Immunization against clostridium difficile disease

Abstract

The invention provides active and passive immunization methods for preventing and treating Clostridium difficile infection, which involve percutaneous administration of C. difficile toxin-neutralizing polyclonal immune globulin, C. difficile toxoids, or combinations thereof. Also provided by the invention are C. difficile toxoids, C. difficile toxin-neutralizing polyclonal immune globulin, and methods of identifying subjects that produce C. difficile toxin-neutralizing polyclonal immune globulin.

1 . A method of producing a  Clostridium difficile  toxoid, said method comprising the steps of: 
 providing a  Clostridium difficile  bacterium;    culturing said bacterium in medium lacking animal products to generate a culture;    purifying clostridial toxin A and clostridial toxin B from said culture to generate a mixture of purified toxins A and B; and    inactivating said purified toxins A and B by incubation in formaldehyde at a temperature of 25° C. or less to generate said clostridial toxoid.    
   
   
       2 . The method of  claim 1 , wherein said purified toxins A and B are incubated in formaldehyde at a temperature of about 15° C. or less, or a temperature of about 5° C. or less.  
   
   
       3 . The method of  claim 1 , wherein said purified toxins A and B are present in said mixture at a ratio of about 0.1:1 to 10:1 or a ratio of about 2:1.  
   
   
       4 . A  Clostridium difficile  toxoid produced by the method of  claim 1 .  
   
   
       5 . A vaccine composition comprising the  Clostridium difficile  toxoid of  claim 4 .  
   
   
       6 . The vaccine composition of  claim 5 , further comprising 0.012-0.020% formaldehyde.  
   
   
       7 . The vaccine composition of  claim 5 , further comprising an adjuvant.  
   
   
       8 . The vaccine composition of  claim 7 , wherein the adjuvant is an aluminum compound.  
   
   
       9 . A method of producing human immune globulin directed against a  clostridium , said method comprising the steps of: 
 administering a clostridial toxin or toxoid to a human; and    isolating said immune globulin against a  clostridium  from said human.    
   
   
       10 . The method of  claim 9 , wherein the  clostridium  is  Clostridium difficile.    
   
   
       11 . The method of  claim 9 , wherein said clostridial toxoid comprises toxin A and toxin B.  
   
   
       12 .  Clostridium difficile  immune globulin produced by administering a  Clostridium difficile  toxin or toxoid to a human, and isolating said  Clostridium difficile  immune globulin from said human  
   
   
       13 . A method of identifying a human producing a  Clostridium difficile  immune globulin, said method comprising the steps of: 
 obtaining a blood sample from a human vaccinated with a  Clostridium difficile  toxoid;    determining the level of antibodies to  Clostridium difficile  Toxins A and B in said blood sample by an enzyme-linked immunosorbent assay; and    determining the level of in vitro cytotoxicity neutralization activity against  Clostridium difficile  Toxins A and B in said blood sample, wherein detection of increased levels of antibodies to  Clostridium difficile  Toxins A and B in said blood sample, and    detection of in vitro cytotoxicity neutralization activity against  Clostridium difficile  Toxins A and B in said blood sample, indicates identification of a human producing a  Clostridium difficile  immune globulin useful for treatment or prevention of  Clostridium difficile  infection.