sq-23377 and margatoxin

CD4(+) T cells can be divided into three subsets: naive (Tn), central memory (Tcm), and effector memory (Tem) lymphocytes. These subpopulations differ in phenotype, migratory capacity, pattern of secreted cytokines, and activation threshold. T-cell activation is associated with changes in membrane potential, which provide an electrical driving force for calcium entry into the cytosol. These phenomena were shown to precede lymphocyte proliferation, cytokine synthesis, migration, and apoptosis. Hence the aim of the study was the analysis of these early activation events in the subsets of CD4(+) T cells. We measured the membrane potential and intracellular calcium concentration ([Ca(2+)](i)) in CD4(+) Tn, Tcm, and Tem cells as well as the dependency of these parameters in CD4(+) T cells on their cell-to-cell contacts with other leukocyte subsets. The data indicate that membrane potential of CD4(+) T cells is a subset specific feature maintained by direct contact with monocytes. In addition, monocytes were found to control Ca(2+) influx in CD4(+) T cells.

Topics: Adolescent; Adult; Calcium; CD4-Positive T-Lymphocytes; Cell Communication; Coculture Techniques; Female; Humans; Ionomycin; Leukocytes, Mononuclear; Male; Membrane Potentials; Monocytes; Neutrophils; Potassium Channel Blockers; Scorpion Venoms; T-Lymphocyte Subsets; Young Adult

The voltage gated potassium channel (Kv1.3) has been shown to play a role in immune responsiveness. Blockade of the channel led to diminution of T cell activation and delayed type hypersensitivity. Previous in vitro studies of the blockade were focused on T cell activation and proliferation. In this study we examined other T and monocytic cell mediated events to glean the extent of the immunosuppressive effects of a Kv1.3 specific inhibitor, Margatoxin (MgTX). We found that MgTX inhibited the intracellular production of Th-1 as well as Th-2 cytokines. MgTX can also inhibit IL-2 production and proliferation of T cells upon stimulation with anti-CD3 and VCAM-1. Furthermore, a redirected cytolytic activity was also inhibited by MgTX. However, MgTX did not inhibit generation of CTL to EBV transformed lymphoma cells or antibody-dependent cellular cytolysis mediated by monocytes. It appears that a Kv1.3 blockade does not affect all immune responses, particularly those of innate immunity.

Topics: Antibody-Dependent Cell Cytotoxicity; Cell Division; Cytokines; Flow Cytometry; Humans; Immunosuppressive Agents; Ion Channel Gating; Ionomycin; Ionophores; Kv1.3 Potassium Channel; Monocytes; Neurotoxins; Potassium Channel Blockers; Potassium Channels; Potassium Channels, Voltage-Gated; Scorpion Venoms; T-Lymphocytes, Cytotoxic; Tetradecanoylphorbol Acetate

The role that potassium channels play in human T lymphocyte activation has been investigated by using specific potassium channel probes. Charybdotoxin (ChTX), a blocker of small conductance Ca(2+)-activated potassium channels (PK,Ca) and voltage-gated potassium channels (PK,V) that are present in human T cells, inhibits the activation of these cells. ChTX blocks T cell activation induced by signals (e.g., anti-CD2, anti-CD3, ionomycin) that elicit a rise in intracellular calcium ([Ca2+]i) by preventing the elevation of [Ca2+]i in a dose-dependent manner. However, ChTX has no effect on the activation pathways (e.g., anti-CD28, interleukin 2 [IL-2]) that are independent of a rise in [Ca2+]i. In the former case, both proliferative response and lymphokine production (IL-2 and interferon gamma) are inhibited by ChTX. The inhibitory effect of ChTX can be demonstrated when added simultaneously, or up to 4 h after the addition of the stimulants. Since ChTX inhibits both PK,Ca and PK,V, we investigated which channel is responsible for these immunosuppressive effects with the use of two other peptides, noxiustoxin (NxTX) and margatoxin (MgTX), which are specific for PK,V. These studies demonstrate that, similar to ChTX, both NxTX and MgTX inhibit lymphokine production and the rise in [Ca2+]i. Taken together, these data provide evidence that blockade of PK,V affects the Ca(2+)-dependent pathways involved in T lymphocyte proliferation and lymphokine production by diminishing the rise in [Ca2+]i that occurs upon T cell activation.

Topics: Calcium; Cell Division; Cells, Cultured; Charybdotoxin; Dose-Response Relationship, Drug; Humans; Immunosuppressive Agents; Inositol 1,4,5-Trisphosphate; Ion Channel Gating; Ionomycin; Lymphocyte Activation; Lymphokines; Mitogens; Neurotoxins; Potassium Channels; Scorpion Venoms; T-Lymphocytes