PMID: 8960096Nov 29, 1996Paper

Limiting dilution analysis of virus-specific memory B cells by an ELISPOT assay

Journal of Immunological Methods
Mark K Slifka, R Ahmed

Abstract

Many acute viral infections induce long-term, sometimes even life-long humoral immunity. To characterize this immune response, accurate quantitation of memory B cells and plasma cells is essential. Plasma cells can be quantitated directly ex vivo by virtue of their ability to spontaneously secrete antibody. Memory B cells on the other hand, do not spontaneously secrete antibody but require antigenic stimulation in order to proliferate and differentiate into antibody secreting cells (ASC). In this study, an ELISPOT-based limiting dilution assay (LDA) was developed for quantitating virus-specific B cell memory following acute lymphocytic choriomeningitis virus (LCMV) infection of adult mice. Antiviral memory B cell precursor (MBCp) frequencies were calculated from in vitro stimulated cultures using either a conventional ELISA-based LDA to measure accumulated virus-specific antibody in the culture medium or a new ELISPOT-based LDA that identifies the antibody-secreting daughter cells directly. In terms of sensitivity, the ELISPOT-based LDA and the ELISA-based LDA both calculated LCMV-specific MBCp frequencies to be approximately 1/2 x 10(4) spleen cells. However, compared to the 12 days of in vitro stimulation required to estimate...Continue Reading

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Citations

Apr 17, 2004·Journal of Immunological Methods·Shane CrottyRafi Ahmed
Mar 23, 2004·Annual Review of Immunology·Raymond M WelshEva Szomolanyi-Tsuda
Aug 2, 2008·International Journal of Biomedical Imaging·Jun ZhaoGe Wang
Sep 15, 2007·The Journal of Clinical Investigation·Benyue ZhangRobert S Mittler
Feb 12, 2014·PLoS Neglected Tropical Diseases·Mohammad Murshid AlamEdward T Ryan
Oct 14, 2003·Proceedings of the National Academy of Sciences of the United States of America·Loren D EricksonRandolph J Noelle
Sep 19, 2008·Immunology·Xue Q LiuMichelle N Wykes
Jul 20, 2010·Immunological Reviews·Ian J Amanna, Mark K Slifka
Oct 24, 2006·Journal of Immunological Methods·Ian J Amanna, Mark K Slifka
Dec 3, 2005·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Takeji UmemuraHarvey J Alter
Apr 7, 1998·Immunity·M K SlifkaR Ahmed
Feb 2, 2017·Annual Review of Immunology·Florian Weisel, Mark Shlomchik

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