Modulation of T lymphocyte and eosinophil functions in vitro by natural tetranortriterpenoids isolated from Carapa guianensis Aublet Fausto K.Ferraris a,1 , Rodrigo Rodrigues b,2 , Vagner P.da Silva b , Raquel Figueiredo b , Carmen Penido a,3 , Maria das Graças M.O. Henriques a, ⁎ ,3 a Laboratório de Farmacologia Aplicada, Farmanguinhos, Fundação Oswaldo Cruz, Rua Sizenando Nabuco 100, 21041-250, Rio de Janeiro, RJ, Brazil b Laboratório de Química de Produtos Naturais, Farmanguinhos, Fundação Oswaldo Cruz, Rua Sizenando Nabuco 100, 21041-250, Rio de Janeiro, RJ, Brazil a b s t r a c t a r t i c l e i n f o Article history: Received 28 April 2010 Received in revised form 5 August 2010 Accepted 16 September 2010 Available online 15 October 2010 Keywords: Andiroba Limonoids Allergy CC chemokines T cell activation We have previously described the anti-allergic activities ofa pooled fraction of tetranortriterpenoids (TNTPs) containing 6α-acetoxygedunin, 7-deacetoxy-7-oxogedunin, andirobin and methyl angolensate isolated from the seeds of Carapa guianensis. In the present study, we performed in vitro studies in order to elucidate the mechanisms by which TNTPs present their anti-allergic effects and to identify the bioactive compound(s) present in such fraction. Here, we show that in vitro incubation of eosinophils with the pooled TNTP fraction, as well as with each one of the five isolated tetranortriterpenoids, impaired the adhesion of eosinophils to tumor necrosis factor-α (TNF-α)-primed tEND.1 endothelial cells. Furthermore, the individual or pooled TNTPs impaired CCL11/eotaxin-mediated chemotaxis. By contrast, pooled TNTPs failed to inhibit adhesion and chemotaxis of T lymphocytes. However, TNTPs were able to impair anti-CD3 monoclonal antibody-induced T cell proliferation and the expression of CD25 and CD69. These data suggest that TNTPs prevent T cell activation. Pretreatment of splenocytes with the pooled TNTP fraction, as well as with each one of the five isolated TNTPs, inhibited ovalbumin (OVA)-induced in vitro production of interleukin-2, chemokine (C-C motif) ligand 11 (CCL11) and regulated on activation normal T cell expressed and secreted (RANTES, also known as CCL5).TNTPs (except 6α-acetoxygedunin) also impaired nuclear factor-κB (NFκB) nuclear translocation in OVA-challenged splenocytes.Taken together,these results demonstrate that the anti-allergic effects of TNTPs isolated from C. guianensis might rely on their ability to inhibit eosinophil migration, as well as the activation of T lymphocytes, which is shared by the five isolated TNTPs. © 2010 Elsevier B.V. All rights reserved. 1. Introduction Plant extracts are commonly used in folk medicine to treat diseases. Many plant extracts have been shown to contain pharmaceutically relevant components [1]. Carapa guianensis Aublet, a member of the Meliaceae family,is widely used in popular medicine in Braziland other countries in the vicinity of the Amazon rainforest. C. guianensis contains triterpenes,tetraterpenes,alkaloids and limonoids. The presence of these chemicals is phytochemically characteristic of all members of the Meliaceae family. The proposed medicinal properties of C. guianensis have been attributed to the presence of limonoids, which are tetranortriterpenoids [for review see 2]. We previously demonstrated that a fraction isolated from C. guianensis seeds contains a number of different tetranortriterpenoid (TNTP) compounds, including 6α-acetoxygedunin,7-deacetoxy-7- oxogedunin, andirobin, gedunin and methyl angolensate. This fraction of TNTPs displayed marked anti-allergic and anti-inflammatory properties in rodents [3–5]. Such anti-allergic and anti-inflammatory effects relied on the ability of TNTPs to inhibit edema formation triggered by antigenic challenge [4]. The mechanism by which TNTPs inhibits edema formation is dependent on signaling pathways triggered by histamine, bradykinin and platelet-activating factor (PAF) [3]. Pooled TNTPs also impaired the production ofdifferent inflammatory mediators that trigger leukocyte infiltration into the site of inflammation.Among the inflammatory mediators that are impaired are the eosinophilotactic mediators, interleukin (IL)-5 and chemokine (C-C motif) ligand (CCL11)/eotaxin. Impairment of IL-5 and CCL11 seems to depend on the inhibition of nuclear factor-κB (NFκB) translocation, a phenomenon observed in vitro and in vivo [4]. However, these results were obtained by assaying a fraction of TNTP International Immunopharmacology 11 (2011) 1–11 ⁎ Corresponding author.Departamento de Farmacologia Aplicada, FarManguinhos, FIOCRUZ,R. Sizenando Nabuco, 100,Manguinhos,Rio de Janeiro,RJ, cep 21041-250, Brazil.Tel.: +55 21 39772482; fax: +55 21 25642559. E-mail address: gracahenriques@fiocruz.br (M.G.M.O. Henriques). 1 Ferraris is a PhD student ofPost-Graduation Program in Cellular and Molecular Biology of Oswaldo Cruz Institute. 2 Current Address: Laboratório de Ciências Químicas, CCT, Universidade Estadual do Norte Fluminense Darcy Ribeiro;Avenida Alberto Lamego 2000, Horto 28013-602, Campos dos Goytacazes, RJ, Brazil. 3 These authors contributed equally to this work. 1567-5769/$ – see front matter © 2010 Elsevier B.V. All rights reserved. doi:10.1016/j.intimp.2010.09.010 Contents lists available at ScienceDirect International Immunopharmacology j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / i n t i m p