Equine protozoal myeloencephalitis vaccine

Abstract

The present invention provides an immunogenically active component comprising inactivated Sarcocystis neurona cells and/or inactivated Neospora hughesi cells; antigens derived therefrom; DNA derived therefrom; or a mixture; or in combination with other vaccine components thereof. Further provided are vaccine compositions useful for preventing or ameliorating equine protozoal myeloencephalitis infection and disease and a method for the cell culture propagation of protozoan parasites.

1 . An immunogenically active component for preventing or ameliorating equine protozoal myoencephalitis infection or disease which comprises: 
 tachyzoite antibody-inducing inactivated  Neospora hughesi  cells,    a mixture of merozoite antibody-inducing inactivated  Sarcocystis neurona  cells and tachyzoite antibody-inducing inactivated  Neospora hughesi  cells,    a merozoite antibody-inducing antigen derived from  Sarcocystis neurona  cells,    a tachyzoite antibody-inducing antigen derived from  Neospora hughesi  cells,    DNA derived from  Sarcocystis neurona  cells,    DNA derived from  Neospora hughesi  cells,    or a mixture thereof.    
   
   
       2 . The component according to  claim 1  which comprises the antigen derived from  Sarcocystis neurona  cells, DNA derived from said cells or the mixture thereof.  
   
   
       3 . The component according to  claim 1  which comprises inactivated  Neospora hughesi  cells, the antigen derived from said cells, DNA derived from said cells or the mixture thereof.  
   
   
       4 . The component according to  claim 1  wherein said active component is present in sufficient quantity to provide at least 1×10 4  inactivated cells per dosage unit form.  
   
   
       5 . A vaccine composition which comprises an effective immunizing amount of the immunogenically active component of  claim 1 , a pharmacologically acceptable carrier; and optionally an immunogenically stimulating adjuvant.  
   
   
       6 . The vaccine composition according to  claim 5  wherein said active component is present in sufficient quantity to provide at least 1×10 4  inactivated cells per dosage unit form.  
   
   
       7 . The vaccine composition according to  claim 5  wherein said active component is present in sufficient quantity to provide at least 1×10 6  inactivated cells per dosage unit reform.  
   
   
       8 . The vaccine composition according to  claim 5  wherein said active component comprises the antigen derived from  Sarcocystis neurona  cells, DNA derived from said cells or the mixture thereof and the active component is present in an amount sufficient to produce a merozoite inducing serum neutralizing antibody response which has a neutralizing effect on  Sarcocystis neurona  merozoites.  
   
   
       9 . The vaccine composition according to  claim 5  wherein said active component comprises inactivated  Neospora hughesi  cells, the antigen derived from said cells, DNA derived from said cells or the mixture thereof and the active component is present in an amount sufficient to produce a tachyzoite inducing serum neutralizing antibody response which has a neutralizing effect on  Neospora hughesi  tachyzoites.  
   
   
       10 . The vaccine composition according to  claim 5  wherein the immunogenically stimulating adjuvant is present at about 1% to 50% by weight.  
   
   
       11 . The vaccine composition according to  claim 10  wherein said adjuvant is present at about 5% to 20% by weight.  
   
   
       12 . (canceled)  
   
   
       13 . The vaccine composition according to  claim 5  wherein said adjuvant is a metabolizable oil.  
   
   
       14 . The vaccine composition according to  claim 13  wherein the pharmacologically acceptable carrier is a balanced salt solution.  
   
   
       15 . A vaccine composition for the prevention or amelioration of equine protozoal myoencephalitis disease in equines comprising: 
 a first immunogenically active component comprising merozoite antibody-inducing inactivated  Sarcocystis neurona  cells, a merozoite antibody-inducing antigen derived from said cells, DNA derived from said cells capable of inducing a merozoite antibody immune response or a mixture thereof,    a second immunogenically active component comprising tachyzoite antibody-inducing inactivated  Neospora hughesi  cells, a tachyzoite antibody-inducing antigen derived from said cells, DNA derived from said cells capable of inducing a tachyzoite antibody immune response, or a mixture thereof,    a pharmacologically acceptable carrier and optionally an immunogenically stimulating adjuvant.    
   
   
       16 . The vaccine composition according to  claim 15  wherein said first immunologically active component comprises the inactivated  Sarcocystis neurona  cells and said second immunologically effective component comprises the inactivated  Neospora hughesi  cells.  
   
   
       17 . The vaccine composition according to  claim 15  wherein said first immunologically active component is present in an amount sufficient to produce a merozoite inducing serum neutralizing antibody response which has a neutralizing effect on  Sarcocystis neurona  merozoites, and wherein said second immunologically active component is present in an amount sufficient to produce a tachyzoite inducing serum neutralizing antibody response which has a neutralizing effect on  Neospora hughesi  tachyzoites.  
   
   
       18 . A method for the prevention or amelioration of equine protozoal myoencephalitis disease in equines which comprises administering to said equine the immunogenically active component of  claim 1 .  
   
   
       19 . A method for the prevention or amelioration of equine protozoal myoencephalitis disease in equines which comprises administering to said equine a therapeutically effective amount of the vaccine composition of  claim 5  comprises.  
   
   
       20 . A method for the prevention or amelioration of equine protozoal myoencephalitis disease in equines which comprises administering to said equine the vaccine composition of  claim 15 .  
   
   
       21 . The method according to  claim 19  wherein said vaccine is administered parenterally.  
   
   
       22 . The method according to  claim 19  wherein said vaccine is administered intramuscularly.  
   
   
       23 . A method for the cell culture propagation of  Sarcocystis neurona  or  Neospora hughesi  protozoan parasite which comprises: 
 a) growing a monolayer of cells having a confluency of 80%-100%;    b) refeeding said cells with supplemented growth media;    c) inoculating said cells with merozoites or tachyzoites;    d) holding the inoculated cells for 4-12 days;    e) decanting the supplemented growth media from the inoculated cells; and    f) refeeding said cells a second time with supplemented growth media.    
   
   
       24 . The method according to  claim 23  wherein the cells are selected from the group consisting of Equine Dermal cells; Maiden Darby Bovine Kidney cells; African Green Monkey Kidney cells; Canine Monocyte cells; Mouse Monocyte cells; Fetal Rhesus Monkey Kidney cells; Feline Kidney cells, Maiden Darby Canine Kidney cells; and Baby Hamster Kidney cells.  
   
   
       25 . The method according to  claim 23  wherein the cells are Equine Dermal cells or African Green Monkey Kidney cells.