Development of a novel in vitro assay (ALS assay) for evaluation of vaccine-induced antibody secretion from circulating mucosal lymphocytes

Clinical and Diagnostic Laboratory Immunology
H S Chang, D A Sack

Abstract

We describe here a novel method for measuring in vitro antibody secretion from the tissue culture of human B lymphocytes in peripheral blood mononuclear cells (PBMC) after oral vaccination with a killed cholera vaccine. Enzyme-linked immunosorbent assay (ELISA) titers of the antibody secreted in the cell supernatant were determined. The validation results demonstrated that human PBMC remained viable and continued to secrete antibodies (total immunoglobulin A [IgA] and IgG) for up to 4 days of incubation at 37 degrees C with 5% CO(2) in cell cultures. The secreted antibody concentration correlated positively with the PBMC concentration and incubation time in the tissue culture and correlated negatively with the storage time of the whole blood at room temperature. In vitro assay of secreting antibody in the lymphocyte supernatant (i.e., the ALS assay) is capable of the detecting specific antibody response after oral vaccination with a killed whole-cell-plus-B-subunit cholera vaccine (WC-B) in healthy adults in a phase I clinical trial. Postimmunization PBMC secreted antibodies to cholera toxin in the cell supernatants. Antibody production did not require any in vitro antigen stimulation. In the ALS assay, antigen-specific antibod...Continue Reading

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Citations

Aug 26, 2009·Infection and Immunity·Ana A WeilStephen B Calderwood
Jan 16, 2013·Journal of Immunological Methods·Kimberly A LivingstonCharles B Stephensen
Oct 2, 2014·Contemporary Clinical Trials·Shaikh Meshbahuddin AhmadCharles B Stephensen
Dec 25, 2010·Journal of Immunological Methods·Xiao-Song HeHarry B Greenberg
Dec 29, 2013·Clinical and Vaccine Immunology : CVI·Richelle C CharlesEdward T Ryan
Oct 12, 2007·Clinical and Vaccine Immunology : CVI·Odd OdinsenJean-Pierre Allain
Jul 11, 2006·Expert Review of Vaccines·Regina C LaRocqueStephen B Calderwood
Oct 23, 2018·Medicina·Olumide AjibolaAnthonius A Eze
Jun 18, 2017·Frontiers in Immunology·Michael J CarterJohannes Trück
Feb 11, 2020·Frontiers in Cellular and Infection Microbiology·Michael J CarterShrijana Shrestha
Jul 28, 2021·Nature Reviews. Immunology·Ed C Lavelle, Ross W Ward

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