Abstract
This study aimed to comprehend the largely unknown role of voltage-gated potassium channel 1.3 (Kv1.3) in the phagocytic function of macrophages. We found that blocking of the Kv1.3 channel with 100 pmol L-1Stichodactyla helianthus neurotoxin (ShK) enhanced the phagocytic capacities of both resting and lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages in the chicken erythrocyte system. In the fluorescein isothiocyanate (FITC)-labeled Escherichia coli k-12 system, ShK increased the phagocytic capacities of resting RAW264.7 cells, but not of the LPS-stimulated cells, as LPS alone stimulated almost saturated phagocytosis of the macrophages. ShK increased the nitric oxide (NO) production in LPS-activated cells, but not in resting RAW264.7 cells. There was no effect of ShK alone on the cytokine secretions in resting RAW264.7 cells, but it suppressed IL-1β secretion in LPS-stimulated RAW264.7 cells. At a concentration of 100 pmol L-1, ShK did not affect the viability of the tested cells. Kv1.3 was expressed in RAW264.7 cells; this expression was downregulated by LPS, but significantly upregulated by disrupting caveolin-dependent endocytosis with filipin III. In addition, cytochalasin D, an inhibitor of actin polymerization, did not affect the Kv1.3 expression. Thus, blocking of the Kv1.3 channel enhances the phagocytic capacity and NO production of this cell line. Our results suggest that Kv1.3 channel serves as a negative regulator of phagocytosis in macrophages and can therefore be a potential target in the treatment of macrophage dysfunction.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Cahalan MD, Chandy KG. The functional network of ion channels in T lymphocytes. Immunol Rev, 2009, 231: 59–87
Qiu M, Campbell T, Breit S. A potassium ion channel is involved in cytokine production. Clin Exp Immuonl, 2002, 130: 67–74
Villalonga N, David M, Bielanska J, Vicente R, Comes N, Valenzuela C, Felipe A. Immunomodulation of voltage-dependent K+ channels in macrophages: molecular and biophysical consequences. J Gen Physiol, 2010, 135: 135–147
Wang J, Huang WL, Wang C, Liu RY. Dynamic process of phagocytosis and forms of macrophage cell death induced by ingestion of apoptotic neutrophils. Sci China Life Sci, 2014, 57: 1018–1023
Shirai T, Hilhorst M, Harrison DG, Goronzy JJ, Weyand CM. Macrophages in vascular inflammation—from atherosclerosis to vasculitis. Autoimmunity, 2015, 48: 139–151
Yan L, Tan XQ, Chen WX, Zhu H, Cao JM, Liu HR. Enhanced vasoconstriction to α1 adrenoceptor autoantibody in spontaneously hypertensive rats. Sci China Life Sci, 2014, 57: 681–689
Vicente R, Escalada A, Coma M, Fuster G, Sánchez-Tilló E, López-Iglesias C, Soler C, Solsona C, Celada A, Felipe A. Differential voltage-dependent K+ channel responses during proliferation and activation in macrophages. J Biol Chem, 2003, 278: 46307–46320
Vicente R, Escalada A, Villalonga N, Texidó L, Roura-Ferrer M, Martín-Satué M, López-Iglesias C, Soler C, Solsona C, Tamkun MM, Felipe A. Association of Kv1.5 and Kv1.3 contributes to the major voltage-dependent K+ channel in macrophages. J Biol Chem, 2006, 281: 37675–37685
Lam J, Wulff H. The lymphocyte potassium channels Kv1.3 and KCa3.1 as targets for immunosuppression. Drug Dev Res, 2011, 72: 573–584
Gao YD, Hanley PJ, Rinné S, Zuzarte M, Daut J. Calcium-activated K+ channel (KCa3.1) activity during Ca2+ store depletion and store-operated Ca2+ entry in human macrophages. Cell Calcium, 2010, 48: 19–27
Tano JY, Lee RH, Vazquez G. Macrophage function in atherosclerosis: potential roles of TRP channels. Channels (Austin), 2012, 6: 141–148
Demaurex N, El Chemaly A. Physiological roles of voltage-gated proton channels in leukocytes. J Physiol, 2010, 588: 4659–4665
Moreno C, Prieto P, Macías Á, Pimentel-Santillana M, de la Cruz A, Través P G, Boscá L, Valenzuela C. Modulation of voltage- dependent and inward rectifier potassium channels by 15-epi-lipoxin-A4 in activated murine macrophages: implications in innate immunity. J Immunol, 2013, 191: 6136–6146
Vicente R, Escalada A, Soler C, Grande M, Celada A, Tamkun MM, Solsona C, Felipe A. Pattern of Kvβ subunit expression in macrophages depends upon proliferation and the mode of activation. J Immunol, 2005, 174: 4736–4744
Yang Y, Wang YF, Yang XF, Wang ZH, Lian YT, Yang Y, Li XW, Gao X, Chen J, Shu YW, Cheng LX, Liao YH, Liu K. Specific Kv1.3 blockade modulates key cholesterol-metabolism-associated molecules in human macrophages exposed to ox-LDL. J Lipid Res, 2013, 54: 34–43
Leanza L, Zoratti M, Gulbins E, Szabò I. Induction of apoptosis in macrophages via Kv1.3 and Kv1.5 potassium channels. Curr Med Chem, 2012, 19: 5394–5404
Villalonga N, David M, Bielanska J. Immunomodulatory effects of diclofenac in leukocytes through the targeting of Kv1.3 voltage- dependent potassium channels. Biochem Pharmacol, 2010, 80: 858–866
Ell Wulff H, Castle NA, Pardo LA. Voltage-gated potassium channels as therapeutic targets. Nat Rev Drug Discov, 2009, 8: 982–1001
Pardo LA. Voltage-gated potassium channels in cell proliferation. Physiology, 2004, 19: 285–292
Bradding P, Wulff H. The K+ channels KCa3.1 and Kv1.3 as novel targets for asthma therapy. Brit J Pharmacol, 2009, 157: 1330–1339
Lei X, Ma A, Xi Y, Zhang W, Yao Y, Du Y. Inhibitory effects of blocking voltage-dependent potassium channel 1.3 on human monocyte-derived macrophage differentiation into foam cells. J Peking Univ (Health Sci), 2006, 38: 257–261
Pennington MW, Harunur Rashid M, Tajhya RB, Beeton C, Kuyucak S, Norton RS. A C-terminally amidated analogue of ShK is a potent and selective blocker of the voltage-gated potassium channel Kv1.3. FEBS Lett, 2012, 586: 3996–4001
Matheu MP, Beeton C. Imaging of effector memory T cells during a delayed-type hypersensitivity reaction and suppression by Kv1.3 channel block. Immunity, 2008, 29: 602–614
Datta K, Soni JL, Awadhiya RP, Datta IC. Erythrophagocytosis in phenylhydrazine induced acute anaemia in chickens. Res Vet Sci, 1989, 47: 136–137
Wang M, Zhao D, Yang Y, Liu J, Wang J, Yin X, Yang L, Zhou X. The cellular prion protein negatively regulates phagocytosis and cytokine expression in murine bone marrow-derived macrophages. PLoS One, 2014, 9: e102785
Tartaro K, VanVolkenburg M, Wilkie D, Coskran TM, Kreeger JM, Kawabata TT, Casinghino S. Development of a fluorescence-based in vivo phagocytosis assay to measure mononuclear phagocyte system function in the rat. J Immunotoxicol, 2014, 16: 1–8
Zhang M, Behrens EM, Atkinson TP, Shakoory B, Grom AA, Cron RQ. Genetic defects in cytolysis in macrophage activation syndrome. Curr Rheumatol Rep, 2014, 16: 439
Hsieh CH, Nickel EA, Chen J, Schwacha MG, Choudhry MA, Bland KI, Chaudry IH. Mechanism of the salutary effects of estrogen on Kupffer cell phagocytic capacity following trauma-hemorrhage: pivotal role of Akt activation. J Immunol, 2009, 182: 4406–4414
Zhou X, Yang W, Li J. Ca2+- and protein kinase C-dependent signaling pathway for nuclear factor-κB activation, inducible nitric-oxide synthase expression, and tumor necrosis factor-α production in lipopolysaccharide- stimulated rat peritoneal macrophages. J Biol Chem, 2006, 281: 31337–31347
Turner MD, Nedjai B, Hurst T, Pennington DJ. Cytokines and chemokines: at the crossroads of cell signalling and inflammatory disease. Biochim Biophys Acta, 2014, 1843: 2563–2582
Schulert GS, Grom AA. Macrophage activation syndrome and cytokine- directed therapies. Best Pract Res Clin Rheumatol, 2014, 28: 277–292
Striz I, Brabcova E, Kolesar L, Sekerkova A. Cytokine networking of innate immunity cells: a potential target of therapy. Clin Sci (Lond), 2014, 126: 593–612
Gu J, Xu H, Han Y, Dai W, Hao W, Wang C, Gu N, Xu H, Cao J. The internalization pathway, metabolic fate and biological effect of superparamagnetic iron oxide nanoparticles in the macrophage-like RAW264.7 cell. Sci China Life Sci, 2011, 54: 793–805
Kong L, Ge BX. MyD88-independent activation of a novel actin- Cdc42/Rac pathway is required for Toll-like receptor-stimulated phagocytosis. Cell Res, 2008, 18: 745–755
Martínez-Mármol R, Villalonga N, Solé L, Vicente R, Tamkun MM, Soler C, Felipe A. Multiple Kv1.5 targeting to membrane surface microdomains. J Cell Physiol, 2008, 217: 667–673
Vicente R. Differential voltage-dependent K+ channel responses during proliferation and activation in macrophages J Biol Chem, 2003, 278: 46307–46320
Author information
Authors and Affiliations
Corresponding author
Additional information
This article is published with open access at link.springer.com
Rights and permissions
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0), which permits use, duplication, adaptation, distribution, and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
About this article
Cite this article
Zhu, H., Yan, L., Gu, J. et al. Kv1.3 channel blockade enhances the phagocytic function of RAW264.7 macrophages. Sci. China Life Sci. 58, 867–875 (2015). https://doi.org/10.1007/s11427-015-4915-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11427-015-4915-3