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Titolo:
Development of a novel in vitro assay (ALS assay) for evaluation of vaccine-induced antibody secretion from circulating mucosal lymphocytes
Autore:
Chang, HS; Sack, DA;
Indirizzi:
Johns Hopkins Univ, Sch Hyg & Publ Hlth, Dept Int Hlth, Vaccine Testing Unit, Baltimore, MD 21205 USA Johns Hopkins Univ Baltimore MD USA 21205 g Unit, Baltimore, MD 21205 USA
Titolo Testata:
CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY
fascicolo: 3, volume: 8, anno: 2001,
pagine: 482 - 488
SICI:
1071-412X(200105)8:3<482:DOANIV>2.0.ZU;2-D
Fonte:
ISI
Lingua:
ENG
Soggetto:
ORAL CHOLERA VACCINATION; VIBRIO-FLUVIALIS; ELISPOT ASSAY; MIMICUS; CELLS; TOXIN; ENUMERATION; RESPONSES; HOLLISAE; MEMORY;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
24
Recensione:
Indirizzi per estratti:
Indirizzo: Chang, HS 10326 Champions Way, Laurel, MD 20723 USA 10326 Champions Way Laurel MD USA 20723 y, Laurel, MD 20723 USA
Citazione:
H.S. Chang e D.A. Sack, "Development of a novel in vitro assay (ALS assay) for evaluation of vaccine-induced antibody secretion from circulating mucosal lymphocytes", CL DIAG LAB, 8(3), 2001, pp. 482-488

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 Igc) for up to 4 days of incubation at 37 degreesC with 5% CO2 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 lymphocytesupernatant (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 antibody titers of prevaccination samples were barely detectable, whereas serum antitoxin ELISA titers in background of prevaccine samples were significantly higher than the ALS titers, We conclude that, without any in vitro antigen stimulation after vaccination, PBMC secrete antibodies into the supernatants in the ALS assay. This assay can quantitatively measure the antigen-specific antibody production from the PBMC culture in postvaccination blood samples.

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Documento generato il 07/04/20 alle ore 02:48:38