Mediators of Inflammation

Mediators of Inflammation / 2014 / Article
Special Issue

Immunology and Infection by Protozoan Parasites

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Research Article | Open Access

Volume 2014 |Article ID 506450 | 12 pages |

TRPV1 Antagonism by Capsazepine Modulates Innate Immune Response in Mice Infected with Plasmodium berghei ANKA

Academic Editor: Mauricio Martins Rodrigues
Received03 Jun 2014
Accepted08 Jul 2014
Published24 Aug 2014


Thousands of people suffer from severe malaria every year. The innate immune response plays a determinant role in host’s defence to malaria. Transient receptor potential vanilloid 1 (TRPV1) modulates macrophage-mediated responses in sepsis, but its role in other pathogenic diseases has never been addressed. We investigated the effects of capsazepine, a TRPV1 antagonist, in malaria. C57BL/6 mice received 105 red blood cells infected with Plasmodium berghei ANKA intraperitoneally. Noninfected mice were used as controls. Capsazepine or vehicle was given intraperitoneally for 6 days. Mice were culled on day 7 after infection and blood and spleen cell phenotype and activation were evaluated. Capsazepine decreased circulating but not spleen F4/80+Ly6G+ cell numbers as well as activation of both F4/80+and F4/80+Ly6G+ cells in infected animals. In addition, capsazepine increased circulating but not spleen GR1+ and natural killer (NK) population, without interfering with natural killer T (NKT) cell numbers and blood NK and NKT activation. However, capsazepine diminished CD69 expression in spleen NKT but not NK cells. Infection increased lipid peroxidation and the release of TNFα and IFNγ, although capsazepine-treated group exhibited lower levels of lipid peroxidation and TNFα. Capsazepine treatment did not affect parasitaemia. Overall, TRPV1 antagonism modulates the innate immune response to malaria.

1. Introduction

Malaria is an infectious disease caused by intracellular protozoans of the genus Plasmodium and transmitted from person to person through bites of infected mosquitoes. It affects millions of people annually and is a leading cause of child mortality in underdevelopment countries [1]. Severe malaria such as cerebral malaria is frequently fatal and outcome of infection depends on host’s immune response, with innate immunity playing a determinant role in it [2, 3]. Available antimalarial therapy targets the Plasmodium. However, elimination of the parasite does not halt the clinical consequences of disease as surviving patients from severe malaria can develop a range of neurological deficits [4, 5]. In this context, an effective immune response is essential for patient’s recovery. Innate response is the major immunity component of patients who have been infected with Plasmodium for the first time, being essential to the development of an effective acquired immune response [6].

Recently, a protective role for transient receptor vanilloid 1 (TRPV1), a nonselective cation channel found on both neuronal and nonneuronal cells, was suggested in bacteria-induced sepsis [710]. Indeed, in the absence of TRPV1 activation, macrophage functions such as their ability to phagocytose and to release inflammatory mediators (nitric oxide (NO), reactive oxygen species (ROS), and cytokines) are impaired [9]. Also, TRPV1 has been linked to macrophage survival [9]. Evidence suggests a feedback between TRPV1 activation and ROS production may exist; in addition to modulating oxidative stress by downregulating ROS generation, this receptor can be directly activated by hydrogen peroxide (H2O2) [11] and regulated by superoxide anion release [1214]. Oxidative stress generation has a direct impact on macrophage-erythrocyte-endothelium interactions and imbalances of this pathway may trigger excessive damage and impaired host’s immune response to malaria [15, 16].

Herein, the role of TRPV1 in malaria was investigated for the first time. We used the TRPV1 antagonist, capsazepine, to assess whether TRPV1 is able to modulate the innate immune response to malaria in animals infected with Plasmodium berghei ANKA.

2. Materials and Methods

2.1. Animals

Inbred male C57BL/6 mice (8 weeks old) were used. Mice were obtained from the animal’s facility of the Department of Parasitology, Institute of Biomedical Sciences, University of São Paulo. Mice were kept in a climatically controlled environment and given food and water ad libitum. All procedures were approved by the Ethics Committee of the University of São Paulo and carried out in accordance with the Brazilian society for animal welfare (SBCAL).

2.2. Malaria Induction

Malaria was induced by a single intraperitoneal (i.p.) injection of 105 red blood cells (RBCs) infected with Plasmodium berghei ANKA (clone 1.49L) as described by Elias et al. [17]. Parasitaemia was assessed daily in a blood smear stained by Giemsa, by microscopy, from day 5 to day 7 following infection and was expressed as % of infected RBCs. Mice were terminally anaesthetised with a mixture of ketamine (75 mg/kg; Dopalen, Ceva, Brazil) and xylazine (1 mg/kg; i.p.; Sigma-Aldrich, Brazil), and exsanguinated by cardiac puncture on day 7 after infection (premortality end point; [18, 19]). Their blood and spleen were collected for further analysis. The plasma was separated and stored at −70°C for further quantitation of plasma aldehydes and cytokines. Cell population phenotype and activation were evaluated by flow cytometry. Noninfected mice were used as controls.

2.3. Pharmacological Treatment

In order to assess the role of TRPV1 in malaria, animals received the TRPV1 antagonist, capsazepine (Sigma-Aldrich, Brazil; , uninfected (control) group and , infected group), intraperitoneally from 24 h following infection, for 6 days (2x day, 50 μg/animal; [9]). Vehicle (10% DMSO in saline, 120 μL/animal; , uninfected (control) group and , infected group) was used as controls.

2.4. Flow Cytometry Analysis

Blood and spleen samples from infected and uninfected mice and single-cell suspensions were prepared. Peripheral blood cells were isolated by Percoll gradient (Sigma-Aldrich, Brazil). Spleens were homogenized and passed through a nylon mesh of 70 μm to create a single-cell suspension. Cells were stained with Trypan blue (Sigma-Aldrich, Brazil) and assessed for viability in a haemocytometer. Cells (106) were washed, resuspended in flow cytometry buffer (2% foetal calf serum (Invitrogen, Brazil) in phosphate buffered saline-PBS (Sigma-Aldrich, Brazil)), and stained with directly conjugated monoclonal antibodies (BD Biosciences or eBiosciences, Brazil): anti-F4/80 FITC, anti-IAb PE, anti-Ly6G PerCP, anti-GR1 Pe-Cy7, anti-CD3 PE and APC, anti-NK1.1 FITC, and anti-CD69 PECy5. Events were acquired on a BD FACSCanto (BD Biosciences-Immunocytometry Systems) and analyzed using FlowJo software (Tree Star Inc.). In order to analyze monocyte/macrophage and neutrophil populations, cells expressing CD3+, CD4+, CD8+, and CD19+ were gated out and the phenotypic F4/80, GR1, and Ly6G lineage markers were evaluated. Results are expressed as representative two-colour dot-plots as well as number of cells (×106), except for IAb, expressed as mean fluorescence.

2.5. Plasma Cytokine Levels

The plasma levels of TNFα, IFNγ, IL-4, IL-6, IL-2, IL-10, and IL-17 were evaluated by using a cytometric bead array (CBA) mouse Th1/Th2/Th17 cytokine kit (BD Biosciences, Brazil) according to manufacturer’s instructions. Analysis was performed on a Facscalibur cytometer flow cytometer (BD Biosciences-Immunocytometry Systems). Results were calculated in CBA FCAP Array software (BD Biosciences, Brazil) as pg/mL and are expressed as fold-increase relative to uninfected controls.

2.6. Plasma Aldehydes

Total plasma aldehyde (mainly malondialdehyde) concentrations were quantified as an index of lipid peroxidation and oxidative stress, as previously described [20, 21]. Briefly, 100 μL of sample were incubated with 100 μL of PBS and 400 μL of thiobarbituric acid (0.67%; Sigma-Aldrich, Brazil), at 90°C for 45 min. Samples were then centrifuged at 1,000 xg for 10 min. Three hundred μL of the supernatant was incubated with 300 μL of butanol (Sigma-Aldrich, Brazil) and 30 μL of a saturated solution of sodium chloride (Sigma-Aldrich, Brazil). Samples were mixed in a vortex, centrifuged at 1,000 xg for 2 min and then added to a 96-well plate (200 μL/well). Absorbance was read at 535 and 572 nm and the difference between the absorbance was used to calculate the aldehyde concentrations, using the molar extinction coefficient of the chromophore (1.56 × 105 M−1/cm−1). Results are expressed as fold-increase relative to uninfected controls.

2.7. Data Analysis

The results are presented as the mean ± standard deviation (SD). The percentages of inhibition are reported as mean ± SD of inhibitions obtained in each individual experiment compared with control samples. Statistical comparisons of the data were performed by ANOVA followed by Bonferroni and unpaired -test when appropriate. The values < 0.05 were considered significant.

3. Results

3.1. Capsazepine Does Not Affect Parasitaemia

Figure 1 shows parasitaemia levels up to day 7 after infection, in mice treated with either vehicle or capsazepine. Parasitaemia progressively increased in both groups. Repeated treatment with capsazepine had no effect on parasitaemia. Experiments were performed with a premortality end-point. Thus, no signs of cerebral malaria such as reduced responsiveness to stimulation, ataxia, respiratory distress or prostration, paralysis, and convulsions were observed in either capsazepine- or vehicle-infected mice. However, both groups of animals displayed piloerection and/or abnormal posture, as a result of infection.

3.2. Capsazepine Alters Circulating Monocyte but Not Spleen Macrophage Population Number and Activation

As represented in Figures 2(a)2(e), three distinct populations of peripheral blood leukocytes were detected in all groups of uninfected and infected animals: F4/80+, F4/80+Ly6G+ and Ly6G+ cells. Malaria induction had no effect on Ly6G+ cell population as no statistical significance was found between any of the evaluated groups. Mean ± SD values for Ly6G+ populations are as follows: vehicle-uninfected group , capsazepine uninfected group , vehicle-infected group , and capsazepine infected group . On the other hand, P. berghei ANKA infection increased F4/80+Ly6G+ cell numbers in both vehicle- (3.9-fold increase) and capsazepine- (1.9-fold increase) treated groups when compared to uninfected controls, whilst no effects were observed on F4/80+ cell population (Figures 2(b) and 2(c)). In addition, malaria caused increased activation of both F4/80+ (5.1-fold increase) and F4/80+Ly6G+ (6.6-fold increase) circulating cells when compared to uninfected animals, as denoted by expression of IAb on these cells (Figures 2(d) and 2(e)). Repeated administration of capsazepine in infected animals caused reduction of F4/80+Ly6G+ population expansion by % but did not affect F4/80+ cell numbers (Figures 2(b) and 2(c)). In addition, P. berghei ANKA-induced activation of F4/80+ and F4/80+Ly6G+ cells was halted by capsazepine. As depicted in Figures 2(d) and 2(e), capsazepine treatment reduced by % and %, the expression of IAb on F4/80+ and F4/80+Ly6G+ cells, respectively, when compared to vehicle-treated infected controls. As infection raised the number of circulating F4/80+Ly6G+ cells, a decline was noticed in the GR1+ cell population (%). However, infected mice treated with capsazepine exhibited a higher number of these cells when compared with both their uninfected- (1.8-fold increase) and infected- (2.4-fold increase) control animals. Mean ± SD values for GR1+ populations are as follows: vehicle-uninfected group , capsazepine uninfected group , vehicle-infected group , and capsazepine infected group . Capsazepine treatment in uninfected animals had no effects in regard to expansion or activation of both F4/80+ and F4/80+Ly6G+ cells (Figures 2(a)2(e)) and neither on GR1+ cells.

Similarly, F4/80+, F4/80+Ly6G+ and Ly6G+ cells were detected in spleen samples obtained from both infected and uninfected mice whether or not they were treated with capsazepine (Figures 3(a)3(e)). As observed for circulating Ly6G+ cells, malaria induction had no effects on spleen Ly6G+ cells. Mean ± SD values for spleen Ly6G+ populations are as follows: vehicle-uninfected group , capsazepine uninfected group , vehicle-infected group , and capsazepine infected group . However, P. berghei ANKA injection raised the numbers of F4/80+ and F4/80+Ly6G+ cells in both vehicle- (2.0- and 5.8-fold increase, resp.) and capsazepine- (1.5- and 4.9-fold increase, resp.) treated groups when compared to their respective uninfected controls (Figures 3(b) and 3(c)). Also, P. berghei ANKA infection augmented F4/80+ and F4/80+Ly6G+ cells in both vehicle- (5.5- and 2.0-fold increase, resp.) and capsazepine- (2.3- and 1.8-fold increase, resp.) treated groups when compared to their respective uninfected controls (Figures 3(d) and 3(e)). Capsazepine had no effects on the number or activation of spleen F4/80+ and F4/80+Ly6G+ cells (Figures 3(a)3(e)). Spleen GR1+ cell population remained unaltered irrespective of treatments and mean ± SD values are as follows: vehicle-uninfected group , capsazepine uninfected group , vehicle-infected group , and capsazepine infected group . Moreover, capsazepine had no effects on F4/80+ and F4/80+Ly6G+ cells (Figures 3(a)3(e)) when administered to uninfected mice.

3.3. Capsazepine Modulates Blood and Spleen NK and NKT Population Number and Activation

We also evaluated the effects of capsazepine on circulating and spleen NK (CD3NK1.1+) and NKT (CD3+NK1.1+) cells. Peripheral blood NK and NKT cells were detected in all groups of uninfected and infected animals (Figures 4(a)4(e)). Malaria induction had no effects on either NK or NKT cell numbers when compared to uninfected control mice (Figures 4(b) and 4(d)). On the other hand, NK, but not NKT activation via CD69 expression, was increased in both vehicle- (11.1-fold increase) and capsazepine- (11.5-fold increase) treated groups when compared to their respective uninfected controls (Figure 4(c)). However, capsazepine treatment significantly increased NK cell population (Figure 4(b)) when compared to either its uninfected control group (1.8-fold increase) or vehicle-treated infected animals (1.5-fold increase). Also, capsazepine was able to enhance activation of NKT-infected cells (2.7-fold increase; Figure 4(e)). Importantly, NK cells account for the majority of circulating and activated NK1.1+ cells in infected animals (Figure 4).

Similarly, spleen NK and NKT cells were detected in all groups of uninfected and infected animals (Figures 5(a)5(e)). Malaria induction caused spleen NKT but not NK cell population expansion when compared to uninfected control mice (Figures 5(b) and 5(d)). This was observed for both vehicle- (5.6-fold increase) and capsazepine- (4.1-fold increase) treated mice (Figure 5(d)). Figures 5(c) and 5(e) demonstrate that CD69 expression was augmented on both spleen NK and NKT cells obtained from infected mice treated with either vehicle (5.3- and 6.9-fold increase, resp.) or capsazepine (5.6- and 3.1-fold increase, resp.). However, NKT activation was % lower in capsazepine-treated mice when compared to vehicle-infected controls (Figure 5(e)). Capsazepine treatment in infected mice had no effects on spleen NK and NKT cell numbers or NK activation (Figure 5). Similarly, capsazepine did not alter spleen NK and NKT profile (cell number and activation) in samples obtained from uninfected mice (Figure 5).

3.4. Capsazepine Reduces Lipid Peroxidation and Plasma TNFα Levels in P. berghei ANKA-Infected Mice

P. berghei ANKA infection increased lipid peroxidation in both vehicle- and capsazepine-treated mice, as demonstrated by the levels of plasma aldehydes (Figure 6(a)). However, this increase was less pronounced in the capsazepine-treated group (% reduction). Malaria also triggered the release of TNFα and IFNγ in both infected groups (Figures 6(b) and 6(c)). TNFα production was markedly reduced by capsazepine treatment (%; Figure 6(b)). On the other hand, capsazepine treatment did not affect IFNγ release triggered by malaria (Figure 6(c)). Vehicle- and capsazepine-treated uninfected mice exhibited similar levels of plasma aldehydes, TNFα, and IFNγ (data not shown). Production of IL-4, IL-6, IL-2, IL-10, and IL-17 was not detected in any of the evaluated groups.

4. Discussion

Since its discovery, evidence has accumulated that TRPV1 has the potential to play a key role in a variety of pathologies, especially those associated with imbalances of the immune and inflammatory response, such as asthma [2224] and rheumatoid arthritis [2527]. Recent reports demonstrated that TRPV1 is expressed on immune cells such as macrophages and peripheral-blood mononuclear cells in inflammatory conditions [9, 2830]. More recently, TRPV1 was suggested to modulate a range of macrophage-mediated responses to bacterial infection [9]. Indeed, TRPV1 deletion or antagonism has been associated with poorer outcome of experimental sepsis as TRPV1 blockade increases pathogen load and also facilitates the transition from a local to a systemic inflammatory response to bacteria [7, 9, 10, 3134]. In addition, TRPV1 knockout (TRPV1 KO) mice challenged with intestinal bacteria or LPS present with a dysregulated production of inflammatory mediators, including NO, ROS, and cytokines such as TNFα, IL-10, and IL-6 [79]. So far, there are no reports of TRPV1 playing any roles in the immune response to other pathogens.

Here, we show for the first time that TRPV1 antagonism by capsazepine, a nonselective antagonist, modulates the innate immune response to malaria. Reports have shown that capsazepine presents species- and modality-specific activity on TRPV1 and also inhibits acetylcholine receptors, voltage-gated calcium channels, and hyperpolarization-activated cyclic nucleotide-gated channels, in addition to TRPV1 [3538]. However, it is important to highlight that capsazepine was administered in this study, as described by Fernandes and collaborators [9] who showed that repeated treatment with this drug in vivo produces a similar profile to that of TRPV1 KO mice in response to bacterial infection [9]. Herein, we induced malaria by injecting P. berghei ANKA in a susceptible strain of mice known to develop cerebral malaria-like symptoms and to present with 60–100% mortality within the second week following injection (for review see [19]). Our studies were carried out at a premortality end-point, that is, 7 days following infection. At this time point, parasitaemia had reached 11%, in agreement with previous studies [17, 22].

Evidence suggests that the innate immune response plays an important role during the early phase of infection, with activated monocytes and neutrophils releasing nonspecific inflammatory mediators such as ROS and cytokines [2, 39] and exerting their roles as phagocytes and antigen-presenting cells when in contact with circulating infected RBCs [40]. Indeed, phagocytosis of infected RBCs by peripheral blood and tissue phagocytes is suggested to be the major mechanism of Plasmodium removal [41]. As a result of this interaction, phagocytes may damage the endothelium, thus contributing to the collapse of the circulation and in the case of cerebral malaria, damaging the brain microvasculature [4, 42]. During infection, monocytes differentiate into macrophages in the spleen and also in the brain, becoming available in the brain microvasculature [4]. Activated macrophages contribute to pathogen clearance and activation of lymphocytes, in an attempt to stimulate the generation of an acquired immune response capable of improving parasite removal and fighting a secondary infection [2, 43].

As previously mentioned, TRPV1 can be found on immune cells. It is then expected that antagonism of TRPV1 channels expressed on these cells would affect their phenotypes. Our results show that at 7 days after infection, there is activation of both monocytes and spleen macrophages (F4/80+). Interestingly, we detected a group of circulating F4/80+Ly6G+ cells which became markedly expanded and activated (as denoted by IAb expression) following infection. This is the first report to our knowledge of their contribution to malaria. However, it was recently suggested in a model of infection caused by vaccinia virus inoculation in C57BL/6 mice that F4/80+Ly6G+ cells are indeed monocytes with a great capacity of producing ROS and IFNγ whilst Ly6G monocytes produce NO and TNFα [44]. In addition, the same study suggested that F4/80+Ly6G+ cells replace Ly6G monocytes as infection progresses. Herein, we show the existence of F4/80+Ly6G+ cells also in the spleen and that they exhibit a similar profile (in terms of number and activation pattern) to that of Ly6G cells in this tissue. As the population of F4/80+Ly6G+ cells rises, GR1+ cells (myeloid-derived suppressor cells) decline with infection. This is expected as an increase of mature monocytes and neutrophils normally occurs as a result of infection [45, 46]. However, at 7 days after infection, these changes in the balance and/or activation of F4/80+, F4/80+Ly6G+, and GR1+ cells could not be noticed at spleen level. However, it is possible that these alterations may occur in this organ at a different time point not addressed by this study.

Repeated treatment with capsazepine caused inhibition of activation of both F4/80+ and F4/80+Ly6G+ cells as well as reduction of the number of circulating F4/80+Ly6G+ cells in infected mice. Interestingly, GR1+ cell number is markedly increased in the same group of animals, suggesting that, as capsazepine shuts down F4/80+- and F4/80+Ly6G+-mediated responses, either by deactivating them or decreasing their availability in the circulation, these cells are progressively replaced by GR1+ cells in order to restore the immune responses that may be dependent on F4/80+- and F4/80+Ly6G+ monocytes. Indeed, GR1+ cells are suggested to be recruited in order to compensate “the loss” of monocytes that undergo polarization following Plasmodium or bacterial infection [47, 48]. These cells are also called regulatory monocytes/macrophages and have a potent ability to suppress T cell proliferation and Th1 responses [49]. On the other hand, the definite role of these cells in critical illness is still of debate (for review see [47]).

To evaluate the impact this shift on monocyte profile had on inflammatory mediator release, we measured the levels of plasma aldehydes (index of lipid peroxidation secondary to oxidative stress) and cytokines such as TNFα and IFNγ. Indeed, high systemic levels of inflammatory mediators such as TNFα, IFNγ, and aldehydes are correlated with severe malaria in humans [50]. In fact, lipid peroxidation is produced during malaria as a result of the interactions between monocytes/macrophages and infected erythrocytes, and also the endothelium [51, 52]. In addition, cytokines are also released in response to malaria, triggering suppression of erythropoiesis and activation of a variety of circulating and spleen cells [2, 53, 54]. We found that whilst diminished levels of lipid peroxidation-derived aldehydes and TNFα were detected in animals with malaria that had been treated with capsazepine, IFNγ production remained similar to that observed for samples obtained from vehicle-infected mice. A reduction of oxidative stress was expected as capsazepine was previously suggested to inhibit oxidative stress in cultured RAW264 monocytes/macrophages in a TRPV1 independent manner, although this data was obtained from cells that had not been challenged with any pathogen product [55]. Later, TRPV1 deletion was shown to decrease ROS production by macrophages in sepsis [9] and to modulate the release of these mediators in other inflammatory conditions [14, 56]. However, loss of TRPV1 function has been associated with increased production of TNFα upon bacterial infection [7, 9, 32]. It is possible that TRPV1 differently modulates monocytes and macrophages with respect to their ability to produce TNFα. Also, TRPV1 effects on TNFα production may vary at different stages of malaria.

NK and NKT cells are important effectors of the innate immune response to malaria, directly recognizing Plasmodium-infected RBCs and malarial antigens, in addition to producing IFNγ in order to contain parasitaemia [2, 18, 40]. Indeed, these cells rise early during malaria and have been suggested to mediate the differentiation of Th1/Th2 responses and to be essential for the trafficking of leukocytes to the brain in cerebral malaria (for review see [2]). In addition, similarly to macrophages, they can accumulate into the brain during cerebral malaria, contributing to a worse outcome [18]. NK cells have also been linked to dendritic cell maturation and T cell activation in the spleen by releasing of cytokines [53, 57]. Also, evidence has shown that during malaria, NK cells may undergo an intense turnover or even migrate out of the spleen [18]. This was investigated at a similar time-point to that used in our study (1 week following infection). Here, we assessed the dynamics between circulating and spleen NK and NKT cells. We found that P. berghei ANKA-induced infection caused expansion of spleen NKT but not NK cell population. This was not accompanied by any change on circulating NK and NKT cell numbers. It is possible that at this time-point, NK and NKT cells have already migrated to the brain. Indeed, Hansen and collaborators showed that NK cells accumulate into the brain of C57BL/6 mice as early as 4 days following infection with P. berghei ANKA, triggering the migration of T cells to the brain microvasculature [18]. On the other hand, we show that both NK and NKT cells became activated in response to infection, with NK cells representing the majority of activated cells. Disappointingly, expansion of spleen NKT cells did not translate in their activation as only few of these cells expressed CD69. Capsazepine treatment in infected mice led to a further expansion of the circulating NK population, but its activation was similar to that of vehicle-infected group. In addition, capsazepine treatment reduced by half the activation of spleen NKT cells when compared to its infected-control. We would expect that the reduction of activation of spleen NKT cells and F4/80+Ly6G+ monocytes by capsazepine would impair IFNγ release. However, we show capsazepine inhibitory effects on these populations do not affect IFNγ production in malaria, indicating that either TRPV1 may not play a role on IFNγ production or NK cells become the sole source of IFNγ once TRPV1 is blocked. In addition, the lack of effect of capsazepine on parasitaemia may be related to the similar levels of IFNγ detected in both groups of animals.

Our study provides the first evidence that TRPV1 modulates malaria by mediating innate immune response, specifically by interfering with the expansion and activation of effector cells, especially monocytes. We also show that TRPV1 regulates the immunological balance between different monocyte populations in addition to modulating mediator release by them. It is possible that blocking TRPV1 may be either beneficial, as a reduction of oxidative stress may reflect on reduced vascular dysfunction, or deleterious, as impairment of innate response may lead to an inefficient removal of the parasite in addition to an inefficient acquired immune response to malaria. However, the impact TRPV1 antagonism may have on severe malaria outcome is of importance and remains to be investigated.

Conflict of Interests

The authors declare no conflict of interests.


This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq; Brazil), Coordenação de Aperfeiçoamento de Pessoal de Nivel Superior (CAPES; Brazil), Fundação de Amparo à Pesquisa e Desenvolvimento Científico do Maranhão (FAPEMA; Brazil), and Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP; Brazil).


  1. World Health Organization, World Malaria Report: 2013, 2013.
  2. L. Schofield, “Intravascular infiltrates and organ-specific inflammation in malaria pathogenesis,” Immunology and Cell Biology, vol. 85, no. 2, pp. 130–137, 2007. View at: Publisher Site | Google Scholar
  3. V. Pradhan and K. Ghosh, “Immunological disturbances associated with malarial infection,” Journal of Parasitic Diseases, vol. 37, no. 1, pp. 11–15, 2013. View at: Publisher Site | Google Scholar
  4. H. J. Shikani, B. D. Freeman, M. P. Lisanti, L. M. Weiss, H. B. Tanowitz, and M. S. Desruisseaux, “Cerebral malaria: we have come a long way,” The American Journal of Pathology, vol. 181, no. 5, pp. 1484–1492, 2012. View at: Publisher Site | Google Scholar
  5. A. Moulick, S. Maitra, B. S. Sarkar, A. Jana, and S. Sarkar, “Vivax malaria presenting with myelitis: a rare complication,” Journal of Clinical and Diagnostic Research, vol. 7, no. 5, pp. 914–916, 2013. View at: Publisher Site | Google Scholar
  6. M. M. Stevenson and E. M. Riley, “Innate immunity to malaria,” Nature Reviews Immunology, vol. 4, no. 3, pp. 169–180, 2004. View at: Publisher Site | Google Scholar
  7. I. A. Clark, “How TNF was recognized as a key mechanism of disease,” Cytokine and Growth Factor Reviews, vol. 18, no. 3-4, pp. 335–343, 2007. View at: Publisher Site | Google Scholar
  8. E. S. Fernandes, M. A. Fernandes, and J. E. Keeble, “The functions of TRPA1 and TRPV1: moving away from sensory nerves,” British Journal of Pharmacology, vol. 166, no. 2, pp. 510–521, 2012. View at: Publisher Site | Google Scholar
  9. E. S. Fernandes, L. Liang, S. Smillie et al., “TRPV1 deletion enhances local inflammation and accelerates the onset of systemic inflammatory response syndrome,” Journal of Immunology, vol. 188, no. 11, pp. 5741–5751, 2012. View at: Publisher Site | Google Scholar
  10. V. Guptill, X. Cui, A. Khaibullina et al., “Disruption of the transient receptor potential vanilloid 1 can affect survival, bacterial clearance, and cytokine gene expression during murine sepsis,” Anesthesiology, vol. 114, no. 5, pp. 1190–1199, 2011. View at: Publisher Site | Google Scholar
  11. K. Susankova, K. Tousova, L. Vyklicky, J. Teisinger, and V. Vlachova, “Reducing and oxidizing agents sensitize heat-activated vanilloid receptor (TRPV1) current,” Molecular Pharmacology, vol. 70, no. 1, pp. 383–394, 2006. View at: Publisher Site | Google Scholar
  12. E. S. Fernandes, C. T. Vong, S. Quek et al., “Superoxide generation and leukocyte accumulation: key elements in the mediation of leukotriene B4-induced itch by transient receptor potential ankyrin 1 and transient receptor potential vanilloid 1,” The FASEB Journal, vol. 27, no. 4, pp. 1664–1673, 2013. View at: Publisher Site | Google Scholar
  13. P. Puntambekar, D. Mukherjea, S. Jajoo, and V. Ramkumar, “Essential role of Rac1/NADPH oxidase in nerve growth factor induction of TRPV1 expression,” Journal of Neurochemistry, vol. 95, no. 6, pp. 1689–1703, 2005. View at: Publisher Site | Google Scholar
  14. A. Starr, R. Graepel, J. Keeble et al., “A reactive oxygen species-mediated component in neurogenic vasodilatation,” Cardiovascular Research, vol. 78, no. 1, pp. 139–147, 2008. View at: Publisher Site | Google Scholar
  15. C. K. B. Ferrari, P. C. S. Souto, E. L. França, and A. C. Honorio-França, “Oxidative and nitrosative stress on phagocytes' function: from effective defense to immunity evasion mechanisms,” Archivum Immunologiae et Therapiae Experimentalis, vol. 59, no. 6, pp. 441–448, 2011. View at: Publisher Site | Google Scholar
  16. S. Percário, D. R. Moreira, B. A. Q. Gomes et al., “Oxidative stress in Malaria,” International Journal of Molecular Sciences, vol. 13, no. 12, pp. 16346–16372, 2012. View at: Publisher Site | Google Scholar
  17. R. M. Elias, M. Correa-Costa, C. R. Barreto et al., “Oxidative stress and modification of renal vascular permeability are associated with acute kidney injury during P. berghei ANKA infection,” PLoS ONE, vol. 7, no. 8, Article ID e44004, 2012. View at: Publisher Site | Google Scholar
  18. D. S. Hansen, N. J. Bernard, C. Q. Nie, and L. Scholeld, “NK cells stimulate recruitment of CXCR3+ T cells to the brain during Plasmodium berghei-mediated cerebral malaria,” Journal of Immunology, vol. 178, no. 9, pp. 5779–5788, 2007. View at: Publisher Site | Google Scholar
  19. L. Rénia, S. W. Howland, C. Claser et al., “Cerebral malaria Mysteries at the blood-brain barrier,” Virulence, vol. 3, no. 2, pp. 193–201, 2012. View at: Publisher Site | Google Scholar
  20. R. P. Bird and H. H. Draper, “Comparative studies on different methods of malonaldehyde determination,” Methods in Enzymology, vol. 105, pp. 299–305, 1984. View at: Publisher Site | Google Scholar
  21. L. Sharma, J. Kaur, and G. Shukla, “Role of oxidative stress and apoptosis in the placental pathology of plasmodium berghei infected mice,” PLoS ONE, vol. 7, no. 3, Article ID e32694, 2012. View at: Publisher Site | Google Scholar
  22. R. Nassini, P. Pedretti, N. Moretto et al., “Transient receptor potential ankyrin 1 channel localized to non-neuronal airway cells promotes non-neurogenic inflammation,” PLoS ONE, vol. 7, no. 8, Article ID e42454, 2012. View at: Publisher Site | Google Scholar
  23. F. Tsuji and H. Aono, “Role of transient receptor potential vanilloid 1 in inflammation and autoimmune diseases,” Pharmaceuticals, vol. 5, no. 8, pp. 837–852, 2012. View at: Publisher Site | Google Scholar
  24. J. Yang, H. M. Yu, X. D. Zhou, V. P. Kolosov, and J. M. Perelman, “Study on TRPV1-mediated mechanism for the hypersecretion of mucus in respiratory inflammation,” Molecular Immunology, vol. 53, no. 1-2, pp. 161–171, 2013. View at: Publisher Site | Google Scholar
  25. E. S. Fernandes, F. A. Russell, D. Spina et al., “A distinct role for transient receptor potential ankyrin 1, in addition to transient receptor potential vanilloid 1, in tumor necrosis factor α-induced inflammatory hyperalgesia and Freund's complete adjuvant-induced monarthritis,” Arthritis and Rheumatism, vol. 63, no. 3, pp. 819–829, 2011. View at: Publisher Site | Google Scholar
  26. J. Keeble, F. Russell, B. Curtis, A. Starr, E. Pinter, and S. D. Brain, “Involvement of transient receptor potential vanilloid 1 in the vascular and hyperalgesic components of joint inflammation,” Arthritis and Rheumatism, vol. 52, no. 10, pp. 3248–3256, 2005. View at: Publisher Site | Google Scholar
  27. F. A. Russell, E. S. Fernandes, J. Courade, J. E. Keeble, and S. D. Brain, “Tumour necrosis factor α mediates transient receptor potential vanilloid 1-dependent bilateral thermal hyperalgesia with distinct peripheral roles of interleukin-1β, protein kinase C and cyclooxygenase-2 signalling,” Pain, vol. 142, no. 3, pp. 264–274, 2009. View at: Publisher Site | Google Scholar
  28. T. K. Finney-Hayward, M. O. Popa, P. Bahra et al., “Expression of transient receptor potential C6 channels in human lung macrophages,” The American Journal of Respiratory Cell and Molecular Biology, vol. 43, no. 3, pp. 296–304, 2010. View at: Publisher Site | Google Scholar
  29. C. I. Saunders, R. G. Fassett, and D. P. Geraghty, “Up-regulation of TRPV1 in mononuclear cells of end-stage kidney disease patients increases susceptibility to N-arachidonoyl-dopamine (NADA)-induced cell death,” Biochimica et Biophysica Acta: Molecular Basis of Disease, vol. 1792, no. 10, pp. 1019–1026, 2009. View at: Publisher Site | Google Scholar
  30. J. Zhao, L. Ching, Y. R. Kou et al., “Activation of TRPV1 prevents OxLDL-induced lipid accumulation and TNF-α-Induced inflammation in macrophages: role of liver X receptor α,” Mediators of Inflammation, vol. 2013, Article ID 925171, 14 pages, 2013. View at: Publisher Site | Google Scholar
  31. P. Bryant, M. Shumate, G. Yumet, C. H. Lang, T. C. Vary, and R. N. Cooney, “Capsaicin-sensitive nerves regulate the metabolic response to abdominal sepsis,” The Journal of Surgical Research, vol. 112, no. 2, pp. 152–161, 2003. View at: Publisher Site | Google Scholar
  32. N. Clark, J. Keeble, E. S. Fernandes et al., “The transient receptor potential vanilloid 1 (TRPV1) receptor protects against the onset of sepsis after endotoxin,” The FASEB Journal, vol. 21, no. 13, pp. 3747–3755, 2007. View at: Publisher Site | Google Scholar
  33. T. Iida, I. Shimizu, M. L. Nealen, A. Campbell, and M. Caterina, “Attenuated fever response in mice lacking TRPV1,” Neuroscience Letters, vol. 378, no. 1, pp. 28–33, 2005. View at: Publisher Site | Google Scholar
  34. Y. Wang, M. Novotný, V. Quaiserová-Mocko, G. M. Swain, and D. H. Wang, “TRPV1-mediated protection against endotoxin-induced hypotension and mortality in rats,” American Journal of Physiology: Regulatory Integrative and Comparative Physiology, vol. 294, no. 5, pp. R1517–R1523, 2008. View at: Publisher Site | Google Scholar
  35. R. J. Docherty, J. C. Yeats, and A. S. Piper, “Capsazepine block of voltage-activated calcium channels in adult rat dorsal root ganglion neurones in culture,” British Journal of Pharmacology, vol. 121, no. 7, pp. 1461–1467, 1997. View at: Publisher Site | Google Scholar
  36. L. Liu and S. A. Simon, “Capsazepine, a vanilloid receptor antagonist, inhibits nicotinic acetylcholine receptors in rat trigeminal ganglia,” Neuroscience Letters, vol. 228, no. 1, pp. 29–32, 1997. View at: Publisher Site | Google Scholar
  37. C. H. Gill, P. J. Strijbos, S. A. Bates, W. Cairns, D. Owen, and C. H. Davies, “Capsazepine inhibits a recombinant human HCN1-mediated current,” British Journal of Pharmacology, vol. 135, no. 5, p. 251, 2001. View at: Google Scholar
  38. P. McIntyre, L. M. McLatchie, A. Chambers et al., “Pharmacological differences between the human and rat vanilloid receptor 1 (VR1),” British Journal of Pharmacology, vol. 132, no. 5, pp. 1084–1094, 2001. View at: Google Scholar
  39. U. Frevert and A. Nacer, “Immunobiology of Plasmodium in liver and brain,” Parasite Immunology, vol. 35, no. 9-10, pp. 267–282, 2013. View at: Google Scholar
  40. C. Coban, K. J. Ishii, T. Horii, and S. Akira, “Manipulation of host innate immune responses by the malaria parasite,” Trends in Microbiology, vol. 15, no. 6, pp. 271–278, 2007. View at: Publisher Site | Google Scholar
  41. V. Gallo, O. A. Skorokhod, E. Schwarzer, and P. Arese, “Simultaneous determination of phagocytosis of Plasmodium falciparum-parasitized and non-parasitized red blood cells by flow cytometry,” Malaria Journal, vol. 11, article 428, 2012. View at: Publisher Site | Google Scholar
  42. F. M. S. de Leoratti, S. C. Trevelin, F. Q. Cunha et al., “Neutrophil paralysis in Plasmodium vivax malaria,” PLoS Neglected Tropical Diseases, vol. 6, no. 6, Article ID e1710, p. e1710, 2012. View at: Publisher Site | Google Scholar
  43. C. L. L. Chua, G. Brown, J. A. Hamilton, S. Rogerson, and P. Boeuf, “Monocytes and macrophages in malaria: protection or pathology?” Trends in Parasitology, vol. 29, no. 1, pp. 26–34, 2013. View at: Publisher Site | Google Scholar
  44. M. A. Fischer, M. L. Davies, I. E. Reider et al., “CD11b+, Ly6G+ cells produce type I interferon and exhibit tissue protective properties following peripheral virus infection,” PLoS Pathogens, vol. 7, no. 11, Article ID e1002374, 2011. View at: Publisher Site | Google Scholar
  45. J. A. Villadangos, “Hold On, the Monocytes Are Coming!,” Immunity, vol. 26, no. 4, pp. 390–392, 2007. View at: Publisher Site | Google Scholar
  46. Q. Yang, P. Ghose, and N. Ismaila, “Neutrophils mediate immunopathology and negatively regulate protective immune responses during fatal bacterial infection-induced Toxic Shock,” Infection and Immunity, vol. 81, no. 5, pp. 1751–1763, 2013. View at: Publisher Site | Google Scholar
  47. J. N. Fullerton, A. J. O'Brien, and D. W. Gilroy, “Pathways mediating resolution of inflammation: when enough is too much,” The Journal of Pathology, vol. 231, no. 1, pp. 8–20, 2013. View at: Google Scholar
  48. J. A. van Ginderachter, A. Beschin, P. D. Baetselier, and G. Raes, “Myeloid-derived suppressor cells in parasitic infections,” European Journal of Immunology, vol. 40, no. 11, pp. 2976–2985, 2010. View at: Publisher Site | Google Scholar
  49. D. I. Gabrilovich and S. Nagaraj, “Myeloid-derived suppressor cells as regulators of the immune system,” Nature Reviews Immunology, vol. 9, no. 3, pp. 162–174, 2009. View at: Publisher Site | Google Scholar
  50. K. E. Lyke, R. Burges, Y. Cissoko et al., “Serum levels of the proinflammatory cytokines interleukin-1 beta (IL-1β), IL-6, IL-8, IL-10, tumor necrosis factor alpha, and IL-12(p70) in Malian children with severe Plasmodium falciparum malaria and matched uncomplicated malaria or healthy controls,” Infection and Immunity, vol. 72, no. 10, pp. 5630–5637, 2004. View at: Publisher Site | Google Scholar
  51. S. L. Hazen, “Oxidized phospholipids as endogenous pattern recognition ligands in innate immunity,” Journal of Biological Chemistry, vol. 283, no. 23, pp. 15527–15531, 2008. View at: Publisher Site | Google Scholar
  52. E. Schwarzer, H. Kühn, E. Valente, and P. Arese, “Malaria-parasitized erythrocytes and hemozoin nonenzymatically generate large amounts of hydroxy fatty acids that inhibit monocyte functions,” Blood, vol. 101, no. 2, pp. 722–728, 2003. View at: Publisher Site | Google Scholar
  53. M. Vitale, M. Della Chiesa, S. Carlomagno et al., “NK-dependent DC maturation is mediated by TNFα and IFNγ released upon engagement of the NKp30 triggering receptor,” Blood, vol. 106, no. 2, pp. 566–571, 2005. View at: Publisher Site | Google Scholar
  54. F. Gimenez, S. B. De Lagerie, C. Fernandez, P. Pino, and D. Mazier, “Tumor necrosis factor α in the pathogenesis of cerebral malaria,” Cellular and Molecular Life Sciences, vol. 60, no. 8, pp. 1623–1635, 2003. View at: Publisher Site | Google Scholar
  55. M. J. Garle, A. Knight, A. T. Downing, K. L. Jassi, R. H. Clothier, and J. R. Fry, “Stimulation of dichlorofluorescin oxidation by capsaicin and analogues in RAW 264 monocyte/macrophages: lack of involvement of the vanilloid receptor,” Biochemical Pharmacology, vol. 59, no. 5, pp. 563–572, 2000. View at: Publisher Site | Google Scholar
  56. T. Schilling and C. Eder, “Importance of the non-selective cation channel TRPV1 for microglial reactive oxygen species generation,” Journal of Neuroimmunology, vol. 216, no. 1-2, pp. 118–121, 2009. View at: Publisher Site | Google Scholar
  57. F. Gerosa, B. Baldani-Guerra, C. Nisii, V. Marchesini, G. Carra, and G. Trinchieri, “Reciprocal activating interaction between natural killer cells and dendritic cells,” The Journal of Experimental Medicine, vol. 195, no. 3, pp. 327–333, 2002. View at: Publisher Site | Google Scholar

Copyright © 2014 Elizabeth S. Fernandes et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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