Purified nontypable Haemophilus influenzae P5 protein as a vaccine for nontypable Haemophilus influenzae infection

Abstract

The present invention relates to P5 outer membrane protein of the non-typable Haemophilus influenzae bacterial strain and antibodies directed to P5 protein. The invention also relates to a method of isolating P5 protein and a vaccine composition for use in the treatment of non-typable Haemophilus influenzae infection.

What is claimed is:  
     
       1. P5 outer membrane protein of nontypable Haemophilus influenzae bacterial strains which is substantially free of bacterial endotoxin, which is purified without the use of denaturing detergents or reducing agents and which elicits bactericidal antibodies. 
     
     
       2. The P5 protein of  claim 1  wherein the amino acid sequence is provided in FIG. 2 (SEQ ID NO:1)  SEQ ID NO: 1 , wherein the P5 protein elicits bactericidal antibodies against nontypable Haemophilus influenzae in a host. 
     
     
       3. A preparation of P5 protein of nontypable Haemophilus influenzae which is substantially free of bacterial endotoxin, wherein the protein is purified without the use of denaturing detergents or reducing agents and wherein the protein elicits bactericidal antibodies against nontypable Haemophilus influenzae in a host. 
     
     
       4. A vaccine composition for use in the treatment of nontypable Haemophilus influenzae infections which comprises an effective amount of nontypable Haemophilis influenzae P5 protein which is substantially free of bacterial endotoxin, which is purified without the use of denaturing detergents or reducing agents and which elicits bactericidal antibodies against nontypable Haemophilus influenzae in a host; in a pharmaceutically acceptable vehicle, and an optional adjuvant. 
     
     
       5. The vaccine composition of  claim 4 , further comprising at least one additional antigen or one or more organisms. 
     
     
       6. The vaccine composition of  claim 5 , wherein the organism is selected from the group consisting of a bacterium, virus, parasite and fungus. 
     
     
       7. The vaccine composition of  claim 5 , wherein the organism or organisms are etiologic agents of otitis media, or sinusitis or chronic pulmonary obstructive disease. 
     
     
       8. The vaccine composition of  claim 5 , wherein the organism or organisms are etiologic agents of meningitis. 
     
     
       9. The vaccine composition of  claim 8 , wherein the organism is selected frog  from the group consisting of: Streptococcus pneumonia, Streptococcus pyogenes, Group A, Staphylococcus aureus, Branhamella catarrhalis, and respiratory syncytial virus. 
     
     
       10. The vaccine composition of  claim 5 , wherein the additional antigen is an outer membrane protein of nontypable Haemophilus influenzae. 
     
     
       11. The vaccine composition of  claim 10 , wherein the outer membrane protein is selected from the group consisting of peptidoglycan-associated outer membrane lipoprotein Haemophilus lipoprotein, PAL or  of Haemophilus ( PAL )  and  PAL cross-reacting protein (PCP). 
     
     
       12. A vaccine composition comprising an antigen conjugate comprising nontypable Haemophilus influenzae P5 protein which is substantially free of bacterial endotoxin, which is purified without the use of denaturing detergents or reducing agents and which elicits bactericidal antibodies against nontypable Haemophilus influenzae in a host, conjugated to an antigen of an organism or an epitope or epitopes of that antigen, in a pharmaceutically acceptable vehicle. 
     
     
       13. The vaccine composition of  claim 12 , wherein the organism is nontypable Haemophilus influenzae. 
     
     
       14. The vaccine composition of  claim 13  wherein the antigen is an oligo  oligo- or polysaccharide antigen of nontypable Haemophilus influenzae. 
     
     
       15. The vaccine composition of  claim 12 , wherein the antigen is an outer membrane protein of nontypable Haemophilus influenzae. 
     
     
       16. A  The vaccine composition of  claim 15  wherein the outer membrane protein is peptidoglycan-associated outer membrane lipoprotein Haemophilus lipoprotein  of Haemophilus. 
     
     
       17. A method of treating against nontypable Haemophilus influenzae, comprising administering to an individual an immunogenic amount of P5 protein of nontypable Haemophilus influenzae which is substantially free of bacterial endotoxin, wherein said P5 protein is purified without the use of denaturing detergents or reducing agents and which elicits bactericidal antibodies against nontypable Haemophilus influenzae in a host. 
     
     
       18. A  The method of  claim 17  wherein P5 protein has the amino acid sequence shown in FIG. 2 (SEQ ID NO:1)  SEQ ID NO: 1 . 
     
     
       19. The method of  claim 18  wherein the P5 protein is administered in conjunction with at least one other antigen of nontypable Haemophilus influenzae. 
     
     
       20. The method of  claim 19 , wherein the antigen is an oligo- or polysaccharide or an outer membrane protein of nontypable Haemophilus influenzae. 
     
     
       21. The method of  claim 20 , wherein the outer membrane protein is peptidoglycan-associated outer membrane lipoprotein Haemophilus lipoprotein  of Haemophilus. 
     
     
       22. A method of reducing susceptibility to acute otitis media, sinusitis or chronic pulmonary obstructive disease comprising administering to an individual an immunogenic amount of P5 protein of nontypable Haemophilus influenzae which is substantially free of bacterial endotoxin, wherein said P5 protein in  is purified without the use of denaturing detergents or reducing agents and which elicits bactericidal antibodies against nontypable Haemophilus influenzae in a host. 
     
     
       23. The vaccine composition of  claim 12  which further comprises an adjuvant. 
     
     
       24. The preparation of  claim 3  which further comprises an adjuvant.