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. Author manuscript; available in PMC: 2011 Jan 22.
Published in final edited form as: Vaccine. 2009 Nov 21;28(4):907. doi: 10.1016/j.vaccine.2009.11.002

Figure 1. Memory B cell antibody-secreting assay against influenza hemagglutinin antigens.

Figure 1

PBMC were cultured as described in Material and Methods to reactivate MBC. The cells were then plated, at several dilutions, onto 96-well nitrocellulose dishes coated with 1µg/well of recombinant influenza HA antigens, KLH (negative control), or antibody against human IgG. After a 4-hour culture, the cells were washed off and the plates developed with a second anti-human IgG antibody conjugated to alkaline phosphatase, and TCIP/BCP substrate. Spots were counted on an ImmunoSpot automated plate reader. Plates were set up such that PBMC from all the time points from a single subject were on the same plate. The wells shown are from a single subject, though representative of all subjects responding in the assay.