Intestinal Diseases Research Program, Department of Medicine, McMaster University, Hamilton, Ontario L8N3Z5, Canada
![]() |
ABSTRACT |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
T helper
2 (Th2) cytokines interleukin (IL)-4 and IL-13, which activate signal
transducer and activator of transcription 6 (STAT6) are expressed in
the muscularis externa during nematode infection and are candidate
mediators of the associated hypercontractility. To determine the locus
of action of these cytokines, we examined the IL-4- and IL-13-induced
hypercontractility of the isolated muscle cells from STAT6 +/+ and
STAT6 /
mice. We compared the results with cells isolated from
Trichinella spiralis-infected STAT6 +/+ and STAT6
/
mice. Carbamylcholine chloride (Carbachol) induced the contraction
of jejunal muscle cells in a concentration-dependent manner maximal
contraction (Rmax 26.7 ± 1.9%). Cells from T. spiralis-infected STAT6
/
mice showed the hypertrophy (cell
lengths 41.4 ± 0.8 to 89.0 ± 8.7 µm) and
hypercontractility (Rmax 37.5 ± 1.3%) induced by
infection. IL-4R
mRNA was detected in dispersed smooth muscle cells.
Incubation of longitudinal muscle-myenteric plexus (LMMP) with IL-4 and
IL-13 enhanced Carbachol-induced muscle contraction (Rmax
35.5 ± 1.9 and 32.4 ± 2.9%, respectively). Incubation of LMMP from STAT6
/
mice with IL-4 did not enhance the contraction. The hypercontractility in T. spiralis-infected mice was
attenuated in STAT6
/
mice (P < 0.02). These
results indicate both IL-4 and IL-13 induce hypercontractility of
muscle cells via the STAT6 pathway, and this is the basis for
hypercontractility observed in T. spiralis-infected mice.
intestine; motility; cytokine
![]() |
INTRODUCTION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
INFLAMMATION OF THE PROXIMAL gastrointestinal tract and airways is accompanied by changes in smooth muscle contractility. This contributes to the clinical expression of diseases such as inflammatory bowel disease and asthma. Studies in asthma have identified T helper 2 (Th2) cells and their cytokines in the development of airway hyperreactivity in atopic and intrinsic asthmatics (4, 35). A previous study of tissue from Crohn's disease patients also revealed hypercontractility of intestinal muscle (33), and preliminary data identify the Th2 cytokines interleukin (IL)-4 and IL-13 as putative mediators (2, 3).
To explore mechanisms underlying inflammation-associated muscle hypercontractility in the gut, we have used mice infected with the nematode parasite Trichinella spiralis. Our studies to date show that the hypercontractility of intestinal muscle strips to carbamylcholine chloride (Carbachol) is T cell dependent (31, 32) and is attenuated in mice deficient in signal transducer and activator of transcription 6 (STAT6) (21), again implicating IL-4 and IL-13 as putative mediators.
Exact locus and mechanisms underlying the actions of these cytokines remains unclear. Goldhill et al. (18) reported that IL-4 administration in vivo enhanced the response to cholinergic nerve stimulation in murine small intestinal longitudinal muscle via a leucotriene D4-mediated effect. This, however, does not exclude a direct effect of Th2 cytokines on smooth muscle cells, as has recently been shown for IL-4 and IL-13 on airway smooth muscle cells (22). Thus inflammation-associated increase in contractility of intestinal muscle strips could reflect a direct effect on smooth muscle cells, an indirect action of IL-4 and IL-13 on enteric nerves (18), or an indirect action mediated via another cell type or via a component of the extracellular matrix.
IL-4 exerts its biological effects by binding to the IL-4 receptor
-chain, a component of both the type 1 and type 2 IL-4 receptor
(IL-4R) (14, 16, 37). In the type 2 IL-4R, IL-4R
is
paired with IL-13R
1, which also binds IL-13 (19, 34). Signal transduction may occur by two separate pathways, phosphorylation and activation of STAT6 by janus kinase 1 (JAK1) and JAK3, which, once
activated, dimerizes, translocates to the nucleus, and binds to
specific promoter regions to regulate gene transcription (24, 25,
36). Our previous work has implicated the STAT6 pathway in the
Carbachol-induced hypercontractility of jejunal muscle strips from
T. spiralis-infected mice (21).
Thus the purpose of this study was to determine the locus of action of
IL-4 and IL-13 defined as their abilities to alter intestinal muscle
contractility by performing studies on single muscle cells freshly
isolated from the intestine of control or Trichinella
spiralis-infected mice, in the presence or absence of the STAT6
pathway. Specifically, we examined whether the IL-4R is expressed on
these cells, whether IL-4 and IL-13 induce hypercontractility, and
whether this is STAT6 mediated.
![]() |
MATERIALS AND METHODS |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Materials. The following materials were used in this study: collagenase (CLS type l), trypsin inhibitor, BSA, acrolein, leflunomide, protease, and Mayer's hematoxylin from Sigma (St. Louis, MO); HEPES from BioShop Canada (Burlington, ON, Canada); IL-4 and IL-13 from R&D Systems (Minneapolis, MN); DMEM and antibiotic-antimycotic from GIBCO-BRL Life Technologies (Gaithersburg, MD); rabbit anti-human polyclonal antibody anti-CD3, normal swine serum (NSS), biotinylated swine anti-rabbit, streptavidin-peroxidase conjugate, and Faramount aqueous mounting medium from DAKO Diagnostics (Mississauga, ON, Canada); and diaminobenzidine (DAB) from Zymed.
Mice.
Studies were performed on male C57BL/6 mice with or without T. spiralis infection and STAT6 /
mice between 6 and 10 wk of age. STAT6
/
mice on a C57BL/6 background were originally produced by the gene mutation as described by Takeda et al. (27).
Breeding pairs of STAT6
/
mice and their wild-type littermates
(STAT6 +/+) were obtained from the John Curtin School of Medical
Research, Australian National University, Canberra, Australia. Mice
were kept in filter-isolated cages in groups of four to five in
positive-pressure rooms with a constant ambient temperature and a
14:10-h light-dark cycle. All experiments were approved by the Animal
Care Commmittee at McMaster University and were conducted in accordance
with the guidelines of the Canadian Council on Animal Care.
Trichinella infection. Mice were infected by the administration of 0.1 ml of phosphate-buffered saline containing 375 T. spiralis larvae by gavage. The larvae were obtained from infected rodents 60-90 days after infection using a modification of the technique described by Castro and Fairbairn (11). The T. spiralis culture originated in the Department of Zoology at the University of Toronto, and the colony was maintained through serial infections alternating between male Sprague-Dawley rats and male CD1 mice.
Immunohistochemistry for histological analysis. Muscle layers from mice infected with T. spiralis were fixed in 10% formalin and embedded in paraffin for histological analysis. Sections were cut at a thickness of 3 µm and immunostaining for T-lymphocytes was performed with a rabbit anti-human polyclonal antibody in a technique similar to the three-step immunoperoxidase method (26). Slides were washed with xylene and ethanol, placed in freshly prepared methanol solution for 30 min, transferred to protease solution (protease 125 units in 50 ml TBS pH 7.6 with 50 µl of CaCl2) for 15 min, and washed with water and Tris-buffered saline (TBS). NSS (1%) was dropped on the slides as a protein-blocking agent for 15 min. Slides were incubated with 1:400 CD3 antibody in 1% NSS overnight in a humid chamber. Slides were rinsed in TBS, and incubated with 1:600 biotinylated swine anti-rabbit in 1% NSS for 90 min, rinsed in TBS, incubated with 1:1,200 streptavidin-peroxidase conjugate in NSS for 90 min. Slides were rinsed in TBS, incubated with DAB for 8 min, counter-stained with Mayer's hematoxylin for 10 min, and mounted in glycerin gelatin.
Preparation of dispersed smooth muscle cells. Muscle cells were isolated from longitudinal muscle-myenteric plexus (LMMP) of the C57BL/6 mice jejunum by a method similar to that used by Bitar and Makhlouf (7) to prepare smooth muscle cells from the guinea pig stomach. The uninfected mice and the mice infected 8 days previously with T. spiralis were killed by cervical dislocation. The jejunum was removed and placed in DMEM with 1% antibiotic-antimycotic. LMMP was peeled carefully from jejenum. The LMMP were preincubated with or without cytokines (10 ng/ml IL-4, 10 ng/ml IL-13) for 16 h in the 5% CO2 incubator. LMMP was incubated for two successive 10-min periods at 31°C in 10 ml of HEPES medium containing the following (in mM): 98.1 NaCl, 3.87 KCl, 2.42 NaH2PO4H2O, 4.86 L-glutamic-acid, 4.86 fumaric acid, 4.86 pyruvate, 11.17 glucose, 1.79 CaCl2, 1.2 MgSO47H2O, and 23.5 HEPES, pH 7.4, containing 10 mg/ml of collagenase, BSA, and trypsin inhibitor. After incubation, the partly digested LMMP was washed with enzyme-free HEPES medium and reincubated in 10 ml of fresh HEPES medium to allow the cells to disperse spontaneously. Cells were then harvested by filtration through a 210-µm polyester mesh.
Detection of IL-4R in muscle by RT-PCR.
Expression of mRNA of IL-4R in dispersed longitudinal single smooth
muscle cells with or without T. spiralis infection was investigated by a method described previously (30). Total
cellular RNA was isolated based on previously described guanidium
isothiocyanate method (12). The concentration of RNA was
determined by measuring absorbance at 260 nm and its purity was
confirmed using the ratio of absorbency at 260:280 nm. RNA was stored
at
70°C until used for RT-PCR. mRNA was then reversed transcribed
as described previously to yield cDNA, and the cDNA was amplified by
PCR using gene-specific primers. cDNA (0.1 µg in 50 µl aliquots)
were then mixed with 20 pmol each of upstream (5'-GAGT GAG TGG AGT CCT
AGC ATC-3') and downstream (5'-GCT GAA GTA ACA GAA CAG GC-3') primers
for IL-4R
(13). PCR was performed in 50-µl volumes
containing 200 µM 2-deoxynucleotide 5'-triphosphate, 1.5 mM
MgCl2, and 2.5 units Taq polymerase with corresponding
buffer and distilled water. Messages for IL-4R
were coamplified
using the following parameters: denaturation 94°C for 30 s,
annealing 55°C for 30 s, and extension at 72°C for 60 s.
PCR products [soluable IL-4R (sIL-4R) 241 bp; membrane IL-4R (mIL-4R)
127 bp] were separated by 2.5% agarose gel electrophoresis and then
visualized under ultraviolet light after ethidium bromide staining.
Measurement of contraction and relaxation in dispersed cells. Dispersed cells were stimulated by the addition of a 0.8 ml aliquot of the cell suspension to 0.1 ml of the test agent and then incubated at room temperature for 30 s, because we previously found that Carbachol induced the maximal contractile response in jejunal longitudinal smooth muscle cells after 30 s of incubation. The reaction was interrupted by the addition of acrolein in a final concentration of 1%. The median cell length of 50 cells on each slide was measured with a microscope using image-splitting micrometry, and the percent decrease from control in the mean cell length was determined.
Statistics. Each experiment was performed at least four times and results are presented as means ± SE. Statistical analyses were performed using Student's t-test for comparison of two means or one-way ANOVA for the comparison of more than two means. A P value <0.05 was considered statistically significant.
![]() |
RESULTS |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Immunohistochemistry revealed the presence of CD3+
cells in the longitudinal muscle layer from mice infected with T. spiralis 8 days previously (Fig.
1a); no lymphocytes were seen
in the muscle layers of noninfected mice (data not shown). As shown in
Fig. 1b, both mIL-4R and sIL-4R forms of the -chain of
the Il-4R were expressed on single muscle cells isolated from control
and T. spiralis-infected mice. At 29 cycles, there was no
change in the PCR products for either the mIL-4R (127 bp) or the sIL-4R
(241 bp) between control and infected mice (Fig. 1).
|
We then examined the characteristics of single muscle cells isolated
from the longitudinal muscle layer from control and T. spiralis-infected mice. In muscle from control mice, the mean cell
length was 42.4 ± 6.2 µm and this increased by 110% to
89.0 ± 8.7 µm in cells from infected mice (P < 0.05). Reduction in mean cell length after stimulation by 10 nM
Carbachol was 26.7 ± 1.9% in control cells compared with
37.5 ± 1.3% in cells isolated from infected mice, reflecting a
40% increase in contractility (P < 0.01). In
addition, the half-maximal effective dose (ED50) for
Carbachol-induced contraction was 1,000-fold less in cells from
infected mice (0.3 pM vs. 0.3 nM in infected and control mice,
respectively) (Fig. 2).
|
To determine the effects of the Th2 cytokines IL-4 and IL-13 on muscle
contractility, strips of muscle were preincubated with the cytokine
before dispersion of the cells and subsequent stimulation by Carbachol.
Preincubation with IL-4 or IL-13 did not alter mean cell length; cell
length was 43.1 ± 1.7 and 44.5 ± 3.4 µm in IL-4 and IL-13
exposed cells, respectively, compared with 42.4 ± 6.2 µm in
control cells. Carbachol-induced contraction of cells exposed to IL-4
was 35.5 ± 1.9%, reflecting a 33% increase in contractility compared with control cells. In addition, the ED50 for
Carbachol-induced contraction was ~22-fold less in IL-4-exposed cells
compared with control (13 pM vs. 0.3 nM, respectively). A similar
change in profile was seen in cells preexposed to IL-13.
Carbachol-induced contractility was 32.4 ± 2.9% in IL-13-exposed
cells, representing a 21% increase above that of control cells. In
addition, the ED50 for Carbachol-induced contraction was
~120-fold less in IL-13-exposed cells compared with control (2.5 pM
vs. 0.3 nM) (Fig. 3).
|
In STAT6 /
mice, mean cell length was 41.4 ± 0.8 µm and was
similar to that of wild-type mice. Carbachol-induced contraction of
cells from STAT6
/
mice was 23.2 ± 1.1% compared with that of 26.7 ± 1.9% observed in wild-type mice. As expected,
preincubation with IL-4 did not alter Carbachol-induced contractility
with values of 22.4 ± 1.3% and 0.2 nM for maximal contraction
(Rmax) and ED50, respectively, being not
significantly different from those of wild-type mice. In IL-13 exposed
cells from STAT6
/
mice, Rmax was increased at
27.7 ± 0.8% (P < 0.05) and the ED50
was significantly less than that of control cells at 40 pM (Fig.
4).
|
To evaluate the role of these cytokines in the muscle changes observed
in infected mice, we examined the characteristics of single muscle
cells isolated from STAT6-deficient mice in the absence or presence of
T. spiralis infection. The mean length of cells from
uninfected STAT6 /
mice was 41.4 ± 0.8 µm and this value
increased significantly by 15% to 47.4 ± 2.1 µm in infected
STAT6
/
mice (P < 0.05). However, this
infection-induced increment in cell length in STAT6
/
mice was
substantially less than that of 110% seen in wild-type mice. In
infected STAT6
/
mice, Carbachol-induced contraction was 25.5 ± 1.4 compared with 23.2 ± 1.1% in noninfected STAT6
/
mice. However, the ED50 fell from 0.1 nM in uninfected
STAT6
/
mice to 0.8 pM in infected mice (Fig.
5).
|
![]() |
DISCUSSION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Results of this study show that cells isolated from the longitudinal muscle layer of T. spiralis-infected mice exhibit a substantially greater degree of shortening on exposure to Carbachol than cells isolated from control mice. This finding indicates that the inflammation-induced hypercontractility is a property of the muscle cells, rather than the syncytium. Thus muscle cells isolated from this model may be used to examine changes in excitation-contraction coupling that underlie this hyperresponsiveness to Carbachol. The presence of CD3+ cells in the muscularis externa, taken together with the observation that the increased shortening of isolated cells is attenuated in STAT6-deficient infected mice, is consistent with the hypothesis that Th2 cytokines mediate these effects at the level of the muscle cell.
We also show that intestinal muscle cells from control or infected mice
express the IL-4R and are therefore, able to respond to IL-4 and
IL-13. Indeed, we show that when these cells are preincubated with IL-4
or IL-13, they exhibit a greater degree of shortening on subsequent
exposure to Carbachol. These findings are similar to those observed in
T. spiralis-infected mice and provide further support for
the hypothesis that IL-4 and IL-13, acting directly on muscle cells via
the STAT6 pathways, mediate muscle hypercontractility during nematode infection.
We believe muscle hypercontractility contributes to the increased propulsive forces seen in extrinsically denervated intestinal segments during T. spiralis infection (5). The hypercontractility observed in muscle from the proximal intestine contrasts with the hypocontractility observed in the distal small intestine (23), thus generating an aboral gradient of force-generation by muscle that likely contributes to worm expulsion (29). Because the successful expulsion of nematode parasites is also STAT6 dependent (6, 21, 28), these data provide further support for a linkage between increased contractility of muscle and the successful eviction of the parasite from the gut. This generates the concept that the muscle cell acts as an extension of the mucosal immune system and, under appropriate signaling from lymphocytes, can alter its physiology to contribute to host defence.
Our results indicate there are two components to the increased
responsiveness to Carbachol in muscle from T. spiralis-infected mice. There is an increase in the maximum
shortening of muscle cell length, as well as a decrease in the
ED50 of Carbachol-induced contraction. Our findings suggest
that different mechanisms underlie these changes. In muscle cells from
Trichinella-infected mice, there is an ~1,000-fold
increase in affinity for Carbachol. This was reduced by >50% in
infected STAT6-deficient mice. Preincubation of muscle from control
mice with IL-4 or with IL-13 resulted in smaller decreases in
ED50. Changes were more marked in cells treated with IL-13
than with IL-4 and were largely STAT6 dependent. These observations
suggest that the increased affinity of the muscarinic receptor for
Carbachol on murine muscle cells is increased by Th2 cytokines
(IL-13 > IL-4) and that these, together with STAT6-independent mechanisms, contribute to the increased responsiveness of these cells
to Carbachol during inflammation induced by T. spiralis. Inflammation-induced changes in muscarinic receptors have also been
reported in the airways. Fryer et al. (17) reported that in asthma, there is decreased M2 muscarinic receptor expression on
cholinergic nerves. These neuronal M2 receptors inhibit acetylcholine release. Thus the reduced expression of these receptors induces bronchoconstriction via increased release of acetylcholine acting on M3
receptors on muscle. Furthermore, Fryer et al. (17) showed that this downregulation of the M2 receptor could also be induced by
exposing the tissue to the Th1 cytokine interferon-.
Increased shortening of cells induced by Carbachol was 40% in cells
isolated from T. spiralis-infected mice and was only 10% in
cells from STAT6-deficient-infected mice, indicating that ~75% of
the increased contractility is mediated via the STAT6 pathway. IL-4-induced hypercontractility was 33%, and this was completely STAT6
dependent. In contrast, that induced by IL-13 was only 21% and was
largely STAT6 independent. These findings are compatible with a model
in which the large STAT6-dependent component of muscle hypercontractility is mediated by IL-4, whereas the minor
STAT6-independent component is IL-13 mediated. Increased force
generation, resulting in a greater shortening of muscle cells, may
reflect changes in the contractile protein content of the cells.
Blennerhassett et al. (9) reported that T. spiralis-induced inflammation caused a fivefold increase in the
amount of and
-isoforms of smooth muscle actin per cell. These
increases in the smooth muscle-specific actins may affect force
production and further demonstrate the plasticity of smooth muscle in
the inflamed intestine.
In previous studies in the rat, we reported trophic changes in
smooth muscle during nematode infection. These changes were a
combination of hypertrophy and hyperplasia (10). These
findings are reflected, in part, in the present studies where we show
that the length of smooth muscle cells isolated from T. spiralis-infected mice is 110% greater than that of cells
isolated from control mice. Although resting cell length was unchanged
in uninfected STAT6-deficient mice compared with wild-type mice, there
was only a 15% increase in cell length after infection in
STAT6-deficient mice. This observation indicates that the trophic
effect of inflammation is largely STAT6-dependent. This could not be
reproduced by preincubating tissue with either IL-4 or IL-13 for
16 h. Although a longer exposure time might be required to induce
the degree of hypertrophy seen after 8 days of infection, it is also
possible that the hypertrophy could represent the effects of mediators
elaborated downstream from the STAT6 dependent generation of the Th2
response in this model. Transforming growth factor- (TGF-
) is
known to exert trophic effects on smooth muscle (8) and is
known to be upregulated by IL-4 (15). Furthermore, we have
preliminary evidence that TGF-
is expressed in the muscularis
externa during T. spiralis infection in the mouse
(1).
From the results of this study, we hypothesize that T cells
infiltrating the muscularis externa produce IL-4 and IL-13, which act
directly on muscle to 1) increase contractility largely
through STAT6-dependent mechanisms; 2) increase the affinity
of the muscarinic receptor through STAT6 dependent and independent
pathways, and 3) to induce hypertrophy of muscle, most
likely via the induction of TGF-. This scenario, in terms of trophic
and contractile changes in smooth muscle, is similar to that portrayed
in asthma (20) and further emphasizes similarities between
inflammatory conditions of the lung and gut.
![]() |
ACKNOWLEDGEMENTS |
---|
This study was supported by a grant from the Canadian Insitutes of Health Research (to S. M. Collins), and from a Research Initiative Award (to H. Akiho) from the Canadian Association of Gastroenterology and AstraZeneca.
![]() |
FOOTNOTES |
---|
Address for reprint requests and other correspondence: S. M. Collins, Rm. 4W8, HSC, Medicine, McMaster Univ., 1200 Main St. West, Hamilton, Ontario L8N3Z5, Canada (E-mail: scollins{at}fhs.csu.mcmaster.ca).
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Received 27 August 2001; accepted in final form 12 October 2001.
![]() |
REFERENCES |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
1.
Akiho, H,
Degiorgio R,
Barbara G,
Blennerhassett PA,
Deng Y,
and
Collins SM.
The role of TGF in muscle hypercontractility in a murine model of post-infectious irritable bowel syndrome (Abstract).
Gastroenterology
120:
A714,
2001[ISI].
2.
Akiho, H,
Deng Y,
Lovato P,
Ceponis PJM,
Blennerhassett PA,
and
Collins SM.
Evidence for cytokine-induced activation of signal transducer and activator of transcription 6 (STAT6) in human intestinal muscle cells (Abstract).
Gastroenterology
120:
A315,
2001[ISI].
3.
Akiho, H,
Lovato P,
Blennerhassett PA,
and
Collins SM.
Interleukin-4-induced hypercontractility of human intestinal muscle cells is mediated via a Stat-6: implications for motility changes in Crohn's disease (Abstract).
Gastroenterology
118:
A873,
2000[ISI].
4.
Akimoto, T,
Numata F,
Tamura M,
Takata Y,
Higashida N,
Takashi T,
Takeda K,
and
Akira S.
Abrogation of bronchial eosinophilic inflammation and airway hyperreactivity in signal transducers and activators of transcription (STAT) 6-deficient mice.
J Exp Med
187:
1537-1542,
1998
5.
Alizadeh, H,
Weems WA,
and
Castro GA.
Intrinsic jejunal propulsion in the guinea pig during parasitism with Trichinella spiralis.
Gastroenterology
93:
784-790,
1987[ISI][Medline].
6.
Bancroft, AJ,
McKenzie ANJ,
and
Grencis RK.
A critical role for IL-13 in resistance to intestinal nematode infection.
J Immunol
160:
3453-3461,
1998
7.
Bitar, KN,
and
Makhlouf GM.
Receptors on smooth cells: characterization by contraction and specific antagonists.
Am J Physiol Gastrointest Liver Physiol
242:
G400-G407,
1982
8.
Black, PN,
Young PG,
and
Skinner SJM
Response of airway smooth muscle cells to TGF-1: effects on growth and synthesis of glycosaminoglycans.
Am J Physiol Lung Cell Mol Physiol
271:
L910-L917,
1996
9.
Blennerhassett, MG,
Bovell FM,
Lourenssen S,
and
McHugh KM.
Characteristics of inflammation-induced hypertrophy of rat intestinal smooth muscle cell.
Dig Dis Sci
44:
1265-1272,
1999[ISI][Medline].
10.
Blennerhassett, MG,
Vignjevic P,
Vermillion DL,
and
Collins SM.
Inflammation causes hyperplasia and hypertrophy in smooth muscle of rat small intestine.
Am J Physiol Gastrointest Liver Physiol
262:
G1041-G1046,
1992
11.
Castro, GA,
and
Fairbairn DF.
Carbohydrates and lipids in Trichinella spiralis larvae and their utilization.
J Parasitol
55:
51-58,
1969[ISI][Medline].
12.
Chomczynski, P,
and
Sacchi N.
Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction.
Anal Biochem
162:
156-159,
1987[ISI][Medline].
13.
Chilton, PM,
and
Fernandez-Botran R.
Regulation of the expression of the soluble and membrane forms of the murine IL-4 receptor.
Cell Immunol
180:
104-115,
1997[ISI][Medline].
14.
De Vries, JE.
The role of IL-13 and its receptor in allergy and inflammatory responses.
J Allergy Clin Immunol
102:
165-169,
1998[ISI][Medline].
15.
Elovic, AE,
Ohyama H,
Sauty A,
McBride J,
Tsuji T,
Nagai M,
Weller PF,
and
Wong DTW
IL-4-dependent regulation of TGF- and TGF-
expression in human eosinophils.
J Immunol
160:
6121-6127,
1998
16.
Finkelman, FD,
Wynn TA,
Donaldson DD,
and
Urban JF, Jr.
The role of IL-13 in helminth-induced inflammation and protective immunity against nematode infections.
Curr Opin Immunol
11:
420-426,
1999[ISI][Medline].
17.
Fryer, AD,
Adamko DJ,
Yost BL,
and
Jacoby DB.
Effects of inflammatory cells on neuronal M2 muscarinic receptor function in the lung.
Life Sci
64:
449-455,
1999[ISI][Medline].
18.
Goldhill, J,
Morris S,
Maliszewski C,
Urban J,
Funk C,
Finkelman F,
and
Donohue T.
Interleukin-4 modulates cholinergic neural control of mouse small intestinal longitudinal muscle.
Am J Physiol Gastrointest Liver Physiol
272:
G1135-G1140,
1997
19.
Hilton, DJ,
Zhang JG,
Metcalf D,
Alexander WS,
Nicola NA,
and
Wilson TA.
Cloning and characterization of a binding subunit of the interleukin 13 receptor that is also a component of the interleukin 4 receptor.
Proc Natl Acad Sci USA
93:
497-501,
1996
20.
Hirst, SJ.
Airway smooth muscle as a target in asthma.
Clin Exp Allergy
30, Suppl:
54-59,
2001[ISI].
21.
Khan, WI,
Vallance BA,
Blennerhassett PA,
Deng Y,
Verdu EF,
Matthaei KI,
and
Collins SM.
Critical role for signal transduction and activation of transcription factor 6 in mediating intestinal muscle hypercontractility and worm expulsion in Trichinella spiralis-infected mice.
Infect Immun
69:
838-844,
2001
22.
Laporte, JC,
Moore PE,
Baraldo S,
Jouvin MH,
Church TL,
Schwartzman IN,
Panettieri RA,
Kinet JP,
and
Shore SA.
Direct effect of interleukin-13 on signaling pathways for physiological responses in cultured human airway smooth muscle cells.
Am J Respir Crit Care Med
164:
141-148,
2001
23.
Marzio, L,
Blennerhassett P,
Chiverton S,
Vemillion DL,
Langer J,
and
Collins SM.
Altered smooth muscle function in worm-free regions in Trichinella infected rats.
Am J Physiol Gastrointest Liver Physiol
259:
G306-G313,
1990
24.
Schindler, C,
and
Darnell JJ.
Transcriptional responses to polypeptide ligands: the JAK-STAT pathway.
Annu Rev Biochem
64:
621-651,
1995[ISI][Medline].
25.
Smerz-Bertling, C,
and
Duschl A.
Both interleukin 4, and interleukin 13 induce tyrosine phosphorylation of the 140 kDa subunit of the interleukin 4 receptor.
J Biol Chem
270:
966-970,
1995
26.
Tak, PP,
van der Lubbe PA,
Cauli A,
Daha MR,
Smeets TJM,
Kluin PM,
Meinders AE,
Yanni G,
Panayi GS,
and
Breedveld FC.
Reduction of synovial inflammation after anti-CD4 monoclonal antibody treatment in early rheumatoid arthritis.
Arthritis Rheum
38:
1457-65,
1995[ISI][Medline].
27.
Takeda, K,
Tanaka T,
Shi W,
Matsumoto M,
Minami M,
Kashiwamura S,
Nakanishi K,
Yoshida N,
Kishimoto T,
and
Akira S.
Essential role of STAT6 in IL-4 signaling.
Nature
380:
627-630,
1996[ISI][Medline].
28.
Urban, JF, Jr,
Noben-Trauth N,
Donaldson DD,
Madden KB,
Morris SC,
Collins M,
and
Finkelman FD.
IL-13, IL-4R, and Stat6 are required for the expulsion of the gastrointestinal nematode parasite Nippostrongylus brasiliensis.
Immunity
8:
255-264,
1998[ISI][Medline].
29.
Vallance, BA,
Blennerhassett PA,
and
Collins SM.
Increased intestinal muscle contractility and worm expulsion in nematode infected mice.
Am J Physiol Gastrointest Liver Physiol
35:
G321-G327,
1997.
30.
Vallance, BA,
Blennerhassett PA,
Deng Y,
Matthaei KI,
Young IG,
and
Collins SM.
IL-5 contributes to worm expulsion and muscle hypercontractility in a primary T. spiralis infection.
Am J Physiol Gastrointest Liver Physiol
277:
G400-G408,
1999
31.
Vallance, BA,
Croitoru K,
and
Collins SM.
T lymphocyte-dependent and -independent intestinal smooth muscle dysfunction in the T. spiralis-infected mouse.
Am J Physiol Gastrointest Liver Physiol
275:
G1157-G1165,
1998
32.
Vallance, BA,
Galeazzi F,
Collins SM,
and
Snider DP.
CD4 T cells and major histocompatibility complex class II expression influence worm expulsion and increased intestinal muscle contraction during Trichinella spiralis infection.
Infect Immun
67:
6090-6097,
1999
33.
Vermillion, DL,
Huizinga JD,
Riddell RH,
and
Collins SM.
Altered small intestinal smooth muscle function in Crohn's disease.
Gastroenterology
104:
1692-1699,
1993[ISI][Medline].
34.
Welham, MJ,
Learmonth L,
Bone H,
and
Schrader JW.
Interleukin-13 signal transduction in lymphohemopoietic cells. Similarities and differences in signal transduction with interleukin-4 and insulin.
J Biol Chem
270:
12286-12296,
1995
35.
Ying, S,
Humbert M,
Barkans J,
Corrigan CJ,
Pfister R,
Menz G,
Larche M,
Robinson D,
Durham SR,
and
Kay AB.
Expression of IL-4 and IL-5 mRNA and protein product by CD4+ and CD8+ T cells, eosinophils, and mast cells in bronchial biopsies obtained from atopic and nonatopic (intrinsic) asthmatics.
J Immunol
158:
3539-3544,
1997[Abstract].
36.
Yu, CR,
Kirken RA,
Malabarba MG,
Yound HA,
and
Ortaldo JR.
Differential regulation of the Janus kinase-STAT pathway, and biologic function of IL-13 in primary human NK and T cells: a comparative study with IL-4.
J Immunol
161:
218-227,
1998
37.
Zurawski, SM,
Chomarat P,
Djossou O,
Bidaud C,
McKenzie AN,
Miossec P,
Banchereau J,
and
Zurawski G.
The primary binding subunit of the human interleukin-4 receptor is also a component of the interleukin-13 receptor.
J Biol Chem
270:
13869-13878,
1995