirus and further allows for differentiation between vaccinated, non-vaccinated and wild-type exposed mammals.  
     
     
         10 . The vaccine according to  claim 1  wherein said lentivirus is selected from the group consisting of equine infectious anemia virus (EIAV), human immunodeficiency virus (HIV), feline immunodeficiency virus (FIV), bovine immunodeficiency virus (BIV) and simian immunodeficiency virus (SIV).  
     
     
         11 . The vaccine according to  claim 2  wherein said lentivirus is selected from the group consisting of equine infectious anemia virus (EIAV), human immunodeficiency virus (HIV), feline immunodeficiency virus (FIV), bovine immunodeficiency virus (BIV) and simian immunodeficiency virus (SIV).  
     
     
         12 . The vaccine according to  claim 11  wherein the lentivirus EIAV lacks the ability to express p.26 antigen and is safe and effective.  
     
     
         13 . The vaccine according to  claim 12  wherein the lack of the ability to express p26 antigen results from a non-functional gag gene.  
     
     
         14 . The vaccine according to  claim 13  wherein the non-functional gag gene results from a deletion in the gag gene.  
     
     
         15 . The vaccine according to  claim 13  wherein the deletion in the gag gene results from one or more deletions in a gene region selected from the group consisting of a portion of the gag gene, all of the gag gene and a gene regulating expression of the gag gene or an insertion of a stop codon in a gene affecting the expression of the gag gene.  
     
     
         16 . The vaccine according to  claim 11  wherein the gene-deleted EIAV further lacks the ability to replicate in vitro.  
     
     
         17 . A vaccine for effectively and safely immunizing equines from EIA, said vaccine comprising a gene-deleted EIAV wherein said gene-deleted EIAV lacks the ability to express p26 and allows differentiation of vaccinated from wild type exposed equines.  
     
     
         18 . The vaccine of  claim 17  further comprising an adjuvant.  
     
     
         19 . The vaccine of  claim 17  wherein the EIAV is inactivated.  
     
     
         20 . The vaccine of  claim 17  comprising an inactivated EIAV and an adjuvant  
     
     
         21 . A method of immunizing mammals against disease produced by an EIAV comprising, administering to said mammals the vaccine of  claim 11 .  
     
     
         22 . An EIAV vaccine that allows equines to be safely vaccinated and protected from disease and/or infection without converting to a seropositive status on the Coggins Test or any other test which measures p26 antibodies.  
     
     
         23 . A diagnostic to detect all or a portion of the gag gene of EIAV comprising a PCR probe for said gene.  
     
     
         24 . A diagnostic to differentiate between a vaccinated and wild type exposed equine comprising the PCR probe of  claim 23 .  
     
     
         25 . A method of preparing a lentivirus vaccine comprising: 
   1  ) deleting all or a portion of a gag gene from the lentivirus;      2 ) transfecting a tissue culture with the resulting gene-deleted lentivirus to produce a persistently transfected cell culture;      3 ) growing the persistently transfected cell culture;      4 ) harvesting the persistently-transfected cell culture;      5 ) optionally inactivating the harvested cell culture; and    optionally adjuvantiing the harvested cell culture.