Parasite Immunology, 2009, 31, 347356
Review Article
Teladorsagia circumcincta in the sheep abomasum: defining
the role of dendritic cells in T cell regulation and protective
immunity
T. N. MCNEILLY,1 E. DEVANEY2 & J. B. MATTHEWS1,3
1
Moredun Research Institute, Pentlands Science Park, Midlothian, UK, 2Parasitology Group, Veterinary Infection and Immunity,
Faculty of Veterinary Medicine, University of Glasgow, Glasgow, UK, 3Department of Veterinary Clinical Studies, Royal (Dick)
School of Veterinary Studies, University of Edinburgh, Midlothian, UK
SUMMARY
Parasitic nematodes of the small-ruminant gastrointestinal
tract pose a problem worldwide. The impact of these patho-
gens is worsened by the emergence of anthelmintic resistance
to all three available classes of drugs. In addition to causing
considerable economic loss, these parasites are detrimental
to the health and welfare of sheep and goats. Vaccination
offers an alternative approach to drug-based control and a
great deal of investment has gone into the investigation of
protective antigens for some of these nematode species.
However, attempts at vaccination are hindered by a lack of
understanding of how best to promote protective immunity
to nematode species, such as Teladorsagia circumcincta,
which inhabits the abomasum of sheep. This situation con-
trasts with that in murine models of gastrointestinal nema-
tode infection, where the basis of protective immunity is
increasingly well understood. In this review, we discuss the
current knowledge of the immune effector mechanisms elic-
ited by T. circumcincta and consider the probable role of
dendritic cells in the initiation of both effector and regula-
tory responses in the abomasum.
Keywords abomasum, dendritic cells, immune response, regu-
latory T cells, sheep, Teladorsagia circumcincta
Correspondence: Jacqui Matthews, Division of Parasitology,
Moredun Research Institute, Pentlands Science Park, Midlothian
EH25 0PZ, UK (e-mail: jacqui.matthews@moredun.ac.uk).
Received: 18 December 2008
Accepted for publication: 17 February 2009
 2009 Blackwell Publishing Ltd
DOI: 10.1111/j.1365-3024.2009.01110.
INTRODUCTION
Teladorsagia circumcincta is the major cause of ovine
parasitic gastroenteritis in temperate regions: almost all
grazing sheep in these areas are infected and many ani-
mals harbour thousands of worms. This nematode is
currently controlled by using a combination of anthel-
mintic treatments and pasture management; however,
drug resistance is widespread and a number of isolates
have been reported that exhibit phenotypic resistance to
all three classes of anthelmintics (13). T. circumcincta
resides within the abomasum (the fourth division of the
ruminant stomach, analogous to the monogastric stom-
ach) and is primarily a cause of disease in lambs. In
these animals, clinical signs range from suppressed appe-
tite and reduced liveweight gain, to diarrhoea, dehydra-
tion and death. Reductions in appetite and weight gain
have been attributed to the nutritional cost of generating
an anti-parasite immune response (4). Third-stage larvae
(L3) penetrate abomasal glands within 24 h of infection
and grow rapidly, undergoing two moults before emerg-
ing into the lumen approximately 10 days post-infection
(dpi). The damage that the larvae cause and the host
inflammatory responses that they generate result in a
protein-losing gastropathy, which is subsequently exacer-
bated by the effects of adult nematodes at the abomasal
mucosal surface (5). Protective immunity against chal-
lenge with T. circumcincta L3 does not occur following
primary infection, but develops after trickle or continual
infection over a number of weeks (6). The level of
immunity that develops varies with the age (7) and
genotype (8) of the animal and is compromised in ewes
during the period around parturition and early lactation
(9).
347
T. N. McNeilly et al. Parasite Immunology

Strong correlations have been observed between mean

IMMUNE RESPONSES TO TELADORSAGIA
worm length in individual animals and peak IgA levels [at
CIRCUMCINCTA: CURRENT KNOWLEDGE
6 days post-challenge (dpc)] measured in gastric efferent
A number of studies have investigated the immunological lymph (7). Likewise, significant inverse correlations have
basis of resistance to T. circumcincta. Despite this research, been observed between IgA concentrations in mucosal
the precise effector mechanisms involved in immunity are scrapings and the length of T. circumcincta harvested from
not fully understood. Expression of resistance to T. cir- both naturally and experimentally infected sheep (17).
cumcincta involves changes in the abomasal microenviron- High levels of immunity are thought to lead to L4 arrest
ment that affect the parasite in a number of ways: these (or inhibition) and such larvae are termed early L4 (EL4).
include immune exclusion of incoming L3 (10), slowing of Phenotypically, EL4 in previously infected, immune sheep
larval development (7,1013) and effects on adult worm are <15 mm in length and are equivalent to those nema-
length and fecundity (12). A schematic overview detailing todes recovered from a primary infection of less than
the current status of knowledge with regard to immunity 5 days duration (7). That the arrest of L4 has an immuno-
to T. circumcincta is shown in Figure 1. logical basis is supported by experiments where larvae
A hypersensitivity reaction is proposed to mediate were noted to resume development following cortico-
exclusion and or expulsion of incoming L3 in previously steroid treatment of experimentally infected sheep (13). In
infected, immune sheep (4,14,15). This effect has been the absence of further L3 infection, arrest of L4 is short-
observed as early as 2 days after challenge with T. circum- lived (13), indicating that continued exposure to parasite
cincta L3 in sheep rendered immune by trickle infection antigen is critical to maintaining this check in develop-
(10). Local IgA levels may have an effect on L3 establish- ment. The proportion of EL4 that develop after L3 chal-
ment: significant negative correlations have been observed lenge of previously infected animals varies with age. For
between anti-larval excretory secretory (ES)-specific example, 45-month-old sheep administered with a trickle
mucus IgA levels and the number of nematodes that estab- infection are able to respond immunologically to subse-
lish after L3 challenge of previously infected ewes (16). quent challenge and, while such animals have a higher per-
Locally produced nematode-specific IgA is also thought to centage of EL4 than challenge control sheep, the
have a role in affecting the rate of development of fourth- proportions of EL4 are substantially less than those
stage larvae (L4) in the gastric gland of immune sheep. observed in similarly treated 10-month-old sheep (7). Lar-
vae displaying the phenotypic characteristics of EL4 have
also been recovered 5, 10 and 21 dpi from the abomasal
wall of sheep given a primary infection; however, these
Initiating factors that direct
adaptive immune response
constitute a low proportion (i.e. <10%) of the total larval
UNKNOWN burden (18). The number of EL4 observed after 5 days in
a primary infection is dose-dependent (19) and has been
proposed to be mediated by chemokines that are rapidly
Genetic influence on ability to upregulated via an innate immune response to incoming
mount adaptive response
(especially impact on FEC) L3 (13).
Immunity to T. circumcincta also operates through
Factors that effects on adult worm length and female fecundity (12):
regulate the IgA has been identified as having a role in this effect (20).
adaptive immune Immune response operates at different
response T. circumcincta-specific IgE responses have also been pro-
levels: dependent on animal age
UNKNOWN posed to have an impact on faecal egg output. For exam-
Characterised well in natural and ple, in one study where faecal egg counts (FEC) were
experimental infection
Influence on ability to respond to compared with antibody responses in lambs during their
vaccination currently unknown first grazing season, IgE levels specific to a high-molecu-
lar-weight L3 antigen were associated with low cumulative
FEC (21). In Soay sheep, reduced FEC have been linked
Responses directed at Responses directed at to an allele at a microsatellite locus positioned in the first
nematode speed of excluding or expelling larvae intron of the IFN-c gene and this allele was also associ-
development and female from the mucosa
fecundity older animals (>8 mo) ated with increased parasite-specific IgA (22). Here, the
younger animals (< 8 mo) authors proposed that polymorphisms that affected the
Figure 1 Schematic overview detailing the current status of expression of IFN-c could enhance activity of Th2
knowledge with regard to immunity to Teladorsagia circumcincta. responses (see below). Subsequent studies have identified

348  2009 Blackwell Publishing Ltd, Parasite Immunology, 31, 347356

Volume 31, Number 7, July 2009
additional polymorphisms in the same region of the IFN-
c gene and showed that one of these was associated with a
reduced FEC in Texel, but not in Suffolk sheep (23).
Taking into account the above observations, under field
conditions, where animals of different ages are likely to be
exposed to prolonged L3 challenge, it is likely that incom-
ing and developing T. circumcincta would be subjected to
immune responses that lead to adult worm and larval
retardation as well as larval exclusion from the mucosa.
The level of immunity that is expressed gradually improves
over time as animals age. Much of the work described
above details the role that humoral immune responses may
have in resistance to T. circumcincta. Experiments in which
the efferent gastric lymph ducts of sheep were cannulated
have provided insights into the dynamics of the local cellu-
lar response during infection. In these experiments, large
numbers of lymphoblasts were detected in efferent lymph
within 5 days of challenge, especially in previously infected
sheep (10,11). The role that these cells may have in protec-
tive immunity was indicated in a set of experiments
whereby gastric lymph, pooled from immune sheep at 1
5 dpc, was used to successfully transfer immunity to nave
sheep (7,24). In agreement with these findings, the cellular-
ity of the abomasal mucosa has also been observed to
increase substantially in immune sheep subjected to L3
challenge: most of the cells that infiltrate the mucosa are
T cells and eosinophils (25). Mast cells may also affect
T. circumcincta at this site: worm burdens negatively corre-
late with levels of abomasal mast cell protease, released
from activated mast cells (26), and numbers of globule leu-
cocytes (14). The latter are thought to be degranulated
mast cells (27). These findings indicate that, similar to gas-
trointestinal nematodes in other host species, the main
immune response elicited by T. circumcincta is Th2-like.
Confirming this, recent experiments demonstrated a pre-
dominance of transcripts encoding Th2-associated cyto-
kines (i.e. IL-4, IL-5 and IL-13) in abomasal lymph node
(ALN) cells harvested after challenge with L3 (28). Inter-
estingly, mRNA expression of these cytokines was greater
at early time-points (day 5) postinfection in previously
infected, immune sheep compared with nave sheep, con-
firming that these Th2-associated cytokines may be associ-
ated with a protective immune response. This study also
demonstrated an upregulation of ALN cell IL-10 and
TGF-b mRNA, cytokines associated with regulatory T
(Treg) cell function in humans and rodents (29,30).
Increases in these cytokines were observed following infec-
tion of both nave and previously infected, immune sheep.
The cytokine profiles described suggest that T. circum-
cincta induces local immune responses typical of a Th2
type in concert with the production of Treg cells, not dis-
similar to observations made in nematode infections of
 2009 Blackwell Publishing Ltd, Parasite Immunology, 31, 347356
T. circumcincta in the sheep abomasum
other species (31). The interplay between Th2 effector and
Treg cell populations in the abomasal wall may be a criti-
cal determinant of the outcome of infection with T. cir-
cumcincta in sheep. An optimal vaccination strategy may
thus depend upon the correct balance of these cell popula-
tions in the abomasum.
INITIATION OF IMMUNE RESPONSES IN THE
ABOMASUM: A CRITICAL ROLE FOR
ABOMASAL DC?
It is becoming increasingly clear that the type of adap-
tive immune response generated following either infection
or immunization is influenced by initial interactions
between antigens and cells of the innate immune system.
More specifically, dendritic cells (DC), cells specialized
in presenting antigen to T cells, appear to play a central
role in polarizing T cell responses towards either Th1,
Th2 or Treg cell responses (32,33). As stated previously,
the balance between these different T cell responses is
likely to be a key factor in the development of protec-
tive immunity against T. circumcincta infection. In order
to dissect the factors that influence this balance, an
understanding of the basic mechanisms by which aboma-
sal DC present T. circumcincta antigens to T cells is cru-
cial. Despite the importance of the abomasum as a site
of infection, virtually nothing is known of the induction
of immune responses at the mucosal location in this
organ.
From observations in other species, and from studies of
afferent lymph DC in sheep, it is likely that following cap-
ture of T. circumcincta antigen by DC within the abomasal
mucosa, these cells migrate to inter-follicular T cell zones
within organized lymphoid tissue where they present anti-
gen to nave Th cells via major histocompatibility complex
(MHC) class II molecules, thus initiating antigen-specific
adaptive immune responses. Conditioning of DC by fac-
tors released by either the parasite or other host cells
within the mucosa is likely to determine the type of T cell
response subsequently induced (see below). Within the
gastrointestinal tract of sheep and other species, DC are
found primarily in two locations: scattered below the epi-
thelium or concentrated within areas of mucosa-associated
lymphoid tissue (MALT) (34,35). However, unlike other
regions of the ovine gastrointestinal tract, the abomasal
mucosa does not appear to possess any MALT (36).
Therefore, antigen uptake at this site is presumably medi-
ated primarily by sub-epithelial DC populations, which
then migrate to draining ALN to interact with nave T
cells, although direct interaction of sub-epithelial DC with
lymphocytes within the mucosa may also be possible (37).
As yet, no information exists regarding abomasal DC
349
T. N. McNeilly et al.
(a) (b)
populations in sheep; however, our group recently
identified MHC class II-positive cells within abomasal
mucosa that, judging by morphology and distribution, are
likely to be DC (T. N. McNeilly unpublished data,
Figure 2).
Despite the lack of information on abomasal DC in
sheep, there is a growing amount of data on ovine DC in
other tissues. Dentritic cells in the respiratory tract, orona-
sal cavity and intestinal tissues of sheep have been pheno-
typed in situ by using a variety of markers (35,38,39)
including CD1b, CD11c CD205, CD209 (DC-SIGN) and
signal regulatory protein alpha (SIRPa, CD172a). These
markers appear to identify different subsets of DC at vari-
ous sites. Dentritic cells in intestinal and respiratory tis-
sues have been shown to lie in close proximity to the
epithelial layer, ideally positioned for antigen sampling
and distribution to local lymph node (LN). The potential
function of ovine DC subsets has been explored using DC
isolated from afferent lymph (ALDC) following cannula-
tion of pseudoafferent lymphatics draining oronasal
mucosa and skin. These studies indicate that ALDC can
be distinguished on the basis of expression of SIPRa, a
membrane-bound inhibitory receptor capable of regulating
T cell function via binding to CD47 (40). SIRPa+ and
SIRPa) subsets appear to have distinct functions: SIRPa)
DC synthesize high levels of IL-12 and minimal IL-10,
whereas SIRPa+ DC express high levels of IL-10 and
virtually no IL-12, suggesting that SIRPa) DC are more
likely to promote Th1-biased responses and SIRPa+ DC,
Th2-type responses (41). In addition, SIRPa) ALDC con-
tain a higher proportion of cytoplasmic apoptotic DNA
and cytokeratin-positive inclusions compared with SIRPa+
DC, suggesting that SIRPa) ALDC are more involved in
self-antigen presentation and tolerance induction (42).
These two subsets of ALDC also differentially express a
variety of other surface receptors including Toll-like recep-
tors (TLRs), b2 integrins and C-type lectins (4143),
implying that each subset uses different sets of receptors
to interact with antigen. At the functional level, ovine DC
have been shown to migrate from peripheral sites in vivo
(42,44), express co-stimulatory molecules including CD40,
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Parasite Immunology
Figure 2 Immunohistochemical (IHC)
staining for major histocompatibility
complex (MHC) class II in ovine
abomasal mucosa. (a) MHC class II-posi-
tive cells are present in a sub-epithelial
location; (b) isotype control.
CD80 (B7.1) and CD86 (B7.2) (38,42) and are capable of
presenting antigen to T cells (38,45). However, the ability
of different ovine DC subsets to polarize T-cell responses
has not been studied directly. Defining the relevant DC
subsets present in the abomasum and how they interact
with T cells following infection in vivo are important to
understanding the role that these cell types have in immu-
nity to T. circumcincta.
REGULATION OF THE IMMUNE RESPONSE IN
THE ABOMASUM
In addition to inducing Th2-type responses, antigen-
primed DC are capable of eliciting Treg cells (46). These
cells have been proposed to play an important role in the
chronicity of infections by downregulating immune
responses and thus assisting parasite survival in the host
(31). A number of different types of Treg have been identi-
fied in humans and mice, including naturally occurring
CD4+ Treg [which differentiate in the thymus, express high
levels of IL-2R a-chain (CD25) and constitute a major
population of Treg], and Th1 and Th3 cells which differen-
tiate from nave T cells in the periphery (47). Expansion
of Treg subsets in the periphery requires antigenic stimula-
tion by DC; although once expanded, Treg cells can
suppress in a non-antigen-specific manner (48,49).
Suppression of effector T cell responses occurs via three
major mechanisms, all of which require close proximity
between Treg and effector cells, either within the target tis-
sue or the regional lymph nodes: (i) direct killing of effec-
tor T cells; (ii) inhibition of cytokine production,
particularly IL-2, by effector T cells; and (iii) secretion of
regulatory cytokines, such as IL-10 and TGF-b (30). Fur-
thermore, Treg cells are thought to directly inhibit DC
function, for example by downregulating the expression of
DC co-receptors CD80 and CD86 (50), or by impairing
the establishment of stable contacts between DC and effec-
tor T cells (51). The transcription regulator Foxp3 (fork-
head box P3) is a key gene for the generation and
maintenance of natural CD4+CD25+ Treg cells in rodents
and humans (52), and is the most widely used marker for
 2009 Blackwell Publishing Ltd, Parasite Immunology, 31, 347356
Volume 31, Number 7, July 2009
these cells, despite some reports indicating that it can be
transiently expressed at low levels in non-Treg cells in
humans. Foxp3 is highly conserved between species and is
likely to identify T cells with regulatory functions in sheep,
although this has yet to be formally proven.
We have recently identified numerous Foxp3+ T cells
(Foxp3+ CD3+ cells) within the abomasal mucosa and
ALN of T. circumcincta-infected sheep (T. N. McNeilly
unpublished data; Figure 3). This preliminary observa-
tion, together with the previous report of upregulated IL-
10 and TGF-b expression in ALN cells postinfection
(28), suggests that, analogous with other nematode infec-
tions (53,54), T. circumcincta also induces Treg-like cells.
If this is indeed the case, then this observation poses a
number of interesting questions. For example, what is the
role of DC in the induction of abomasal Treg cells? Are
variations in host susceptibility to T. circumcincta related
to differences in Treg responses? Do T cell responses
change from a more regulatory to an effector response
(or phenotype) during continual or trickle exposure to
T. circumcincta (i.e. breakdown of tolerance to the para-
site)? Could this explain why protective immunity takes
time to develop? A greater understanding of Treg
responses following T. circumcincta infection, focusing on
the parasite and host factors involved in induction of
these cells and the interaction between Treg and effector
cell populations will be relevant to understanding the
mechanisms by which protective immunity evolves. Such
information will have practical applications for the devel-
opment of vaccines to control this important nematode
of sheep.
(a)
Figure 3 Immunohistochemical (IHC)
staining for Foxp3 and pan-T-cell marker
CD3 in ovine abomasal lymph node
(ALN) and abomasal mucosa from a
sheep infected with Teladorsagia circum-
cincta. Positive nuclear Foxp3 staining is
present within ALN T-cell areas (a) and
within the abomasal mucosa (b). Note the (c)
presence of a T. circumcincta larva within
the abomasal mucosa in (b) indicated by
the arrow. Double staining for Foxp3
(3,3-diaminobenzidine, dark brown) and
CD3 (3-amino-9-ethylcarbazole, red)
within the abomasal mucosa (cd)
identifies Foxp3+CD3+ cells within the
abomasal mucosa (examples indicated by
arrowheads). (d) A high-power view of
the boxed region in (c).
 2009 Blackwell Publishing Ltd, Parasite Immunology, 31, 347356
T. circumcincta in the sheep abomasum
HELMINTH PARASITES ACTIVATE DC IN
NONCONVENTIONAL WAYS
As described previously, infection of sheep with T. circum-
cincta results in a Th2-dominated response but how that
response is initiated and, indeed, which component(s) of
that response might be responsible for protection remains
unresolved. While the role of DC in polarizing immune
responses to Th1 is well documented, how DC handle hel-
minth antigens to promote Th2 responses is much less
clear. Most of our information on the interactions of DC
with helminth antigens comes from studies carried out
using bone marrow (Bm) DC derived from mice exposed
to Ag in vitro. In contrast to the activation of DC follow-
ing exposure to bacterial antigens, stimulation with hel-
minth products results in DC that display few of the
classical activation markers, but which retain the ability to
drive T cells down the Th2 pathway. This is best exempli-
fied from studies using soluble egg antigen (SEA) from
Schistosoma mansoni. With the exception of a small
increase in MHC II expression, murine Bm-DC exposed
to SEA do not activate classical markers or produce cyto-
kines but still polarize Th cells to Th2 (55). A consistent
finding of these studies was the failure of SEA to drive
IL-12 expression and, indeed, to inhibit bacterial antigen-
induced IL-12 (56). A comparison of genes upregulated in
DC following exposure to SEA or to LPS demonstrated
the distinct profile elicited by each stimulus, while co-
exposure of DC to both SEA and LPS resulted in the
downregulation of multiple pro-inflammatory genes by
both IL-10-dependent and -independent mechanisms (57).
(b)
200 m
(d)
100 m
351
T. N. McNeilly et al.
In a nematode model more comparable with T. cir-
cumcincta, transfer to nave mice of DC stimulated with
Nippostrongylus brasiliensis excretory secretory products
(NES), resulted in a Th2-type response in the draining
LN (58); NES-matured DC showed upregulated expres-
sion of CD86 and OX40L, but not CD80 or MHC II,
demonstrating the selective maturation driven by nema-
tode products. These cells also produced only certain
cytokines, for example IL-6 and IL-12p40, but not IL-
12p70. Previous studies had demonstrated that NES had
a strong adjuvant capacity in that it promoted Th2
responses when administered alone or in the presence of
Freunds complete adjuvant which would normally
favour Th1 induction (59). This phenomenon was likely
mediated via DC, as pre-incubation of DC with NES
was shown to inhibit LPS-induced Th1 polarization by
inhibition of IL-12p70 (58). Whether T. circumcincta ES
products similarly polarize T cell responses has yet to
be determined.
Helminth parasites are complex multicellular organisms
containing a myriad of antigens that can influence
immune responses both positively and negatively. Attempts
to define the molecular components of helminths that
interact with DC have identified a role for glycan moieties
in polarizing T cell responses. Schistosome glycolipids, in
particular the carbohydrate epitope GalNAcb1-4(Fuca1-
2Fuca1-3)GlcNAc (LDN-DF), which is abundant in schis-
tosome eggs but not adult worms, trigger the secretion of
IL-10, IL-6 and TNF-a from human monocytes (60). Sub-
sequent studies showed that the C-type lectin DC-SIGN
acts as a receptor for both GalNAcb1-4(Fuca1-3)GlcNAc
(LDNF) and the Lewisx antigen found in schistosomes
(61). Interestingly, immunization of sheep with ES of Hae-
monchus contortus was shown to induce a strong antibody
response against LDNF epitopes present on H. contortus
glycoproteins, which was correlated with levels of protec-
tion (62). However, subsequent studies confirmed that,
although H. contortus LDNF stimulated high levels of
antibody, it was unlikely to be responsible for the protec-
tion mediated by the well-characterized H-gal-GP fraction
(63). A different lipid fraction of S. mansoni eggs or adult
worms, phosphatidylserine (PS), was shown to inhibit IL-
12 production by DC and to result in T cell populations
that produced IL-4 and IL-10, and which were capable of
suppressing the proliferation of other T cells, i.e. had the
characteristics of Treg cells. The active fraction was shown
to signal through TLR-2 (46). The specificity of this
response was demonstrated by HPLC separation of the
schistosome PS fraction; the IL-10-inducing fraction was
shown to be monacetylated lyso-PS while, diacetylated PS
was responsible for Th2 induction. Nematodes, too, con-
tain many molecules that modulate immune responses by
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Parasite Immunology
affecting DC function. Of these, ES-62, the phosphorylch-
oline-containing glycoprotein from the filarial nematode
Acanthocheilonema viteae, is the best characterized. In vitro,
ES-62 has been shown to modulate the function of DC
and other cells of the immune system (64). Exposure to
ES-62 in vivo resulted in a population of Bm-DC that pro-
duced lower levels of IL-12 and TNF-a in response to
LPS (65), an effect that could be mimicked using phos-
phorylcholine alone.
These observations highlight the potential influence of
glycanDC interactions in polarizing T cell responses in
helminth infections, and have implications regarding vac-
cine design. The failure of recombinant nematode vaccines
may reflect differences in important structural T- and B-
cell epitopes on recombinant proteins compared with
native antigens, with the result that immune responses are
not directed towards protective epitopes. However, it is
also possible that differences in glycan structures between
recombinant and native antigens affect the type of
immune response generated (Th1 Th2 Treg) via altera-
tions in DC function, ultimately influencing vaccine
efficacy.
Although much useful data have been obtained from
in vitro models of DC function, in vivo, DC do not act on
T cells in isolation. Several recent studies have identified a
network of interacting cells and immune factors acting on
DC to influence T cell responses. For example, eosinophil-
derived neurotoxin (EDN), a secretory product of eosin-
ophils was shown to induce Th2 responses via its ability
to promote the maturation and migration of DC (66).
EDN is an alarmin, a member of a diverse group of
endogenous molecules that activate cells of the innate
immune system in response to damage or trauma (67).
EDN activates DC by signalling through TLR-2 and, in
addition, functions as an anti-microbial peptide and as a
chemoattractant for eosinophils. Immunization of mice
with OVA in the presence of EDN resulted in a higher
ratio of IgG1 : IgG2 and production of increased levels of
IL-5, IL-6, IL-10 and IL-13, but not IL-4, compared with
mice immunized with OVA alone, an outcome that was
dependent upon TLR-2 ligation. Cells other than eosin-
ophils, such as neutrophils, activated macrophages and
some epithelial cells in the placenta can also produce
EDN. The penetration of T. circumcincta into, and subse-
quent emergence from, the mucosal glands is undoubtedly
associated with tissue damage, and it is conceivable that
this may result in the release of molecules that function as
alarmins to prime DC and or other cells of the innate
immune system. High levels of eosinophils are also found
in the abomasal mucosa of immune sheep challenged with
T. circumcincta (25) but whether ovine eosinophils express
EDN-like molecules or, indeed, whether other endogenous
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Volume 31, Number 7, July 2009
TLR ligands exist in the abomasum is completely unex-
plored.
It is now appreciated that the tissue environment of a
DC population plays a major role in the phenotype and
functional capacity of DC. This is particularly so in the
intestine, where multiple mechanisms exist to promote
homeostasis and prevent overt inflammatory responses
[reviewed in Ref. (68)]. In both humans and mice, DC iso-
lated from the gut default to a Th2 response, even in the
presence of strong Th1-promoting stimuli (69), a phenom-
enon that appears to be associated with their conditioning
by intestinal epithelial cells (IEC). For example, DC
derived from human colon stimulated with Salmonella
typhimurium polarized allogeneic naive T cells to Th2.
However, this feature was not restricted to colonic DC, as
monocyte-derived (mo) DC incubated with supernatants
from Caco-2 cells (a human colonic epithelial cell line)
also drive Th2 responses. In contrast, moDC activated by
direct contact with bacteria secrete both IL-10 and IL-12
with the subsequent induction of both Th1 and Th2
responses (70). The cross-talk between IEC and DC is in
part mediated by thymic stromal lymphopoeitin (TSLP),
an IL-7-like cytokine, released by IEC and other cell types
such as basophils. When levels of TLSP were reduced by
small interfering RNA (siRNA), the ability of IEC to
drive Th2 responses in nave T cells was suppressed and
instead Th1 cytokines were produced (69). These data
imply that in the intestinal environment, resident DC are
conditioned to elicit anti-inflammatory responses via their
interaction with IEC and or other cell types. The signifi-
cance of these findings to nematode infection was recently
demonstrated in a mouse model of Trichuris muris. Here,
TSLP produced by IEC inhibited the production of pro-
inflammatory cytokines by DC (71); mice with a specific
deletion in IjB kinase (IKK)-b expressed significantly
lower levels of TSLP resulting in increased frequency of
IFN-c-secreting CD4+ T cells, while in TSLP receptor KO
mice, pro-inflammatory and Th1 responses were upregulat-
ed. Consistent with their impaired Th2 response, these
mice had significantly higher worm burdens than did wild-
type mice.
If Th2 responses are indeed protective against T. circum-
cincta, and if the default in the abomasum is also Th2, it
is interesting to speculate how these parasites are able to
colonize the abomasum and survive to adulthood in the
face of a potentially harmful immune response. What fac-
tors might then tip the balance in favour of worm expul-
sion? As alluded to above, the balance between effector
and Treg cells may play a key role in worm survival. A
specialized population of DC has been shown to be
important in Treg cell induction in the murine small intes-
tine and mesenteric lymph nodes. These cells bear the aE
 2009 Blackwell Publishing Ltd, Parasite Immunology, 31, 347356
T. circumcincta in the sheep abomasum
integrin CD103+ and are important for promoting the
conversion of nave T cells into Foxp3+ Treg cells, while
CD103) DC produced pro-inflammatory cytokines (72).
TGF-b and retinoic acid were key to the induction of Treg
cells by CD103+ DC, and inhibitors of the retinoic acid
receptor were sufficient to block conversion. Based on the
kinetics of accumulation of CD103+ DC and CD103) DC,
it was proposed that CD103+ cells derive from the tissues
where they may have been conditioned by exposure to
IEC, for example, while CD103) cells derive from blood
precursors (73).
These findings from studies in mice and humans serve to
illustrate the complexities of the initial interactions between
pathogen-derived molecules and DC. Over the past
10 years, much information has been obtained about recep-
tors on DC that respond to pathogen-derived molecules and
of the signalling cascades that are initiated upon ligation.
Recent studies have shown that the functional capacity of a
DC population is influenced by both anatomical location
and interactions with other cell types and their products.
Based on work in other systems, it is interesting to speculate
whether during a primary, non-protective infection with
T. circumcincta, abomasal DC may adopt an anti-inflam-
matory regulatory phenotype, while during a protective
response, a switch to an inflammatory DC phenotype may
occur. This switch could be driven by changes within the
abomasal microenvironment and result in differing T cell
responses to the parasite. The challenge for studies in rumi-
nants is now to define the nature of the interactions occur-
ring in the abomasum between cells of the innate immune
system and parasitic nematodes and to apply that informa-
tion to the rational development of novel vaccines.
CONCLUDING REMARKS
Research into this area will inform future vaccine strate-
gies by providing an insight into those factors involved in
initiating protective immune responses to T. circumcincta.
Addressing questions such as how protective immune
responses develop following infection, why most recombi-
nant antigens fail to protect and how to select appropriate
adjuvants to stimulate optimal activation of DC will be
key to the development of successful vaccines. Significant
progress has been made in defining many of the compo-
nents of the ES released by T. circumcincta (16) but addi-
tional work is needed to determine how these molecules
interact with immune cells either to modulate or to acti-
vate responses and how best to present immunostimulato-
ry molecules to the immune system in sheep. Given that
many reagents are now available for studying ovine
immune responses, the next few years should bring a much
clearer picture of how parasite ES products interact with
353
 T. N. McNeilly et al.
ovine DC and other cell types in the abomasum and how
these interactions can be manipulated to optimize adaptive
T-cell responses.
ACKNOWLEDGEMENTS
JBM and TMcN acknowledge the Scottish Government
for funding their research. The authors would like to
thank Jeanie Finlayson and the Histopathology Labora-
tory at Moredun Research Institute for assistance with
immunohistochemical staining.
CONFLICTS OF INTEREST
The authors have no conflicts of interest concerning the
work reported in this paper.
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