text:
Page 1
Porcine T lymphocytes and NK cells – An update
Wilhelm Gerner, Tobias Ka ¨ser, Armin Saalmu ¨ller*
Institute of Immunology, Department of Pathobiology, University of Veterinary Medicine Vienna, Veterina ¨rplatz 1, 1210 Vienna, Austria
1. Introduction
During the last three decades the importance of swine in
agricultural and biomedical research has resulted in a substantial
increase in the research efforts on the porcine immune system. In
agricultural research the interaction of pathogens with the
immune system plays the most important role, and detailed
knowledge of the host immune system is required to identify
protective immune mechanisms and to define immuno-correlates
of protection. Knowledge about protective immune mechanisms
gives the chance for a better monitoring of vaccination efforts and
for an improvement of vaccination strategies. Studies on the
and the identification of pathogen-specific epitopes are often
aimed to design new or more efficient vaccines.
The adaptive immune system based on the function of T and B
lymphocytes is therefore one of the major targets of research.
Stimulated by an interaction of cells belonging to the innate
immune system, like dendritic cells and monocytes, B and T
lymphocytes generate pathogen-specific reactions and, further-
more, develop long-lasting immunological memory.
The activity of B lymphocytes can be easily detected by their
soluble products, specific antibodies, whereas monitoring of
antigen-specific T-cell responses needs a detailed knowledge
about the phenotypes of responding T lymphocytes and the
respective T-cell subpopulations.
Monoclonal antibodies (mAbs) directed against porcine leuko-
cyte differentiationantigens have beendeveloped (summarized by
[1,2]), and especially mAbs directed against antigens selectively
expressed on T lymphocytes represent now appropriate tools for a
Developmental and Comparative Immunology 33 (2009) 310–320
Article history:
Available online 2 July 2008
T cells
NK cells
Natural killer (NK) cells represent an important cell population of the innate immune system with the
ability to attack spontaneously pathogen-infected and malignant body cells as well as to produce
immune-regulatorycytokines. Tlymphocytesbelongtotheadaptiveimmunesystem and performawide
array of functions in immune regulation, inflammation and protective immune responses. In this review
we summarize the current knowledge about the phenotype and functional characteristics of these two
cell populations in swine.
Porcine NK cells can be distinguished from T cells by the complex phenotype perforin+
CD3?CD4?CD5?CD6?CD8a+CD8b?CD11b+CD16+. Investigations so far show that these cells have the
capacity to lyse virus-infected target cells and respond to various regulatory cytokines. Such cytokines
can induce interferon-g (IFN-g) production in porcine NK cells, as well as the up-regulation of effector/
activation molecules like perforin and CD25. Porcine T cells can be divided into a number of
subpopulations, including a prominent fraction of T cells expressing T-cell receptors (TCR) with gd-
chains.LikeTCR-abTcells, these TCR-gdTcells canexpressCD8aandMHCclass II,twomoleculeswhich
in swine seem to be correlated with an activation status of T cells. Functional properties of these cells
seem to include cytolytic activity as well as antigen presentation; however, both aspects require further
investigation. Like in other species, TCR-ab T cells in swine comprise MHC class-I restricted cytolytic T
cells, T-helper cells and recently identified regulatory T cells. We summarize data on the phenotype and
function of these cells including memory cell formation. Current knowledge suggests that MHC class-I
restricted cytolytic T cells can be identified by the expression of CD8ab heterodimers. T-helper cells
express CD4 as well as other activation-related markers, including CD8a, MHC class II and CD45RC.
Porcine regulatory T cells have a phenotype similar to that of mouse and humans: CD4+CD25+Foxp3+.
First results indicate that these cells can suppress proliferation of other T cells and produce IL-10. Finally,
the abundant expression of swine-specific activation markers CD8a and MHC class II on T cells and NK
cells is discussed in more detail.
? 2008 Elsevier Ltd. All rights reserved.
* Corresponding author. Tel.: +43 1 25077 2750; fax: +43 1 25077 2791.
E-mail address: firstname.lastname@example.com (A. Saalmu ¨ller).
Contents lists available at ScienceDirect
Developmental and Comparative Immunology
journal homepage: www.elsevier.com/locate/dci
0145-305X/$ – see front matter ? 2008 Elsevier Ltd. All rights reserved.
Page 2
detailed identification of T cells and T-cell subpopulations.
Combined with functional analyses, functionally distinct T-cell
subsets which play a role in various immune reactions were
In this article we aim to summarize the current knowledge
about porcine T lymphocytes and porcine T-lymphocyte sub-
populations (Table 1). In addition, we will discuss some
peculiarities of porcine T lymphocytes with regard to unusual
and unexpected T-cell phenotypes.
Additional chapters of the article deal with the current
knowledge about natural killer (NK) cells in swine. This cell
population distributed to the innate immune system gained in
importance during recent years because of its role as a central
player of the immune system, emerging as a leukocyte population
with key functions in the interaction between cells of the innate
and adaptive immune system.
2. Porcine NK cells
2.1. Phenotype of porcine NK cells
Porcine NK cells were firstly described as CD2+CD4?CD5?CD8a+
cells [3,4]. Other studies identified a NK-inhibitory antigen, PNK-1,
was later defined as CD18, a differentiation antigen involved in NK-
cell function but without any selective expression on NK cells.
but CD16 is also expressed on cells of the myeloid lineage, e.g. on
monocytes. So far selective markers forporcine NKcells are missing
and NK cells can be defined only by distinct marker combinations,
e.g. CD8a vs. CD16, CD8a vs. CD3 (Gerner, unpublished results),
or as presented in Fig. 1D as CD3?CD4?CD8a+subset of peripheral
blood lymphocytes (PBLs). A recent study from Denyer et al. [8]
defined NK cells of swine as cells with the phenotype perfor-
In our analyses we could confirm these data [9] and further
show that with regard to MHC-class II DR expression (Fig. 2C) this
lymphocyte subset represents a heterogeneous fraction. Porcine
NK cells can be MHC-class II?as depicted in Fig. 2C, but we also
have analyzed NK cells from swine showing a weak to distinct
MHC-class II DR expression, both ex vivo and after in vitro
stimulation ([9] and data not shown). We have to date no
convincing explanation for this phenomenon, but one might
speculate that this MHC-class II DR expression defines an
activation status of the respective NK-cell fractions, since a
correlation of MHC-class II DR expression with activation could
be shown for porcine T cells in previous publications [10,11].
The homogeneous high expression of CD45RC as presented in
Fig. 2D also seems to be unexplained. In contrast to the expression
pattern on T lymphocytes, the expression of CD45RC on NK cells
seems to be constitutive and was not down regulated upon
stimulation with cytokines [9]. Therefore, CD45RC expression on
porcine NK cells appears to be independent of a prior activation
2.2. Functional characterization of porcine NK cells
Experiments characterizing the spontaneous cytolytic activity
of porcine NK cells against various cell lines have been reported by
different groups [3,4,12,13]. Furthermore,cytolyticactivity against
target cells infected with pseu-dorabies virus (PRV) and transmis-
sible gasteroenteritis virus was described [14,15]. Increased exvivo
NK-cell activity in swine with chronical infectious diseases was
also observed [16] and lethal infection with classical swine fever
virus (CSFV) led to a tremendous decrease of NK-cell activity
during the course of infection [17].
Some in vitro studies showed that NK-cell activity could be
enhanced by interferon-g (IFN-g) [18] and interleukin-2 (IL-2)
[19]. Administration of IL-12 resulted in an increased cytolytic
activity and secretion of tumor necrosis factor-a (TNF-a) [20],
whereas addition of IL-4 diminished the NK-cell activity of IL-2-
stimulated NK cells [21]. As demonstrated by Domeika et al. [22], a
combination of IL-12 and IL-18 was able to stimulate NK cells for
an enhanced IFN-g production. Recently, our laboratory showed a
synergistic effect of IL-2, IL-12 and IL-18 on the cytolytic activity,
perforin expression and IFN-g production of NK cells, and,
furthermore, the ability of cytokine-stimulated NK cells for
CD25 and additional MHC-class II DR surface expression [9].
3. Porcine T cells
3.1. T-cell development in thymus
As in other higher vertebrates, the thymus of swine is a highly
specialized lymphatic organ for T-cell maturation. In the early
prenatal period this organ is colonized by T-cell precursors,
originating from the bone marrow. During early ontogeny Sinkora
et al. [23] postulated a sequential colonization of porcine thymus
by CD3e-bearing thymocytes. The early thymocytes appeared on
day 40 of gestation and showed high CD3e expression. They
belonged to the TCR-gd lineage. In a second wave – 15 days later –
mature TCR-ab thymocytes with high CD3e expression could be
observed, but their appearance was preceded by TCR-ab
thymocytes with low CD3e antigen density. In the prenatal period
only a small percentage of thymocytes was positive for CD25 and
belonged mainly to the TCR-ab thymocytes, whereas in the
postnatal period the percentage of CD25+thymocytes continu-
ously increased [23]. Analyses of the TCR-gd thymocyte subset
Table 1
Overview of porcine lymphocyte subsets.
Functional lymphocyte subsetPhenotypeFrequency within PBLReferences
NK cellsPerforin+CD2+CD3?CD4?CD5?CD6?CD8a+
2–10%, age-dependent decrease[8,9]a
NKT cells
TCR-gd T cells
T-helper cells
5–50%, age-dependent decrease
8–21%, up to 40%
19–60%, variable portions of CD4+CD8a?and
Not yet investigated
Not yet investigated
Not yet investigated
CD3+CD4+, poorly investigated so far
CD3+CD4+, poorly investigated so far
Not yet identified
aGerner, unpublished results.
bSaalmu ¨ller, unpublished results.
W. Gerner et al./Developmental and Comparative Immunology 33 (2009) 310–320
Page 3
which developed during prenatal and postnatal ontogeny revealed
a heterogeneous population with regard to expression of other
differentiation antigens. Hirt et al. [24] could define by the
expression of CD2 a panel of TCR-gd thymocytes varying in the
composition of their respective TCR; furthermore, the existence of
additional CD8-defined subsets in the CD2-positive TCR-gd
thymocyte fraction was obvious [24,25]. In more recent work
Sinkora et al. [26,27] defined 12 TCR-gd thymocyte subsets
characterized by the differentiationantigens CD1, CD2, CD4, CD8a,
CD8b and CD45RC. It seemed that CD4+TCR-gd thymocytes
Fig. 1. Analysis of CD4/CD8a expression of porcine peripheral blood lymphocytes by four-color flow cytometry. Peripheral blood lymphocytes (PBLs) gated according to their
in (C). If T and non-T cells are separated according to CD3 expression (B), in the non-T cell group a CD8a+subpopulation is revealed, which represents NK cells (D). The
remaining CD3+T cells still consist of four CD4/CD8a-defined subsets (F). A further separation of CD3+T cells into TCR-ab- and TCR-gd-expressing subsets (E) uncovers that
all peripheralbloodTCR-gdT cellsareCD4?butconsistof aCD8a?anda CD8a+subpopulation (H).Incontrast,TCR-abTcells stillcomprisefour CD4/CD8a-defined subsets:
CD4+T-helper cells, which are either CD8a+or CD8a?, a dominant fraction of CD4?CD8a+cells which contains MHC-class I-restricted cytolytic T lymphocytes and a very
small fraction of CD4?CD8a?TCR-ab T cells of unknown function (G). Presented data are representative for 6-month-old swine.
W. Gerner et al./Developmental and Comparative Immunology 33 (2009) 310–320
Page 4
bearing CD1 and CD8ab molecules did not further mature and did
not leave the thymus. This was in contrast to the group of CD4?
TCR-gd thymocytes which gave rise to all extrathymic TCR-gd T
cells. These cell populations seemed to contain CD1+CD2+TCR-gd
progenitors which further maturated and down-regulated their
CD1, and in some cases their CD2 expression, before they left
thymus as either CD2?CD8?or CD2+CD8?and CD2+CD8a+TCR-gd
Compared to other vertebrates, porcine thymocytes derived
from juvenile animals show a very similar expression of the
differentiation antigens CD4 and CD8 (mouse: [28]; man: [29];
swine: [30]). CD4?CD8?thymic cells which were described as
early thymocytes showed low expression of CD1 [31,32]. The
CD4+CD8+double-positive population representing the common
thymocytes was described as the most prominent thymic cell
fraction [25,30] with the highest CD1 antigen expression [32].
CD4?CD8+and CD4+CD8?thymocytes seemed to represent more
mature thymocyte subsets, which contained cells showing a
down-regulation and partial loss of CD1 antigen expression [32].
Nearly all of the thymocytes, with the exception of a small fraction
of CD4?CD8?cells, showed a homogeneous CD2 [30] and CD3
expression [25].
3.2. Phenotype and function of porcine T-cell subsets
Research on T cells in mice and humans over the last decades
TCR-ab and TCR-gd T cells to cytolytic vs. T-helper cells, the latter
one including the functional subpopulations of TH1, TH2 and
recently identified TH17 cells. In addition, NKT cells and regulatory
T cells (Tregs) have been identified and are subject of intensive
research activities. In swine efforts have been undertaken to
elucidate these subsets as well, but partially, with limited success,
e.g. data on the Th1/Th2 paradigm are mainly based on studies
using real-time PCR [33–35] or cytokine production was investi-
gated only from entire lymphocyte populations [36,37]. Also, to
our knowledge the Th17 subset has not been described in pigs so
far. Therefore, we will focus in this review on two well-defined T-
cell subsets in porcine immunology: TCR-ab T cells and TCR-gd T
cells, and describe further subpopulations based on this classifica-
tion(see alsoTable 1).Furthermore,Tregsidentified recentlyinthe
pig will be addressed.
3.3. Phenotype of porcine TCR-gd T cells
Like in other ungulates, TCR-gd T cells in swine form a major T-
cell subpopulation within PBLs in young animals [25]. However,
the frequency of this population decreases strongly with age ([25],
Saalmu ¨ller, unpublished results), which is also accompanied by a
T cells can be identified by a mAb directed against the d-chain
(clone PGBL22A [38]), which enables, as presented in Fig. 1E, a
direct identification of this T-cell subset. Another antibody
directed against the TCR-d chain has been described recently
[39]. Also, a mAb named PPT27 was shown to react with the
porcine TCR-gd complex,but this antibody seemed to recognize an
epitope that is not present on all TCR-gd T cells [40]. Furthermore,
mAbsagainstCD3moleculesonlyassociatedwiththe TCR-gd have
been described [41]. One of these antibodies (PPT16) was shown to
react with the vast majority of PGBL22A+ cells within PBL [42].
Antibodies directed against leukocyte differentiation antigens
clustered in swine workshop cluster (SWC) 4, SWC5 and SWC6
have also been reported to react with various subsets of gd T cells
[38,42]. The use of the antibody clone PGBL22A, directed against
the TCR-d chain, in combination with mAbs against CD4 and CD8a
revealed that extrathymic porcine TCR-gd T cells are CD4?but can
be either CD8a+or CD8a?(Figs. 1H and 3A, reviewed in [43,44]). It
has also been shown that a subpopulation of TCR-gd T cells
expresses CD2 and that in combination with CD8a three different
subsets can be defined: CD2+CD8a+, CD2+CD8a?and CD2?CD8a?
[25,45]. The latter subset is dominating within the PBL compart-
ment, whereas CD2+CD8a?cells are enriched in lymphoid tissues
[45]. This subset also expresses a unique 46 kDa TCR-g-chain,
whereas the CD2?CD8a?population has g-chains of 37–38 kDa
Fig. 2. Analysis of CD8b, MHC-class II DR and CD45RC expression in porcine NK cells. CD3?CD4?CD8a+NK cells (A) as outlined in Fig. 1 were analyzed for CD8b, MHCII and
CD45RC expression, respectively. The histograms show that the vast majority of porcine NK cells are CD8b?MHCII?(B and C) but are all CD45RC+(D). Presented data are
representative for blood-derived lymphocytes from 6-month-old swine.
W. Gerner et al./Developmental and Comparative Immunology 33 (2009) 310–320
Page 5
[45,46]. In addition, porcine TCR-gd T cells exhibit a weak
expression of CD5 and are CD6?, which is in contrast to TCR-ab
T cells showing a CD5highCD6+phenotype [4,12].
The expression of CD8b, MHC-class II DR and CD45RC on TCR-
gd T cells is shown in Fig. 3. Blood-derived CD3+PGBL22A+TCR-gd
T cells were analyzed and displayed, as illustrated in Fig. 1E and H.
The contour plots presented in Figs. 1H and 3A display CD4 vs.
CD8a expression and confirm the existence of two major
subpopulations of CD8a?and CD8a+TCR-gd T cells. Fig. 3B
illustrates that all TCR-gd T cells in blood are negative for CD8b.
However, a number of studies reported the existence of CD8b+
intestinal lymph [47], indicating that cells with this phenotype
might only be found in particular lymphatic organs. Fig. 3C
illustrates that MHC-class II DR is expressed by a subpopulation of
TCR-gd T cells. When these cells are further analyzed for CD8a
expression, a co-expression of MHC-class II DR and CD8a is
revealed (Fig. 3F), which has also been described in [26]. In
(Fig. 3E). When TCR-gd T cells are further analyzed for CD45RC
expression (Fig. 3D), again the existence of a positive and a
negative subpopulation becomes visible, with the majority of cells
being CD45RChigh. Interestingly, if these two subpopulations are
investigated for CD8a expression, it is seen that CD45RC?cells are
either CD8a?or CD8adim(Fig. 3G), whereas the CD45RC+
population contains CD8a+and CD8a?cells with an expression
that is either ''on'' or ''off'' (Fig. 3H). This is in contrast to reported
data on gd-thymocytes where the CD45RC?population was much
larger and contained also CD8ahighcells [26]. In addition, these
authors described changes of TCR-gd T cells in blood and spleen
during ontogeny up to an age of 1 year. For both compartments an
increase in the frequency of CD8a+MHC-class II DR+TCR-gd T cells
was described, accompanied by a decrease of CD8a?MHCII?cells
we observed a strong decrease of the overall frequency of
PGBL22A+TCR-gd T cells in animals older than 2 years. This is
illustrated in Fig. 4, comparing the frequency of TCR-gd T cells
within PBLs of a 6-month-old pig (Fig. 4A) with a 2-year-old pig
(Fig. 4E). In addition, it is shown that nearly all remaining TCR-gd T
vast majorityof these cells againexpressed MHC-class II molecules
(Fig. 4G). However, the frequency of CD45RC?TCR-gd T-cell does
seem to be age-independent, since no increase of this cell fraction
was observed in older pigs (Fig. 4D vs. H; Gerner, unpublished
3.4. Function of porcine TCR-gd T cells
Despite this considerable knowledge about phenotypic sub-
populations and their distribution within different lymphatic
organs (also reviewed in [43]), knowledge about the function of
this prominent T-cell subset in swine is still rudimental. For
humans and mice it is known that TCR-gd T cells can respond to
non-peptide antigens (reviewed in [48]). In swine a co-stimulation
Fig. 3. Analysis of CD8b, MHC-class II DR and CD45RC expression in porcine TCR-gd T cells. CD3+TCR-gd+T cells (A) as outlined in Fig. 1 were analyzed for CD8a, CD8b, MHC-
class II DR and CD45RC expression, respectively. The histograms show that TCR-gd T cells are CD8b?(B) but separate into MHC-class II DR+and MHC-class II DR?
subpopulations (C). If these subpopulations are analyzed for CD8a expression, it becomes obvious that the majority of MHC-class II DR?cells are also negative for CD8a (E)
but the entire MHC-class II DR+TCR-gd T cell subpopulation co-expresses CD8a (F). If CD45RC expression is analyzed on porcine TCR-gd T cells a vast population of CD45RC+
cells is found, whereas only a minor subset is CD45RC?(D). If these two subsets are analyzed for CD8a expression, no definite correlation is found: CD45RC?cells show a
negative to dim expression of CD8a (G), whereas CD45RC+TCR-gd T cells subsets can be either CD8a+or CD8a?(H). Presented data are representative for blood-derived
lymphocytes from 6-month-old swine.
W. Gerner et al./Developmental and Comparative Immunology 33 (2009) 310–320
Page 6
of TCR-gd T cells was achieved by use of plant-extracts from
Acanthospermum hispidum; however, which component of the
extract was responsible for this activation was not elucidated [49].
Also, an antigen-specific proliferation and IFN-g response of TCR-
gd T cells derived from young pigs to mycobacteria has been
reported [50]. Interestingly, this response was higher in animals
which had been immunized with a Bacille Calmette Guerin (BCG)
vaccine prior to in vitro re-stimulation, pointing to a potential
memory formation of TCR-gd T cells. This is in accordance with a
recent study in cattle, where the generation of a memory
population of gd-T cells has been described after vaccination
with killed Leptospira borgpetersenii [51], and also in humans,
where the existence of Vg2 Vd2-T cells with memory functions has
been reported (reviewed in [52]). Whether such putative memory
TCR-gd T cells can be identified by a down-regulation of CD45RC,
as it is described for porcine memory T-helper cells (see below), is
currently not known, although TCR-gd T cells with this phenotype
exist as illustrated in Figs. 3D, 4D and H, respectively.
Having outlined the potential existence of TCR-gd T cells with
porcine TCR-gd T cells to the antigen of so-called ''emergency''
foot-and-mouth disease virus (FMDV) vaccines by Takamatsu
et al. [43] was described. These authors report that after in vitro
cultivation of PBLs from FMD naive pigs together with FMDV
antigen a proliferation of CD2+CD8a+TCR-gd T cells was
observed. Additionally, purified TCR-gd T cells responded to
Whether porcine TCR-gd T cells possess cytolytic functions as
reported for mice and humans [53] has been addressed only by a
few studies. Early work by Yang and Parkhouse [54] described
CD3-redirectedcytotoxicityofTCR-gdTcells, andonenewerstudy
reported a spontaneous cytolytic activity of PGBL22A-purified
TCR-gd T cells against K562 cells [55], which was increased when
cells were isolated from BCG vaccinated pigs. Interestingly, these
authors also described a specific lysis of BCG-infected syngeneic
monocytes by TCR-gd T cells isolated from BCG-vaccinated
animals. In contrast, other investigations showed that porcine
TCR-gd T cells do not express perforin [8,43], raising the question
whether the reported cytolytic activity of porcine TCR-gd T cells is
dependent on other effector molecules or whether perforin can
only be found during certain activation stages.
As demonstrated in Fig. 3C, some TCR-gd T cells show MHC-
evidence that MHC-class II DR molecules expressed on gd-T cells
can present antigen and therefore these cells may act as
professional antigen-presenting cells since an expression of the
co-stimulatory molecules CD80/86 was also described. Interest-
ingly, the same function has recently been reported for humangd-
T cells [57] and highlights the functional plasticity of this T-cell
mice and humans, like involvement in wound healing and
regulation of inflammation, have not been addressed in swine
sofar and await elucidation.Also, the prominenceofTCR-gdT cells
in blood circulation of young pigs, which points to an important
role of this cell population in the juvenile immune system,
deserves further investigation and may in turn be rewarded by
approaches for novel intervention strategies in order to improve
the health status of young pigs.
3.5. Phenotype of TCR-ab T cells
Up to now, mAbs specific for porcine TCR-ab molecules are still
missing. Therefore, TCR-ab T cells can only be identified indirectly
by a combination of antibodies against CD3 and TCR-gd, as shown
in Fig. 1. Nevertheless, even before specific mAbs forgd-T cells had
been developed, a combination of functional assays and phenotyp-
ing experiments indicated that porcine TCR-ab T cells are
[4,12,32,58]. However, if a population of
Fig. 4. Comparison of frequency and phenotype of TCR-gd T cells from 6-month- and 2-year-old swine. The frequency of TCR-gd T cells within total T cells decreases strongly
with age as illustrated in histogram (A), which represents a 6-month-old pig, vs. histogram (E), showing TCR-gd expression of T cells from 2-year-old swine. If these TCR-gd T
cells arefurther analyzed forCD8a, MHC-classII DRand CD45RCexpression,itisrevealed thattheremaininggd-Tcells inthe2-year-old pignearly entirelyexpress CD8a(F),
whereas TCR-gd T cells from the 6-month-old pig are in their majority CD8a?(B). As already outlined in Fig. 3, this CD8a expression is often correlated with MHC-class II DR
expression, since all MHC-class II DR+gd-T cells co-express CD8a, regardless of the animals' age (C vs. G). Also, no obvious age-related changes are found for CD45RC
expression, with the majority of TCR-gd T cells belonging to the CD45RC population (D vs. H).
W. Gerner et al./Developmental and Comparative Immunology 33 (2009) 310–320
Page 7
CD3+TCR-gd?PBLs is analyzed for expression of CD4 and CD8a,
four subpopulations can be identified: CD4?CD8a+, CD4+CD8a?,
CD4+CD8a+and a very small fraction of CD4?CD8a?cells (Fig. 1G).
The existence of a considerable fraction of CD4+CD8a+T cells in
extrathymic sites is a peculiarity of the porcine immune system
described already in the late 1980s [30,32,59]. The frequency of
this cell population is quite variable, ranging from less than 5% in
gnotobiotic piglets up to 80% (of CD4+T cells) in adult swine. The
four CD4/CD8a defined subpopulations can be further character-
ized by analysis of the expression of CD8b (Fig. 5B), MHC-class II
DR (Fig. 5C) and CD45RC (Fig. 5D), respectively. For CD8b
expression (Fig. 5B) at least a positive and a negative population
can be found within porcine TCR-ab T cells with considerable
animal to animal variation in regard to fractions of CD8b?and
CD8b+cells as well as in the expression profile of CD8b, even
between animals from the same age (Gerner, unpublished results).
If CD8b+cells are analyzed for CD4/CD8a expression, it is revealed
that nearly all of these cells belong to the CD4?CD8ahighfraction
(Fig. 5F), which contains cytolytic T lymphocytes (see below). In
contrast, CD8b?TCR-ab T cells represent the remaining CD4+and
CD4?CD8a?cells, as well as a varying population of CD4?CD8a+
lymphocytes (Fig. 5E) with currently unknown function. MHC-
classII DR expression (Fig. 5C) is also found in a subpopulation of
porcine TCR-ab T cells with an antigen density comparable to the
antigen density on TCR-gd T cells (Fig. 3C). However, usually no
clear distinction between MHC-classII DR positive and negative
cells is found. If MHC-class II DR+cells are analyzed for their CD4/
CD8a expression, an enrichment of these cells within the
CD4+CD8a+and CD4?CD8a+populations becomes identifiable
(Fig. 5H). In contrast, nearly the entire CD4+CD8a?population is
MHC-class II (Fig. 5G). However, to a considerable extent MHC-
class II cells also exist in the CD4+CD8a+and CD4?CD8a+
populations (Fig. 5G), indicating a heterogeneity in regard to
MHC-class II DR expression within these two subpopulations. A
similar diversity is found for CD45RC expression; however, here
most TCR-ab T cells are positive for this marker and show a high
antigen density (Fig. 5D). CD45RC?lymphocytes are found
primarily in the CD4+CD8a+subset, but to a lesser extent these
cells also belong to CD4?CD8a+and CD4+CD8a?subpopulations
(Fig. 5I).
A further marker for CD4+T-helper cells has been described by
Revilla et al. recently [60]. In this study a mAb named 2E3 was
shown to label a subpopulation of CD3+CD4+T cells but not
myeloid, B cells or NK cells. The antigen was partially expressed on
CD4+CD8a+cells, but the majority of 2E3+-labeled cells belonged
to the CD4+CD8a?subpopulation. In addition, CD4+2E3+T cells
were in their majority CD45RA+MHC-class II DR?CD29?. In regard
to the identification of TCR-ab T cells with memory function in
addition to the already mentioned markers for CD45 isoforms, a
cross-reactive antibody recognizing porcine CD29 is available [61].
Fig. 5. Analysis of CD8b, MHC-class II DR and CD45RC expression in porcine TCR-ab T cells. CD3+TCRgd?T cells (A) as outlined in Fig. 1 were analyzed for CD4, CD8a, CD8b,
MHC-classIIDRandCD45RCexpression,respectively.ThehistogramsshowthatTCR-abTcellsshowaheterogeneous expressionofCD8b(B).Ifthesecellsareseparatedinto
CD8b+and CD8b?subpopulations and analyzed for CD4 and CD8a expression, it is revealed that CD8b+TCR-ab T cells are CD4?CD8ahigh(F), which represent the subset of
MHC-class I restricted cytolytic T cells. The remaining CD8b?cells comprise nearly all CD4+ T-helper cells and small fractions of CD4?CD8a?and CD4?CD8a+cells (E), both
with currently unknown function. MHC-class II DR expression in porcine TCR-ab T cells is also heterogeneous (C). If MHC-class II DR+and MHC-class II DR?T cells are
analyzed for CD4/CD8a expression, it is revealed that MHC-class II DR+cells in their vast majority belong either to the CD4+CD8a+or to the CD4?CD8ahighpopulation (H). In
contrast, the MHC-class II DR?subpopulation comprises nearly all CD4+CD8a?T-helper cells but also subgroups of CD4+CD8a+and CD4?CD8ahighT lymphocytes. CD45RC
expression again varies fromabsence to veryhigh expressionlevels, withthe majorityof cellsbelongingto theCD45RChighgroup(D). IfCD45RChighcells areanalyzed forCD4/
CD8a expression, a dominance of CD4+CD8a+cells within this subpopulation becomes visible, but also CD4?CD8a+T cells can be CD45RC?(I). CD45RC+cells, however,
comprise all four CD4/CD8a defined T-cell subpopulations (J). Presented data are representative for blood-derived lymphocytes from 6-month-old swine.
W. Gerner et al./Developmental and Comparative Immunology 33 (2009) 310–320
Page 8
T cells with a high CD29 expression were shown to be enriched in
the CD4+CD8a+subpopulation.
3.6. Functional characterization of TCR-ab T cells
A number of studies aimed to combine these various
subpopulations with functional analyses. Early studies showed
that a separation of porcine lymphocytes into CD6+and CD6?
subsets leads to an accumulation of cells with spontaneous lytic
activity in the CD6?fraction, which comprises NK cells and TCR-gd
T cells, whereas allo- and antigen-specific lysis of CSFV- and PRV-
infected targets was observed in the CD6+subset, consisting
mainly of CD3+TCR-ab+T lymphocytes [12,62,63]. A separation of
CD4?CD8a+lymphocytes into CD5+and CD5?subsets also led to
an enrichment of allo-reactive cells in the CD5+population [4]. In
addition, antigen specific lysis of CSFV-infected targets was found
to be performed by CD4?cells [62]. This led to the notion that
cytolytic T cells in swine are CD4?CD5+CD6+CD8ahigh[64].
However, in a newer study a lytic activity of CD4+CD6+
lymphocytes against ASFV-infected syngeneic target cells was
described [8]. Interestingly, the specific lysis induced by these cells
was lower when compared to the same number of CD4?CD6+
effector cells used in an identical experimental setup. These
ab T cells, identifying this molecule in the CD3+CD6+CD8b+T-cell
CD4+T-cell subset with the phenotype CD3+CD4+CD6+CD8a+.
Furthermore, this study also described perforin+CD3+CD6?T cells
which showed a spontaneous lytic activity against K562 and P815
target cells after in vitro activation with IL-2, therefore resembling
lymphokine-activated killer cells or activated NKT cells. In
summary, these data indicate a considerable phenotypical
heterogeneity of porcine TCR-ab T-cell subsets capable of
cytolysis, but suggest that the majority of cytolytic TCR-ab T
cells representing the classical MHC-class I-restricted cytolytic
T-cell subsetinswinecan
CD2+CD3+CD4?CD5highCD6+CD8ahighCD8b+T-lymphocyte subpo-
pulation. Conclusive data on the phenotype of cytolytic memory
TCR-ab T cells are lacking so far, but these cells might be CD45RC?
as established for porcine T-helper cells (see below, Fig. 5I).
The CD4+lymphocyte subset has also been investigated in a
number of studies with the aim to identify correlations between
function and the phenotype of these cells. Especially the already-
described CD8a co-expression on a substantial proportion of CD4+
cells has received much attention. Functional studies on sorted
CD4+CD8a+and CD4+CD8a?cells showed that both subpopula-
tions proliferated after stimulation in allogeneic co-cultures but
only the CD4+CD8a+population responded by proliferation in a
recall assay to PRV [65]. In addition, this antigen-specific response
was MHC-class II restricted. This was a first indication that the
porcine memory T-helper cells are included in the CD4+CD8a+T-
cell subpopulation. In a consecutive study it could also be shown
that PRV-specific CD4+CD8a+cells proliferated after stimulation
with a PRV-derived peptide, produced IL-2 and provided help to B
cells for antibody production [66].
Further support for the assumption that the CD4+CD8a+
subpopulation is formed by T-helper memory cells came from
the observation that in newborn piglets this population is nearly
absent but thereafter steadily increases with age [25,61,67–69],
obviously reflecting the gradual build-up of immunological
memory after enduring contact with various antigens. However,
it has also been shown that in vitro stimulation of CD4+CD8a?T
cells by superantigens or polyclonal stimuli led to an up-regulation
both activated and memory T-helper cells in swine belong to the
beallocated to the
CD4+CD8a+population [11]. This is in contrast to reported data
from mice and humans showing that in these species some CD8+
peripheral T cells have the ability to express CD4 upon polyclonal
or antigenic stimulation in vitro and in vivo [71–73]. Functional
analyses suggested that these CD4+CD8+still have a cytolytic
activity since CD4 ligation led to an up-regulation of FasL
expression [71]. As outlined above, in swine some CD4+CD8a+T
cells with cytolytic activity also seem to exist; however, these cells
form only a small subset within the entire CD4+CD8a+T-cell
A combination of functional assays with phenotypic analyses
also showed that T-helper cells responding to polyclonal stimula-
tion become MHC-class II+but lose CD45RC expression, and that
the same phenotype (CD4+CD8a+MHCII+CD45RC?) is found on
proliferating cells in antigen-specific in vitro recall responses
against CSFV [11] and FMDV-derived peptides [74,75]. Whether
the expressed MHC-class II DR molecules on activated/memory T-
helper cells have a functional role (as discussed for gd-T cells, see
above) is currently unknown.
Also, in vitro recall assays showedthat the expression of the 2E3
antigen (see above) seems to be restricted to naive T-helper cells,
since antigen-specific proliferation was found in a sorted
CD4+2E3?cell fraction but not in the CD4+2E3+population [60].
Interestingly, in a consecutive study the assumption that MHC-
class II expression is found on the entire T-helper memory
population has been challenged since it was demonstrated that
sorted CD4+2E3?MHCII?cells contain a minor fraction of cells
which can respond to their specific antigen by proliferation in vitro
[76]. In addition, the same laboratory showed recently that within
PBLs CD4+2E3?cells express mRNA for the chemokine receptors
CXCR3 and CCR4 but not CCR7, whereas the latter was found in
CD4+2E3+cells [77]. CCR7, which is found on human naive and
central memory T-helper cells (reviewed in [78]), could therefore
not be readily detected in a porcine cell population containing
memory T cells. Whether this means that central memory T cells
do not exist in pigs, do not express CCR7, or can only be found in
certain lymphaticcompartmentsand/or during certain time points
after infection awaits clarification. Also, CXCR3, which is usually
found on human TH1 cells, and CCR4 expressed by human TH2 cells
[79,80], were both found on CD4+2E3?cells, providing no further
insight into the potential polarization of T-helper cells in pigs.
In summary, our current knowledge on CD4+porcine T-helper
cells suggests that these cells are MHC-class II-restricted and up-
regulate CD8a and MHC-class II DR upon activation but lose
CD45RC expression. In general, memory T-helper cells seem to
retain this phenotype (CD8a+CD45RC?MHCII+), although may be
not all of them constitutively express MHC-class II DR+.
3.7. Regulatory T cells (Tregs)
Besides the various subpopulations of T lymphocytes with
cytolytic activity and T lymphocytes with the function of T-helper
species, these Tregs seem to have the ability to down-regulate
immune responses. In swine Tregs are primarily defined as
CD4+CD25+T lymphocytes expressing the transcription factor
Foxp3 [82]. In T lymphocytes derived from blood and secondary
lymphoid tissues two main subsets of this minor subpopulation
(5–10% of the CD4+cell population) can be discriminated by their
expression of CD25 detected by the mAb 3B2 [83]: CD4+cells with
low (CD4+CD25dim) and CD4+cells with high CD25 (CD4+CD25high)
antigen density. With regard to other differentiation antigens
(CD8a, CD45RC and MHC-class II DR), both subsets showed very
W. Gerner et al./Developmental and Comparative Immunology 33 (2009) 310–320
Page 9
of further Treg subsets. To distribute specific functions to the
respective subsets will be the content of further research. To date
functional differences between the CD4+CD25-defined Treg sub-
sets – either CD25dimor CD25high– are obvious. Whereas both
CD25+T-cell subsets produced IL-10 after polyclonal stimulation,
only the CD4+CD25highsubset showed a cell-dependent inhibitory
activity against the proliferative response of polyclonal activated T
cells [81].
4. Summary and outlook: peculiarities of porcine T
lymphocytes and NK cells
Analyses of porcine PBLs for the existence of major subpopula-
tions revealed strong similarities to other species, but also some
swine-specific characteristics. The two most prominent, the
abundant surface expression of CD8a and the expression of
MHC-class II DR molecules on resting T lymphocytes, will be
discussed in more detail below.
4.1. Expression of CD8a
a molecular mass of 33–35 kDa under reducing and 66–70 kDa
under non-reducing conditions ([31,84] summarized by [85]).
With a panel of CD8-specific mAbs, different epitopes on the CD8-
dimers could be defined and these epitopes could be distributed to
two different chains of the molecule. They are located either on the
CD8a or on the CD8b chain [54,86]. The expression of CD8ab
heterodimers is comparable to the expression of CD8 on T
lymphocytes of other species. These CD8a heterodimers char-
acterize also in swine MHC-class I-restricted cytolytic T lympho-
cytes (reviewed in [64]). In contrast to all species analyzed so far
stands the expression of the CD8a chain on porcine lymphocytes.
Porcine lymphocytes derived either from blood or from lymphoid
tissue show a high percentage of cells with surface expression of
CD8a (see above, and [30]). These molecules are mostly formed by
CD8a homodimers, but it cannot be excluded that a minor fraction
of CD8a molecules is expressed either as monomer or as
aggregates with more than two chains (Saalmu ¨ller, unpublished
results). Furthermore, these CD8a homodimers might be co-
expressed together with CD8ab heterodimers on the same cell
within distinct populations of CD3+CD4?CD8b+TCR-ab T lym-
phocytes (Gerner, unpublished results). The putative functions of
such CD4?TCR-ab T cells expressing either solely CD8aa
homodimers or a mixture of homo- and heterodimers are
unknown. Clearly, the biological relevance of the abundant
expression of CD8a molecules on a substantial proportion of
T cells [38] and CD3?CD4?NK cells [3,4,12] is an open question.
4.2. Expression of CD8a on NK-cells and TCR-gd T cells
The role of CD8a expression in NK cells is not clear, but the
molecules seem not to be involved in the recognition of target cells
or control of NK-cell function (Saalmu ¨ller, unpublished results).
Also not clear is the role of CD8a expression on TCR-gd T
lymphocytes. With regard to the differentiation, maturation and
the age-dependent changes in the expression of the respective
surface molecules (see Fig. 4), one can speculate that the
expression of CD8a on TCR-gd T lymphocytes is activation-
dependent and that CD8?TCR-gd T cells acquire CD8a during
activationand furthermaturation.Thiseffect couldbe simulatedin
vitro, where CD8-negative TCR-gd T cells stimulated with
Concanavalin A showed an enhanced CD8a expression after
several days of cultivation (Saalmu ¨ller, unpublished results).
4.3. Expression of CD8a on TCR-ab T cells
Similar results were obtained with CD4+T cells which became
CD8a+upon stimulation and in vitro cultivation [11,30]. It seems
that this phenomenon happens also in vivo. This hypothesis could
be verified and confirmed by a panel of publications in which
antigen-specific recall immune responses of memory T-helper
cells were distributed to the CD4+CD8a+T-cell subpopulation (see
above and [11,65,66,69,74,75]). Therefore it seems that the
unusual CD8a expression in swine depends on prior activation
of the respective cells and CD8a surface expression on lympho-
cytes seems to represent a marker correlating with individual
immunological history and at least partially with the acquisition of
memory function. However, the detailed mechanisms and the
immunological function of the CD8a expression are still not clear.
Obviously, these points might be content of further research.
5. Expression of MHC-class II DR on porcine lymphocytes
Another unexpected peculiarity of the immune system of swine
is the expression of MHC-class II DR molecules on a substantial
proportion of resting lymphocytes [10]. Besides the known
expression on B cells with high antigen density, this MHC-class
II DR expression can be found on NK cells, TCR-gd, as well as on
TCR-ab T lymphocytes. The expression of MHC-class II molecules
[87,88], but was not described before for resting T lymphocytes
II DR expression with horse, for which an age-dependent increase
in the MHC-class II positive T-cell population was described [89].
5.1. MHC-class II DR expression on porcine NK cells
Porcine NK cells show a heterogeneous MHC-class II expression
pattern. There are individual differences in the frequency of MHC-
class II DR+NK cells ranging from a very low amount of positive NK
cells (see Fig. 2) up to 80% MHC-class II DR positive NK-cell
fractions (Gerner, unpublished results). Nothing is known about
the detailed functions of the respective cells, but it could be shown
by Pintaric et al. [9] that this MHC-class II DR expression could be
induced in vitro after stimulation of the NK cells with a cocktail of
cytokines. Therefore one can speculate that the heterogeneity
detected ex vivo can be explained by a possible in vivo stimulation
and activation of the NK-cell subset, and that these MHC-class II
defined NK-cell fractions might have different in vivo functions or
are regulated in a different manner. However, data showing these
differences are currently missing.
5.2. MHC-class II DR expression on TCR-gd T lymphocytes
WithregardtoTCR-gdTcells, theexpressionofMHC-classII DR
seems to correlate with the CD8a expression and is influenced by
the age of the respective animals (Figs. 3 and 4). As mentioned
above, resultsfrom infectionexperiments studyingthe roleof TCR-
gd T cells postulated a memory population of TCR-gd T cells
[50,55]. One can speculate that this MHC-class II DR+TCR-gd cell
fraction is responsible for the antigen-specific recall reactivity. A
more detailed elucidation of this phenomenon has to be the
content of further research activities.
MHC-class II DR expression is normally correlated with antigen
presentation and this role in the immune response could also be
demonstrated for MHC-class II DR+TCR-gd T cells by Takamatsu
et al. [56], who assigned this cell population a function as
professional antigen-presenting cells. In conclusion, MHC-class II
DR+TCR-gd T lymphocytes may cover a broad spectrum of
W. Gerner et al./Developmental and Comparative Immunology 33 (2009) 310–320
Page 10
immunological reactivity. They are involved in antigen presenta-
tion, a function combined with the onset of immune responses
mainly distributed to the innate immune system, and they also
seem to play a role in memory T-cell reactions, response
mechanisms of the highly specialized adaptive immune system.
This shows once more the puzzling plasticity of this T-cell
5.3. MHC-class II expression on TCR-ab T lymphocytes
MHC-class II DR expression on TCR-ab T lymphocytes seems to
correlate with the expression of CD8a, but there is no strict
overlap. Indeed MHC-class II DR+cells are mainly located in the
CD4?CD8a+and CD4+CD8a+T-cell subpopulations (Fig. 5H), but,
on the other hand, also MHC-class II DR?subsets of these
subpopulations exist (Fig. 5G). As mentioned for the other
lymphocyte populations, the expression of MHC-class II DR on
TCR-ab T cells can be as well induced by in vitro activation and the
surface expression of already MHC-class II DR+T cells can be
further increased [10,11]. Also interesting is the observation that
highly purified MHC-class II DR+T lymphocytes can act in vitro as
stimulator cells in an allogeneic mixed leukocyte culture, which
might indicate a possible role of TCR-ab T cells as professional
antigen-presenting cells [90], but these preliminary findings need
further elucidation in an antigen-specific immune reaction.
Nevertheless, it can be discussed that MHC-class II DR+T
lymphocyteswhichseem to be generatedduring previous immune
responses by de novo surface expression of MHC-class II DR may
represent a cell fraction containing memory T cells and cells with
immuno-regulatory function with high importance in a secondary
recall immune response.
6. Other T-lymphocyte subpopulations
During the last few years, in various species other minor T-cell
subpopulations have been defined, including Tregs, NKT cells and
TH17 cells. As mentioned above, the existence of Tregs in swine
could be shown and the functional activity of this minor T-cell
subset demonstrated [81,82]. With regard to NKT cells, Denyer
et al. [8] were able to define them phenotypically by their ability of
perforin expression. TH17 cells characterized by their IL-17
synthesis represent a topic for further research. Furthermore,
there are several open questions about the existence and
phenotype of memory T cells and their various subsets with
distinct functional activities. Therefore, the porcine immune
system and especially porcine T lymphocytes still represent a
wide-open field for further research activities.
[1] Lunney JK. Characterization of swine leukocyte differentiation antigens.
Immunol Today 1993;14:147–8.
[2] Saalmu ¨ller A. Characterization of swine leukocyte differentiation antigens.
Immunol Today 1996;17:352.
[3] Pescovitz MD, Lowman MA, Sachs DH. Expression of T-cell associated antigens
by porcine natural killer cells. Immunology 1988;65:267–71.
[4] Saalmu ¨ller A, Hirt W, Maurer S, Weiland E. Discrimination between two
subsets of porcine CD8+ cytolytic T lymphocytes by the expression of CD5
antigen. Immunology 1994;81:578–83.
[5] Dato ME, Kim YB. Characterization and utilization of a monoclonal antibody
inhibiting porcine natural killer cell activity for isolation of natural killer and
killer cells. J Immunol 1990;144:4452–62.
[6] Halloran PJ, Sweeney SE, Strohmeier CM, Kim YB. Molecular cloning and
identification of the porcine cytolytic trigger molecule G7 as a Fc gamma RIII
alpha (CD16) homologue. J Immunol 1994;153:2631–41.
[7] Aller SC, Cho D, Kim YB. Characterization of the cytolytic trigger molecules G7/
PNK-E as a molecular complex on the surface of porcine phagocytes. Cell
Immunol 1995;161:270–8.
[8] Denyer MS, Wileman TE, Stirling CM, Zuber B, Takamatsu HH. Perforin
expression can define CD8 positive lymphocyte subsets in pigs allowing
phenotypic and functional analysis of natural killer, cytotoxic T, natural killer
T and MHC un-restricted cytotoxic T-cells. Vet Immunol Immunopathol
[9] Pintaric M, Gerner W, Saalmu ¨ller A. Synergistic effects of IL-2, IL-12 and IL-18
on cytolytic activity, perforin expression and IFN-g production of porcine
natural killer cells. Vet Immunol Immunopathol 2008;121:68–82.
[10] Saalmu ¨ller A, Weiland F, Reddehase MJ. Resting porcine T lymphocytes
expressingclassII majorhistocompatibility
[11] Saalmu ¨ller A, Werner T, Fachinger V. T-helper cells from naive to committed.
Vet Immunol Immunopathol 2002;87:137–45.
[12] Pauly T, Weiland E, Hirt W, Dreyer-Bux C, Maurer S, Summer-field A, et al.
Differentiation between MHC-restricted and non-MHC-restricted porcine
cytolytic T lymphocytes. Immunology 1996;88:238–46.
[13] Piriou L, Chilmonczyk S, Genetet N, Albina E. Design of a flow cytometric assay
for the determination of natural killer and cytotoxic T-lymphocyte activity in
human and in different animal species. Cytometry 2000;41:289–97.
[14] Martin S, Wardley RC. Natural cytotoxicity detected in swine using Aujeszky's
disease virus infected targets. Res Vet Sci 1984;37:211–8.
[15] Cepica A, Derbyshire JB. Characterisation of the effector cells in antibody-
cells infected with transmissible gastroenteritis virus. Res Vet Sci 1986;41:
[16] Richerson JT, Misfeldt ML. Host environment as a modulating factor of swine
natural killer cell activity. Vet Immunol Immunopathol 1989;23:309–19.
[17] Pauly T, Ko ¨nig M, Thiel HJ, Saalmu ¨ller A. Infection with classical swine fever
virus: effects on phenotype and immune responsiveness of porcine T lym-
phocytes. J Gen Virol 1998;79:31–40.
[18] Charley B, Petit E, La BC. Interferon-induced enhancement of newborn pig
natural killing (NK) activity. Ann Rech Vet 1985;16:399–402.
[19] Takamatsu H, Koide F. Augmentation of porcine spontaneous cell-mediated
cytotoxicity in vitro. Nippon Juigaku Zasshi 1985;47:749–59.
[20] Cho D, Lee WJ, Halloran PJ, Trinchieri G, Kim YB. Enhancement of porcine
natural killer cell activity by recombinant human and murine IL-12. Cell
Immunol 1996;172:29–34.
[21] Knoblock KF, Canning PC. Modulation of in vitro porcine natural killer cell
activity by recombinant interleukin-1 alpha, inter-leukin-2 and interleukin-4.
Immunology 1992;76:299–304.
[22] Domeika K, Berg M, Eloranta ML, Alm GV. Porcine interleukin-12 fusion
protein and interleukin-18 in combination induce interferon-gamma produc-
tion in porcine natural killer and T cells. Vet Immunol Immunopathol
[23] Sinkora M, Sinkora J, Rehakova Z, Butler JE. Early ontogeny of thymocytes in
pigs: sequential colonization of the thymus by Tcell progenitors. J Immunol
[24] Hirt W, Saalmu ¨ller A, Reddehase MJ. Expression of gamma/delta Tcell recep-
tors in porcine thymus. Immunobiology 1993;188:70–81.
[25] Yang H, Parkhouse RM. Phenotypic classification of porcine lymphocyte sub-
populations in blood and lymphoid tissues. Immunology 1996;89:76–83.
[26] Sinkora M, Sinkorova J, Holtmeier W. Development of gd thymocyte subsets
during prenatal and postnatal ontogeny. Immunology 2005;115:544–55.
[27] SinkoraM,SinkorovaJ,CimburekZ,HoltmeierW.TwogroupsofporcineTCRgd+
thymocytes behave and diverge differently. J Immunol 2007;178:711–9.
[28] Scollay R, Bartlett P, Shortman K. T cell development in the adult murine
thymus: changes in the expression of the surface antigens Ly2, L3T4 and B2A2
during development from early precursor cells to emigrants. Immunol Rev
[29] Lanier LL, Allison JP, Phillips JH. Correlation of cell surface antigen expression
on human thymocytes by multi-color flow cytometric analysis: implications
for differentiation. J Immunol 1986;137:2501–7.
[30] Saalmu ¨ller A, Reddehase MJ, Bu ¨hring HJ, Jonjic S, Koszinowski UH. Simulta-
neous expression of CD4 and CD8 antigens by a substantial proportion of
resting porcine T lymphocytes. Eur J Immunol 1987;17:1297–301.
[31] Pescovitz MD, Lunney JK, Sachs DH. Preparation and characterization of mono-
clonal antibodies reactive with porcine PBL. J Immunol 1984;133:368–75.
[32] Saalmu ¨ller A, Hirt W, Reddehase MJ. Phenotypic discrimination between
thymic and extrathymic CD4?CD8?and CD4+CD8+porcine T lymphocytes.
Eur J Immunol 1989;19:2011–6.
[33] Dawson HD, Royaee AR, Nishi S, Kuhar D, Schnitzlein WM,Zuckermann F, et al.
Identification of key immune mediators regulating T helper 1 responses in
swine. Vet Immunol Immunopathol 2004;100:105–11.
[34] Royaee AR, Husmann RJ, Dawson HD, Calzada-Nova G, Schnitzlein WM,
Zuckermann FA, et al. Deciphering the involvement of innate immune factors
in the development of the host response to PRRSV vaccination. Vet Immunol
Immunopathol 2004;102:199–216.
[35] Dawson HD, Beshah E, Nishi S, Solano-Aguilar G, Morimoto M, Zhao A, et al.
Localized multigene expression patterns support an evolving Th1/Th2-like
paradigm in response to infections with Toxoplasma gondii and Ascaris suum.
Infect Immun 2005;73:1116–28.
[36] Aasted B, Bach P, Nielsen J, Lind P. Cytokine profiles in peripheral blood
mononuclear cells and lymph node cells from piglets infected in utero with
porcine reproductiveand respiratory syndromevirus. Clin Diagn Lab Immunol
W. Gerner et al./Developmental and Comparative Immunology 33 (2009) 310–320
Page 11
[37] Sipos W, Duvigneau JC, Pietschmann P, Schilcher F, Hofbauer G, Hartl RT, et al.
Porcine dermatitis and nephropathy syndrome (PDNS) is associated with a
systemic cytokine expression profile indicative of proinflammation and a Th1
bias. Vet Immunol Immunopathol 2005;107:303–13.
[38] Davis WC, Zuckermann FA, Hamilton MJ, Barbosa JI, Saalmu ¨ller A, Binns RM,
et al. Analysis of monoclonal antibodies that recognize gd T/null cells. Vet
Immunol Immunopathol 1998;60:305–16.
[39] Tang WR, Shioya N, Eguchi T, Ebata T, Matsui J, Takenouchi H, et al. Char-
acterization of new monoclonal antibodies against porcine lymphocytes:
molecular characterization of clone 7g3, an antibody reactive with the con-
stant region of the T-cell receptor d-chains. Vet Immunol Immunopathol
[40] Yang H, Parkhouse RM. Characterization of the porcine gammadelta T-cell
receptor structure and cellular distribution by monoclonal antibody PPT27.
Immunology 2000;99:504–9.
[41] Yang H, Parkhouse RM, Wileman T. Monoclonal antibodies that identify the
CD3 molecules expressed specifically at the surface of porcine gd-T cells.
Immunology 2005;115:189–96.
[42] Davis WC, Haverson K, Saalmu ¨ller A, Yang H, Lunney JK, Hamilton MJ, et al.
Analysis ofmonoclonal antibodiesreacting withmolecules expressed ongd T-
cells. Vet Immunol Immunopathol 2001;80:53–62.
[43] Takamatsu HH, Denyer MS, Stirling C, Cox S, Aggarwal N, Dash P, et al. Porcine
gd T cells: possible roles on the innate and adaptive immune responses
following virus infection. Vet Immunol Immunopathol 2006;112:49–61.
[44] Charerntantanakul W, Roth JA. Biology of porcine T lymphocytes. Anim Health
Res Rev 2006;7:81–96.
[45] Saalmu ¨ller A, Hirt W, Reddehase MJ. Porcine g/d T lymphocyte subsets
differing in their propensity to home to lymphoid tissue. Eur J Immunol
[46] Hirt W, Saalmu ¨ller A, Reddehase MJ. Distinct g/d T cell receptors define two
subsets of circulating porcine CD2?CD4?CD8?T lymphocytes. Eur J Immunol
[47] Thielke KH, Hoffmann-Moujahid A, Weisser C, Waldkirch E, Pabst R, Holtmeier
W, et al. Proliferating intestinal g/d Tcells recirculate rapidly and are a major
source of the g/d Tcell pool in the peripheral blood. Eur J Immunol
[48] Hayday AC.gd Cells: a right time and a right place for a conserved third way of
protection. Annu Rev Immunol 2000;18:975–1026.
[49] Summerfield A, Saalmu ¨ller A. Interleukin-2 dependent selective activation of
porcine gd T lymphocytes by an extract from the leaves of Acanthospermum
hispidum. Int J Immunopharmacol 1998;20:85–98.
[50] Lee J, Choi K, Olin MR, Cho SN, Molitor TW. gd T cells in immunity induced by
Mycobacterium bovis bacillus Calmette—Guerin vaccination. Infect Immun
[51] Blumerman SL, Herzig CT, Baldwin CL. WC1+ gd Tcell memory population is
induced by killed bacterial vaccine. Eur J Immunol 2007;37:1204–16.
[52] Morita CT, Jin C, Sarikonda G, Wang H. Nonpeptide antigens, presentation
mechanisms, and immunological memory of human Vg2Vd2 T cells: discri-
minating friend from foe through the recognition of prenyl pyrophosphate
antigens. Immunol Rev 2007;215:59–76.
[53] CardingSR, EganPJ.gdT cells:functional plasticity and heterogeneity. NatRev
Immunol 2002;2:336–45.
[54] Yang H, Parkhouse RM. Differential expression of CD8 epitopes amongst
porcine CD8-positive functional lymphocyte subsets. Immunology 1997;92:
[55] Olin MR, Hwa CK, Lee J, Molitor TW. gd T-lymphocyte cytotoxic activity
against Mycobacterium bovis analyzed by flow cytometry. J Immunol Methods
[56] Takamatsu HH, Denyer MS, Wileman TE. A sub-population of circulating
porcine gd T cells can act as professional antigen presenting cells. Vet
Immunol Immunopathol 2002;87:223.
[57] Brandes M, Willimann K, Moser B. Professional antigen-presentation function
by human gd T cells. Science 2005;309:264–8.
[58] Hammerberg C, Schurig GG. Characterization of monoclonal antibodies direc-
ted against swine leukocytes. Vet Immunol Immunopathol 1986;11(107):21.
[59] Pescovitz MD, Lunney JK, Sachs DH. Murine anti-swine T4 and T8 monoclonal
antibodies: distribution and effects on proliferative and cytotoxic T cells. J
Immunol 1985;134:37–44.
[60] Revilla C, Rodriguez-Carreno MP, Alvarez B, Chamorro S, Alonso LM, Ezquerra
A, et al. 2E3, a new marker that selectively identifies porcine CD4+ naive T
cells. Dev Comp Immunol 2004;28:239–50.
[61] Zuckermann FA, Husmann RJ. Functional and phenotypic analysis of porcine
peripheral blood CD4/CD8 double-positive Tcells. Immunology 1996;87:
[62] Pauly T, Elbers K, Ko ¨nig M, Lengsfeld T, Saalmu ¨ller A, Thiel HJ. Classical swine
fever virus-specific cytotoxic T lymphocytes and identification of a Tcell
epitope. J Gen Virol 1995;76:3039–49.
[63] De Bruin TG, VanRooij EM,De VisserYE, Voermans JJ, Samsom JN, Kimman TG,
et al. Discrimination of different subsets of cytolytic cells in pseudorabies
virus-immune and naive pigs. J Gen Virol 2000;81:1529–37.
[64] Saalmu ¨ller A, Pauly T, Hohlich BJ, Pfaff E. Characterization of porcine T
lymphocytes and their immune response against viral antigens. J Biotechnol
[65] Summerfield A, Rziha HJ, Saalmu ¨ller A. Functional characterization of porcine
CD4+CD8+ extrathymic T lymphocytes. Cell Immunol 1996;168:291–6.
[66] Ober BT, Summerfield A, Mattlinger C, Wiesmu ¨ller KH, Jung G, Pfaff E, et al.
Vaccine-induced, pseudorabies virus-specific, extrathymic CD4+CD8+ mem-
ory T-helper cells in swine. J Virol 1998;72:4866–73.
[67] Pescovitz MD, Sakopoulos AG, Gaddy JA, Husmann RJ, Zuckermann FA. Porcine
peripheral blood CD4+/CD8+ dual expressing T-cells. Vet Immunol Immuno-
pathol 1994;43:53–62.
[68] Grierson SS, King DP, Tucker AW, Donadeu M, Mellencamp MA, Haverson K,
et al. Ontogeny of systemic cellular immunity in the neonatal pig: correlation
with the development of post-weaning multisystemic wasting syndrome. Vet
Immunol Immunopathol 2007;119:254–68.
[69] Zuckermann FA. Extrathymic CD4/CD8 double positive Tcells. Vet Immunol
Immunopathol 1999;72:55–66.
[70] Fachinger V, Schlapp T, Saalmu ¨ller A. Evidence for a parapox ovis virus-
associated superantigen. Eur J Immunol 2000;30:2962–71.
[71] Kitchen SG,Jones NR, LaForge S,Whitmire JK,Vu BA,GalicZ,et al.CD4 onCD8+
Tcells directly enhances effector function and is a target for HIV infection. Proc
Natl Acad Sci USA 2004;101:8727–32.
[72] Kitchen SG, Whitmire JK, Jones NR, Galic Z, Kitchen CMR, Ahmed R, et al. The
CD4 moleculeonCD8+T lymphocytesdirectly enhancestheimmuneresponse
to viral and cellular antigens. Proc Natl Acad Sci USA 2005;102:3794–9.
[73] Zloza A, Sullivan YB, Connick E, Landay AL, Al-Harthi L. CD8+T cells that
express CD4 on their surface (CD4dimCD8brightTcells) recognize an antigen-
specific target, are detected in vivo, and can be productively infected by T-
tropic HIV. Blood 2003;102:2156–64.
[74] GernerW,DenyerMS,TakamatsuHH,WilemanTE,Wiesmu ¨llerKH,PfaffE,etal.
c and d/d haplotype miniature swine. Virus Res 2006;121:223–8.
[75] Blanco E, McCullough K, Summerfield A, Fiorini J, Andreu D, Chiva C, et al.
Interspecies major histocompatibility complex-restricted Th cell epitope on
foot-and-mouth disease virus capsid protein VP4. J Virol 2000;74:4902–7.
[76] Revilla C, Chamorro S, Alvarez B, Perez C, Ezquerra A, Alonso F, et al. Analysis of
functional heterogeneity of porcine memory CD4+ Tcells. Dev Comp Immunol
[77] Revilla C, Alvarez B, Lopez-Fraga M, Chamorro S, Martinez P, Ezquerra A, et al.
Differential expression of chemokine receptors and CD95 in porcine CD4+
Tcell subsets. Vet Immunol Immunopathol 2005;106:295–301.
[78] Sallusto F, Geginat J, Lanzavecchia A. Central memory and effector memory T
cell subsets: function, generation, and maintenance. Annu Rev Immunol
[79] Moser B, Wolf M, Walz A, Lotscher P. Chemokines: multiple levels of leukocyte
migration control. Trends Immunol 2004;25:75–84.
[80] Kim CH, Rott L, Kunkel EJ, Genovese MC, Andrew DP, Wu L, et al. Rules of
chemokine receptor association with T cell polarization in vivo. J Clin Invest
[81] Ka ¨ser T, Gerner W, Hammer SE, Patzl M, Saalmu ¨ller A. Phenotypic and
functional characterisation of porcine CD4+CD25highregulatory T cells. Vet
Immunol Immunopathol 2008;122:153–8.
[82] Ka ¨ser T, Gerner W, Hammer SE, Patzl M, Saalmu ¨ller A. Detection of Foxp3
protein expression in porcine T lymphocytes. Vet Immunol Immunopathol
[83] Bailey M, Stevens K, Bland PW, Stokes CR. A monoclonal antibody recognising
an epitope associated with pig inter-leukin-2 receptors. J Immunol Methods
[84] Jonjic S, Koszinowski UH. Monoclonal antibodies reactive with swine lym-
phocytes. I. Antibodies to membrane structures that define the cytolytic T
lymphocyte subset in the swine. J Immunol 1984;133:647–52.
[85] Saalmu ¨ller A, Aasted B, Canals A, Dominguez J, Goldman T, Lunney JK, et al.
Analyses of mAb reactive with porcine CD8. Vet Immunol Immunopathol
[86] Zuckermann FA,PescovitzMD,Aasted B,DominguezJ,Trebichavsky I,Novikov
international swine CD workshop. Vet Immunol Immunopathol 1998;60:
[87] Evans RL, Faldetta TJ, Humphreys RE, Pratt DM, Yunis EJ, Schlossman SF.
Peripheral human T cells sensitized in mixed leukocyte culture synthesize
and express Ia-like antigens. J Exp Med 1978;148:1440–5.
[88] Bevan DJ, Chisholm PM. Co-expression of CD4 and CD8 molecules and de novo
expression of MHC class II antigens on activated rat Tcells. Immunology
[89] Lunn DP, Holmes MA, Duffus WP. Equine T-lymphocyte MHC II expression:
variation with age and subset. Vet Immunol Immunopathol 1993;35:225–38.
[90] Saalmu ¨ller A, Maurer S. Major histocompatibility antigen class II expressing
resting porcine T lymphocytes are potent antigen-presenting cells in mixed
leukocyte culture. Immunobiology 1994;190:23–34.
[91] Pescovitz MD, Book BK, Aasted B, Dominguez J, Ezquerra A, Trebichavsky I,
et al. Summary of workshop findings for antibodies reacting with porcine T-
cells and activation antigens: results from the Second International Swine CD
Workshop. Vet Immunol Immunopathol 1998;60:251–60.
[92] Saalmu ¨ller A, Aasted B, Canals A, Dominguez J, Goldman T, Lunney JK, et al.
Summary of workshop findings for porcine T-lymphocyte antigens. Vet
Immunol Immunopathol 1994;43:219–28.
W. Gerner et al./Developmental and Comparative Immunology 33 (2009) 310–320


text:
Aidan Maguire's Appearances
115 Plays / 0 Likes / 0 Comments
Tree Light Pictures produced and created this 3 minute event video for Red Bull Ireland. We filmed the construction of the Red Bull Cúl 5 arena over a week long period then the event day itself. Aerial…+ More details
39.3K Plays / 685 Likes / 67 Comments
Needle Exchange tells the story of Spencer and Glenn, two best friends who helped each other swap their heroin habits with a passion for tattoos. Ink is their link, but their bond is tested when a…+ More details
Browse Videos
Here are all of the videos that Aidan Maguire has uploaded to Vimeo. Appearances are videos that Aidan Maguire has been credited in by others.

subset: commoncrawl


text:
Sign in now to view, license and download this image or contact us to see this image without a watermark. Want to embed this image on a non-commercial website or blog? Learn more.
Similar imagesView all
Man Running Stock PhotoRunning,Puddle,Splashing,Sports Shoe,Close-up,Adult,Adults Only,Beauty,Color Image,Day,Horizontal,Human Body Part,Human Leg,Motion,One Man Only,One Person,Outdoors,Part Of,Photography,Recreational Pursuit,Shoe,Silhouette,Sport,Sunlight,The Next StepPhotographer Chris ColeCollection: The Image Bank


text:
Our special coverage of the closest race for years
Explore the personal stories of Syrian refugees in Lebanon
21 Jun 2010 14:47 GMT | Politics, US & Canada, Egypt, Israel, United States
" />
George Mitchell, the US special envoy to the Middle East, met with Hosni Mubarak, the Egyptian president, on Saturday to discuss the ongoing political deadlock in the Middle East.
The meeting also tackled efforts to lift the Israeli siege imposed on Gaza since Hamas took control of the Strip.
Egypt was the latest stop for Mitchell on his current tour of the region. On Friday, he met with Mahmoud Abbas, the Palestinian president, in Ramallah.
The meeting came after what Israel described as a decision to ease the blockade - following an international outcry over the attack on a Gaza-bound flotilla that left nine people dead.
Both Abbas and Ismail Haniya, the deposed prime minister reiterated that the Israeli blockade of the Gaza Strip must be completely lifted. But can those making this demand capitalise on unprecedented international pressure to achieve that end? And will more planned aid flotillas bound for Gaza help or hinder them in achieving their goal?
Inside Story, with presenter Mike Hanna, discusses with Sabri Saidam, an advisor to Mahmoud Abbas; Ephraim Sneh, a retired IDF general and the chairman of the S. Daniel Abraham Centre for Strategic Dialogue; and Rosemary Hollis, a Middle East specialist at London's City University.This episode of Inside Story aired from Sunday, June 20, 2010.
Source: Al Jazeera
Content on this website is for general information purposes only. Your comments
are provided by your own free will and you take sole responsibility for any direct
or indirect liability. You hereby provide us with an irrevocable, unlimited, and
global license for no consideration to use, reuse, delete or publish comments, in
accordance with Community Rules & Guidelines and Terms and Conditions.
Activists in the Czech Republic demand that a pig farm built over a former Nazi concentration camp be shut down.
Politics, War & Conflict, Human Rights
In religiously conservative Brazil, women risk jail time and their lives to undergo the procedure that is illegal but commonplace.
Health, Human Rights, Latin America
Trash is a fast-growing import in the Scandinavian country, which turns it into heat for people's homes.
Environment, Europe, Sweden
Critics say the UK's home secretary is using Muslims as a 'political punchbag' in a bid to shore up right-wing support.
Human Rights, Politics, Racism
It is a source of sustenance but also of tension and new political realities are now increasing uncertainty.
Egypt, Sudan, Middle East
A media post-mortem into the UK's surveillance report. Plus, why five years on in Argentina the media looks so familiar.
Media, Edward Snowden, Security
The untold story of how the Lebanese community overcame the odds and found their place in multicultural Australia.
Asia Pacific, Australia, Lebanon
How to keep a telenovela profitable and demanding sponsors happy while inspiring the production team to stay passionate.
Arts & Culture, Mexico, Latin America


text:
Peppa Pig: Peppa and George's Shiny Sticker Play BookPaperback Peppa Pig
List price $5.92
You save $0.78 13% off
Free delivery worldwide
Dispatched in 2 business days
When will my order arrive?
- Publisher: Ladybird Books Ltd
- Format: Paperback | 16 pages
- Dimensions: 158mm x 293mm x 2mm | 122g
- Publication date: 2 March 2011
- Publication City/Country: London
- Sales rank: 642
Product description
Not suitable for children under 3 years of age "Peppa and George's Extra Special Sticker Book" is a fantastically fun children's activity book featuring your favourite "Peppa Pig" characters. You can help Peppa and George solve puzzles, complete dot-to-dot drawings, find their way through mazes and use your fabulously fluffy fabric Peppa Pig stickers too! "The Peppa Pig" range of books are fun, interactive and educational, ideal for encouraging children to start to read by themselves. Titles available from Ladybird include: "Peppa's Super Noisy Sound Book", "The Story of Peppa Pig", "Peppa's Busy Day" and many more!
Other people who viewed this bought:
Peppa Pig: Peppa and George's Wi...
USD$5.00 - Save $0.92 15% off - RRP $5.92
Peppa Pig: Find-the-hat Sticker ...
USD$5.11 - Save $0.81 13% off - RRP $5.92
Peppa Pig Busy Pack
USD$5.64 - Save $0.28 (4%) - RRP $5.92
Peppa Pig Carry Along Colouring
USD$3.70 - Save $2.22 37% off - RRP $5.92
Peppa Pig: Peppa and the Big Tra...
USD$5.58 - Save $1.83 24% off - RRP $7.41
Peppa Pig: Little Library
USD$7.07 - Save $0.34 (4%) - RRP $7.41
Other books in this category
Peppa Pig: Little Library
USD$7.07 - Save $0.34 (4%) - RRP $7.41
Oh, the Places You'll Go!
USD$6.55 - Save $2.34 26% off - RRP $8.89
Guess How Much I Love You Board ...
USD$7.69 - Save $1.20 13% off - RRP $8.89
Slinky Malinki's Christmas Crack...
USD$6.53 - Save $2.36 26% off - RRP $8.89
Peppa Pig: Marvellous Magnet Boo...
USD$10.02 - Save $0.36 (3%) - RRP $10.38
Where's Spot?
USD$7.26 - Save $1.63 18% off - RRP $8.89
- Categories:
Customer reviews
By Yuzuko Huffman 24 Mar 2014
I got this peppa pig sticker book for my 2 years old daughter who loves to watch peppa pig show. This activity book is cute with stickers and all colored pages but there are 3 cutting activity pages that's just on the other side of different activity page. If you cut the page, other activity pages get cut. They should've put blank page for cutting activity pages. All the activity contents seem to be pretty boring, to me. My daughter hasn't seen this book yet so maybe it's fun for toddler kids...
On the other hand, the book was delivered in 7 days which I expected longer than that. (I live in the States.)
Product safety notice
Not suitable for children under 3 years of age


text:
Top Posts
Latest Posts
Robinson Cano out of lineup with bone bruise
May 2, 2011, 6:01 PM EDT
A bone bruise in his left hand has Robinson Cano out of the Yankees lineup for Monday's game against the Tigers, though the All-Star does hope to play Tuesday. Cano was injured Sunday while trying a catch a pickoff throw from pitcher Ivan Nova. Eduardo Nunez is starting at second base in his place tonight. …
David Freese likely out until late July following hand surgery
May 2, 2011, 5:40 PM EDT
David Freese exited yesterday's game with a broken hand thanks to a Scott Linebrink fastball and today general manager John Mozeliak announced that his return timetable is 9-12 weeks. According to Mozeliak he's unlikely to return until late July, which is a big blow to a Cardinals roster already decimated by injuries. Freese was off…
Curt Schilling and Mariano Rivera each opine on bin Laden. Guess which one was sensible.
May 2, 2011, 5:02 PM EDT
Ultimately what baseball players think about the major issues of the day that aren't baseball-related is rather unimportant, but I found this distinction to be 100% unsurprising. Here's Mariano Rivera when asked about the death of Osama bin Laden by Ben Shpigel: "You do something like that, somewhere along the way you're going to pay.…
Homer Bailey to come off DL, rejoin Reds' rotation Thursday
May 2, 2011, 4:49 PM EDT
Out since mid-March with a shoulder injury, Homer Bailey is scheduled to come off the disabled list Thursday and start against the Astros. Bailey also missed a big chunk of last season with shoulder problems, but when healthy posted a 4.46 ERA and 100/46 K/BB ratio in 109 innings to finally show some of the…
HBT Daily: The good, the bad and the ugly of April
May 2, 2011, 4:32 PM EDT
Tiffany and I wonder if the Twins and White Sox have dug too deep a hole. Er, two holes.
Mets prospect Jenrry Mejia has completely torn MCL in elbow
May 2, 2011, 4:16 PM EDT
Jenrry Mejia was off to a good start at Triple-A, but the 21-year-old top prospect is in danger of missing the rest of the season after the Mets announced today that he has a completely torn MCL in his right elbow. Andy Martino of the New York Daily News reports that Mejia will seek a…
It's Military Appreciation Night at Nationals Park
May 2, 2011, 4:00 PM EDT
This is a coincidence — the event was already planned — but the tonight is Military Appreciation Night at Nationals Park. For the occasion, military personnel, be they active or retired, with a valid military ID can receive up to four free tickets to the game. All you need to do is to go to…
Sammy Sosa enjoyed himself at Heat-Celtics game yesterday
May 2, 2011, 3:27 PM EDT
Mocksession.com (via The Big Lead) grabbed a nice screen shot of Sammy Sosa cracking up in the background while ESPN sideline reporter Doris Burke was on camera during yesterday's Heat-Celtics playoff game. The guy in the black shirt must have told a helluva joke. And apparently they didn't get the memo about the home crowd…
Albert Pujols played third base yesterday for the first time since 2002
May 2, 2011, 2:17 PM EDT
David Freese and Nick Punto leaving yesterday afternoon's game with injuries put the Cardinals in a tough spot and led to Albert Pujols playing third base for the first time since 2002. He moved from first base to third base for the final 1.1 innings of a 6-5 loss to the Braves and handled his…
Pablo Sandoval to have surgery tomorrow
May 2, 2011, 2:00 PM EDT
I love Twitter. Before it was invented, you'd have to be friends with a baseball player in order to have him tell you that he's having surgery tomorrow. As was reported over the weekend, Sandoval likely faces 4 to 6 weeks of recovery time. If that schedule holds, it means he'll be back the first or…
Scott Rolen unlikely to return from DL when eligible Friday
May 2, 2011, 1:50 PM EDT
Scott Rolen is eligible to come off the disabled list Friday, but Tom Groeschen of the Cincinnati Enquirer reports that the "odds are slim" he'll be ready to return from a strained left shoulder by then. Reds trainer Paul Lessard told Groeschen that Rolen is progressing in his recovery, but added: "We don't have a…
Padres say new television deal will lead to increased payroll
May 2, 2011, 1:13 PM EDT
With an Opening Day payroll of just $45 million the Padres rank 27th in player spending, but chief executive officer Jeff Moorad said yesterday that the team is negotiating a new local television contract that could increase the payroll by 10-20 percent as soon as 2012. Cox Communications' current 10-year deal to broadcast Padres games…
Ozzie Guillen calls his two-game suspension for tweeting "a good move" by MLB
May 2, 2011, 12:31 PM EDT
I thought MLB suspending Ozzie Guillen two games for posting on Twitter following his ejection last week was too extreme, but yesterday the White Sox manager told Doug Padilla of ESPNChicago.com that the punishment "was a very fair one": I think it was good for baseball, myself and the integrity of the game. If MLB…
Roger McDowell was lucky to just get a suspension
May 2, 2011, 12:00 PM EDT
I missed this yesterday — which was apparently written before baseball suspended Roger McDowell for two weeks — but Gwen Knapp of the San Francisco Chronicle reports on Roger McDowell's behavior in San Francisco last week from another perspective: that of the fan who didn't hire Gloria Allred to reenact McDowell's behavior at a press…
Phillies activate Domonic Brown from DL, send him to Triple-A
May 2, 2011, 11:46 AM EDT
Rather than keep him on a minor-league rehab assignment for his fractured hamate bone, the Phillies activated Domonic Brown from the disabled list yesterday and then optioned him to Triple-A. That means he'll continue to play in the minors, but will do so as a Triple-A player rather than a rehabbing big leaguer. While perhaps…
Carl Pavano loses to Royals, but defeats garbage can
May 2, 2011, 11:19 AM EDT
Francisco Liriano's struggles are more extreme and have gotten more attention during the Twins' awful first month, but Carl Pavano has been plenty terrible himself with a 5.84 ERA through six starts. More worrisome than the bloated ERA is that Pavano's strikeout rate continues to plummet, dropping from 7.2 strikeouts per nine innings in 2009…
Must-click link: Will Leitch on Bud Selig
May 2, 2011, 11:00 AM EDT
Great column over at New York Magazine from Will Leitch today, in which Will assesses Bud Selig: the most effective commissioner in professional sports. I can't help but agree. His failures have been exceedingly high-profile and photo-worthy, but as Will notes, Selig has learned from them and hasn't made the same mistake twice. Meanwhile, his…
Rays say Alex Cobb was tipping pitches in debut yesterday
May 2, 2011, 10:47 AM EDT
Called up from Triple-A to make a spot start for the Rays yesterday, Alex Cobb allowed four runs while failing to make it out of the fifth inning against the Angels and afterward pitching coach Jim Hickey told Marc Topkin of the St. Petersburg Times that the 23-year-old right-hander was tipping his pitches: It was…
R.A. Dickey to slay Grendel's mother
May 2, 2011, 10:30 AM EDT
And he looks like he could do it too, no? As if there aren't enough reasons to love R.A. Dickey, Tyler Kepner of the New York Times gives us another: Dickey names his bats after weapons from epic poems and fantasy fiction: One bat is called Orcrist the Goblin Cleaver and the other is Hrunting.…
Andrew McCutchen talking long-term extension with Pirates
May 2, 2011, 10:15 AM EDT
Andrew McCutchen declined to give any specifics when asked yesterday, but Dejan Kovacevic of the Pittsburgh Post Gazette reports that the Pirates are discussing a long-term contract extension with the 24-year-old center fielder. McCutchen won't be arbitration eligible until 2013, so to buy out any of his free agent years a contract extension would have…
More Top Posts
Most Commented
- Ex-Cardinals outfielder Curt Ford was assaulted in St. Louis and told to "go back to Ferguson" (122)
- David Ortiz: "Nobody in MLB history has been tested for PEDs more than me" (118)
- Rob Manfred says it would be hard to reinstate Pete Rose in a limited way (91)
- The MLBPA releases a statement on Kris Bryant, mentions possible litigation (90)
- Did David Ortiz admit to more than he realized with his Players' Tribune editorial? (88)
- J. Fernandez (5027)
- A. Rendon (4434)
- Y. Tomas (4186)
- C. Martinez (4043)
- K. Jansen (4037)
- J. Verlander (3960)
- C. Davis (3822)
- H. Pence (3739)
- A. Cobb (3635)
- C. Rodon (3608)


text:
Right Where You Are Sitting Now
Now Playing
Coincidence Control Network: File #020
This week: Nicholas Pell goes to charm school, Life on Mars, Pell on politics, the Tupac hologram, The beat goes on, SkyNet, and why you shouldn't support the God Save The Queen re-release! Personnel – Joe Nolan, Nicholas Pell, Joseph Matheny, and Ken Eakins. Linky Poos Neil Young reinvents digital audio format - Link Mars Viking Robots 'Found Life' - Link Boehner Endorses Romney - Link Tupac back from the dead as a hologram at Coachella - Link Kerouac Estate Controversy - Link What's After Autonomous Cars? Humanoid Robots - Link Don't support the re-release of God Save The Queen - Link Musical Interludes Yppah - They Know What Ghost Know Serge Gainsbourg - Requeim Pour Un Con M.C. Lars - Hot Topic is not PunkRock
Subscribe & Follow
- Arts
- Business
- Comedy
- Culture & Travel
- Education
- Entertainment
- Hobbies & Gaming
- General
- Lifestyle & GLBT
- Music
- Religion
- Sports & Recreation
- Technology
- Health & Science
- News & Politics
- Kids & Family
- Government & Organizations
- Show All


text:
Engagement Rachel Polk and Dustin Parker
Rachel Polk and Dustin Parker announce their engagement.
Rachel is the daughter of Michael and Kathy Polk of Hastings. She is a 2012 graduate from the College of St. Benedict and is employed as a registered nurse at St. Cloud Hospital.
Dustin is the son of Rick and Tammy Parker of Sartell. He is a 2010 graduate from St. Cloud State and is employed as a district sales manager for KLN Family Brands/Barrel O'Fun Foods.


text:
Woodworking is a dying art that the current generation of DIYers just aren't taking the time to learn. Woodworking is often a skill passed down from generation to generation and as technology creeps more and more into our everyday lives our dads will more likely know how to setup a POP email server than cut a dovetail joint.
I'm in the camp of wishing I knew more or anything about woodworking. Yes I can cut and measure lumber but there is no art to it. The beauty in woodworking (from what I've heard) is getting to a point where you can go away from plans and start customizing your own furniture or other project.
Here are 13 woodshops that any woodworker or not will love. The clean floors and flawless tools will make any tool junky envious.
Check out the rest of our woodworking articles and if you are into awesome looking workshops from notable makers, builders, DIYers and craftsman, then you must visit Tool Crave.
Timothy Dahl
Latest posts by Timothy Dahl (see all)
- Camp in Style with a DIY Folding Tripod Stool - March 31, 2015
- Magnetic Fire Hose® Work Pants: The Next Generation in Work Wear - March 30, 2015
- Spring is the Perfect Time Change Out Your Burnt Out Bulbs - March 30, 2015


text:
Puzzle & Dragons Z ships over 1.5 million on 3DS
Puzzle & Dragons Z shipments have exceeded the 1.5 million mark, GungHo Online announced today.
The Nintendo 3DS iteration of GungHo Online's trendy role-playing puzzle game launched last December, topping charts in its opening weekend and scored more than 800,000 sales in its first fortnight on the Japanese market.
Prior to the title's release, GungHo Online boss Kazuki Morishita announced his company endeavored to sell at least one million copies of Puzzle & Dragons Z. Mission accomplished, I'd say.
Of course, the success of the mobile version absolutely dwarfs that of its 3DS companion. As of November 2013, the original Puzzle & Dragons had been downloaded more than 23 million times across North America, Japan, and Korea, making ludicrous amounts of money along the way.
A western release of Puzzle & Dragons Z is confirmed to be in the cards, but no concrete details have emerged regarding the localization or when it might arrive.
3DS「パズドラZ」の国内累計出荷本数が150万本を突破 [4Gamer]
Beard View impressions of Puzzle & Dragons Z on 3DS
11:00 PM on 01.23.2015
Puzzle & Dragons: Super Mario Bros. Edition looks awesome, coming in May
8:04 AM on 01.14.2015
GungHo: Puzzle & Dragons Z is still coming west
12:00 AM on 01.13.2015
Puzzle & Dragons Z ships over 1.5 million on 3DS
1:30 PM on 07.17.2014
Wii U sells 109K, Puzzle & Dragons Z tops charts in Japan
10:30 PM on 12.26.2013
TGS: She doesn't look very Gung Ho about her job
9:24 AM on 09.20.2013
GungHo CEO: 'I want to top Nintendo's sales'
1:00 PM on 05.31.2013
GungHo wants Puzzle & Dragons to sell one million on 3DS
11:30 PM on 05.13.2013
Puzzle & Dragons Z Updates, Screenshots, & Videos
Comments not appearing? Anti-virus apps like Avast or some browser extensions can cause this.
Easy fix: Add
[*].disqus.com to your software's white list. Tada! Happy comments time again.
Did you know? You can now get daily or weekly email notifications when humans reply to your comments.


text:
Help please with counter top choices
First time poster, long time lurker!
We are about to start our kitchen remodel process and I am completely stuck on the counter top choice! Hoping to get some help here. I absolutely LOVE carerra marble and at the same time know that it is not suited for our family of 5- bummer:(
I am doing a creamy white painted cabinets with stainless appliances. My kitchen is on the small side with not a lot of natural light. I want to do as much white/ cream as possible and keep the perimeter counters and island the same to brighten and open up the space. I have thought many times that I "found it" and then a few days later I begin to second guess myself and question if another material would be better. arrrrr!
This has been my counter top choice path taken so far:
1. Carerra marble- LOVE, after much research have come the conclusion it probably is not best for our family. I would constantly worry about sratches and stains.
2. Corian Rain Cloud- very impressed when I saw it locally in a large piece. The kitchen showroom employee said it can be orderd "glossed" or shined so it has even more the appearance of marble. BUT, he also said, that if it is shined it will show scratches.
3. Princess White granite- LOVE! Saw this material on this site and truly feel this would be the ticket ! After searching high and low for it in my state of Wisconsin and finding it in a stone yard near Milwuakee I about fell off my chair when I was told the price! $9,000! Wow! considering the corian and other materials I have been quoted have been around $3500-$5000. Cant do the granite.
4. Finally, I have most recently come across what I think is a brand new shade by Silestone called Lagoon. I love this! I brought it home and compared it to the large sample of Coran Rain Cloud and felt the Lagoon had more if a subtle look to it. I guess I would say more of the look of marble with no veining if that makes sense.
So that is where I am at. I am leaning towards the Silestone Lagoon, but have found very little images of it on the internet. I really would love to see it in person but can't find a big piece on display anywhere in my area.
Please note, I realize I can't mimick carerra marble, Im not trying to fool anyone with another product I just want the simple, clean, feel of marble if that makes sense?
Any images of Lagoon or any other ideas would be helpful! thank you!
Wow, I could have posted this! In the same boat... love Cararra so much but know how I'd fret over it. Also looking at rain cloud. I don't have any help to offer, just wanted to commiserate :-) Good luck with your search! Interested to hear what you settle on.
I also love the look of marble, but could not handle the staining and etching. I went with a whitish Delicatus.(they vary in color from yellowish to whitish). I have creamy white cabinets and love the look of them together. The kitchen has seen serious use and I have not had any staining or etching on the Delicatus.
Well, Home Depot's sale ended today so I ended up ordering Silestone Lagoon- I sure hope and pray it looks good. If anyone has any photos or links where I can see finished kitchens with this product I would love it.
I did go with the Ogee (sp?) edge which seems to be what many marble kitchens use.
5 weeks from now I guess I will know! :)
Also, still welcome any suggestions of other alternatives- I have a few days to cancel my order.
thank you!
@enright5 - I know this is a few months after your initial posts, but I came across your posts while I was looking for photos of real life installations of Lyra and Lagoon Silestone. I can total relate to the original post. I love marble too and the look it provides, but it is not a good fit for my family. Knowing this I am not looking to trick people into thinking I have marble - just LOVE the general look and feel. SO what did you finally decide on?


text:
Dianna Fuemana Wikipedia
Related Headline News and Top Stories
Recomended News to Read
LG L80 Dual Now Available in India
INDIAPOST – Earlier in April this year, LG had officially announced the L80 dual SIM smartphone for Indonesia. Now LG India has quietly listed the handset on its official website for Rs 17,500. Note that the other two L III series handsets, LG L70 and the L90 are already available in India Rs 14,500 and […]
BRICS Set Up Bank to Counter Western Hold on Global Finances
INDIAPOST – Leaders of the BRICS emerging market nations launched a $100 billion (£58.36 billion) development bank and a currency reserve pool on Tuesday in their first concrete step towards reshaping the Western-dominated international financial system. The bank aimed at funding infrastructure projects in developing nations will be based in Shanghai, and India will preside over […]


text:
Zachary Rodgers | April 12, 2011 | Comments
Buick, Olay, and Visa are first to use a new "sponsored screensaver" large display ad on the device.
Amazon is offering a cheaper version of its Kindle e-book reader subsidized with ads. Aghast book lovers may be comforted to know the new ads will not interrupt the reading experience, and Amazon appears to be taking steps to safeguard the quality of the ad experience.
The product is named Kindle with Special Offers and retails for $114 - $25 off the list price for the standard Kindle product. Ads take two forms. The highest impact is a "sponsored screensavers" display, the first of which will be offered by Buick, Olay, Visa, and Amazon.com Reward Visa Card from Chase. The second ad type displays on the bottom of the home screen.
Craig Bierley, Buick's director of advertising and promotions, praised the device's ad potential in a statement. "Kindle's high contrast e-ink display eliminates glare and is perfect for emotionally engaging and impactful brand imagery," he said.
To enhance ad quality, Amazon is soliciting user feedback on individual ads via a Kindle app called AdMash. Kindle customers who have installed the free app will preview pairs of sponsored screensaver ads, and the one with the most votes will go live to a wider audience of kindle owners.
Users can also set preferences for the screensaver ads, for instance requesting more travel, architecture, or landscape images.
"Together, AdMash voting and Kindle Screensaver Preferences help us select the most attractive and engaging sponsored screensavers to display on your Kindle," Amazon said.
Kindle with Special Offers is available for pre-order in the U.S. and will ship on May 3.
No stranger to digital media sales, Amazon appears poised to become a larger player in interactive advertising. The company has lately begun advertising a number of advertising related posts, as observed by Ben Schachter, an analyst with Macquarie Group.
In a recent job posting it noted, "Third party advertising on Amazon's retail websites is one of the company's fastest growing and most profitable businesses, and Amazon itself is making significant investments in display advertising across the web."
On the heals of a fantastic event in New York City, ClickZ Live is taking the fun and learning to Toronto, June 23-25. With over 15 years' experience delivering industry-leading events, ClickZ Live offers an action-packed, educationally-focused agenda covering all aspects of digital marketing. Register today!
Until March 2012, Zach Rodgers was managing editor of ClickZ's award-winning coverage of news and trends in digital marketing. He reported on the rise of web companies, data markets, ad technologies, and government Internet policy, among other subjects.
June 23-25, 2015
July 9, 2015
August 10-12, 2015
September 8-10, 2015
October 21-22, 2015
Hong Kong, May 5-6, 2015
A Buyer's Guide to Affiliate Management Software
Manage your performance marketing with the right solution. Choose a platform that will mutually empower advertisers and media partners!
Google My Business Listings Demystified
To help brands control how they appear online, Google has developed a new offering: Google My Business Locations. This whitepaper helps marketers understand how to use this powerful new tool.
May 6, 2015
12:00pm ET/9:00am PT


text:
To print this article open the file menu and choose Print.
Click here to return to previous page
Article published May 13, 2014
Dump truck driver cited in Route 20 accident
AUBURN — A dump truck driver drove with an upright dumping bed into a utility pole, taking down power lines on Route 20 near Prospect Street and TJMAXX about 3:30 Tuesday morning. The truck rolled over, and the driver was trapped in the vehicle with live wires on top of it until National Grid cut the power.
The driver was not injured, Police Chief Andrew J. Sluckis said.
Route 20 was closed for about six hours while the accident was investigated by Auburn police and the state police truck team, and while power, cable and phone crews repaired equipment to restore service.
Heather M. Hennessey, 34, of 10 Railroad Ave., Millbury, was driving the dump truck owned by Jamie Remillard of 4 Peter St., Uxbridge, at the time of the accident. Mrs. Remillard is a subcontractor for the state Department of Transportation repaving project on Route 20.
Trooper Scott Maguire cited Ms. Hennessey for a commercial vehicle height violation, a license restriction (corrective lens), and defective equipment. The truck was supposed to have an alarm to let the driver know the dumper was in the upright position, but the alarm was disconnected, according to authorities.
The morning commute in the area was a "mess and a half" because of the accident, police said.
National Grid restored power by about 8 a.m., and two other utility companies were wrapping up their work by 9.
Contact Linda Bock via email at firstname.lastname@example.com. Follow her on Twitter @LindaBockTG
Order the Telegram & Gazette, delivered daily to your home or office! www.telegram.com/homedelivery
Copyright Worcester Telegram & Gazette Corp.


text:
Hadaki City Sink-Sidekick Dopp Toiletry Kit - Gray CirclesItem # HN-KAL624
- Measures 11.75W x 4.25D x 6.75H inches
- Features thick stitching and quality craftsmanship
- Features exterior and interior zippered pockets
- 2 mesh pockets for extra organization
Similar Travel Accessories
KitchenAid KSM150PSGC 5 Qt. Artisan Series Stand Mixer - Gloss Cinnamon
Celestron AstroMaster 114 EQ Reflector Telescope
Bowflex SelectTech 1090 Adjustable Dumbbells- 10-90 lbs.
$279.99 - $829.98
Pearl Mantels Shenandoah Traditional Fireplace Mantel Shelf
$169.98 - $329.98
Precision Cape Cod Chicken Coop
Palram Nature Greenhouse - Twin-Wall - Mythos Series
$425.98 - $718.49
The Hadaki by Kalencom City Gray Circles Collection
for reviewing this product when you join My Hayneedle Rewards
Write a review
200 points for each review = $2
300 points for reviews including an image or video = $3 Learn More
Customer Q & A
Enter your question, and one of our Customer Care experts will respond via email and also post the answer here.
Your Valid Email
Your Question


text:
Stock Photo:
cute girl bothered by something from the side
- Image ID: 106378865
- Release Information: Signed model release filed with Shutterstock, Inc
- Copyright: swissmacky
High quality images curated from over 150 countries.
Over 40 million photos, vectors, illustrations, and more.
Simple and flat pricing by image or by month.
Over 230,000 images downloaded every day.


text:
Coming up in the next {{countdown}} {{countdownlbl}}
Coming up next:
Skip to this video now
Now Playing:
Related Videos
Now Playing: {{itm.title}}
Play Video
This transcript has been automatically generated and may not be 100% accurate.
Dennis Rodman Holds Court in North Korea
Former NBA star watched an exhibition basketball game with Kim Jong Un.
Dennis Rodman's Financial Trouble
Former basketball star's third ex-wife claims he owes child support.
Now Playing: Dennis Rodman Holds Court in North Korea
Now Playing: Dennis Rodman's Financial Trouble
Now Playing: US-Cuba Relations: Stage Set for Historic Meeting Between Obama and Castro
Now Playing: Iran's Influence: Far Beyond Its Borders
Now Playing: Iran Negotiation Countdown Begins
Now Playing: Turkey Paralyzed by Nationwide Blackout
Now Playing: One Dead and One Wounded at Fort Meade; Germanwings Copilot Was Treated for 'Suicidal Tendencies,' Man Overboard
Now Playing: Protestors Clash With Israeli Police On Land Day, Germanwings Crash Victim's DNA Identified, Dedication of Edward M. Kennedy Institute
Now Playing: Prosecutor Says Germanwings Co-Pilot Had Suicidal Thoughts in the Past
Now Playing: Amanda Knox Weighs Next Move After Italian Court Decision
Now Playing: Germanwings Co-Pilot's Problematic Past Becomes Focus
Now Playing: Iran Nuclear Talks Approaching Deadline
Now Playing: Dozens Injured in AirCanada Crash Landing
Now Playing: Iran Nuclear Negotiations: Deal or No Deal?
Now Playing: Rep. Ed Royce Says Military Action on the Table if Nuclear Deal Fails
Now Playing: Germanwings Plane Crash Prompts Changes
Now Playing: Investigation Into the Final Moments of Germanwings Flight
Now Playing: Germanwings Crash Timeline Updated After Reported Andreas Lubitz Revelations
Now Playing: Air Canada Flight Crash Lands, Skids Off Runway
Now Playing: Two Torn Up Doctor's Notes Found in Germanwings Co-Pilot's Home
Now Playing: Amanda Knox 'Relieved and Grateful' After Italian Court's Ruling


text:
|U.S. Constitution Online
|Quick Links: FAQ Topics Forums Documents Timeline Kids Vermont Constitution Map Citation
<<Previous Question | Question Index | Subject Index | Constitutional Index | Next Question>>
Q71. "I cannot find anywhere in the Constitution that refers to separation of church and state."
A. Though many people assume the 1st Amendment sets out some separation, the phrase does not appear in the Constitution. The phrase "wall of separation" appears to have been coined by Jefferson, in speaking of the religious liberties granted by the Constitution and the Bill of Rights. Madison, however, said that there is a line between church and state, not a wall — the distinction may or may not be significant.
In practice the separation is more theoretical than actual. In a truly separate society, we would not invoke the name of God on our currency, nor would we speak so highly of our Judeo-Christian values. But we do — the fact of the matter is, completely separating religion and government is probably impossible, so long as religion is an important part of the lives of the citizenry. The best we can hope for, and what I think the Constitution tries to protect, is to ensure that there is no discrimination on the basis of religious belief — that there be no religion litmus test.
Web site designed and maintained by
Steve Mount.
© 1995-2010 by Craig Walenta. All rights reserved.
Contact the Webmaster.
Site Bibliography.
How to cite this site.
Please review our privacy policy.
Last Modified: 16 Aug 2010
Valid HTML 4.0