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Clinical and Vaccine Immunology, September 2007, p. 1108-1116, Vol. 14, No. 9
1071-412X/07/$08.00+0     doi:10.1128/CVI.00004-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Ex Vivo Monitoring of Antigen-Specific CD4⁺ T Cells after Recall Immunization with Tetanus Toxoid

Division of Immunology and Allergy, Department of Medicine, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland

Received 3 January 2007/ Returned for modification 7 February 2007/ Accepted 2 July 2007

To monitor antigen-specific CD4⁺ T cells during a recall immune response to tetanus toxoid (TT), a sequential analysis including ex vivo phenotyping and cytokine flow cytometry, followed by cloning and T-cell-receptor (TCR) spectratyping of cytokine-positive CD4⁺ T cells, was performed. Grossly, twice as many TT-specific CD4⁺ T-cell clones, ex vivo derived from the CCR7^+/– CD69⁺ interleukin-2-positive (IL-2⁺) CD4⁺ subsets, belonged to the central memory (T_CM; CD62L⁺ CD27⁺ CCR7⁺) compared to the effector memory population (T_EM; CD62L^– CD27^– CCR7^–). After the boost, a predominant expansion of the T_CM population was observed with more limited variations of the T_EM population. TCR beta-chain-variable region (BV) spectratyping and sequencing confirmed a large concordance between most frequently expressed BV TCR-CDR3 from ex vivo-sorted CCR7^+/– CD69⁺ IL-2⁺ CD4⁺ subsets and BV usage of in vitro-derived TT-specific CD4⁺ T-cell clones, further demonstrating the highly polyclonal but stable character of the specific recall response to TT. Taken together, ex vivo flow cytometry analysis focused on the CCR7^+/– CD69⁺ IL-2⁺ CD4⁺ subsets appears to target the bulk of antigen-specific T cells and to reach an analytical power sufficient to adequately delineate in field trials the profile of the antigen-specific response to vaccine.

Published ahead of print on 18 July 2007.

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