Acta Tropica 210 (2020) 105548

Contents lists available at ScienceDirect

Acta Tropica
journal homepage: www.elsevier.com/locate/actatropica

Fasciola hepatica-derived molecules as potential immunomodulators T

1 ⁎,2
Gerardo Manuel Corral-Ruiz , Luvia Enid Sánchez-Torres
Departamento de Inmunología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prolongación de Carpio y Plan de Ayala, s/n, 11340 Ciudad de
México, México

A R T I C LE I N FO A B S T R A C T

Keywords: Through the years, helminths have co-existed with many species. This process has allowed parasites to live
Fasciola hepatica within them for long periods and, in some cases, to generate offspring. In particular, this ability has allowed
Immunoregulation Fasciola hepatica to survive the diverse immunological responses faced within its wide range of hosts. The vast
Helminth repertoire of molecules that are constantly secreted in large quantities by the parasite, acts directly on several
Trematode
cells of the immune system affecting their antiparasitic capacities. Interestingly, these molecules can direct the
Excretory-secretory molecules
Inflammation
host immune response to an anti-inflammatory and regulatory phenotype that assures the survival of the parasite
with less harm to the host. Based on these observations, some of the products of F. hepatica, as well as those of
other helminths, have been studied, either as a total extract, extracellular vesicles or as purified molecules, to
establish and characterize their anti-inflammatory mechanisms. Until now, the results obtained encourage
further research directed to discover new helminth-derived alternatives to replace current therapies, which can
be useful for people suffering from inflammatory diseases like autoimmunity or allergy processes that affect their
life quality. In this review, some of the most studied molecules derived from F. hepatica and their modulating
capacities are discussed.

1. Introduction The clinical manifestations and the consequent pathology of the

helminth infection depend on several factors. Two of them are the
Helminths are a large group of multicellular organisms that can moment that a certain parasitic load threshold is exceeded, and the
parasite diverse hosts. Currently, soil-transmitted helminthiases are the organ or the tissue they are located. An important aspect is the ability
most important in human epidemiological terms, since there are 1.5 that the helminths have to move through different organs such as the
billion infected people, equivalent to almost 24% of the world popu- colon, small intestine, lymph nodes, liver, lung, and brain (Riet et al.,
lation. Despite the cases of coinfection, this percentage increases if 2007). This situation has arisen the theory that their migration through
other helminthiases, such as schistosomiasis and filariasis are con- the different organs, besides being a maturation process, could also be
sidered (World Health Organization, 2018). These infections cause assumed as a survival mechanism (Meeusen, 1999; Mulcahy et al.,
important health problems including malnutrition (Cattadori et al., 2004; Mulcahy et al., 2005).
2016), low physical development, and cognitive problems Helminths have accompanied man throughout his history; both
(Pabalan et al., 2018). organisms have experienced intimate coevolution and despite the wide
These parasites possess great biological variability. They have a diversity and heterogeneity of the parasites, they induce a similar
wide range of intermediary and definitive hosts that go from snails and pattern of immune responses in their hosts (McSorley and
flies to mammals. Helminths can enter the host in several ways, mainly Maizels, 2012). In the acute phase of the infection, an initial in-
by ingestion of contaminated water or food, by penetration through the flammatory response is established however, the response directed to
skin or by the bite of an arthropod. Another feature that defines them is larval stages is not always effective and the effector mechanisms in-
the diversity of the phases they present inside the host, which may volved can even cause damage to the host. Later in the infection, a
include eggs, larval stages, and even adults; each phase represents a protective Th2 response against the adult stages is detected, but the
complex antigenic mosaic, to which the host immune system must re- modulation of the immune response exerted by the parasites favored
spond (Riet et al., 2007). their persistence inside the host. The coadaptation experienced between

⁎
Corresponding author.
E-mail address: [email protected] (L.E. Sánchez-Torres).
1
ORCID ID Gerardo Manuel Corral-Ruiz: 0000-0001-6496-8190
2
ORCID ID Luvia Enid Sánchez-Torres: 0000-0002-7128-849X

https://doi.org/10.1016/j.actatropica.2020.105548
Received 7 January 2020; Received in revised form 21 April 2020; Accepted 18 May 2020
Available online 04 June 2020
0001-706X/ © 2020 Published by Elsevier B.V.
G.M. Corral-Ruiz and L.E. Sánchez-Torres Acta Tropica 210 (2020) 105548

the host and the parasite has favored the chronicity of the infections. In For this purpose, several studies have been carried out in which the
most of the cases, the infection becomes asymptomatic and a great immune modulation and even the suppressive capacity of F. hepatica are
parasitic load is required for the development of the clinical entity evaluated against the development of autoimmune diseases. A series of
(Maizels et al., 2014; Wammes et al., 2014; Yazdanbakhsh et al., 2002). different immunological mechanisms that reduce the pathogenic effect
It means that during the asymptomatic phase, the patient does not ask and, in some cases, even totally prevent the development of the disease
for medical advice if available, and the parasites remain for a long time. have been identified, and strategies like infection of animals with F.
Interestingly, there is an association between the high prevalence of hepatica or the administration of the diverse molecules from this hel-
helminthiases in certain populations, with the low prevalence of im- minth are been used (Walsh et al., 2009; Lund et al., 2016). The mo-
mune disorders associated with inflammation like autoimmunities or lecules with the most interesting immunomodulatory capacity de-
allergies (Maizels et al., 2014; Wammes et al., 2014; scribed are tegument-derived, molecules present in F. hepatica derived-
Yazdanbakhsh et al., 2002). Additionally, the spontaneous amelioration extracellular vesicles (EVs) or those released as secretory/excretory
of patients with autoimmune pathology when naturally infected with (ES) products. In some cases, the precise signaling pathways employed
one helminth has been documented (Correale and Farez, 2011). by them are still under extensive and deeper analysis.
The host interaction with these parasites is globally considered as a The tegument consists of a syncytial layer of approximately 15 μm
not reciprocal metabolic dependent relationship, where only the para- of thickness, which covers the entire soma of the parasite. The anato-
site results benefited. More recently, it has been pointed out that, for a mical architecture of this organ is formed by a plasma membrane that
successful relationship to be developed, this must be on the edge of serves as a support for the outermost area composed of the glycocalyx.
commensalism, where harm is not generated to the host and the con- Below the plasma membrane, there is a rich mitochondria region in-
tinued transmission of the parasite is guaranteed. However, in certain cluded in an interstitial region followed by a basement membrane that
cases, this type of interaction approaches mutualism, benefiting also the is connected through channels; these structures allow the passage of the
host, all this as a process of adaptation resulting from a natural selec- components needed for the replacement of the tegument, which takes
tion (Jouvin and Kinet, 2012; Navarro et al., 2013). place every 2 to 3 h during the infection (Hacarız et al., 2012;
Hamilton et al., 2009; Wilson et al., 2011). This structure constitutes
2. Fasciola hepatica the interface where the interaction and communication between the
host and the parasite take place. The tegument exerts vital functions in
Fascioliasis is a zoonotic disease caused by the liver fluke F. hepa- the development of F. hepatica that implies the maintenance of a
tica, which is considered as one of the most successful parasites due to homeostatic state that involves nutrient absorption, waste excretion,
its wide distribution. In comparison with other helminths, it possesses and sensory functions. Of utmost importance is the interaction of the
the capacity to infect the largest variety of hosts, but mammals behave tegument with the immune system due to the antigenic complexity and
as definitive hosts (Dalton et al., 2013). They become parasitized by the the ability of several molecules to modulate the cells of the immune
ingestion of plants contaminated with the infective phase, the meta- system. This capacity is considered as a parasite defense mechanism; it
cercaria. Once ingested, the juvenile phase hatches in the duodenum, has been shown that an enormous amount of molecules are released
migrates through the intestinal epithelium into the peritoneal cavity constantly through the tegument and its vesicular content, favoring a
until it reaches the liver, where it continues migrating through the continuous interaction with the immune cells of the host (Hacarız et al.,
parenchyma; finally, the parasite arrives at the bile ducts where it 2012; Alba et al., 2016; Anuracpreeda et al., 2006).
achieves its final stage of maturation and the eggs are released The great complexity of the tegument, composed by at least 369
(Robinson et al., 2013; Rokni, 2014). It has been estimated that ap- proteins, has made difficult to characterize its physiological and im-
proximately 180 million people are at risk of being infected and 17 mune regulatory functions; additionally, the presence of abundant N-
million people are already infected with this helminth (Cwiklinski et al., glycosylated proteins and glycolipids that constitute the glycocalyx of
2016). the tegument makes more complex the composition of this structure
F. hepatica has one of the largest genomes sequenced within pa- that coats the parasite (Ravida et al., 2016; Ravida et al., 2016).
thogenic organisms, implying a great capacity of synthesis of a vast F. hepatica releases constantly EVs. These vesicles allow the dialog
repertoire of proteins that support the complex life cycle and the great between the parasite and the host by the exportation of proteins and
adaptation it possesses (Rasouli et al., 2019; Robinson et al., 2009). microRNAs (miRNAs). They actively participate in the parasites´ life
This feature also allows this helminth to have a wide range of definitive cycle, pathogenicity, and modulating the immune response towards the
hosts; this adaptive ability is magnified, since, as mentioned above, development of tolerance to the parasite and non-related antigens. The
within its definitive host it must modify itself, migrate and respond to content of the EVs can be internalized by the host cells, and once inside,
very different physiological environments. An example of this capacity they deliver the cargo molecules directly. It was demonstrated that the
it is noted when the adult phase lodges inside the bile ducts changing its vesicle content has a distinctive role in ensuring the parasite infection
metabolism from an aerobic to an anaerobic one (Cwiklinski et al., since they possess 52% of the until now identified proteins contained in
2015). Additionally, the ability to establish successful infections is re- the parasite secretome (Escudero et al., 2019; Marcilla et al., 2012). As
lated to the great diversity of molecules with immune regulatory mentioned, these vesicles contain miRNAs, molecules with repressing
properties that it possesses making chronicity one of the consequences gene expression capacity. It has been shown that the miRNAs profile
of the regulated milieu induced by the parasite. In this review, some of detected in the vesicles is different when released from the adult or
these molecules needed to ensure the establishment of F. hepatica juvenile parasite, leading to a different outcome in the receiving cell.
within the host are described depending on their location and functions. Data support the hypothesis that miRNAs are the mediators of the
The capacity of some of these molecules to direct or re-direct the im- previously demonstrated immune modulatory function of the EVs
mune response of the host to a regulatory profile has been postulated (Fromm et al., 2017; Fromm et al., 2015; Vidigal and Ventura, 2015).
and exploited to modify the course of diseases both infectious and of This mechanism highlights the role played by the miRNAs in the
immunological origin, with a potent inflammatory and pathogenic parasite adaptation and the parasite modulation exerted in the host.
background (McSorley and Maizels, 2012; Dalton et al., 2013; Another mechanism used by helminths to survive, migrate, obtain
Flynn et al., 2007). The helminth-derived products that induce im- nutrients and, to evade the immune response of the host, is the release
munological environments that prevent or reverts these inflammatory of the ES proteins (Cancela et al., 2008). The exact composition of this
states are an interesting starting point for the discovery, characteriza- complex mixture of molecules depends on the helminth's biological
tion, and generation of new prophylactic and/or therapeutic im- phase. It includes actively secreted components and those released as
munological strategies. metabolic by-products that comprise proteins, glycoproteins,

2
G.M. Corral-Ruiz and L.E. Sánchez-Torres Acta Tropica 210 (2020) 105548

polysaccharides, glycolipids, etc. Most of them are also delivered via has been seen that the promotion of this phenotype could be induced by
the EVs as mention above. In the case of F. hepatica ES molecules the suppression of the MAPK pathway and the increase of the inhibitory
(FhESM), these components are released by the oral and ventral protein SOCS3 which down-regulate TLR4 signaling (Hamilton et al.,
suckers, as well as through the tegument (Jefferies et al., 2001). Ro- 2009; Flynn et al., 2010; Vukman et al., 2013; Vukman et al., 2013).
binson et al. described 160 different proteins in the secretions of the Likewise, it has been described that the components of the tegument
adult phase, including proteases such as cathepsins B and L (CB and also induce the suppression of the NFκ-B and MAPK pathways as well as
CL), leucine aminopeptidase and carboxypeptidase, along with the fatty the increase of SOCS3 expression in mast cells (Vukman et al., 2013).
acid-binding protein (FABP) and the F. hepatica saposin-like protein It has been shown that the 12 kDa FABP isoform (FABP1), a mole-
(FhSAP), all of them necessary for its metabolism (Robinson et al., cule found in the tegument, can interact with the co-receptors CD14
2009). Additionally, antioxidant enzymes, which efficiently protect the and MD2, preventing the formation of the TLR4-LPS complex; in a
parasite from reactive oxygen species released by eosinophils, such as sepsis model, the administration of FABP1 provoked a decrease in IFN-
the superoxide dismutase (SOD), glutathione-S-transferase (GST), γ, TNF-α, GM-CSF, IL-12p70, IL-3 and IL-15 in serum (Martin et al.,
thioredoxin peroxidase and peroxiredoxin are also molecules that are 2015; Santiago and Espino, 2014). Benitez et al. showed that both
part of the FhESM [20,21,25,37,38]. The most abundant components isoforms of this protein (12 kDa and 15kDA) suppress TLR-stimulation
are the cysteine proteases FhCB and FhCL, two major virulence factors by multiple bacterial ligands, like whole extracts from Enterococcus
of the parasite. They participate in diverse processes such as the excyst faecalis, Klebsiella pneumoniae or heat-killed Listeria monocytogenes
of the metacercariae, tissue invasion, migration feeding, tegument (Benítez et al., 2017). The 15kDA isoform (Fh15) was also tested in a
shedding, and immune system evasion (Cancela et al., 2008; lethal sepsis model where it averted the serum concentration increase of
Deniziak et al., 2013; McGonigle et al., 2008; McNulty et al., 2017). The different cytokines and chemokines such as IL-1β, TNF-α, IFN-γ, IL-6 IL-
shared protein content between tegument, EVs, and FhESM, reflects the 12p70, MCP-1, MIP1α and, MIP1β; all these molecules related to the
importance of these molecular mediators in the constant host-parasite activation of endothelial cells, macrophages and other leucocytes
interaction (Fromm et al., 2015). (Benitez et al., 2018).
One of the fields not so widely studied during the infection with F. The glycoproteins and glycolipids that constitute the glycocalyx of
hepatica is the role played by eggs and their molecules, as well as the the tegument possess a mixture of oligosaccharide motifs that can in-
parasite's microbiota. One important aspect that should not be forgotten teract with innate immune receptors, like C-type lectins receptors.
is the fact that helminths are usually very prolific, characteristic that Ravida et al. reported that the predominant glycoproteins contain oli-
confers them a great evolutionary advantage by being able to release a gomannose oligosaccharides (Ravida et al., 2016). Aldridge and O'Neill
huge quantity of fertile eggs per day. It is estimated that a single adult showed that after incubating DCs with antigens derived from the te-
of F. hepatica releases around 50,000 eggs per day. This represents a gument, the expression of the mannose receptor enhanced. When DCs
great antigenic challenge for the host. If the infection becomes chronic, were then co-cultured with T lymphocytes, their interaction caused the
this condition remains for a long time since this parasite can survive for latter to become anergic. The anergic phenotype was established by the
more than a decade in the host (Moxon et al., 2010). expression of Grail, Ctla-4 and PD-1, reduced cell proliferation and the
The tegument-, the EVs-, the FhESM- and the eggs-derived mole- lack of cytokine production (IL-2, IL-4, IL-5, and IFN-γ). They also ob-
cules represent a complex antigenic mosaic that participates in diverse served a similar effect in F. hepatica-infected mice and animals injected
physiological functions during the biological cycle of the parasite. with tegumental antigens (Aldridge and O'Neill, 2016). In the same
Besides, some of these molecules are also responsible for the main- line, Rodriguez et al. showed that LPS-mature monocytes-derived DCs
tenance of the chronicity of the infection. The diverse and interesting (mo-DCs) pulsed with a tegument extract, polarized T cells to a Th2
immunomodulatory effects exerted by the parasite detailed below, profile since these cells produced IL-4; the interaction between mo-DCs
favor the last point. and the tegument was given by the recognition of the Tn antigen
(αGalNAc-Thr/Ser) through the Macrophage Gal/GalNAc lectin (MGL)
3. Molecular mechanisms of immunoregulation of F. hepatica receptor. Besides, purified CD11b+ MGL2+ cells from the peritoneal
cavity of F. hepatica infected mice acquired common markers associated
To understand the mechanisms through which F. hepatica and its with a type 2 macrophages (M2) phenotype and expressed inhibitory
antigens exert their immunomodulatory effects previously mentioned, molecules such as Programmed Death-Ligand (PD-L) 1 and PD-L2.
it is of utmost importance the knowledge of the communication es- Moreover, they showed that mo-DCs were able to recognize mannosy-
tablished between the parasite and the host immune system since the lated glycoconjugates through CD209 (or DC-SIGN), favoring anergic
beginning. Their first interaction is through the innate immune system and/or Tregs cells that silence the immune system of the host
cells, especially with antigen-presenting cells (APC) like dendritic cells (Rodríguez et al., 2017a,b).
(DCs) and macrophages. Despite the lack of knowledge about the exact The peroxiredoxin and GST released by the parasite through EVs
mechanisms by which helminths can modulate the immune response, into the bloodstream are capable to induce the expression of Ym-1 in
the role played by this branch of the immune system is decisive in the splenic macrophages which correlates with their low activation medi-
establishment of chronic infection. The consequences of the initial ated by TLRs. The induction of M2 macrophages and splenic DCs al-
dialog established, impact on the mechanisms exerted by the adaptive tered the expression levels of these receptors, generating an environ-
branch of the immune response, also involved in the elimination/ ment that promoted the survival of the parasite and the establishment
clearance of the parasite. The direct or the APC-mediated effect of the of chronic infection (Cwiklinski et al., 2015; Donnelly et al., 2008;
molecules of the parasite on T and B lymphocytes determines the cy- Flynn and Mulcahy, 2008).
tokine milieu and the spectrum of antibodies produced. The immunosuppression provoked by the interaction of the tegu-
Hamilton et al. observed that in bone marrow-derived DCs cultured mental and EVs antigens is reinforced by those contained in the FhESM.
with tegument derived extract, the maturation, and function of these Falcón et al. reported that murine bone marrow-derived DCs pulsed
cells were suppressed; additionally, their phagocytic activity was in- with FhESM did not induce the maturation of these cells, as they were
hibited and the expression of co-stimulation molecules (such as CD80, unable to produce cytokines. Additionally, the FhESM impaired the
CD86, and CD40) was decreased as well as the synthesis of in- ability of the DCs to be activated by TLR ligands as LPS, poly I:C, and to
flammatory cytokines (like IL-6, IL-12p70, and TNF-α) when stimulated stimulate allospecific responses (Falcón et al., 2010). In 2018, Guasconi
with peptidoglycan and lipopolysaccharide (LPS). Therefore, these DCs et al. showed that molecules contained in the FhESM are sensed by the
were unable to activate T lymphocytes, favoring the chronicity of the Dectin-1 receptor. The signaling pathway activated induced increased
infection. The exact mechanisms are not completely known; however, it arginase activity, as well as IL-10 and TGF-β production in BALB/c

3
G.M. Corral-Ruiz and L.E. Sánchez-Torres Acta Tropica 210 (2020) 105548

macrophages in vitro (Guasconi et al., 2018). clinical symptoms in an EAE model when animals were infected with F.
A protein that is part of the FhESM, is the F. hepatica helminthic hepatica; the positive effects were correlated with changes in innate
defense molecule (FhHDM-1). FhHDM-1 is a protein conserved amongst cells such as DCs and macrophages. In the case of the DCs, they were
trematodes; it has biochemical properties similar to those of the able to produce IL-10 and a decreased expression of co-stimulatory
mammal antimicrobial cathelicidin. This molecule when released needs molecules, while macrophages had a low expression of MHC-II and
to be proteolytically processed by FhCL, originating a 34 amino acid produced TGF-β; this study also showed that IL-10 was not necessary
residue protein fragment that possesses an amphipathic helix region for this outcome, since IL-10 KO mice infected with F. hepatica, also
responsible for its biological activity; it binds to the LPS preventing its showed a decrease in EAE symptoms and severity (Walsh et al., 2009).
recognition by macrophages and DCs. The lack of signaling avoids cell In the same experimental model, Finlay et al. found that the FhESM
activation and the release of inflammatory mediators. This situation avoid the infiltration of Th1, Th17 (CD4+ IFN-γ+ and IL-17+), and
allows the newly excysted juvenile to penetrate through the lamina neutrophils to the brain; they also reported that the FhESM directed the
propia of the intestine. FhHDM-1 is expressed in all the biological immune response towards a Th2 profile evidenced by infiltrating T cells
phases of F. hepatica inside its definitive host; it is not only able to limit (CD4+ IL-4+ and IL-13+) and eosinophils in the brain, peritoneal
the inflammation originated by the microbiota, but also interfere with cavity, blood, and spleen (Finlay et al., 2016). Quinn et al. confirmed
the innate response towards other bacterial entities that might be co- the capacity of F. hepatica to attenuate the clinical symptoms in the EAE
infecting. This protein has a major immunologic relevance because it model. They showed that the innate immune cells are susceptible to
can suppress the ability of macrophages to process and present antigens immune training through an anti-inflammatory phenotype when ex-
by avoiding the acidification of the lysosomes (Robinson et al., 2011). posed to a total F. hepatica extract, both in vitro and in vivo systems
The inhibition of the lysosomal V-ATPase, necessary for the H+ efflux (Quinn et al., 2019).
into the lysosome, impair the activation of the CB of the host within the Similarly, Lund et al. using a T1D model, prevented the develop-
lysosomal lumen (Robinson et al., 2012). Additionally, FhHDM-1 pre- ment of the disease in most of the animals administered with FhESM.
vents the activation of the NLRP3 inflammasome and the synthesis of They identified in the pancreatic lymph node, the participation of TGF-
IL-1β in response to particulate activators of the inflammasome β and IL-10 secreting M2 macrophages and IL-10 secreting Breg lym-
(Alvarado et al., 2016). In this context, Alvarado et al., have proposed phocytes; both in synergy inhibited the activation and secretion of IFN-
the use of FhHDM as a potential therapeutic option in autoimmune γ from autoreactive T cells and favored the decrease of the insulitis in
pathologies (Alvarado, 2014). NOD mice (Lund et al., 2014).
On the other hand, FhCL, when internalized by macrophages, is FhHDM-1 has been widely used in different immune-mediated dis-
transported to the endolysosomal compartments, where it resists the ease models because of its anti-inflammatory properties; recent studies
degradation by host proteases. Once inside, FhCL degrades TLR3, pre- showed that the administration of this protein in different murine
venting it from recognizing other antigens and inhibiting the down- models of inflammation, like T1D, EAE, asthma and, rheumatoid ar-
stream signaling cascade of this receptor. This event impairs cell acti- thritis, induced a reduction in the severity of those diseases (Lund et al.,
vation and the consequent synthesis of proinflammatory cytokines such 2016; Tanaka et al., 2018). In both cases, T1D and EAE, this protein
as IFN-γ, favoring an immunological switch to a Th2 profile increased the survival rate and reduced the percent of NOD mice with
(Donnelly et al., 2010). Prowse et al. described that FhCL can also de- insulitis or clinical symptoms, inflammation, and demyelination in
grade the CD4 co-receptor, avoiding an adequate antigenic presentation Central Nervous System respectively (Lund et al., 2016). Besides, Ta-
and allowing the generation of anergic T lymphocytes (Prowse et al., naka et al. showed that, in an asthma model, this protein reduced the
2002). numbers of neutrophils, lymphocytes, and eosinophils in bronch-
The immune response directed towards the eggs or the antigens oalveolar lavage fluid as well as the inflammation and mucus produc-
released by them initiates an anti-inflammatory response orchestrated tion in the lung (Tanaka et al., 2018). Finally, in a murine model of
by IL-10 and TGF-β, which can induce the activation of Treg lympho- rheumatoid arthritis induced by collagen type II, Khan et al. evidenced
cytes. This response allows the eggs to be protected during the passage that this F. hepatica-derived molecule protects against cartilage loss and
from and through the bile ducts to the intestine, and finally being re- the architecture destruction. These improvements could be in vitro-as-
leased through the feces without interrupting the biological cycle. sociated to the maintenance of the active function of the molecular
Within the molecules secreted by the eggs of F. hepatica, Moxon et al. player Sequestosome 1 which is a negative regulator of the NF- κB
demonstrated the presence of proteases like the leucine aminopepti- (Khan et al., 2019).
dase, suggesting that the activity of this protease favors the passage Likewise, it has been established that EVs can have a therapeutic
through the digestive system and later, in the appropriate conditions, potential relying on his contain proving its immunomodulatory capa-
allows the hatching of the eggs with the release of the miracidium. city. Roig et al. showed that previous treatment with F. hepatica derived
Nonetheless, there is still no information about the mechanisms by EVs mitigate the disease activity index in a DSS-induced colitis model,
which these proteins are released by the egg (Moxon et al., 2010). hampering the production of pro-inflammatory cytokines such as IL-6,
In 2015, Japa et al. demonstrated that F. hepatica possesses a family IL-17, and, TNF-α. The molecules responsible for this activity are still
of TGF-like molecules (TLM). One of them, named FhTLM is highly under investigation (Roig et al., 2018).
expressed in newly excysted juveniles, and unembryonated egg, and it Despite the encouraging results obtained with F. hepatica derived
was established that is necessary for the parasite development molecules to modulate inflammatory processes, it must take into ac-
(Japa et al., 2015). Later, the same group showed that FhTLM influ- count that the induced regulatory state also affects the immune re-
ences the immune response of the host as this molecule signals through sponses to other antigens. It has been reported an increased suscept-
the Smad2/3 pathway. The presence of FhTLM induces the differ- ibility to suffering diseases whose protective inflammatory response is
entiation of the monocyte-derived macrophages to a regulatory phe- based on a Th1 profile (Lucena et al., 2017). An example of this was
notype with the production of IL-10, arginase-1, the mannose receptor, shown by O'Neill et al., when mice co-infected with Bordetella pertussis
and PD-L1 (Sulaiman et al., 2016). and F. hepatica or infected and later inoculated with FhESM, showed a
The mentioned mechanisms above are illustrated in Fig. 1. decreased production of IFN-γ and a delayed resolution of the bacterial
The information obtained from in vitro experiments has been ad- infection (O'Neill et al., 2001). Additionally, Brady et al. showed that in
dressed in several in vivo models of inflammation as a proof of concept. the infection with F. hepatica in immunized mice with whole-cell B.
The most employed are murine models of asthma, sepsis, type 1 dia- pertussis vaccine (Pw), the production of IFN-γ was inhibited when
betes (D1T), and experimental autoimmune encephalomyelitis (EAE). spleen cells were stimulated with Pw (Brady et al., 1999). Furthermore,
Walsh et al. showed a delay and decrease in the severity of the Fasciola infection or only the FhESM administration has been proved to

4
G.M. Corral-Ruiz and L.E. Sánchez-Torres Acta Tropica 210 (2020) 105548

Fig. 1. Immune modulation directed by Fasciola hepatica. a) F. hepatica infection induces TGF-β secretion by Tregs avoiding the development of EAE. b) During
chronic infections, eggs are laid, promoting IL-10 and TGF- β secretion and thus inducing Tregs activation. c) Tegumental antigens, composed by glycoproteins, lead
to an enhanced CD206 expression in DCs which promotes lymphocyte anergy after being co-cultivated; furthermore, tegument antigens decrease co-stimulatory
molecule expression ensuring DC hyporesponsiveness. d) Microvesicles containing miRNAs and ES products can suppress DSS-induced colitis by diminishing pro-
inflammatory cytokines that mediate the pathology. e) ES products, composed by a diversity of proteins, favor the activation of TGF- β and IL-10 producers cells, such
as M2 and Bregs avoiding the development of T1D; this antigen mixture also induces Th2 polarization in lymphocytes due to type 2 cytokines, preventing EAE.
Proteins such as FhTLM enhance expression of Arg-1, PD-L1, CD206 and induces IL-10 secretion by M2. FABP bind with MD2 or CD14 preventing TLR4 activation
induced by LPS, suppressing proinflammatory cytokines. FhCL1 degrades CD4 and TLR3 promoting lymphocyte anergy and avoids macrophage activation. FhHDM-1
binds to LPS counteracting macrophage and DC activation and preventing T1D and EAE development. GST and PRX induce YM-1 expression in macrophages and
alter the levels of expression of TLR's in DCs. f) The role played by the microbiota remains unknown, but it cannot be underestimated. M2 Alternative activated
macrophages, Bregs Regulatory B cells, DCs Dendritic cells, EAE Experimental autoimmune encephalomyelitis, ES Excretory-secretory, FABP Fatty acid-binding
protein, FhCL1 F. hepatica cathepsin L, FhHDM-1 F. hepatica defense molecule, GST Glutathione-S-transferase, PBMC Peripheral blood mononuclear cell, PRX
Peroxiredoxin, T1D Type 1 diabetes, Treg Regulatory T cell.

induce apoptosis in peritoneal eosinophils, macrophages, and blood Finally, the study of the host-parasite interaction must be complete
mononuclear cells thus inhibiting the inflammatory functions of these and integrative to determine all the factors that interfere in the result of
cells (Guasconi et al., 2012; Serradell et al., 2007; Serradell et al., 2009; the infection; so, the study of the composition of the trematode mi-
Chen et al., 2019). crobiota and the role it plays should be studied in detail shortly.
Reinforcing the aforementioned suppressive activity, Flynn et al. McNulty et al. described the presence of the endobacteria Neocrickettsia
reported that the infection with F. hepatica affected the predictive ca- in the adult parasite and mature eggs, which's localization covers the
pacity of two diagnoses tests of bovine tuberculosis (Flynn et al., 2007), reproductive tissues and to a lesser extent the somatic ones. They de-
nonetheless, future studies regarding these adverse events are needed as monstrated a vertical transmission towards the offspring of the parasite,
this type of interaction acquires public importance considering the high as well as a horizontal transmission towards the host, since the en-
risk of co-infection due to the co-existence of F. hepatica with other dobacteria are found both in the oral suckers as in the tegument and, by
infectious agents like viruses, bacteria, and even other parasites and the generating host tissue damage, it allows the entry of this microorganism
negative impact on vaccination (Lucena et al., 2017; Howell et al., to the host, conferring to F. hepatica the role of a vector. It is still un-
2018). known in which terms the relationship between the trematode and this

5
G.M. Corral-Ruiz and L.E. Sánchez-Torres Acta Tropica 210 (2020) 105548

symbiont stands, but it is suggested that the latter synthesize vitamins Anuracpreeda, P., Wanichanon, C., Chaithirayanon, K., Preyavichyapugdee, N., Sobhon,
and other nutrients that may be beneficial for the parasite P., 2006. Distribution of 28.5kDa antigen in the tegument of adult Fasciola gigantica.
Acta Trop. 100, 31–40.
(McNulty et al., 2017). Similarly, the role that it plays within the im- Baska, P., Norbury, L.J., Deniziak, A.Z., Wiśniewski, M., Januszkiewicz, K., 2017.
mune system is not known, but it should not surprising if it had an Excretory/secretory products from two Fasciola hepatica isolates induce different
important role in the induced response, like in the case of Wolbachia transcriptional changes and IL-10 release in LPS-activated bovine “BOMA” macro-
phages. Parasitol. Res. 116, 2775–2782.
and Onchocerca volvulus (Taylor et al., 2001). Other important aspects Benítez, M.J.R., Jiménez, C.R., Aguayo, V., Espino, A.M, 2017. Recombinant Fasciola
that should be considered are the existence of a co-infection and the hepatica fatty acid binding protein suppresses toll-like receptor stimulation in re-
type of pathogen co-infecting, the specific composition of the micro- sponse to multiple bacterial ligands. Nat. Sci. Rep. 7, 5455.
Benitez, M.J.R., Jimenez, C.R., Franco, J.J.R., Pérez, W.D.R., Mendez, L.B., Osuna, A.,
biota, and the type of mammal in which the adult is housed. Since F. Espino, A.M., 2018. Fh15 blocks the lipopolysaccharide-induced cytokine storm
hepatica possesses a large number of definitive hosts, the interaction while modulating peritoneal macrophage migration and CD38 expression within
that takes place in a llama or a mouse will not be necessary the same spleen macrophages in a mouse model of septic shock. mSphere 3 e00548-18.
Brady, M.T., O'Neill, S.M., Dalton, J.P., Mill, K.G.H., 1999. Fasciola hepatica suppresses a
(Baska et al., 2017; Piedrafita et al., 2010).
protective Th1 response against Bordetella pertussis. Infect. Immun. 62, 5372–5378.
Cancela, M., Acosta, D., Rinaldi, G., Silva, E., Durán, R., Roche, L., Zaha, A., Carmona, C.,
4. Concluding remarks Tort, J.F., 2008. A distinctive repertoire of cathepsins is expressed by juvenile in-
vasive Fasciola hepatica. Biochimie 90, 1461–1475.
Cattadori, I.M., Sebastian, A., Hao H Katani, R., Albert, I., Eilertson, K.E., Kapur, V.,
The secretome of all the helminth parasites contains a great quantity Pathak, A., Mitchell, S., 2016. Impact of helminth infections and nutritional con-
and diversity of functional products; the immunomodulatory capacity straints on the small intestine microbiota. PLoS ONE. https://doi.org/10.1371/
of helminth-derived molecules is a common characteristic, and it can journal.pone.0159770.
Chen, D., Tian, A.L., Hou, J.L., Li, J.X., Tian, X., Yuan, X.D., Li, X., Elsheikha, H.M., Zhu,
play for or against the host as mention before. As the nematodes, ces- X.-.Q., 2019. The multitasking Fasciola gigantica cathepsin B interferes with various
todes, and others trematodes, F. hepatica possesses a broad antigenic functions of goat peripheral blood mononuclear cells in vitro. Front. Immunol. 10,
spectrum that activates various signaling pathways, most of them fo- 1707.
Correale, J., Farez, M.F., 2011. The impact of parasite infections on the course of multiple
cused on the activation of redundant mechanisms that modulate the sclerosis. J. Neuroimmunol. 233, 6–11.
host´s inflammatory immune response (Quinn et al., 2019; Ditgen et al., Cwiklinski, K., Dalton, J.P., Dufresne, P.J., La Course, J., Williams, D.J.L., Hodgkinson, J.,
2014; Kahl et al., 2018). This elicited response allows F. hepatica to Paterson, S., 2015. The Fasciola hepatica genome: gene duplication and polymorphism
reveals adaptation to the host environment and the capacity for rapid evolution.
survive and to persist for a long time inside the host; besides, it avoids Genome Biol. https://doi.org/10.1186/s13059-015-0632-2.
bystander damage to the host, since the parasite directs a non-in- Cwiklinski, K., O'Neill, S.M., Donnelly, S., Dalton, J.P., 2016. A prospective view of an-
flammatory-regulatory environment. The consequences of the mole- imal and human Fasciolosis. Parasite Immunol. 38, 558–568.
Dalton, J.P., Robinson, M.W., Mulcahy, G., O'Neill, S.M., Donnelly, S., 2013.
cular communication established between the F. hepatica, as well as
Immunomodulatory molecules of Fasciola hepatica: candidates for both vaccine and
other helminths and its hosts, have opened the opportunity to identify immunotherapeutic development. Vet. Parasitol. 195, 272–285.
new pharmacological entities to counteract inflammation-related Deniziak, A.Z., Wasyl, K., Norbury, L.J., Wesołowska, A., Bién, J., Grodzik, M.,
pathologies, like autoimmunity and allergies. Based on the information Wísniewski, M., Baska, P., Wedrychowicz, H., 2013. Characterization and differential
expression of cathepsin L3 alleles from Fasciola hepatica. Mol. Biochem. Parasitol.
described above, the products of F. hepatica are a source of molecules 190, 27–37.
with modulatory capacities that can be considered directly or as a Ditgen, D., Anandarajah, E.M., Meissner, K.A., Brattig, N., Wrenger, C., Liebau, E., 2014.
template for the development of new drugs. Detailed information about Harnessing the helminth secretome for therapeutic immunomodulators. BioMed Res.
Int doi.org/10.1155/2014/964350.
the exact mechanisms elicited is still needed. Donnelly, S., Stack, C.M., O'Neill, S.M., Sayed, A.A., Williams, D.L., Dalton, J.P., 2008.
We apologize to authors whose work has not been cited due to space Helminth 2-Cys peroxiredoxin drives Th2 responses through a mechanism involving
limitations. alternatively activated macrophages. FASEB J. 22, 4022–4032.
Donnelly, S., O'Neill, S.M., Stack, C.M., Robinson, M.W., Turnbull, L., Whitchurch, C.,
Dalton, J.P., 2010. Helminth cysteine proteases inhibit TRIF-dependent activation of
Declaration of Competing Interest macrophages via degradation of TLR3. J. Biol. Chem. 285, 3383–3392.
Escudero, T.E., Gerlach, J.Q., Bennett, A.P.S., Cwiklinski, K., Jewhurst, L., Huson, K.M.,
Joshi, L., Kilcoyne, M., O´Neill, S., Dalton, J.P., Robinson, M.W., 2019. Surface mo-
The authors declare that they have no known competing financial lecules of extracellular vesicles secreted by the helminth pathogen Fasciola hepatica
interests or personal relationships that could have appeared to influ- direct their internalization by host cells. PLoS Negl. Trop. Dis. 13 (1), e0007087.
Falcón, C., Carranza, F., Martínez, F.F., Knubel, C.P., Masih, D.T., Motrán, C.C., Cervi, L.,
ence the work reported in this paper.
2010. Excretory-secretory products (ESP) from Fasciola hepatica induce tolerogenic
properties in myeloid dendritic cells. Vet. Immunol. Immunopathol. 137, 36–46.
Acknowledgments Finlay, C.M., Stefanska, A.M., Walsh, K.P., Kelly, P.J., Boon, L., Lavelle, E.C., Walsh, P.T.,
Mills, K.H.G., 2016. Helminth products protect against autoimmunity via innate type
2 cytokines il-5 and il-3, which promote eosinophilia. J. Immunol. 196, 703–714.
GMC received a fellowship from the Consejo Nacional de Ciencia y Flynn, R.J., Mulcahy, G., 2008. Possible role for toll-like receptors in interaction of
Tecnología (CONACyT) and from BEIFI. LES is fellow of the Comisión Fasciola hepatica excretory/secretory products with bovine macrophages. Infect.
de Operación y Fomento de Actividades Académicas–IPN (COFAA-IPN) Immun. 2, 678–684.
Flynn, R.J., Irwin, J.A., Olivier, M., Sekiya, M., Dalton, J.P., Mulcahy, G., 2007a.
and fellow of the Estímulo al Desempeño de los Investigadores–IPN Alternative activation of ruminant macrophages by Fasciola hepatica. Vet. Immunol.
(EDI-IPN). The authors thank MSc Rodolfo Soria-Castro for the design Immunopathol. 120, 31–40.
of the figure. The figure was created with BioRender.com Flynn, R.J., Mannion, C., Golden, O., Hacariz, O., Mulcahy, G., 2007b. Experimental
Fasciola hepatica infection alters responses to tests used for diagnosis of bovine tu-
berculosis. Infect. Immun. 75, 1373–1381.
References Flynn, R.J., Mulcahy, G., Elsheikha, H.M., 2010. Coordinating innate and adaptive im-
munity in Fasciola hepatica infection: implications for control. Vet. Parasitol. 169,
235–240.
Alba, A., Sánchez, J., Hernández, H., Mosqueda, M., Rodríguez, S.Y., Capó, V., Otero, O.,
Fromm, B., Trelis, M., Hackenberg, M., Cantalapiedra, F., Bernal, D., Marcilla, A, 2015.
Alfonso, C., Marcet, R., Sarracent, J., 2016. Insights into the biological features of the
The revised microRNA complement of Fasciola hepatica reveals a plethora of over-
antigenic determinants recognized by four monoclonal antibodies in redia and adult
looked microRNAs and evidence for enrichment of immuno-regulatory microRNAs in
stages of the liver fluke Fasciola hepatica. Exp. Parasitol. 168, 39–44.
extracellular vesicles. Int. J. Parasitol. 45, 697–702.
Aldridge, A., O'Neill, S.M., 2016. Fasciola hepatica tegumental antigens induce anergic-
Fromm, B., Ovchinnikov, V., Høye, E., Bernal, D., Hackenberg, M., Marcilla, A., 2017. On
like T cells via dendritic cells in a mannose receptor-dependent manner. Eur. J.
the presence and immunoregulatory functions of extracellular microRNAs in the
Immunol. 46, 1180–1192.
trematode Fasciola hepatica. Parasite Immunol. 39. https://doi.org/10.1111/pim.
Alvarado, R., To, J., Lund, M., Pinar, A., Mansell, A., Robinson, M.W., O'Brien, B.A.,
12399.
Dalton, J.P., Donnelly, S., 2016. The immune modulatory peptide FhHDM-1 secreted
Guasconi, L., Serradell, M.C., Masih, D.T., 2012. Fasciola hepatica products induce
by the helminth Fasciola hepatica prevents NLRP3 inflammasome activation by in-
apoptosis of peritoneal macrophages. Vet. Immunol. Immunopathol. 148, 359–363.
hibiting endolysosomal acidification in macrophages. FASEB J. 31, 85–95.
Guasconi, L., Burstein, V.L., Beccacece, I., Mena, C., Chiapello, L.S., Masih, D.T., 2018.
Alvarado, R., 2014. Thesis Submitted for the Degree of Doctor of Philosophy in Science.
Dectin-1 on macrophages modulates the immune response to Fasciola hepatica pro-
University of Technology Sydney.
ducts through the ERK signaling pathway. Immunobiology 223, 834–838.

6
G.M. Corral-Ruiz and L.E. Sánchez-Torres Acta Tropica 210 (2020) 105548

Hacarız, O., Sayers, G., Baykal, A.T., 2012. A proteomic approach to investigate the attenuates the induction of T cell-mediated autoimmune disease. Front. Immunol. 10,
distribution and abundance of surface and internal Fasciola hepatica proteins during 1109.
the chronic stage of natural liver fluke infection in cattle. J. Proteome Res. 11, Rasouli, A., Farahnak, A., Zali, H., Rezaeian, M., Golestani, A., Molaei-Rad, M.B., 2019.
3592–3604. Protein detection of excretory-secretory products and somatic extracts from Fasciola
Hamilton, C.M., Dowling, D.J., Loscher, C.E., Morphew, R.M., Brophy, P.M., O'Neill, S.M., hepatica and F. gigantica using two-dimensional electrophoresis. Iran J. Parasitol. 14,
2009. The Fasciola hepatica tegumental antigen suppresses dendritic cell maturation 379–386.
and function. Infect. Immun. 77, 2488–2498. Ravida, A., Aldridge, A.M., Driessen, N.N., Heus, F.A., Cornelis, H.H., O´Neil, S, 2016a.
Howell, A.K., Tongue, S.C., Curriec, C., Evans, J., Williams, D.J.L., McNeillyc, T.N., 2018. Fasciola hepatica surface coat glycoproteins contain mannosylated and phosphory-
Co-infection with Fasciola hepatica may increase the risk of Escherichia coli O157 lated N-glycans and exhibit immune modulatory properties independent of the
shedding in British cattle destined for the food chain. Prev. Vet. Med. 150, 70–76. mannose receptor. PLoS Negl. Trop. Dis. 10 (4), e0004601.
Japa, O., Hodgkinson, J.E., Emes, R.D., Flynn, R.J., 2015. TGF-β superfamily members Ravida, A., Cwiklinski, K., Aldridge, A.M., Clarke, P., Thompson, R., Gerlach, J.Q.,
from the helminth Fasciola hepatica show intrinsic effects on viability and develop- Kilcoyne, M., Hokke, C.H., Dalton, J.P., O'Neill, S.M., 2016b. Fasciola hepatica surface
ment. Vet. Res. 46, 29. tegument: glycoproteins at the interface of parasite and host. Mol. Cell. Proteom. 15,
Jefferies, J.R., Campbell, A.M., van Rossum, A.J., Barrett, J., Brophy, P.M, 2001. 3139–3153.
Proteomic analysis of Fasciola hepatica excretory-secretory products. Proteomics 1, Riet, E., Hartgers, F., Yazdanbakhsh, M., 2007. Chronic helminth infections induce im-
1128–1132. munomodulation: consequences and mechanisms. Immunobiology 212, 475–490.
Jouvin, M.H., Kinet, J.P., 2012. Trichuris suis ova: testing a helminth-based therapy as an Robinson, M.W., Menon, R., Donnelly, S.M., Dalton, J.P., Ranganathan, S., 2009. An in-
extension of the hygiene hypothesis. J. Allergy Clin. Immunol. 130, 03–10. https:// tegrated transcriptomics and proteomics analysis of the secretome of the helminth
doi.org/10.1016/j.jaci.2012.05.028. pathogen Fasciola hepatica. Mol. Cell. Proteom. 8, 1891–1907.
Kahl, J., Brattig, N., Liebau, E., 2018. The untapped pharmacopeic potential of helminths. Robinson, M.W., Donnelly, S., Hutchinson, A.T., To, J., Taylor, N.L., Norton, R.S.,
Trends Parasitol. https://doi.org/10.1016/j.pt.2018.05.01. Perugini, M.A., Dalton, J.P., 2011. A family of helminth molecules that modulate
Khan, Y.A., Maurya, S.K., Kulkarni, C., Tiwari, M.C., Nagar, G.K., Chattopadhyay, N., innate cell responses via molecular mimicry of host antimicrobial peptides. PLoS
2019. Fasciola helminth defense molecule-1 protects against experimental arthritis by Pathog. https://doi.org/10.1371/journal.ppat.1002042.
inhibiting osteoclast formation and function without modulating the systemic im- Robinson, M.W., Alvarado, R., To, J., Hutchinson, A.T., Dowdell, S.N., Lund, M.,
mune response. FASEB J. https://doi.org/10.1096/fj.201901480RR. Turnbull, L., Whitchurch, C.B., O'Brien, B.A., Dalton, J.P., Donnelly, S., 2012. A
Lucena, A.N., Cuartero, L.G., Mulcahy, G., Zintl, A., 2017. The immunoregulatory effects helminth cathelicidin-like protein suppresses antigen processing and presentation in
of co-infection with Fasciola hepatica: from bovine tuberculosis to Johne's disease. macrophages via inhibition of lysosomal vATPase. FASEB J. 26.
Vet. J. 222, 9–16. Robinson, M.W., Dalton, J.P., O'Brien, B.A., Donnelly, S., 2013. Fasciola hepatica: the
Lund, M.E., O'Brien, B.A., Hutchinson, A.T., Robinson, M.W., Simpson, A.M., Dalton, J.P., therapeutic potential of a worm secretome. Int. J. Parasitol. 43, 283–291.
Donnelly, S., 2014. Secreted proteins from the helminth Fasciola hepatica inhibit the Rodríguez, E., Carasi, P., Frigerio, S., Costa, V., Vliet, S., Noya, V., Brossard, N., Kooyk, Y.,
initiation of autoreactive T cell responses and prevent diabetes in the NOD mouse. Vallejo, J.J., Freire, T., 2017a. Fasciola hepatica immune regulates CD11c+ cells by
PLoS ONE. https://doi.org/10.1371/journal.pone.0086289. interacting with the Macrophage Gal/GalNAc Lectin. Front. Immunol. 8. https://doi.
Lund, M.E., Greer, J., Dixit, A., Alvarado, R., McCauley-Winter, P., To, J., Tanaka, A., org/10.3389/fimmu.2017.00264.
Hutchinson, A.T., Robinson, M.W., Simpson, A.M., O'Brien, B.A., Dalton, J.P., Rodríguez, E., Kalay, H., Noya, V., Brossard, N., Giacomini, C., Kooyk, Y., Vallejo, J.J.,
Donnelly, S., 2016. A parasite-derived 68-mer peptide ameliorates autoimmune dis- Freire, T., 2017b. Fasciola hepatica glycoconjugates immune regulate dendritic cells
ease in murine models of Type 1 diabetes and multiple sclerosis. Nat. Sci. Rep. through the dendritic cell-specific intercellular adhesion molecule-3- grabbing non-
https://doi.org/10.1038/srep37789. integrin inducing T cell anergy. Nat. Sci. Rep. 7. https://doi.org/10.1038/srep46748.
Maizels, R.M., McSorley, H.J., Smyth, D.J., 2014. Helminths in the hygiene hypothesis: Roig, J., Saiz, M.L., Galiano, A., Trelis, M., Cantalapiedra, F., Monteagudo, C., Giner, E.,
sooner or later? Clin. Exp. Immunol. 177, 38–46. Giner, R.M., Recio, M.C., Bernal, D., Sánchez-Madrid, F., Marcilla, A., 2018.
Marcilla, A., Trelis, M., Cortés, A., Sotillo, J., Cantalapiedra, F., Mingue, M.T., Valero, Extracellular vesicles from the helminth Fasciola hepatica prevent DSS-induced acute
M.L., Sánchez del Pino, M.M., Antoli, C.M., Toledo, R., Bernal, D., 2012. Extracellular ulcerative colitis in a T-lymphocyte independent mode. Front. Microbiol. 9, 1036.
vesicles from parasitic helminths contain specific excretory/secretory proteins and Rokni, M.B., 2014. Fasciola hepatica and Fasciola gigantica. Encycl. Food Saf. (Reference
are internalized in intestinal host cells. PLoS ONE. https://doi.org/10.1371/journal. Module in Food Science) 2, 140–145.
pone.0045974. Santiago, O.F., Espino, A.M., 2014. Fasciola hepatica fatty acid binding protein induces the
Martin, I., Hernández, K.C., Santiago, O.F., Espino, A.M., 2015. Fasciola hepatica fatty acid alternative activation of human macrophages. Infect. Immun. 82, 5005–5012.
binding protein inhibits tlr4 activation and suppresses the inflammatory cytokines Serradell, M.C., Guasconi, L., Cervi, L., Chiapello, L.S., Masih, D.T., 2007. Excretory-se-
induced by lipopolysaccharide in vitro and in vivo. J. Immunol. 15, 3924–3936. cretory products from Fasciola hepatica induce eosinophil apoptosis by a caspase-
McGonigle, L., Mousley, A., Marks, N.J., Brennan, G.P., Dalton, J.P., Spithill, T.W., Day, dependent mechanism. Vet. Immunol. Immunopathol. 117, 197–208.
T.A., Maule, A.G., 2008. The silencing of cysteine proteases in Fasciola hepatica newly Serradell, M.C., Guasconi, L., Masih, D.T., 2009. Involvement of a mitochondrial pathway
excysted juveniles using RNA interference reduces gut penetration. Int. J. Parasitol. and key role of hydrogen peroxide during eosinophil apoptosis induced by ex-
38, 149–155. cretory–secretory products from Fasciola hepatica. Mol. Biochem. Parasitol. 163,
McNulty, S.N., Tort, J.F., Rinaldi, G., Fischer, K., Rosa, B.A., Smircich, P., Fontenla, S., 95–106.
Choi, Y.J., Tyagi, R., Pepin, K.H., Mann, V.H., Kammili, L., Latham, P.S., Dell'Oca, N., Sulaiman, A.A., Zolnierczyk, K., Japa, O., Owen, J.P., MaddisonBC, Emes RD,
Dominguez, F., Carmona, C., Fischer, P.U., Brindley, P.J., Mitreva, M., 2017. Hodgkinson, J.E., Gough, K.C., Flynn, R.J., 2016. A trematode parasite derived
Genomes of Fasciola hepatica from the Americas reveal colonization with Neorickettsia growth factor binds and exerts influences on host immune functions via host cytokine
endobacteria related to the agents of Potomac horse and human Sennetsu fevers. receptor complexes. PLoS Pathog. https://doi.org/10.1371/journal.ppat.1005991.
PLOS Genetics. https://doi.org/10.1371/journal.pgen.1006537. Tanaka, A., Allam, V.S.R., Simpson, J., Tiberti, N., Shiels, J., To, J., Lund, M., Combes, V.,
McSorley, H.J., Maizels, R.M., 2012. Helminth infections and host immune regulation. Weldon, S., Taggart, C., Dalton, J.P., Phipps, S., Sukkar, M.B., Donnelly, S., 2018. The
Clin. Microbiol. Rev. 25, 585–608. parasitic 68-mer peptide FhHDM-1 inhibits mixed granulocytic inflammation and
Meeusen, E.N.T., 1999. Immunology of helminth infections, with special reference to airway hyperreactivity in experimental asthma. J. Allergy Clin. Immunol. 141,
immunopathology. Vet. Parasitol. 84, 259–273. 2316–2319.
Moxon, J.V., Flynn, R.J., Golden, O., Hamilton, J.V., Mulcahy, G., Brophy, P.M., 2010. Taylor, M.J., Cross, H.F., Ford, L., Makunde, W.H., Prasad, J.B.K.S., Bilo, K., 2001.
Immune responses directed at egg proteins during experimental infection with the Wolbachia bacteria in filarial immunity and disease. Parasite Immunol. 23, 401–409.
liver fluke Fasciola hepatica. Parasite Immunol. 32, 111–124. Vidigal, J., Ventura, A., 2015. The biological functions of miRNAs: lessons from in vivo
Mulcahy, G., O'Neill, S., Donnelly, S., Dalton, J.P., 2004. Helminths at mucosal barriers studies. Trends Cell Biol. 25, 137–147.
interaction with the immune system. Adv. Drug Deliv. Rev. 56, 853–868. Vukman, K.V., Adams, P.N., Metz, M., Maurer, M., O'Neill, S.N., 2013a. Fasciola hepatica
Mulcahy, G., O'Neill, S., Fanning, J., McCarthy, E., Sekiya, M., 2005. Tissue migration by tegumental coat impairs mast cells’ ability to drive th1 immune responses. J.
parasitic helminths an immunoevasive strategy? Trends Parasitol. 21, 273–277. Immunol. 190, 2873–2879.
Navarro, S., Ferreira, I., Loukas, A., 2013. The hookworm pharmacopoeia for in- Vukman, K.V., Adams, P.N., O'Neill, S.M., 2013b. Fasciola hepatica tegumental coat an-
flammatory diseases. Int. J. Parasitol. 43, 225–231. tigen suppresses MAPK signalling in dendritic cells and up-regulates the expression of
O'Neill, S.M., Mills, K.H.G., Dalton, J.P., 2001. Fasciola hepatica cathepsin L cysteine SOCS3. Parasite Immunol. 35, 234–238.
proteinase suppresses Bordetella pertussis-specific interferon-γ production in vivo. Walsh, K.P., Brady, M.T., Finlay, C.M., Boon, L., Mills, K.H.G., 2009. Infection with a
Parasite Immunol. 23, 541–547. helminth parasite attenuates autoimmunity through TGF-β mediated suppression of
Pabalan, N., Singian, E., Tabangay, L., Jarjanazi, H., Boivin, M.J., Ezeamama, A.E., 2018. Th17 and Th1 responses. J. Immunol. 183, 1577–1586.
Soil-transmitted helminth infection, loss of education and cognitive impairment in Wammes, L.J., Mpairwe, H., Elliott, A.M., Yazdanbakhsh, M., 2014. Helminth therapy or
school-aged children: a systematic review and meta-analysis. PLoS Negl. Trop. Dis. elimination: epidemiological, immunological, and clinical considerations. Lancet
https://doi.org/10.1371/journal.pntd.0005523. Infect. Dis. 11, 1150–1162.
Piedrafita, D., Spithill, T.W., Smith, R.E., Raadsma, H.W., 2010. Improving animal and Wilson, R.A., Wright, J.M., Borges, W.C., Manuel, S.J.P., Dowle, A.A., Ashton, P.D.,
human health through understanding liver fluke immunology. Parasite Immunol. 32, Young, N.D., Gasser, R.B., Spithill, T.W., 2011. Exploring the Fasciola hepatica tegu-
572–581. ment proteome. Int. J. Parasitol. 41, 1347–1359.
Prowse, R.K., Chaplin, P., Robinson, H.C., Spithill, T.W., 2002. Fasciola hepatica cathepsin World Health Organization (2018) Soil-transmitted helminth infections. https://www.
L suppresses sheep lymphocyte proliferation in vitro and modulates surface CD4 ex- who.int/es/news-room/fact-sheets/detail/soil-transmitted-helminth-
pression on human and ovine T cells. Parasite Immunol. 24, 57–66. infectionsAccessed 06 January 2019.
Quinn, S.M., Cunningham, K., Raverdeau, M., Walsh, R.J., Curham, L., Malara, A., Mills, Yazdanbakhsh, M., Kremsner, P.G., Ree, V.R, 2002. Allergy, parasites, and the hygiene
K.H.G., 2019. Anti-inflammatory trained immunity mediated by helminth products hypothesis. Science 292, 490–494.