title: Comparative evaluation of the antibody in lymphocyte supernatant (ALS) and enzyme-linked immunospot (ELISPOT) assays for measuring mucosal immune responses to Shigella antigens.
abstract: Accurately assessing mucosal immune responses to candidate vaccines remains a technical challenge. ELISPOT is widely used as a surrogate of mucosal immune response by directly enumerating circulating antibody secreting cells (ASCs), while antibody in lymphocyte supernatant (ALS) titers the total amount of antibody secreted by ASC ex vivo using ELISA. ALS is more practical than ELISPOT because the ASC supernatant is frozen for ELISA that can be conducted at any time, with any antigen, and in any laboratory. We compared IgA and IgG responses to serotype-specific Shigella LPS using ELISPOT and ALS in subjects following vaccination or infection with Shigella. ALS results correlated well with ELISPOT results, and the ALS method was both sensitive and specific for the detection of antibody responses against Shigella LPS. Based on these observations, the ALS assay is a practical and flexible alternative to ELISPOT for measuring mucosal IgA responses to Shigella LPS antigen.
authors.affiliation: Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA. afeller@jhsph.edu
authors.forename: Andrea J
authors.initials: AJ
authors.lastname: Feller
authors.forename: Robin
authors.initials: R
authors.lastname: McKenzie
authors.forename: David N
authors.initials: DN
authors.lastname: Taylor
authors.forename: Colleen C
authors.initials: CC
authors.lastname: Woods
authors.forename: Shannon L
authors.initials: SL
authors.lastname: Grahek
authors.forename: Dilara
authors.initials: D
authors.lastname: Islam
authors.forename: Malabi M
authors.initials: MM
authors.lastname: Venkatesan
authors.forename: Thomas L
authors.initials: TL
authors.lastname: Hale
authors.forename: August L
authors.initials: AL
authors.lastname: Bourgeois
mesh_terms: D000907:Antibodies, Bacterial / Q000276:immunology*; D000942:Antigens, Bacterial / Q000276:immunology*; D019411:Clinical Laboratory Techniques / Q000379:methods*; D006801:Humans; D018928:Immunity, Mucosal; D007124:Immunoenzyme Techniques / Q000379:methods; D007070:Immunoglobulin A / Q000032:analysis; D007074:Immunoglobulin G / Q000032:analysis; D008214:Lymphocytes / Q000276:immunology*; D012760:Shigella / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D007070:Immunoglobulin A; D007074:Immunoglobulin G