Optimization and qualification of a memory B-cell ELISpot for the detection of vaccine-induced memory responses in HIV vaccine trials.

Journal Article (Journal Article)

Various aspects of the human immune system can be analyzed to determine the efficacy of a vaccine. We have developed a B-cell ELISpot to measure HIV-specific antibody-secreting B cells in the peripheral blood as a result of vaccination or natural infection. Our method includes stimulating peripheral blood mononuclear cells with interleukin-2 and a polyclonal activator, R848, to induce memory B cells to differentiate into antibody-secreting cells. Total immunoglobulin-secreting as well as antigen-specific B cells are then quantified. We have tested several HIV Env gp120 and gp140 proteins from different HIV subtypes, as well as a sensitive consensus group M Env gp140. Our findings indicate that the B-cell ELISpot provides a sensitive and specific tool to detect antigen-specific memory B-cell responses, and it is equally suited to detect antibody-secreting plasmablasts present in the circulation shortly after infection or vaccination.

Full Text

Duke Authors

Cited Authors

  • Walsh, PN; Friedrich, DP; Williams, JA; Smith, RJ; Stewart, TL; Carter, DK; Liao, H-X; McElrath, MJ; Frahm, N; NIAID HIV Vaccine Trials Network,

Published Date

  • August 30, 2013

Published In

Volume / Issue

  • 394 / 1-2

Start / End Page

  • 84 - 93

PubMed ID

  • 23707324

Pubmed Central ID

  • PMC3736720

Electronic International Standard Serial Number (EISSN)

  • 1872-7905

Digital Object Identifier (DOI)

  • 10.1016/j.jim.2013.05.007


  • eng

Conference Location

  • Netherlands