en
Scientific article
English

Comparison of a limiting dilution assay and ELISpot for detection of memory B-cells before and after immunisation with a protein-polysaccharide conjugate vaccine in children

Published inJournal of Immunological Methods, vol. 358, no. 1-2, p. 46-55
Publication date2010
Abstract

In the present study, the limiting dilution assay (LDA) and the ELISpot are compared in their ability to detect serogroup C meningococcal (MenC)-specific memory B-cells in peripheral blood of 12month old children, after 3-dose priming with serogroup C meningococcal conjugate vaccine (MenCV) in infancy. At 12 months of age, MenC-memory B-cells were detected by ELISpot in 61% of children and in 5% by LDA. In contrast, carrier-specific memory B-cells were measurable in 36% of children by LDA and 78% by ELISpot. One month after a booster dose of MenCV given at 12 months of age, MenC-specific memory B-cells were detected in 89% of children by ELISpot and 65% of children by LDA, while diphtheria toxoid-specific memory B-cells were detected in 88% of children by ELISpot and in 74% of children by LDA. Two statistical methods were applied to enumerate memory B-cells with the LDA assay; the Poisson method allowed determination of very low frequencies that could not be determined by the Reed and Muench method. These data are examples of alternative methods to assess long-term protection after protein-polysaccharide conjugate vaccines, through direct measurement of memory B-cells in peripheral blood shortly after immunisation.

Keywords
  • B-Lymphocyte Subsets/immunology
  • Cell Separation
  • Diphtheria Toxoid/immunology
  • Enzyme-Linked Immunosorbent Assay/methods
  • Humans
  • Immunoassay/methods
  • Immunoglobulin G/immunology
  • Immunologic Memory/immunology
  • Infant
  • Lymphocyte Count
  • Meningococcal Vaccines/immunology
  • Polysaccharides, Bacterial/immunology
  • Vaccination/methods
Citation (ISO format)
BLANCHARD ROHNER, Géraldine et al. Comparison of a limiting dilution assay and ELISpot for detection of memory B-cells before and after immunisation with a protein-polysaccharide conjugate vaccine in children. In: Journal of Immunological Methods, 2010, vol. 358, n° 1-2, p. 46–55. doi: 10.1016/j.jim.2010.03.014
Main files (1)
Article (Published version)
accessLevelRestricted
Identifiers
ISSN of the journal0022-1759
119views
0downloads

Technical informations

Creation10/16/2020 5:59:00 PM
First validation10/16/2020 5:59:00 PM
Update time03/16/2023 12:44:32 AM
Status update03/16/2023 12:44:32 AM
Last indexation01/17/2024 1:28:37 PM
All rights reserved by Archive ouverte UNIGE and the University of GenevaunigeBlack