See discussions, stats, and author profiles for this publication at: https:///publication/262189657 Modulation Of Human Basophil Degranulation By Geranylgeranyl CompoundsArticle in Allergology International · May 2014DOI: 10.2332/allergolint.13-LE-0637 · Source: PubMedCITATIONS 0READS 147 authors, including:Masao YamaguchiTeikyo University135 PUBLICATIONS 4,204 CITATIONSSEE PROFILE Hiroyuki NagaseTeikyo University114 PUBLICATIONS 1,654 CITATIONSSEE PROFILEKen OhtaNational Hospital Organization Tokyo Nation…350 PUBLICATIONS 8,606 CITATIONSSEE PROFILEAll content following this page was uploaded by Hiroyuki Nagase on 25 August 2015.The user has requested enhancement of the downloaded file. All in-text references underlined in blue are added to the original document and are linked to publications on ResearchGate, letting you access and read them immediately.Allergology International Vol 63,Suppl 1,2014www.jsaweb.jp !49Dear EditorModulation of Human Basophil Degranulation by GeranylgeranylCompoundsRecent experimental studies have shown that the ba-sophil is critical for a subtype of anaphylaxis or IgE-mediated very late-phase skin inflammation.1Motility and activation of basophils are known to be regulated by various endogenous and !or exogenous sub-stances.The panel of known such basophil-directed molecules is expanding.One important candidate may be geranylgeranyl compounds,which were in-itially demonstrated to be protectors of the gastric mucosa.2These compounds have also demonstrated protective actions in situations that are often hazard-ous to the host.In rodent studies,geranylgeranylace-tone (GGA)suppressed the development of several inflammatory reactions and enhanced tissue regen-eration in vivo .2,3However,the effects of GGA on al-lergic effectors and immunologic reactions have not been fully clarified.In this study,we assessed the pharmacological ac-tions of GGA and related compounds on basophil de-granulation,detected as release of histamine.Baso-phils were obtained from non allergic volunteers by dextran sedimentation of whole blood.Cells were preincubated with GGA (Wako Pure Chemicals,Osaka,Japan)for 15min at 37 ,washed and then stimulated with a secretagogue for 45min.4Degranulation of basophils by polyclonal anti-IgE antibody (MBL,Nagoya,Japan )and by phorbol myristate acetate (PMA )(Sigma,St.Louis,MO,USA)was significantly enhanced by preincubation of cells for 15min with GGA at 1.4or 2.7mM (Fig.1a).Similar results were observed for highly pure baso-phils (purity >95%;prepared by Percoll gradient cen-trifugation followed by negative MACS selection),in-dicating that GGA acts directly on basophils.On the other hand,GGA showed no clear effect on basophil degranulation by a chemokine,monocyte chemoat-tractant protein (MCP)-1or Ca ionophore A23187.As shown in Figure 1b,up to 15min of incubation of ba-sophils with GGA alone at 2.7mM did not induce re-lease of histamine.However,30min or longer incuba-tion resulted in higher,15to 20%histamine release,suggesting that GGA at this concentration might damage basophils in a time-dependent manner.The extent of enhancement of IgE-mediated basophil de-granulation by GGA was mild compared to that by IL-3,and GGA did not show additive augmentation of histamine release by IL-3-treated basophils (Fig.1c).As shown in Figure 1d,another geranylgeranyl com-pound,geranylgeraniol (Sigma),also upregulated ba-sophil histamine release evoked by anti-IgE antibodyor PMA.On the other hand,compounds havingsmaller structures,such as geranylacetone and farne-sol,showed no effect on basophil degranulation (data not shown).Statins inhibit intracellular geranylger-anylation,resulting in suppression of the activation profiles of inflammatory cells,5-7and basophil de-granulation in response to anti-IgE antibody or PMA was significantly suppressed by preincubation with simvastatin at 50μM (Fig.1e).Geranylgeranyl compounds are reported to be in-volved in intracellular signal cascades in various cells,including mast cell activation evoked by IgE crosslinkage.5-7Our present finding that exogenously added GGA can enhance basophil activation suggests that GGA enters basophils and then behaves as a sub-strate in the cell activation pathway.Simvastatin,which can inhibit intracellular geranylgeranylation,suppressed basophil degranulation triggered by anti-IgE antibody and PMA,but not A23187.This finding coincides with our results that GGA augmented baso-phil degranulation evoked by anti-IgE antibody and PMA,but not A23187or MCP-1,suggesting that pro-tein kinase C or related molecule(s)may be the tar-get of GGA.It appears that geranyl and farnesyl com-pounds are not involved in the above pathway in ba-sophils.GGA’s various novel actions are being unveiled through recent experimental approaches.Mostly based on murine studies,this compound is able to suppress vicious inflammatory disorders of the skin or visceral organs,including pulmonary inflammation induced by gefitinib,an inhibitor of epidermal growth factor receptor-mediated signals.3It is thought that one of main in vivo effects of GGA is induction of tissue-stabilizing heat shock proteins.2,3In vivo stud-ies have so far tested GGA at μM to mM concentra-tions and have found increases in those proteins and changes in cell fate.8,9Our present findings suggest that GGA at relatively high concentrations can also exert acute effects on cellular functions and enhance the activation of basophils evoked by certain secre-tagogues.Although the in vivo significance of our findings and the whole aspect of the biologic actions of GGA remain unclear,elucidation of the precise roles of this compound in allergies and other tissue-damaging disorders is of special interest in light of its characteristic host-protecting properties.ACKNOWLEDGEMENTSThe authors thank Ms.Sayaka Igarashi for her excel-lent technical help.Yuko Nakase 1,Masao Yamaguchi 1,Naoya Sugimoto 1,Maho Suzukawa 2,Hidenori Arai 1,Hiroyuki Nagase 1and Ken Ohta 1,2Allergology International.2014;63(Suppl 1):49-51DOI:10.2332!allergolint.13-LE-0637Nakase Y et al.50Allergology International Vol 63,Suppl 1,2014www.jsaweb.jp !Fi g . 1 Mod u lation of b asophil histamine release b y geranylgeranyl and related compo u nds. (a ) Basophil preparations w ere preinc ub ated w ith and w itho u t GGA and then stim u lated w ith vario u s secretagog u es. GGA at 1.4 and 2.7 mM corresponds to 0.5 and 1 μl/ml, respectively. Histamine release w as expressed as a percentage of the total cell u lar histamine after s ub tracting spontaneo u s release (u s u ally <5%). Data are the mean ± SEM of three to six separate experiments. *p ≤ 0.05, vers u s corre-sponding val u es of cells not preinc ub ated w ith GGA. (b ) Baseline histamine release b y b asophils after treatment w ith GGA. Cells w ere preinc ub ated w ith GGA at 1 μl/ml for the indicated times, w ashed and then inc ub ated w itho u t any secretagog u e for 45 min. Data sho w n are representative of three separate experiments that generated similar res u lts. (c ) Priming effects of GGA and IL-3 on b asophil degran u lation. Cells w ere preinc ub ated w ith and w itho u t GGA for 15 min, w ashed and then inc ub ated w ith and w itho u t IL-3 b efore stim u lation w ith anti-IgE anti b ody. Data are the mean ± SEM (n = 3). *p < 0.05 vers u s corresponding val u e of cells not preinc ub ated w ith GGA. p < 0.05 vers u s corresponding val u e of IL-3-u ntreated cells. (d ) Effect of geranyl-geraniol on b asophil histamine release. The indicated concentration of this compo u nd (0.5 μl/ml, 1.5 mM) did not ind u ce non-specifi c histamine release. Data are the mean ± SEM (n = 3). *p < 0.05 vers u s corresponding val u e of cells not preinc ub ated w ith geranylgeraniol. (e ) Simvastatin s u ppressed b asophil degran u lation. Basophils w ere preinc ub ated w ith and w itho u t simvas-tatin for 15 min at 37˚C, w ashed and then stim u lated w ith anti-IgE anti b ody or P MA for 45 min. Data are the mean ± SEM (n = 4). *p < 0.05, vers u s corresponding val u e of cells u ntreated w ith simvastatin.100a 80604020010080604020GGA (mM) - 2.7 - 2.7 IL-3 (pM) - - 300 300 a nti-IgE (P g/ml) 1.4 141.4 141.4 14 1.4 141.414401000.10.210100PMA (ng/ml)Pretreatment with GGA GGA (-)GGA 1.4 mM GGA 2.7 mMA23187 (P g/ml)anti-IgE (P g/ml)MCP-1 (nM)H i s t a m i n e r e l e a s e (%)c H i s t a m i n e r e l e a s e (%)80604020e100806040200d H i s t a m i ne r e l e a s e (%)3020101.4141000.2MCP-1(nM)A23187(P g/ml)10anti-IgE (P g/ml)PMA(ng/ml)100Statin (P M)-50-501.4 14 1.4 1410 100anti-IgE (P g/ml)PMA (ng/ml)10 100515Pretreatment with GGA (min)Pretreatment withgeranylgeraniol 30(-)1.5 mM60120b B a s e l i n e h i s t a m i n e r e l e a s e (%)GGA Modulates Basophil ActivationAllergology International Vol 63,Suppl 1,2014www.jsaweb.jp !511Divisionof Respiratory Medicine and Allergology,Department of Medicine,Teikyo University School of Medicine and 2National Hospital Organization Tokyo National Hospital,Tokyo,Japan Email:myama@med.teikyo−u.ac.jpConflict of interest:No potential conflict of interest was disclosed.REFERENCES1.Karasuyama H,Mukai K,Obata K,Tsujimura Y,Wada T.Nonredundant roles of basophils in immunity.Annu Rev Immunol 2011;29:45-69.2.Ishihara T,Suemasu S,Asasno T,Tanaka K,Mizushima T.Stimulation of gastric ulcer healing by heat shock pro-tein 70.Biochem Pharmacol 2011;82:728-36.3.Namba T,Tanaka K,Hoshino T,Azuma A,Mizushima T.Suppression of expression of heat shock protein 70by ge-fitinib and its contribution to pulmonary fibrosis.PLoS One 2011;6:e27296.4.Koketsu R,Yamaguchi M,Suzukawa M et al .Pretreat-ment with low levels of Fc εRI-crosslinking stimulation en-hances basophil mediator release.Int Arch Allergy Immu-nol 2013;161:S23-31.5.Fujimoto M,Oka T,Murata T,Hori M,Ozaki H.Fluvasta-tin inhibits mast cell degranulation without changing the cytoplasmic Ca 2+level.Eur J Pharmacol 2009;602:432-8.6.Majlesi Y,Samorapoompichit P,Hauswirth AW et al .Cerivastatin and atorvastatin inhibit IL-3-dependent differ-entiation and IgE-mediated histamine release in human basophils and downmodulate expression of the basophil-activation antigen CD203c !E-NPP3.J Leukoc Biol 2003;73:107-17.7.Graham TE,Pfeiffer JR,Lee RJ et al .MEK and ERK acti-vation in ras-disabled RBL-2H3mast cells and novel roles for geranylgeranylated and farnesylated proteins in Fc εRI -mediated signaling.J Immunol 1998;161:6733-44.8.Endo S,Hiramatsu N,Hayakawa K et al .Geranylgerany-lacetone,an inducer of the 70-kDa heat shock protein (HSP70),elicits unfolded protein response and coordi-nates cellular fate independently of HSP70.Mol Pharma-col 2007;72:1337-48.9.Nanke Y,Kawamoto M,Yago T,Chiba J,Yamanaka H,Kotake S.Geranylgeranylacetone,a non-toxic inducer of heat shock protein,induces cell death in fibroblast-like synoviocytes from patients with rheumatoid arthritis.Mod Rheumatol 2009;19:379-83.。