The JI
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     
 

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Tsuchiya, T.
Right arrow Articles by Yamamoto, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tsuchiya, T.
Right arrow Articles by Yamamoto, H.
Right arrowPubmed/NCBI databases
*Gene*GEO Profiles
*HomoloGene*UniGene
*Substance via MeSH
The Journal of Immunology, 2003, 171: 5507-5513.
Copyright © 2003 by The American Association of Immunologists

Role of {gamma}{delta}T Cells in the Inflammatory Response of Experimental Colitis Mice 1

Takahiro Tsuchiya2,*, Sumiaki Fukuda*, Hiromasa Hamada*,{dagger}, Akihiro Nakamura*, Yasuhiro Kohama*, Hiromichi Ishikawa{dagger}, Kazutake Tsujikawa* and Hiroshi Yamamoto3,*

* Department of Immunology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; and {dagger} Department of Microbiology, Keio University School of Medicine, Tokyo, Japan


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
We examined the severity of experimental colitis induced by dextran sulfate sodium (DSS) using immunologically manipulated mice. C57BL/6 mice showed more severe colitis than BALB/c mice, but mice of both strains recovered fully from the disease after the removal of DSS from their drinking water. The infiltrated cells at the lesions were mainly granulocytes in normal littermates. However, C.B-17 scid, IL-7R{alpha} deficient, and TCR-C{beta}{delta} double-deficient mice showed severe colitis and did not recover from the disease even after the removal of DSS. It was found that the infiltrated cells at the lesions in the lethal strains were monocytes. Although both TCR-C{delta}-/- and TCR-C{beta}-/- mice showed severe colitis phenotypes, infiltration in the former is monocyte-dominant while that in the latter is granulocyte-dominant. Thus the type of cells that infiltrate at the lesions of DSS-induced experimental colitis may be controlled by functional T cell subsets. Immunohistological and RT-PCR analyses of the inflamed colon revealed that the murine homologue of human GRO{alpha} released by some cells under the control of {gamma}{delta}T cells is a possible candidate determining the severity of DSS-induced experimental colitis.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
In the intestinal epithelia, lymphocytes and epithelial cells function as the major host defense against viral and bacterial infections, and against food Ags. Mouse intestinal intraepithelial T lymphocytes comprise {alpha}{beta} (40–70%) and {gamma}{delta}T (30–60%) cells (1, 2). {gamma}{delta}T cells are a minor T cell subset present in the spleen and lymph nodes, but represent a major subset within the epithelia of intestine and skin (3, 4, 5, 6). {gamma}{delta}T cells play active roles in the regulation and resolution of inflammatory processes associated with infectious agents (7, 8, 9, 10, 11, 12, 13) and autoantigens (14). They inhibit exaggerated tissue inflammation and necrosis following infection with bacteria, such as Listeria monocytogenes (7, 8, 9, 10, 11) and Mycobacterium tuberculosis (12), or parasites, such as Nippostrongylus braziliensis (10) and Eimeria vermiformis (13).

Homeostasis of intestinal epithelia is regulated by a variety of mechanisms. Recent studies have demonstrated cellular interactions between intestinal epithelial cells (IECs) 4 and intestinal intraepithelial lymphocytes (IELs). Komano et al. (15) reported that the absence of {gamma}{delta}T cells is associated with a reduction in the turnover of IECs, and a down-regulation of the expression of class II molecules. Boismenu and Havran (16) have shown that the keratinocyte growth factor produced by activated {gamma}{delta}T cells stimulates epithelial cell growth. Very recently, homeostasis of IECs was also found to be regulated by B lymphocytes (17, 18). Nishiyama et al. (18) reported that the homeostasis of IECs is negatively regulated by B lymphocytes, whereas it is positively regulated by {gamma}{delta}T cells (15). Moreover, it was suggested that cellular interactions among {gamma}{delta}IELs, {alpha}{beta}IELs, and IECs occur via cell surface molecules, suggesting the existence of an intranet to prevent potential inflammatory responses at the intestinal mucosal surface (19). Thus cellular interactions among IECs and IELs play an important role in the homeostatic regulation of intestinal epithelia.

Inflammatory bowel diseases (IBD), such as ulcerative colitis or Crohn’s disease, are caused by excessive tissue-damaging, chronic inflammatory responses in the intestinal epithelia. The down-regulation of immune responses restores mucosal function to normal, but the precise cellular mechanism for the induction is not well understood. There have been several reports demonstrating the involvement of {gamma}{delta}T cells in patients with IBD (20, 21, 22). McVay et al. (20) showed that chronic inflammatory responses with the characteristics of IBD are associated with distinct changes in the number, distribution, composition, and function of mucosal {gamma}{delta}T cells. The characteristics of various animal models of IBD have been extensively reviewed by Boismenu and Chen (23). The oral administration of dextran sulfate sodium (DSS) induces IBD in experimental animals including mice (24), hamsters (25), rats (26), and guinea pigs (27). Affected animals manifest weight loss, bloody diarrhea, and a hunched posture, and the histological phenotypes (24) are similar to those of human IBD. It is interesting that the removal of DSS from the drinking water initiates recovery from inflammation, and this feature makes it a useful model in which to study the cellular mechanisms involved in tissue repair.

To elucidate the role of T cells in the intestinal epithelia, we examined DSS-induced experimental colitis (EC) using various immunologically manipulated mice. We demonstrate here that T cells, not only {alpha}{beta}T but also {gamma}{delta}T cells, participate in the recovery phase from DSS-induced EC, and that {gamma}{delta}T cells play unique roles in the repair mechanisms from EC.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Mice

Specific pathogen-free BALB/c and C57BL/6 mice aged 5 wk were purchased from Charles River Japan (Yokohama, Japan). TCR-C{delta} deficient mice ({delta}-/-) of a C57BL/6 background (28) were generously provided by Dr. Y. Itohara (Institute of Physical and Chemical Research, Wako, Saitama, Japan) and backcrossed eight times with BALB/c in our animal facility. Background strains of mutant mice are shown in parentheses (BALB/c, B/c; C57BL/6, B6) hereafter. TCR-C{beta} deficient ({beta}-/-) (B6) mice have been described previously (29, 30). TCR-{beta}{delta} double mutant ({beta}-/-{delta}-/-) (B6) mice were created by brother-sister matings of {beta}+/-{delta}-/- (B6) mice in our animal facility. Mutant genotypes or phenotypes were determined by the methods described previously (15). IL-7R {alpha}-chain deficient (IL-7R-/-) (B6) mice (31) were generously provided by Dr. K. Ikuta (Kyoto University, Kyoto, Japan). C.B-17scid (SCID) mice aged 5 wk were purchased from CLEA Japan (Tokyo, Japan). All mice used were between 5 and 8 wk of age in our experiments.

Antibodies

Anti-Mac-1 (M1/70) mAb was purified from hybridoma culture supernatants in our laboratory. Anti-Gr-1 (RB6–8C5) mAb and biotin-conjugated goat anti-rat IgG were purchased from BD PharMingen (San Diego, CA) and Cedarlane Labs (Ontario, Canada), respectively. Anti-KC (GRO-1) mAb is a product of Genzyme (Cambridge, MA).

Induction of experimental colitis

Experimental colitis was induced by the oral administration of DSS (Mr 5000; Wako Pure Chemicals, Osaka, Japan). BALB/c and C.B-17 background mice received of 5% (w/v) DSS and C57BL/6 background mice received 2.5% DSS in their drinking water for 7 days. The intensity of colitis was monitored by hemoccult test (Hemoccult Test Wako; Wako Pure Chemicals) and changes in body weight.

Immunohistochemistry

Samples of colon, 10 mm in length, were embedded directly in OCT; compound (Tissue-Tek; Miles, Elkhart, IN) and frozen at -80°C. The tissue segments were sectioned with a cryostat at 6 µm and the sections were fixed in acetone. The sections were washed three times with PBS, preincubated with 5% BSA (BSA, fraction V, Sigma-Aldrich, St. Louis, MO) to block nonspecific binding of mAbs, and then incubated with each mAb for 30 min at 37°C. The sections were then washed three times with PBS, and incubated with biotin-conjugated goat anti-rat IgG. Endogenous peroxidase activity was blocked with 0.3% H2O2 and 0.1% NaN3 in distilled water for 10 min at room temperature. Next, the sections were washed three times with PBS and incubated with avidin-biotin peroxidase complexes (Vectastain Elite ABC kit; Vector Laboratories, Burlingame, CA). Histochemical color development was achieved by 3,3'-diaminobenzidine (Dojindo, Kumamoto, Japan). Finally, the sections were counterstained with hematoxylin for microscopy. Some sections were stained with H&E.

In vivo labeling and in situ staining of proliferating intestinal epithelial cells

Cells undergoing DNA replication were determined by the methods described earlier (32) with modifications. {delta}+/- and {delta}-/- (B/c) mice, either untreated or after 7 days of DSS administration, received intraperitoneal injections of 5-bromo-2'-deoxyuridine (BrdU, 20 mg/kg body weight) five times at 6-h intervals. One hour after the last injection, the colons were removed and embedded in OCT compound and frozen at -80°C. The 6-µm thick tissue sections were fixed in 4% paraformaldehyde, treated with 2 N HCl containing 0.5% Tween 20, neutralized with 0.1 M boric acid (pH 10), and finally washed with 5% BSA. The tissue sections were further incubated with anti-BrdU mAb (BD PharMingen) followed by incubation with rabbit anti-mouse IgG+A+M (H+L)-HRP (Zymed Laboratory, San Francisco, CA). Endogenous peroxidase activity was blocked by the methods used for immunohistochemistry (see above), and the sections were counterstained with hematoxylin.

In vitro neutrophil migration assay

The migration of neutrophils to inflammatory lesions was examined in vitro using chemotaxi-cell chambers (Kurabo, Osaka, Japan). {delta}-/- or heterozygous {delta}+/- (B/c) mice were given 5% DSS (or water) for 3 days, and the colons were obtained. Colon samples, 10 mm in length, were opened longitudinally and washed. A polycarbonate membrane (0.45 µm) (Corning, New York, NY) was applied to the serosal surface of the colon, and incubated 24 h in colon organ culture medium (5% FCS, 10 mM HEPES, and antibiotics, such as penicillin, streptomycin, kanamycin, and gentamicin, in RPMI 1640) according to the method of Imada et al. (33). After incubation, the culture medium was centrifuged and transferred into the lower-chambers of a chemotaxi-cell. Peritoneal exudate cells were obtained from the peritoneal cavity by injecting 12% casein i.p. to normal BALB/c mice. Six hours after the injection of casein, peritoneal exudate cells were obtained, washed, and added into the upper-chamber of a chemotaxic cell at a dose of 5 x 105 cells/200 µl/chamber. The cells were found to comprise >70% neutrophils by differential cell-staining (DiffQuik kit; Kokusai Shiyaku, Osaka, Japan). The chambers were incubated for 1 h, after which the migrated cells in the lower-chambers were collected and counted. Most (>90%) of the migrated cells were found to be neutrophils.

RT-PCR

{delta}+/- or {delta}-/- (B/c) mice were given 5% DSS for 7 days, and their colons were obtained 0, 3, 7, and 11 days later. IECs from the colons were prepared according to the methods described previously (15). The IECs preparation contained no detectable CD3+ cells. Total RNA was isolated from the IECs with an RNeasy Mini kit (Qiagen, Valencia, CA), and treated with DNaseI (PerkinElmer, Branchburg, NJ). RNA was transcribed to cDNA using an M-MLV Reverse Transcriptase kit (Invitrogen, San Diego, CA), and the cDNA was amplified by AmpliTaq (PerkinElmer). Primer pairs for the murine homologue of human GRO{alpha} (KC), LPS-induced CXC chemokine (LIX), macrophage inflammatory protein-2 (MIP-2), macrophage migration inhibitory factor (MIF), and hypoxanthine phosphorybosyl transferase (HPRT), are shown below. KC: 5'-TAT CGC CAA TGA GCT GCG C-3' and 5'-AAG CCA GCG TTC ACC AGA C-3; LIX: 5'-CTT CCT CAG TCA TAG CCG CAA C-3' and 5'-CCT TTC TTC TCT TCA CTG GGG TC-3'; MIP-2: 5'-CAC ACT TCA GCC TAG CGC CA-3' and 5'-TCA GAC AGC GAG GCA CAT CAG G-3'; MIF: 5'-CCG CAC AGT ACA TCG CAG TG-3' and 5'-ACA GCG GTG CAG GTA AGT GG-3'; HPRT: 5'-CTG TAG ATT TTA TCA GAC TGA AGA G-3' and 5'-GTC AAG GGC ATA TCC AAC AAC AAA-3'.


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
DSS-induced EC in various mutant mice

We first determined the effective dose of DSS for the induction of EC. As shown in Fig. 1A, BALB/c mice showed no symptoms of colitis at a dose of 2.5% DSS, but showed weight loss, bloody diarrhea, and a hunched posture at a dose of 5% DSS for 7 days. The symptoms disappeared after the removal of DSS from the drinking water, and the body weight returned to a normal level. In contrast, C57BL/6 mice showed symptoms at the dose of 2.5%, and recovered from colitis after the removal of DSS (Fig. 1B). However, none of the C57BL/6 mice receiving 5% DSS recovered from EC even after the removal of DSS, and all died between days 10 and 12. The results indicate that C57BL/6 mice have a higher sensitivity to DSS-induced EC than BALB/c mice. Thereafter, we used 2.5 and 5% DSS for mice having C57BL/6 and BALB/c backgrounds, respectively. (C57BL/6 x BALB/c)F1 mice showed intermediate responsiveness to DSS (data not shown).



View larger version (29K):
[in this window]
[in a new window]
  		FIGURE 1. Survival rates of DSS-induced experimental colitis mice. BALB/c (A) and C57BL/6 (B) mice were administered 2.5% (•) or 5.0% ({blacksquare}) dextran sulfate sodium (DSS) in their drinking water for 7 days. Body weight changes were monitored, and the values are expressed as percentage of original body weight. {dagger}, Died.

 
To examine the functional role of lymphocytes in the induction of DSS-induced EC, the sensitivity of severe combined immunodeficiency (C.B-17scid) mice was compared with that of BALB/c mice. As shown in Fig. 2B, SCID mice did not recover from EC and died between days 9 and 22, whereas wild-type (WT) BALB/c mice recovered (Fig. 2A). Similar to SCID mice, IL-7R-/- mice, which have no {gamma}{delta}T cells and severely reduced {alpha}{beta}T cells (24), died on days 11 and 12, but heterozygous littermates recovered from EC after the removal of DSS (Fig. 2C). Inflammatory lesions in SCID and IL-7R-/- mice were examined immunohistologically. Both strains exhibited a loss of epithelial cells, a drop out of crypts, and a decrease of goblet cells (data not shown). There were large numbers of Mac-1+/Gr-1+ cells infiltrating at the lesions of BALB/c mice (Fig. 3, C and D). In contrast, SCID mice showed an infiltration of large numbers of Mac-1+ cells but fewer Gr-1+ cells (Fig. 3, G and H). IL-7R-/- mice showed similar histological results to SCID mice (data not shown). The numbers of infiltrating cells in BALB/c and SCID mice were counted on the immunohistological sections and the results are summarized in Table I.



View larger version (22K):
[in this window]
[in a new window]
  		FIGURE 2. Survival rates of C.B-17scid mice and IL-7R{alpha} deficient mice after DSS administration. A and B, BALB/c (A; five mice) and C.B-17scid (B; six mice) mice were administered 5.0% DSS in their drinking water for 7 days. Body weight changes in BALB/c (A; • with solid lines) and C.B-17 scid (B; {blacksquare}) mice, and occult blood levels of BALB/c (• with dashed line; mean of individual scores) were monitored. C, IL-7R{alpha}+/- (•; three mice of B6 background) and IL-7R{alpha}-/- mice ({blacksquare}; three mice of B6 background) were administered 2.5% DSS in their drinking water for 7 days, and their body weight changes were monitored. {dagger}, Died.

 


View larger version (127K):
[in this window]
[in a new window]
  		FIGURE 3. Immunohistochemical analyses of the colons of EC mice. BALB/c (A–D) and C.B-17 scid (E–H) mice were administered 5.0% DSS in their drinking water for 7 days. On day 7, some mice were killed and serial cryosections of their colons were prepared. H-E staining (A and E) and magnified views of the boxed areas are shown (B and F). Serial sections were stained with anti-Mac-1 (C and G) or with anti-Gr-1 (D and H). Bars, 50 µm.

 

View this table:
[in this window]
[in a new window]
  		Table I. Infiltrated cell types in the inflammatory sites of DSS-induced colitis micea

 
DSS-induced experimental colitis in {delta}-/- mice

The above results suggest that functional lymphocytes may not be necessary for the induction of DSS-induced EC, but play some roles in the recovery from EC. As has been well characterized, {gamma}{delta}T cells locate in intestinal, vaginal, and bronchoalveolar epithelial tissues, and function in epithelial cell homeostasis (15). Next we examined further the role of {gamma}{delta}T cells in the recovery phase from DSS-induced EC. {delta}-/- and {delta}+/- (B/c) mice received 5% DSS for 7 days, and their survival was monitored. All mice exhibited occult blood from days 2 or 3 with bloody diarrhea, and weight loss beginning around days 5-6. However, 4 out of 7 {delta}-/- mice died on days 10 and 11 while the rest recovered (Fig. 4B), whereas {delta}+/- mice all recovered (Fig. 4A). Similar to mice on a BALB/c background, 5 of 7 {delta}-/- (B6) mice responded to DSS-induced EC and died, whereas 9 of 10 {delta}+/- (B6) mice recovered EC (data not shown)



View larger version (25K):
[in this window]
[in a new window]
  		FIGURE 4. Survival rates of TCR mutant mice after DSS administration. {delta}+/- and {delta}-/- (B/c) mice (A and B) were administered 5.0%, and {beta}-/-, {delta}-/-, {beta}-/-{delta}-/- (B6) mice (C) were administered 2.5% DSS in their drinking water for 7 days. A and B, Body weight changes of individual {delta}+/- (A) and {delta}-/- (B/c) (B) mice (eight mice) are shown. C, Body weight changes of individual {beta}-/- (•, six mice), {delta}-/- ({blacksquare}, 6 mice), and {beta}-/-{delta}-/- (B6) ({blacktriangleup}, five mice) are shown. {dagger}, Died.

 
Histological examination of the colons of {delta}-/- (B/c) mice on day 7 was then performed (Fig. 5). The infiltrated cells were found to comprise mainly of Mac-1+ cells (Fig. 5H), with smaller numbers of Gr-1+ cells in the lesions (Fig. 5I). CD4+ and CD8+ cells were scarcely detected (data not shown). It is interesting to note that in {delta}+/- (B/c) mice there were not only Mac-1+ cells (Fig. 5C), but also large numbers of Gr-1+ cells in the mucosal lesions (Fig. 5D). Since polymorphonuclear cells express both Mac-1 and Gr-1, whereas monocytes express Mac-1 but not Gr-1, the infiltrated cells in the {delta}+/- lesions are thought to be granulocytes. In contrast, the infiltrated cells in the lesions of {delta}-/- mice are monocytes (Fig. 5, H and I). In agreement with the immunohistochemical observations, infiltrated cells are thought to be granulocytes in {delta}+/- mice (Fig. 5E) and mononuclear cells in {delta}-/- mice (Fig. 5J) (Table I). The infiltrated cells in {delta}+/- mice were found to be neutrophils by differential staining methods (DiffQuik staining using single cells obtained from the lesions) (data not shown). In the small intestine, inflammation was not observed in either {delta}-/- or {delta}+/- mice (data not shown). {delta}+/- and {delta}-/- mice on a C57BL/6 background showed similar histologies their counterparts on a BALB/c background (data not shown).



View larger version (102K):
[in this window]
[in a new window]
  		FIGURE 5. Immunohistochemical analyses of the colons of TCR mutant mice. {delta}+/- (A–E), {delta}-/- (F–J), {beta}-/- (K–N), and {beta}-/-{delta}-/- (B6) (O–R) mice were administered 2.5% DSS in their drinking water for 7 days. On day 7, some mice were killed and serial cryosections of their colons were prepared. H&E staining (A, F, K, O) and magnified views of the boxed areas are shown (B, E, G, J, L, P). Serial sections were stained with anti-Mac-1 (C, H, M, Q) or anti-Gr-1 (D, I, N, R). Bars, 50 µm.

 
DSS-induced experimental colitis in {beta}-/- mice

We then examined the role of {alpha}{beta}T cells in recovery from EC. {beta}-/- (B6) mice were compared with either {beta}-/-{delta}-/- (B6) or {delta}-/- (B6) mice in terms of recovery from DSS-induced EC. {beta}-/-{delta}-/- double mutants responded similarly to SCID or IL-7R-/- mice (Fig. 2), in that two died on days 4 and 7, two were severely impaired and died on day 21, and one was mildly impaired and the disease progressed. Both {beta}-/- and {delta}-/- mice responded partly to DSS. Three of 6 {beta}-/- mice died around days 6 and 7, and the other three recovered. Three of 6 {delta}-/- mice died on days 5 and 7, one was severely impaired, and two recovered from the disease (Fig. 4C). Histological examination of the colons of {beta}-/- mice at day 7 was then performed (Fig. 5, K–R). The infiltrated cells comprised mainly of Mac-1+ Gr-1+ cells (Fig. 5, M and N) (Table I). In contrast, similar to SCID, IL-7R-/-, and {delta}-/- mice, the infiltrated cells in the {beta}-/-{delta}-/- mice comprised mainly of Mac-1+ Gr-1- cells (Fig. 5, Q and R) (Table I).

Collectively, the results indicated that, although some {beta}-/- and {delta}-/- mice subjected to DSS-induced EC die, the infiltrated cells in the former mice are mainly neutrophils whereas they are monocytes in the latter mice. The evidence suggests that {gamma}{delta} T cells are responsible for the migration of neutrophils.

Proliferating intestinal epithelial cells in {delta}-/- mice

Because the {delta}-/- mice responded differently than {delta}+/- mice to DSS, we examined the effect of DSS on proliferating intestinal epithelial cells. Before DSS treatment, there were no significant differences of the number of BrdU-incorporating cells between {delta}-/- and {delta}+/- (B/c) mice (Fig. 6, A–D), but on day 7, a significant decrease in proliferating IECs was seen in the DSS-treated {delta}-/- mice (Fig. 6, F and H). This suggests that {gamma}{delta}T cells positively regulate the maintenance of epithelial homeostasis during the recovery phase from DSS-induced EC.



View larger version (94K):
[in this window]
[in a new window]
  		FIGURE 6. Epithelial cell proliferation and their expression of KC. The proliferation of epithelial cells of mice with DSS-induced colitis were examined by anti-BrdU staining. {delta}+/- (A, C, E, G) and {delta}-/- (B/c) (B, D, F, H) mice were administered 5.0% DSS in their drinking water for 7 days. On day 0 or 7 aftr treatment, mice received five intraperitoneal injections of BrdU at 6-h intervals. They were then killed and serial cryosections of their colons were prepared. The incorporation of BrdU was visualized by staining the sections with anti-BrdU mAb and second Abs. The expression of KC was examined immunohistochemically in each strain on day 4 of 5.0% DSS administration (I–L). KC was found to be strongly expressed in the colons from {delta}+/- mice (I, K) whereas the colons of {delta}-/- mice showed almost no (J, L) KC expression. The results are representative of four separate experiments. Bars, 50 µm.

 
In vitro examination of cell migration and detection of chemokines by RT-PCR

We examined the in vitro migration of neutrophils to inflammatory colon lesions. Casein-induced peritoneal exudate cells, comprising >70% neutrophils, were incubated in the upper chambers of a chemotaxi-cell with colon culture supernatants in the lower chambers. As summarized in Fig. 7, the colon-cultured supernatant obtained from DSS-administered {delta}+/- (B/c) mice showed higher chemotactic activity to neutrophils than that from DSS-administered {delta}-/- (B/c) mice. Colon culture supernatants obtained from mice not treated with DSS, either {delta}+/- or {delta}-/-, showed no difference in chemotactic activity. Although the baseline was high, the results suggest that the migration of neutrophils into DSS-induced inflammatory lesions is dependent on factors derived at least from {gamma}{delta}T cells.



View larger version (11K):
[in this window]
[in a new window]
  		FIGURE 7. Migration of neutrophils in response to colon culture supernatants. {delta}+/- and {delta}-/- (B/c) mice were administered 5.0% DSS in their drinking water for 3 days. On day 0 or 3, mice were killed, colons were taken, and colon culture supernatants were prepared as described in Materials and Methods. Casein-induced peritoneal cells comprising mainly neutrophils (>70%) were incubated with culture medium in a chemotaxis-cell chamber. One hour after incubation, the migrated cells were collected, enumerated, and their cell types determined. The migrated cells were mainly (>90%) neutrophils. Data are means ± SD of triplicate determinations using the colons from two mice. The results are representative of four separate experiments. Statistical significance between groups was determined by Student’s t test. *, p < 0.05.

 
To detect possible CXC chemokines, which have neutrophil chemotactic activity, RT-PCR was performed using mRNAs from IECs. As shown in Fig. 8, IECs from {delta}+/- mice consistently produced the mRNA for KC (GRO-1) even after DSS administration, however, IECs from {delta}-/- mice produced less mRNA on days 3 and 7 of DSS treatment with the level returning to normal after the removal of DSS (day 11). The levels of the mRNAs for other CXC chemokines (LIX and MIP-2) and MIF mRNAs levels were not changed by DSS treatment. To confirm that the epithelial cells from the colons of {delta}-/- mice produce less KC than those of {delta}+/- mice, the expression of KC was examined immunohistochemically on day 4 of treatment with 5.0% DSS in each strain. As shown in Fig. 6, KC was strongly expressed in the epithelial perinuclear and cytoplasmic areas of the TCR{delta}+/- colons (Fig. 6, I and K) with almost no expression (Fig. 6, J and L) in TCR{delta}-/- colons. These results indicate that IECs from {delta}+/- mice produce enough KC to recruit neutrophils to the lesions. In contrast to {delta}+/- mice, IECs from {delta}-/- mice do not generate KC, resulting in less neutrophil migration.



View larger version (52K):
[in this window]
[in a new window]
  		FIGURE 8. RT-PCR study of the chemokines expressed in damaged or undamaged colon. {delta}+/- and {delta}-/- (B/c) mice were administered 5.0% DSS in their drinking water for the indicated number of days. On days 0 to 11, mice were killed, colons were removed, and intestinal epithelial cells of the colons were examined for their expression of various CXC (KC, LIX, macrophage-inflammatory protein-2) chemokines and MIF. HPRT was used as an internal control.

 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
Recently, Mahler et al. (34) reported major differences in genetic susceptibility to DSS-induced EC among inbred strains of mice. Our results showing C57BL/6 mice to be highly sensitive to DSS-induced EC while BALB/c mice are less sensitive (Fig. 1) are consistent with the earlier observations (34). Although the fact that C57BL/6 and BALB/c mice behave differently in inducing helper T cell subsets has been well described, IL-4 or IFN-{gamma}-R mutant mice show no significant differences from their WT littermates in terms of DSS-induced EC (data not shown in this report). In contrast, both SCID (Ref.35 , and this report) and IL-7R-/- mice (this report) respond to DSS-induced EC and all mice die even after the removal of DSS (Fig. 2). This suggests that the induction of EC is not dependent on functional lymphocytes, while functional lymphocytes do play a role in the recovery from EC. DSS-induced EC may be due to the direct cytotoxicity of DSS on epithelial cells, and functional lymphocytes are not involved in this process.

Immunohistological analyses of SCID and IL-7R-/- mice revealed that the infiltrated cells in these mutants are mainly Mac-1+ Gr-1- cells, i.e., monocytes, whereas the infiltrated cells in their WT or heterozygous littermates are Mac-1+Gr-1+ cells, i.e., granulocytes (neutrophils) (Fig. 3). It can be speculated that functional lymphocytes may release cytokines/chemokines that act to maintain epithelial homeostasis. We then examined which type of T cells, {alpha}{beta} or {gamma}{delta}T cells, participate in the recovery from DSS-induced EC. As shown in Figs. 5 and 7, the mortalities of both strains of mutant mice are similar, however, the infiltrated cell types are quite different, i.e., monocyte (Mac-1+Gr-1-) infiltration was evident in {delta}-/- mice whereas granulocyte (Mac-1+Gr-1+) infiltration was seen in {beta}-/- mice. That {delta}-/- mice show a more severe phenotype than WT littermates is in good agreement with the earlier observations of Boismenu and Chen (23).

Komano et al. (15) reported that the absence of {gamma}{delta}T cells is associated with a reduction in epithelial cell turnover. No such effects were observed in {alpha}{beta}T cell-deficient mice. In IBD patients, such as those with ulcerative colitis or Crohn’s disease, the absolute numbers of {gamma}{delta}T cells increase in the peripheral blood, mainly as a result of an increase in {gamma}{delta}T cells (21) and the V{delta}1/V{gamma}8 T cell subset (22). In mucosal lesions, the number of {gamma}{delta}T cells increases with an altered {gamma}{delta}T cell repertoire (20). It is possible to speculate that {gamma}{delta}T cells act in the repair of disabled epithelium. Boismenu and Havran (16) reported that {gamma}{delta}T cells express keratinocyte growth factor, but that intraepithelial {alpha}{beta}T cells neither produce this factor nor promote the growth of cultured epithelial cells. During the course of the present studies, Chen et al. (36) reported observations similar to ours. They showed that {gamma}{delta}IELs in DSS-treated mice express keratinocyte growth factor, and that {delta}-/- mice are more prone than WT to DSS-induced mucosal injury. Keratinocyte growth factor produced by {gamma}{delta}T cells also functions during wound repair in the skin (37). These findings indicate that intraepithelial {gamma}{delta}T cells regulate the generation and differentiation of intestinal epithelial cells and maintenance of homeostasis in intestinal epithelia. Although the overall mortalities of {delta}-/- and {beta}-/- mice in response to DSS are similar, the mechanisms involved in their recovery from EC may differ.

Judging from the results, inflammation associated with the infiltration of monocytes shows a more severe phenotype (SCID, IL-7R-/-, {delta}-/-, or {beta}-/-{delta}-/- mice) than that associated with granulocyte infiltration (heterozygous or WT littermates, except for {beta}-/- mice). It is not yet known how the infiltrating cell type determines the severity of inflammation; however, it is speculated that macrophages secrete more inflammatory cytokines than granulocytes. Elastase secreted by neutrophils may also act as potent mucus secretagogues in colon epithelial cells (38). The lack of neutrophil infiltration may cause less efficient epithelial cell function. It is generally accepted that infection-induced inflammation is associated with the first wave of granulocyte infiltration and that these are short-lived. The replacement of early granulocyte (neutrophil) infiltration by a second wave of macrophages occurs and acquired immunity is established (39, 40). However, macrophage infiltration to the lesions is not dependent on the infiltration of granulocytes (41). DiTirro et al. (40) reported that {delta}-/- mice secrete less macrophage-attracting chemokines, such as monocyte chemotactic protein-1, upon bacterial infection resulting in a delayed recruitment of macrophages. This observation is not consistent with our results, and the detailed molecular mechanisms of {gamma}{delta}T cell functions remain to be elucidated.

The assay of in vitro cell migration using the culture-supernatants of colon of DSS-administered {delta}+/- and {delta}-/- mice was performed against casein-induced peritoneal exudate cells (Fig. 7). That the supernatants from {delta}+/- mice had higher chemotactic activities than those from {delta}-/- mice, suggesting the existence of chemokines under the control of {gamma}{delta}T cells. The interaction of {gamma}{delta}T cells with granulocytes has not yet been clarified. Recently, a human {gamma}{delta}T cell clone was found to express and secrete not only CC chemokines, such as monocyte chemotactic protein-1, macrophage inflammatory protein-1{beta}, and RANTES, but also a CXC chemokine (IL-8) as granulocyte chemotactic factors (42). To investigate the role of granulocyte chemotactic factors, we examined the expression of CXC chemokines by RT-PCR analysis. As shown in Fig. 8, KC (GRO-1) was expressed from an earlier stage of EC. As it has been reported that a {gamma}{delta}T cell clone expresses CC but not CXC chemokines in mice (43), {gamma}{delta}T cells may be implicated in the recruitment of inflammatory cells to disabled epithelium, and {gamma}{delta}T cells may directly or indirectly regulate the infiltration of granulocytes into the mucosa. In our studies, {delta}-/- mice showed less granulocyte infiltration in inflammation sites than {delta}+/- mice (Fig. 3). These results indicate that activated {gamma}{delta}T cells at inflammation sites may secrete KC, and granulocytes may proliferate and migrate to the inflammation sites.

In the present study, we demonstrate that {delta}-/- mice behave differently than other immunologically manipulated mice, such as SCID, {beta}-/-, and IL-7R-/- mice, to DSS-induced EC. Further understanding of the molecular mechanisms of {gamma}{delta}T cell-mediated immune regulation will lead to therapeutic strategies for IBD.


   Acknowledgments
 
We are grateful to Dr. Margaret Dooley Ohto and Dr. Kae Chang Park for the critical reading of this manuscript.


   Footnotes
 
1 This work was supported by grants-in-aid from the Japanese Ministry of Education, Science, Sports, and Culture and the Japanese Ministry of Health and Welfare. This work was also supported partly through a grant of Long-Range Research Initiative by the Japanese Chemical Industry Association. Back

2 Current address: Department of Neurosurgery, Kochi Medical School, Nankoku, Kochi 783-8505, Japan. Back

3 Address correspondence and reprint requests to Dr. Hiroshi Yamamoto, Department of Immunology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamada-oka, Suita, Osaka 565-0871, Japan. E-mail address: hiroshiy{at}phs.osaka-u.ac.jp Back

4 Abbreviations used in this paper: IEC, intestinal epithelial cell; IEL, intestinal intraepithelial lymphocyte; IBD, inflammatory bowel disease; DSS, dextran sulfate sodium; EC, experimental colitis; BrdU, 5-bromo-2'-deoxyuridine; KC, the murine homologue of human GRO{alpha}; LIX, LPS-induced CXC chemokine; MIP-2, macrophage inflammatory protein-2; MIF, macrophage migration inhibitory factor; HPRT, hypoxanthine phosphorybosyl transferase; WT, wild type. Back

Received for publication February 7, 2003. Accepted for publication September 5, 2003.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Rocha, B., P. Vassalli, D. Guy-Grand. 1991. The V{beta} repertoire of mouse gut homodimeric {alpha} CD8+ intraepithelial T cell receptor {alpha}/{beta}+ lymphocytes reveals a major extrathymic pathway of T cell differentiation. J. Exp. Med. 173:483.[Abstract/Free Full Text]
  2. Ishikawa, H., Y. Li, A. Abeliovich, S. Yamamoto, S. H. Kaufmann, S. Tonegawa. 1993. Cytotoxic and interferon {gamma}-producing activities of {gamma}{delta} T cells in the mouse intestinal epithelium are strain dependent. Proc. Natl. Acad. Sci. USA 90:8204.[Abstract/Free Full Text]
  3. Haas, W., P. Pereira, S. Tonegawa. 1993. {gamma}/{delta} cells. Annu. Rev. Immunol. 11:637.[Medline]
  4. Groh, V., S. Porcelli, M. Fabbi, L. L. Lanier, L. J. Picker, T. Anderson, R. A. Warnke, A. K. Bhan, J. L. Strominger, M. B. Brenner. 1989. Human lymphocytes bearing T cell receptor {gamma}/{delta} are phenotypically diverse and evenly distributed throughout the lymphoid system. J. Exp. Med. 169:1277.[Abstract/Free Full Text]
  5. Guy-Grand, D., D. Cerf-Bensussan, B. Malissen, M. Malassis-Seris, >C. Briottet, P. Vassalli. 1991. Two gut intraepithelial CD8+ lymphocyte populations with different T cell receptors: a role for the gut epithelium in T cell differentiation. J. Exp. Med. 173:471.[Abstract/Free Full Text]
  6. Havran, W. L., S. Grell, G. Duwe, J. Kimura, A. Wilson, A. M. Kruisbeek, R. L. O’Brien, W. Born, R. E. Tigelaar, J. P. Allison. 1989. Limited diversity of T-cell receptor {gamma}-chain expression of murine Thy-1+ dendritic epidermal cells revealed by V{gamma}3-specific monoclonal antibody. Proc. Natl. Acad. Sci. USA 86:4185.[Abstract/Free Full Text]
  7. Mombaerts, P., J. Arnoldi, F. Russ, S. Tonegawa, S. H. Kaufmann. 1993. Different roles of {alpha}{beta} and {gamma}{delta} T cells in immunity against an intracellular bacterial pathogen. Nature 365:53.[Medline]
  8. Skeen, M. J., H. K. Ziegler. 1993. Intercellular interactions and cytokine responsiveness of peritoneal {alpha}/{beta} and {gamma}/{delta} T cells from Listeria-infected mice: synergistic effects of interleukin 1 and 7 on {gamma}{delta}T cells. J. Exp. Med. 178:985.[Abstract/Free Full Text]
  9. Fu, Y. X., C. E. Roark, K. Kelly, D. Drevets, P. Campbell, R. O’Brien, W. Born. 1994. Immune protection and control of inflammatory tissue necrosis by {gamma}{delta} T cells. J. Immunol. 153:3101.[Abstract]
  10. Ferrick, D. A., M. D. Schrenzel, T. Mulvania, B. Hsieh, W. G. Ferlin, H. Lepper. 1995. Differential production of interferon-{gamma} and interleukin-4 in response to Th1- and Th2-stimulating pathogens by {gamma}{delta} T cells in vivo. Nature 373:255.[Medline]
  11. Egan, P. J., S. R. Carding. 2000. Downmodulation of the inflammatory response to bacterial infection by {gamma}{delta}T cells cytotoxic for activated macrophages. J. Exp. Med. 191:2145.[Abstract/Free Full Text]
  12. D’Souza, C. D., A. M. Cooper, A. A. Frank, R. J. Mazzaccaro, B. R. Bloom, I. M. Orme. 1997. An anti-inflammatory role for {gamma}{delta} T lymphocytes in acquired immunity to Mycobacterium tuberculosis. J. Immunol. 158:1217.[Abstract]
  13. Roberts, S. J., A. L. Smith, A. B. West, L. Wen, R. C. Findly, M. J. Owen, A. C. Hayday. 1996. T-cell {alpha}{beta}+ and {gamma}{delta}+ deficient mice display abnormal but distinct phenotypes toward a natural, widespread infection of the intestinal epithelium. Proc. Natl. Acad. Sci. USA 93:11774.[Abstract/Free Full Text]
  14. Groh, V., A. Steinle, S. Bauer, T. Spies. 1998. Recognition of stress-induced MHC molecules by intestinal epithelial {gamma}{delta} T cells. Science 279:1737.[Abstract/Free Full Text]
  15. Komano, H., Y. Fujiura, M. Kawaguchi, S. Matsumoto, Y. Hashimoto, S. Obana, P. Mombaerts, S. Tonegawa, H. Yamamoto, S. Itohara, et al 1995. Homeostatic regulation of intestinal epithelia by intraepithelial {gamma}{delta}T cells. Proc. Natl. Acad. Sci. USA 92:6147.[Abstract/Free Full Text]
  16. Boismenu, R., W. L. Havran. 1994. Modulation of epithelial cell growth by intraepithelial {gamma}{delta}T cells. Science 266:1253.[Abstract/Free Full Text]
  17. Golovkina, T. V., M. Shlomchik, L. Hannun, A. Chervonsky. 1999. Organogenic role of B lymphocytes in mucosal immunity. Science 286:1965.[Abstract/Free Full Text]
  18. Nishiyama, Y., H. Hamada, S. Nonaka, H. Yamamoto, M. Nanno, Y. Katayama, H. Takahashi, H. Ishikawa. 2002. Homeostatic regulation of intestinal villous epithelia by B lymphocytes. J. Immunol. 168:2626.[Abstract/Free Full Text]
  19. Yamamoto, M., K. Fujihashi, K. Kawabata, J. R. McGhee, H. Kiyono. 1998. A mucosal intranet: intestinal epithelial cells down-regulate intraepithelial, but not peripheral, T lymphocytes. J. Immunol. 160:2188.[Abstract/Free Full Text]
  20. McVay, L. D., B. Li, R. Biancaniello, M. A. Creighton, D. Bachwich, G. Lichtenstein, J. L. Rombeau, S. R. Carding. 1997. Changes in human mucosal {gamma}{delta} T cell repertoire and function associated with the disease process in inflammatory bowel disease. Mol. Med. 3:183.[Medline]
  21. Giacomelli, R., I. Parzanese, G. Frieri, A. Passacantando, F. Pizzuto, T. Pimpo, P. Cipriani, A. Viscido, R. Caprilli, G. Tonietti. 1994. Increase of circulating {gamma}{delta} T lymphocytes in the peripheral blood of patients affected by active inflammatory bowel disease. Clin. Exp. Immunol. 98:83.[Medline]
  22. Soderstrom, K., A. Buch, E. Halapi, A. Gronberg, I. Magnusson, R. Kiessling. 1996. Increased frequency of abnormal {gamma}{delta} T cells in blood of patients with inflammatory bowel diseases. J. Immunol. 156:2331.[Abstract]
  23. Boimenu, R., Y. Chen. 2000. Insights from mouse models of colitis. J. Leukocyte Biol. 67:267.[Abstract]
  24. Okayasu, I., S. Hatakeyama, M. Yamada, T. Ohkusa, Y. Inagaki, R. Nakaya. 1990. A novel method in the induction of reliable experimental acute and chronic ulcerative colitis in mice. Gastroenterology 98:694.[Medline]
  25. Yamada, M., T. Ohkusa, I. Okayasu. 1992. Occurrence of dysplasia and adenocarcinoma after experimental chronic ulcerative colitis in hamsters induced by dextran sulphate sodium. Gut 33:1521.[Abstract/Free Full Text]
  26. Tamaru, T., H. Kobayashi, S. Kishimoto, G. Kajiyama, F. Shimamoto, W. R. Brown. 1993. Histochemical study of colonic cancer in experimental colitis of rats. Dig. Dis. Sci. 38:529.[Medline]
  27. Iwanaga, T., O. Hoshi, H. Han, T. Fujita. 1994. Morphological analysis of acute ulcerative colitis experimentally induced by dextran sulfate sodium in the guinea pig: some possible mechanisms of cecal ulceration. J. Gastroenterol. 29:430.[Medline]
  28. Itohara, S., P. Mombaerts, J. Lafaille, J. Iacomini, A. Nelson, A. R. Clarke, M. L. Hooper, A. Farr, S. Tonegawa. 1993. TCR {delta} gene mutant mice: independent generation of {alpha}{beta} T cells and programmed rearrangements of {gamma}{delta} TCR genes. Cell 72:337.[Medline]
  29. Mombaerts, P., A. R. Clarke, M. A. Rudnicki, J. Iacomini, S. Itohara, J. J. Lafaille, L. Wang, Y. Ichikawa, R. Jaenisch, M. L. Hooper, S. Tonegawa. 1992. Mutations in T-cell antigen receptor genes {alpha} and {beta} block thymocyte development at different stages. Nature 360:225.[Medline]
  30. Kanamori, Y., K. Ishimaru, M. Nanno, K. Maki, K. Ikuta, H. Nariuchi, H. Ishikawa. 1996. Identification of novel lymphoid tissues in murine intestinal mucosa where clusters of c-kit+ IL-7R+ Thy1+ lympho-hemopoietic progenitors develop. J. Exp. Med. 184:1449.[Abstract/Free Full Text]
  31. Maki, K., S. Sunaga, Y. Komagata, Y. Kodaira, A. Mabuchi, H. Karasuyama, K. Yokomuro, J.-I. Miyazaki, K. Ikuta. 1996. Interleukin-7 receptor-deficient mice lack {gamma}{delta} T cells. Proc. Natl. Acad. Sci. USA 93:7172.[Abstract/Free Full Text]
  32. Gratzner, H. G.. 1982. Monoclonal antibody to 5-bromo- and 5-iododeoxyuridine: a new reagent for detection of DNA replication. Science 218:474.[Abstract/Free Full Text]
  33. Imada, A., K. Ina, M. Shimada, T. Yokoyama, Y. Yokoyama, Y. Nishio, T. Yamaguchi, T. Ando, K. Kusugami. 2001. Coordinate upregulation of interleukin-8 and growth-related gene product-{alpha} is present in the colonic mucosa of inflammatory bowel disease. Scand. J. Gastroenterol. 36:854.[Medline]
  34. Mahler, M., I. J. Bristol, E. H. Leiter, A. E. Workman, E. H. Birkenmeier, C. O. Elson, J. P. Sundberg. 1998. Differential susceptibility of inbred mouse strains to dextran sulfate sodium-induced colitis. Am. J. Physiol. 274:G544.
  35. Dieleman, L. A., B. U. Ridwan, G. S. Tennyson, K. W. Beagley, R. P Bucy, C. O. Elson. 1994. Dextran sulfate sodium-induced colitis occurs in severe combined immunodeficient mice. Gastroenterology 107:1643.[Medline]
  36. Chen, Y., K. Chou, E. Fuchs, W. L. Havran, R. Boismenu. 2002. Protection of the intestinal mucosa by intraepithelial {gamma}{delta} T cells. Proc. Natl. Acad. Sci. USA 99:14338.[Abstract/Free Full Text]
  37. Jameson, J., K. Ugarte, N. Chen, P. Yachi, E. Fuchs, R. Boismenu, W. L. Havran. 2002. A role for skin {gamma}{delta} T cells in wound repair. Science 296:747.[Abstract/Free Full Text]
  38. Leiper, K., B. J. Campbell, M. D. Jenkinson, J. Milton, L. G. Yu, J. Democratis, J. M. Rhodes. 2001. Interaction between bacterial peptides, neutrophils and goblet cells: a possible mechanism for neutrophil recruitment and goblet cell depletion in colitis. Clin. Sci. Lond. 101:395.[Medline]
  39. Conlan, J. W.. 1996. Early pathogenesis of Listeria monocytogenes infection in the mouse spleen. J. Med. Microbiol. 44:295.[Abstract]
  40. DiTirro, J., E. R. Rhoades, A. D. Roberts, J. M. Burke, A. Mukasa, A. M. Cooper, A. A. Frank, W. K. Born, I. M. Orme. 1998. Disruption of the cellular inflammatory response to Listeria monocytogenes infection in mice with disruptions in targeted genes. Infect. Immun. 66:2284.[Abstract/Free Full Text]
  41. Rogers, H. W., E. R. Unanue. 1993. Neutrophils are involved in acute, nonspecific resistance to Listeria monocytogenes in mice. Infect. Immun. 61:5090.[Abstract/Free Full Text]
  42. Biswas, P., P. Rovere, C. De-Filippi, S. Heltai, C. Smith, L. Dagna, G. Poli, A. A. Manfredi, M. Ferrarini. 2000. Engagement of CD30 shapes the secretion of cytokines by human {gamma}{delta} T cells. Eur. J. Immunol. 30:2172.[Medline]
  43. Boismenu, R., L. Feng, Y. Y. Xia, J. C. Chang, W. L. Havran. 1996. Chemokine expression by intraepithelial {gamma}{delta} T cells: implications for the recruitment of inflammatory cells to damaged epithelia. J. Immunol. 157:985.[Abstract]



This article has been cited by other articles:


Home page
 J. Immunol.Home page
J. Holderness, L. Jackiw, E. Kimmel, H. Kerns, M. Radke, J. F. Hedges, C. Petrie, P. McCurley, P. M. Glee, A. Palecanda, et al.
Select Plant Tannins Induce IL-2R{alpha} Up-Regulation and Augment Cell Division in {gamma}{delta} T Cells
J. Immunol., November 15, 2007; 179(10): 6468 - 6478.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Gastrointest. Liver Physiol.Home page
J.-E. Ghia, P. Blennerhassett, R. T. El-Sharkawy, and S. M. Collins
The protective effect of the vagus nerve in a murine model of chronic relapsing colitis
Am J Physiol Gastrointest Liver Physiol, October 1, 2007; 293(4): G711 - G718.
[Abstract] [Full Text] [PDF]

Home page
 GutHome page
N. Eijkelkamp, C. J Heijnen, A. Lucas, R. T Premont, S. Elsenbruch, M. Schedlowski, and A. Kavelaars
G protein-coupled receptor kinase 6 controls chronicity and severity of dextran sodium sulphate-induced colitis in mice
Gut, June 1, 2007; 56(6): 847 - 854.
[Abstract] [Full Text] [PDF]

Home page
 J. Leukoc. Biol.Home page
A. A. Kuhl, N. N. Pawlowski, K. Grollich, C. Loddenkemper, M. Zeitz, and J. C. Hoffmann
Aggravation of intestinal inflammation by depletion/deficiency of {gamma}{delta} T cells in different types of IBD animal models
J. Leukoc. Biol., January 1, 2007; 81(1): 168 - 175.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire