Vaccine compositions and adjuvant

Abstract

The immune response of an animal to a target immunogen may be enhanced by use of a novel adjuvant which includes low concentrations of killed cells of Mycobacterium avium subspecies avium in combination with mineral oil. The adjuvant may be used in vaccine compositions for the immunization of an animal against any target immunogen, and is particularly preferred for use with immunocontraceptive vaccines such as GnRH and PZP immunocontraceptive vaccines.

We claim:  
     
         1 . In a vaccine composition for immunizing an animal which comprises an immunologically effective amount of a target immunogen in combination with an adjuvant effective to enhance the immune response in an animal to said immunogen, wherein the improvement comprises said adjuvant comprising mineral oil and killed cells of  Mycobacterium avium  subspecies  avium , the concentration of said killed cells of Mycobacterium avium being greater than or equal to about 50 μg per ml and less than or equal to about 400 μg per ml, measured as the dry weight of said killed cells per ml of said vaccine composition, and further wherein the amount of said killed cells of  Mycobacterium avium  is not sufficient to elicit a T cell-mediated delayed hypersensitivity response to said  M. avium  by said animal if said adjuvant was administered alone, without said immunogen.  
     
     
         2 . The composition of  claim 1  wherein said amount of said killed cells of  Mycobacterium avium  subspecies  avium  is less than or equal to about 400 μg, measured as the dry weight of said killed cells.  
     
     
         3 . The composition of  claim 2  wherein said amount of said killed cells of  Mycobacterium avium  subspecies  avium  is less than or equal to about 200 μg, measured as the dry weight of said killed cells.  
     
     
         4 . The composition of  claim 1  wherein said adjuvant further comprises a surfactant.  
     
     
         5 . The composition of  claim 1  wherein said immunogen is selected from the group consisting of GnRH or a GnRH immunogenic analog conjugated to a carrier protein, and porcine zona pellucida.  
     
     
         6 . The composition of  claim 5  wherein said immunologically effective amount comprises an amount effective for inducing immunocontraception of said animal.  
     
     
         7 . The composition of  claim 6  wherein said immunologically effective amount comprises an amount effective for inducing immunocontraception of said animal in a single dose.  
     
     
         8 . The composition of  claim 6  wherein said immunogen is mammalian GnRH or a mammalian GnRH immunogenic analog conjugated to a carrier protein.  
     
     
         9 . The composition of  claim 1  wherein said immunogen comprises GnRH or a GnRH immunogenic analog conjugated to KLH carrier protein, and said GnRH or GnRH immunogenic analog is conjugated to said KLH carrier protein through the C-terminal end of said GnRH or GnRH immunogenic analog.  
     
     
         10 . The composition of  claim 9  further comprising physiologically buffered saline, and further wherein the salt NaCl concentration of said vaccine composition is greater than or equal to about 0.7 M and less than or equal to about 1.0 M, and the pH of said vaccine composition is between about 7.0 and 8.0.  
     
     
         11 . In a method for vaccinating an animal against an immunogen of interest which comprises administering a vaccine composition to said animal which said vaccine composition comprises an immunologically effective amount of said immunogen in combination with an adjuvant effective to enhance the immune response in an animal to said immunogen, wherein the improvement comprises said adjuvant comprising mineral oil and killed cells of  Mycobacterium avium  subspecies  avium , the concentration of said killed cells of  Mycobacterium avium  being greater than or equal to about 50 μg per ml and less than or equal to about 400 μg per ml, measured as the dry weight of said killed cells per ml of said vaccine composition, and further wherein the amount of said killed cells of  Mycobacterium avium  in said vaccine composition is not sufficient to elicit a T cell-mediated delayed hypersensitivity response to  M. avium  by said animal if said adjuvant was administered alone, without said immunogen.  
     
     
         12 . The method of  claim 11  wherein the amount of said killed cells of  Mycobacterium avium  subspecies  avium  in said vaccine composition is less than or equal to about 400 μg, measured as the dry weight of said killed cells.  
     
     
         13 . The method of  claim 12  wherein the amount of said killed cells of  Mycobacterium avium  subspecies  avium  in said vaccine composition is less than or equal to about 200 μg, measured as the dry weight of said killed cells.  
     
     
         14 . The method of  claim 11  wherein said animal is selected from the group consisting of porcine, bovine, equine, feline, canine, primates, Rodentia, Cervidae, and Pachydermata.  
     
     
         15 . The method of  claim 11  wherein said animal is selected from the group consisting of domestic dogs, domestic cats, pigs, cattle, deer, horses, zoo animals, elephants, rodents, and reptiles.  
     
     
         16 . The method of  claim 11  wherein said adjuvant further comprises a surfactant.  
     
     
         17 . The method of  claim 11  wherein said immunogen is selected from the group consisting of GnRH or a GnRH immunogenic analog conjugated to a carrier protein, and porcine zona pellucida.  
     
     
         18 . The method of  claim 17  wherein said immunologically effective amount comprises an amount effective for inducing immunocontraception of said animal.  
     
     
         19 . The method of  claim 18  wherein said immunologically effective amount comprises an amount effective for inducing immunocontraception of said animal in a single dose.  
     
     
         20 . The method of  claim 17  wherein said immunogen is mammalian GnRH or a mammalian GnRH immunogenic analog conjugated to a carrier protein.  
     
     
         21 . The method of  claim 11  wherein said immunogen comprises GnRH or a GnRH immunogenic analog conjugated to KLH carrier protein, and said GnRH or GnRH immunogenic analog is conjugated to said KLH carrier protein through the C-terminal end of said GnRH or GnRH immunogenic analog.  
     
     
         22 . The method of  claim 21  wherein said vaccine composition further comprises physiologically buffered saline, and further wherein the salt concentration of said vaccine composition is greater than or equal to about 0.7 M and less than or equal to about 1.0 M, and the pH of said vaccine composition is between about 7.0 and 8.0.  
     
     
         23 . The method of  claim 11  wherein said vaccine composition is administered by parenteral injection.  
     
     
         24 . A vaccine composition for immunization of an animal comprising an immunologically effective amount of a target immunogen in combination with an adjuvant effective to enhance the immune response in an animal to said immunogen, wherein said adjuvant comprises mineral oil and killed cells of  Mycobacterium avium  subspecies  avium , the concentration of said killed cells of  Mycobacterium avium  being greater than or equal to about 50 μg per ml and less than or equal to about 400 μg per ml, measured as the dry weight of said killed cells per ml of said vaccine composition, and further wherein the amount of said killed cells of  Mycobacterium avium  in said vaccine composition is not sufficient to elicit a T cell-mediated delayed hypersensitivity response to  M. avium  by said animal if said adjuvant was administered alone, without said immunogen.  
     
     
         25 . The composition of  claim 24  wherein said amount of said killed cells of  Mycobacterium avium  subspecies  avium  is less than or equal to about 400 μg, measured as the dry weight of said killed cells.  
     
     
         26 . The composition of  claim 24  wherein said immunogen comprises GnRH or a GnRH immunogenic analog conjugated to an intact mollusk hemocyanin protein, and said GnRH or GnRH immunogenic analog is conjugated to said carrier protein through the C-terminal end of said GnRH or GnRH immunogenic analog.  
     
     
         27 . The composition of  claim 26  wherein said immunogen is of the formula: 
       (X-A m -B-L) n -R 
       wherein: 
 X is said GnRH or a GnRH immunogenic analog,  
 A is an amino acid spacer,  
 m is an integer greater than or equal to 0,  
 B is an amino mercaptan,  
 R is an intact immunogenic mollusk hemocyanin protein,  
 L is a bifunctional crosslinking agent effective for simultaneously binding to the thiol of said mercaptan, and to free amine moieties of said immunogenic mollusk hemocyanin protein, and  
 n is an integer greater than or equal to about 200.  
 
     
     
         28 . The composition of  claim 27  wherein said immunogenic mollusk hemocyanin protein is selected from the group consisting of  Concholepas concholepas  hemocyanin protein, Keyhole Lipmet hemocyanin protein, Horseshoe crab hemocyanin protein, and Abalone hemocyanin protein.  
     
     
         29 . The composition of  claim 28  wherein said immunogenic mollusk hemocyanin protein is  Concholepas concholepas  hemocyanin protein.