Microparticles from streptococcus pneumoniae as vaccine antigens

Abstract

An isolated Streptococcus pneumoniae membrane vesicle microparticle (MP), wherein said MP comprises: the protein Ply at the level of ≥0.070 μg/μg total protein in the MP; and/or the protein LytA at the level of ≥0.070 μg/μg total protein in the MP; and/or the protein PspC at the level of ≥0.130 μg/μg total protein in the MP; and/or the protein RrgB at the level of ≥0.020 μg/μg total protein in the MP. Compositions comprising such MPs. Uses thereof in particular in immunization, as well as methods of manufacture thereof.

What is claimed is: 
     
         1 . An isolated  Streptococcus pneumoniae  membrane vesicle microparticle (MP), wherein said MP comprises:
 i. the protein Ply at the level of ≥0.070 μg/μg total protein in the MP;   ii. the protein LytA at the level of ≥0.070 μg/μg total protein in the MP;   iii. the protein PspC at the level of ≥0.130 μg/μg total protein in the MP; or   iv. the protein RrgB at the level of ≥0.020 μg/μg total protein in the MP.   
     
     
         2 . The  Streptococcus pneumoniae  membrane vesicle microparticle according to  claim 1 , further comprising a capsular polysaccharide of a capsular serotype of  Streptococcus pneumoniae.    
     
     
         3 . The  Streptococcus pneumoniae  membrane vesicle microparticle according to  claim 1 , comprising the protein Ply at the level of ≥0.070 μg/μg total protein in the MP. 
     
     
         4 . The  Streptococcus pneumoniae  membrane vesicle microparticle according to  claim 1 , comprising the protein LytA at the level of ≥0.070 μg/μg total protein in the MP. 
     
     
         5 . The  Streptococcus pneumoniae  membrane vesicle microparticle according to  claim 1 , comprising the protein PspC at the level of ≥0.130 μg/μg total protein in the MP. 
     
     
         6 . The  Streptococcus pneumoniae  membrane vesicle microparticle according to  claim 1 , comprising the protein RrgB at the level of ≥0.020 μg/μg total protein in the MP. 
     
     
         7 . The  Streptococcus pneumoniae  membrane vesicle microparticle according to  claim 1 , comprising the protein PhtD at a level being at least 2-fold compared to  Streptococcus pneumoniae  membrane vesicles obtained in liquid culture in terms of μg/μg total protein in the particle or the vesicle, respectively. 
     
     
         8 . The  Streptococcus pneumoniae  membrane vesicle microparticle according to  claim 1 , comprising the protein RrgA at the level of ≥0.02 μg/μg total protein in the MP. 
     
     
         9 . The  Streptococcus pneumoniae  membrane vesicle microparticle according to  claim 1 , comprising the protein IgA1-protease at the level of ≥0.02 μg/μg total protein in the MP. 
     
     
         10 . A composition comprising a  Streptococcus pneumoniae  membrane vesicle microparticle comprising:
 i. the protein Ply at the level of ≥0.070 μg/μg total protein in the MP;   ii. the protein LytA at the level of ≥0.070 μg/μg total protein in the MP;   iii. the protein PspC at the level of ≥0.130 μg/μg total protein in the MP; or   iv. the protein RrgB at the level of ≥0.020 μg/μg total protein in the MP.   
     
     
         11 . The composition according to  claim 10 , wherein the composition is devoid of whole  Streptococcus pneumoniae  cells. 
     
     
         12 . The composition according to  claim 10 , further comprising capsular polysaccharides from  Streptococcus pneumoniae.    
     
     
         13 . The composition according to  claim 10 , comprising  Streptococcus pneumoniae  membrane vesicle microparticles in an amount of 1 μg/ml. 
     
     
         14 . The composition according to  claim 13  comprising  Streptococcus pneumoniae  membrane vesicle microparticles in an amount of 10 μg/ml. 
     
     
         15 . The composition according to  claim 10 , further comprising an adjuvant. 
     
     
         16 . A method for producing an isolated  Streptococcus pneumoniae  microparticle/s, comprising:
 a. providing  Streptococcus pneumoniae  host cell;   b. culturing said host cell under conditions allowing the production of  Streptococcus pneumoniae  membrane vesicle microparticle/s according to by the host cell; and   c. isolating the microparticle/s thus produced, wherein said microparticles comprise:
 i. the protein Ply at the level of ≥0.070 μg/μg total protein in the MP; 
 ii. the protein LytA at the level of ≥0.070 μg/μg total protein in the MP; 
 iii. the protein PspC at the level of ≥0.130 μg/μg total protein in the MP; or 
 iv. the protein RrgB at the level of ≥0.020 μg/μg total protein in the MP. 
   
     
     
         17 . The method according to  claim 16 , wherein the culturing is carried out in solid phase.