Differential pathways for calcium influx activated by concanavalin A and CD3 stimulation in Jurkat T cells
- 1.4k Downloads
Sustained increase in [Ca2+]c (Δ[Ca2+]c) is a critical early signal from T-cell receptor (TCR/CD3). In general, Ca2+-release activated Ca2+ channels (CRAC) are responsible for the Ca2+ influx and Δ[Ca2+]c after TCR/CD3 stimulation. However, T cells also express Ca2+-permeable nonselective cation channels such as TRPM2 and TRPC. Gd3+ is a relatively selective blocker for CRAC at micromolar concentrations. Here, Jurkat T cells were used to investigate the Gd3+-resistant Ca2+ influx (Δ[Ca2+]c,Gd) induced by concanavalin A (ConA, 1 μg/ml), a widely used mitogenic agent for T cells, or by anti-CD3 Ab (αCD3). αCD3-induced Δ[Ca2+]c was partly (~60%) inhibited by 1 μM Gd3+ while thapsigargin-induced Δ[Ca2+] was almost completely abolished. ConA-induced Δ[Ca2+] was mostly inhibited by 1 μM Gd3+ during the early phase (<30 s of ConA application) and became resistant during the late phase (>2 min). Induction of Δ[Ca2+]c,Gd by αCD3 and ConA was inhibited by 2-aminoethoxydiphenyl borate (2-APB) and by N-(p-amylcinnamoyl) anthranilic acid, indicating that TRPM2 and TRPC are involved in this process. Treatment with Pyr-3, a TRPC3-specific inhibitor, potently suppressed Δ[Ca2+]c,Gd by αCD3 (IC50, 0.16 μM). Patch clamp experiments demonstrated that the TRPM2 channels were activated by ConA, and the TRPC-like channels were activated by αCD3. Our present study suggests that TRPM2 and TRPC3 are activated by ConA and TCR/CD3, respectively, in Jurkat T cells and are responsible for the induction of Δ[Ca2+]c,Gd.
KeywordsCa2+ influx Calcium signaling Lymphocyte Nonselective cation channel TRP channels
This work was supported by a Korea Research Foundation Grant funded by the Korean Government (KRF-2008-314-E00008).
- 1.Amina S, Hashii M, Ma WJ, Yokoyama S, Lopatina O, Liu HX, Islam MS, Higashida H (2010) Intracellular calcium elevation induced by extracellular application of cyclic-ADP-ribose or oxytocin is temperature-sensitive in rodent NG108-15 neuronal cells with or without exogenous expression of human oxytocin receptors. J Neuroendocrinol 22:460–466PubMedCrossRefGoogle Scholar
- 10.Gamberucci A, Giurisato E, Pizzo P, Tassi M, Giunti R, McIntosh DP, Benedetti A (2002) Diacylglycerol activates the influx of extracellular cations in T-lymphocytes independently of intracellular calcium-store depletion and possibly involving endogenous TRP6 gene products. Biochem J 364:245–254PubMedGoogle Scholar
- 12.Hong CW, Kim TK, Ham HY, Nam JS, Kim YH, Zheng H, Pang B, Min TK, Jung JS, Lee SN, Cho HJ, Kim EJ, Hong IH, Kang TC, Lee J, Oh SB, Jung SJ, Kim SJ, Song DK (2010) Lysophosphatidylcholine increases neutrophil bactericidal activity by enhancement of azurophil granule–phagosome fusion via glycine.GlyR alpha 2/TRPM2/p38 MAPK signaling. J Immunol 184:4401–4413PubMedCrossRefGoogle Scholar
- 15.Kiyonaka S, Kato K, Nishida M, Mio K, Numaga T, Sawaguchi Y, Yoshida T, Wakamori M, Mori E, Numata T, Ishii M, Takemoto H, Ojida A, Watanabe K, Uemura A, Kurose H, Morii T, Kobayashi T, Sato Y, Sato C, Hamachi I, Mori Y (2009) Selective and direct inhibition of TRPC3 channels underlies biological activities of a pyrazole compound. Proc Natl Acad Sci USA 106:5400–5405PubMedCrossRefGoogle Scholar
- 35.Yamamoto S, Shimizu S, Kiyonaka S, Takahashi N, Wajima T, Hara Y, Negoro T, Hiroi T, Kiuchi Y, Okada T, Kaneko S, Lange I, Fleig A, Penner R, Nishi M, Takeshima H, Mori Y (2008) TRPM2-mediated Ca2+ influx induces chemokine production in monocytes that aggravates inflammatory neutrophil infiltration. Nat Med 14:738–747PubMedCrossRefGoogle Scholar