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BMC Immunology

, 9:63

First Online: 03 November 2008Received: 23 May 2008Accepted: 03 November 2008


BackgroundIon channels are involved in the control of membrane potential ψ in a variety of cells. The maintenance of ψ in human T lymphocytes is essential for T-cell activation and was suggested to depend mostly on the voltage-gated Kv1.3 channel. Blockage of Kv1.3 inhibits cytokine production and lymphocyte proliferation in vitro and suppresses immune response in vivo. T lymphocytes are a heterogeneous cell population and the expression of Kv1.3 varies among cell subsets. Oxonol diBA-C4-3 was used to determine ψ by flow cytometry. The presence of distinct T cell subsets was evaluated by immunophenotyping techniques and the contribution of Kv1.3 channels for the maintenance of ψ was investigated using selective blockers.

ResultsThe distribution of ψ in T lymphocytes varied among blood donors and did not always follow a unimodal pattern. T lymphocytes were divided into CD3-CD45RO and CD3-CD45RO subsets, whose peak channel values of ψ were -58 ± 3.6 mV and -37 ± 4.1 mV, respectively. MgTX specific inhibitor of Kv1.3 channels had no significant effect in the ψ of CD3-CD45RO subsets but depolarized CD3-CD45RO cells to -27 ± 5.1 mV.

ConclusionCombination of optical methods for determination of ψ by flow cytometry with immuophenotyping techniques opens new possibilities for the study of ion channels in the biology of heterogeneous cell populations such as T lymphocyte subsets.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2172-9-63 contains supplementary material, which is available to authorized users.

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Autor: Fernanda Mello de Queiroz - Cristiano G Ponte - Adriana Bonomo - Rosane Vianna-Jorge - Guilherme Suarez-Kurtz


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