Abstract
Lymphoproliferation assay (LPA) is used to test specific T-cell responses. LPA is performed in 96-well plates with 2-5×105 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.
Original language | English |
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Pages (from-to) | 135-142 |
Number of pages | 8 |
Journal | Journal of Immunological Methods |
Volume | 384 |
Issue number | 1-2 |
DOIs | |
Publication status | Published - Oct 31 2012 |
Keywords
- CMV
- HIV
- Lymphoproliferation
- Miniaturization
- T-cell immunity
- T-cell lines
ASJC Scopus subject areas
- Immunology
- Immunology and Allergy