Lymphocyte proliferation specific for recall, CMV and HIV antigens in miniaturized and automated format

Giuseppina Li Pira, Nadia Starc, Antonella Conforti, Alice Bertaina, Sergio Rutella, Franco Locatelli, Fabrizio Manca

Risultato della ricerca: Contributo in rivistaArticolo in rivista

Abstract

Lymphoproliferation assay (LPA) is used to test specific T-cell responses. LPA is performed in 96-well plates with 2-5 x 10(5) PBMC/well. In order to test numerous antigens, as in the case of epitope mapping or screening of antigenic panels from relevant pathogens, PBMC numbers may not be sufficient. We developed a miniaturized and automated procedure to perform LPA in 384- and 1536-well plates with one fourth to one twentieth of PBMC numbers used for standard assays. Here, we demonstrate that the procedure is reliable and robust using recall antigens and protein and peptide antigens from CMV and HIV. By using HIV specific T-cell lines, we also demonstrate that sensitivity ranges overlap with those of standard LPA and that as few as 3 specific cells/well provide a positive signal. This procedure is consistent with our policy to miniaturize assays for specific T-cell immunity, as we have already established for cytokine secretion assays. (C) 2012 Elsevier B.V. All rights reserved.
Lingua originaleEnglish
pagine (da-a)135-142
Numero di pagine8
RivistaJournal of Immunological Methods
Volume384
DOI
Stato di pubblicazionePubblicato - 2012

Keywords

  • Lymphoproliferation
  • Miniaturization
  • HIV
  • T-cell lines
  • CMV
  • T-cell immunity

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