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. 2001 Jan 15;193(2):233–238. doi: 10.1084/jem.193.2.233

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© 2001 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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(A–C) T cell recall assays in culture after DC immunization. Pre- and postimmunization specimens were thawed and cocultured with MP-pulsed DCs (unpulsed DCs as controls) for 7 d. After 7-d culture, MP-specific T cells were quantified by MHC tetramers (A), ELISPOT (B), and CTL assay (C). (A) MHC tetramer assay. Data are expressed as percent CD8⁺ T cells binding A*0201–MP tetramer. (B) ELISPOT assay. On day 7, cells were transferred to an ELISPOT plate and restimulated (Restim) overnight with specific antigen (10 μg/ml MP) or left without restimulation (as controls), and antigen-specific IFN-γ–producing cells were quantified using an ELISPOT. SEM for all measurements is <30%. (C) CTL assay. MP-specific lysis was measured using MP-pulsed T2 targets. Data shown are after subtracting lysis with control unpulsed T2 targets and from cells after expansion using unpulsed DCs.

(A–C) T cell recall assays in culture after DC immunization. Pre- and postimmunization specimens were thawed and cocultured with MP-pulsed DCs (unpulsed DCs as controls) for 7 d. After 7-d culture, MP-specific T cells were quantified by MHC tetramers (A), ELISPOT (B), and CTL assay (C). (A) MHC tetramer assay. Data are expressed as percent CD8⁺ T cells binding A*0201–MP tetramer. (B) ELISPOT assay. On day 7, cells were transferred to an ELISPOT plate and restimulated (Restim) overnight with specific antigen (10 μg/ml MP) or left without restimulation (as controls), and antigen-specific IFN-γ–producing cells were quantified using an ELISPOT. SEM for all measurements is <30%. (C) CTL assay. MP-specific lysis was measured using MP-pulsed T2 targets. Data shown are after subtracting lysis with control unpulsed T2 targets and from cells after expansion using unpulsed DCs.