Hyporesponsiveness following booster immunization with bacterial polysaccharides is caused by apoptosis of memory B cells

J Infect Dis. 2012 Feb 1;205(3):422-30. doi: 10.1093/infdis/jir750. Epub 2011 Dec 7.


Background: Repeated immunizations with polysaccharide (PS) vaccines cause hyporesponsiveness through undefined mechanisms. We assessed the effects of a PS booster on immune responses, frequency, and survival of PS-specific B-cell subpopulations in spleen and bone marrow.

Methods: Neonatal mice were primed with meningococcus serotype C (MenC) conjugate MenC-CRM(197)+CpG1826, boosted with MenC-CRM(197), MenC-PS, or saline; subsequently, bromodeoxyuridine (BrdU) was injected daily intraperitoneally. MenC-PS-specific cells were labeled with fluorescent MenC-PS and phenotyped by flow cytometry.

Results: After MenC-PS booster, proliferating (BrdU(+)) MenC-PS-specific naive B cells (CD138(-)/B220(+); P = .0003) and plasma cells (CD138(+)/B220(-); P = .0002) in spleen were fewer than after saline booster. BrdU(+) MenC-PS-specific plasma cells were also reduced in bone marrow (P = .0308). Compared to saline, MenC-PS booster reduced BrdU(+) IgG(+) MenC-PS-specific B cells in spleen (P = .0002). Twelve hours after the MenC-PS booster, an increased frequency of apoptotic (AnnexinV(+)) MenC-PS-specific B cells in spleen was observed compared with MenC-CRM(197) (P = .0286) or saline (P = .001) boosters.

Conclusions: We demonstrated that the MenC-PS booster significantly reduced the frequency of newly activated MenC-PS-specific B cells-mostly switched IgG(+) memory cells-by driving them into apoptosis. It shows directly that apoptosis of PS-specific memory cells is the cause of PS-induced hyporesponsiveness. These results should be taken into account prior to consideration of the use of PS vaccines.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adjuvants, Immunologic / administration & dosage
  • Animals
  • Animals, Newborn
  • Annexin A5 / analysis
  • Apoptosis*
  • B-Lymphocytes / chemistry
  • B-Lymphocytes / immunology*
  • Bone Marrow / immunology
  • Bone Marrow / pathology
  • Female
  • Flow Cytometry
  • Immunization, Secondary / methods*
  • Immunologic Memory*
  • Immunophenotyping
  • Leukocyte Common Antigens / analysis
  • Meningococcal Vaccines / administration & dosage
  • Meningococcal Vaccines / immunology*
  • Mice
  • Oligodeoxyribonucleotides / administration & dosage
  • Polysaccharides, Bacterial / administration & dosage
  • Polysaccharides, Bacterial / immunology*
  • Spleen / immunology
  • Spleen / pathology
  • Syndecan-1 / analysis
  • Vaccines, Conjugate / administration & dosage
  • Vaccines, Conjugate / immunology


  • Adjuvants, Immunologic
  • Annexin A5
  • CPG-oligonucleotide
  • Meningococcal Vaccines
  • Oligodeoxyribonucleotides
  • Polysaccharides, Bacterial
  • Syndecan-1
  • Vaccines, Conjugate
  • Leukocyte Common Antigens