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. 2003 May;132(2):332–338. doi: 10.1046/j.1365-2249.2003.02155.x

CCR4 is an up-regulated chemokine receptor of peripheral blood memory CD4+ T cells in Crohn's disease

Y JO *, T MATSUMOTO *,, S YADA *, K FUJISAWA *, M ESAKI *, N ONAI †, K MATSUSHIMA †, M IIDA *
PMCID: PMC1808697  PMID: 12699425

Abstract

Several chemokine receptors are expressed selectively on the surface of T cells depending on their polarization. The aim of this study was to characterize chemokine receptor expression in peripheral blood memory T cells in Crohn's disease (CD) and ulcerative colitis (UC), and to correlate the expression with disease activity. Peripheral blood mononuclear cells (PBMCs) were obtained from 24 patients with CD, 30 patients with UC, 24 normal controls and 10 disease controls. PBMCs were stained by anti-CCR3, CCR4, CCR5, CXCR3, CD4, CD8, CD45RO and β 7 integrin, and the expression of the chemokine receptors were determined by flow cytometry. CCR4 expression on memory T cells was significantly lower in UC than in CD or normal controls, and that of memory CD4+ T and β 7high memory CD4+ T cells was significantly higher in CD than in UC or normal controls. CCR4 expression on memory CD4+ T cells exhibited significant positive correlation with disease activity in CD, and this decreased significantly after treatment. Such a decrease was not found in the disease controls. CCR5 and CXCR3 expression on memory CD8+ T cells was significantly lower in CD than in normal controls. CXCR3 expression on β 7high memory CD4+ T and CXCR3 expression on memory CD8+ T cells were lower in UC than in normal controls. These findings suggest that in peripheral blood memory T cells, chemokine receptor expression is different between CD and UC. Enhancement of CCR4 and suppression of CCR5 and CXCR3 seem to be the characteristic chemokine receptor profile in peripheral blood memory T cells of CD.

Keywords: chemokine receptors, Crohn's disease, memory T cells, ulcerative colitis

INTRODUCTION

Crohn's disease (CD) and ulcerative colitis (UC) are both types of chronic inflammatory bowel disease (IBD), which are of obscure aetiology and which are characterized by inflammatory infiltration within the small or large intestine [1,2]. It has been suggested that the cytokine profiles in the inflamed intestine are different between CD and UC. Whereas Th1 cytokines are predominant in CD, Th2 cytokines are considered to be the main cytokines which regulate inflammation in UC [1,2].

Recruitment and retention of lymphocytes are regulated by adhesion molecules [3], various chemokines and their receptors. Th1 and Th2 cells differ, both with regard to chemokine receptor expression and their responsiveness to chemokines [4–8]. Th1 cells express predominantly CXCR3 and CCR5 on their surface, whereas Th2 cells express predominantly CCR4 and CCR8 [4–10]. CCR4 is one of the major chemokine receptors of Th2-mediated cells, and it has been reported that CCR4-positive cells are predominant in the peripheral blood T cells of patients with atomic dermatitis (AD) and systemic lupus erythematosus (SLE) [11,12]. On the other hand, CCR5- and CXCR3-positive T cells are increased in multiple sclerosis (MS), a disease in which activated peripheral blood T cells are known to produce higher levels of interferon (IFN)-γ[13]. These findings suggest that the chemokine receptor expression of peripheral blood T cells is associated closely with inflammation and cytokine balance in a certain proportion of diseases. It thus seems plausible to assess the chemokine receptor expression of peripheral blood T cells in patients with IBD.

Cell adhesion molecules are also crucial for the adherence and retention of lymphocytes in various tissues. α4β 7 is an important adhesion molecule, which interacts with MAdCAM-1 on intestinal endothelial cells [14]. αEβ 7 is another adhesion molecule, which interacts with E-cadherin [15]. Because β 7 is a common subunit of these adhesion molecules, peripheral blood T cells bearing the β7 integrin are characteristic of intestinal homing cells [14].

In this study, we analysed chemokine receptor expression of peripheral blood T cells in IBD patients, while paying special attention to intestinal homing cells, in order to clarify possible differences in the profile of chemokine receptor expression between CD and UC, and between active and inactive disease. Flow cytometry was applied for the determination of chemokine receptor expression.

SUBJECTS AND METHODS

Subjects

Twenty-four patients with CD, 30 patients with UC, 24 normal controls (NC) and 10 disease controls (DC) treated by total parenteral nutrition (TPN), comprised the subjects of the present investigation. A diagnosis of CD or UC had already been made, based upon confirmation of clinical, endoscopic and histological findings compatible with those diseases. The clinical characteristics of the subjects are summarized in Table 1. Informed consent from each subject was obtained before enrolment into this study, and the study had the approval of the institutional committee.

Table 1.

Clinical characteristics of subjects and controls

Crohn's disease Ulcerative colitis Normal controls
Gender (males/females) 19/5 17/13 14/10
Age (years)  15–54 (28·4) 10–51 (31·4) 22–55 (29·9)
Duration (years) 0·5–16 (4·4) 0·5–19 (5·8)
Affected site
 I + C  7 0
 I 12 0
 C 5 30
SASP/5-ASA (mg/day)  0–3000 (548)   0–4500 (2667)
PSL (mg/day)  0  0–80 (18·4)
Disease activity* 150–486 (217) 94–257 (189)
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I + C, intestine and colon; I, intestine; C, colon; SASP, salazosulphapyridine; 5-ASA, mesalazine;. PSL, peroral prednisolone.

*

Disease activity is expressed as values calculated by the formula for the Crohn's disease activity index [16] and the ulcerative colitis activity index [17]. Figures in parentheses refer to mean values.

The disease activity in each patient was assessed by the Crohn's disease activity index (CDAI) [16] for CD and by the ulcerative colitis disease activity index (UCAI) [17] for UC. In the case of both indices, a value>150 was considered to be indicative of active disease. Fifteen patients with active CD were admitted, and treated by TPN for 4 weeks. In addition, three of them were treated with peroral mesalazine (2250–3000 mg/day). Nine patients with UC were free from any medication at the time of recruitment, while the remaining patients were being treated with prednisolone and mesalazine. In the latter group of UC patients, the therapy was altered by administering larger doses of oral prednisolone (six patients) or by carrying out total colectomy (three patients). In 15 patients with CD and nine patients with UC, the PBMCs were assessed both before and after treatment. Ten DC without Crohn's disease or UC underwent TPN for 2–4 weeks (mean 2·9 weeks). In the DC, chemokine receptor expression was assessed before and after TPN treatment.

Preparation of peripheral blood T cells

A blood sample was obtained from each subject and PBMCs were isolated by the density gradient separation method using lymphocyte-separation medium (ICN Biomedical, Aurora, OH, USA). The viability of the separated cells was confirmed to be greater than 98% by trypan blue staining. The PBMCs were washed twice by RPMI-1640 containing 10% bovine serum albumin, and then they were used for fluorescence-activated cell sorter (FACS) analysis.

FACS analysis

A monoclonal antibody (MoAb) against CCR4 (KM2160, mouse IgG1) was generated, according to the method described previously [18]. The other MoAbs were commercially available. Purified anti-CCR3 (61828·111, rat IgG2A) was purchased from R&D Systems (Minneapolis, MN, USA). Purified anti-CXCR3 (1C6, mouse IgG1), fluorescein isothiocyanate (FITC)-conjugated anti-CCR5 (2D7, mouse IgG2a), purified anti-integrin β 7 (FIB504, rat IgG2a), perCP-conjugated anti-CD4 (SK3, mouse IgG1), perCP-conjugated anti-CD8 (SK1, mouse IgG1), phycoerythrin (PE)-conjugated anti-CD45RO (UCHL-1, mouse IgG2a), FITC-conjugated antimouse IgG1 (rat IgG1), biotin antirat mouse IgG2a (RG7, mouse IgG2b), streptavidin (SA)-allophycocyanin (APC), FITC-conjugated antimouse rat IgG (R19-5, rat IgG1) and isotype-matched control MoAbs conjugated with FITC were obtained from Becton Dickinson (San Jose, CA, USA).

PBMCs were exposed to anti-β 7 integrin for 30 min on ice. After being washed in phosphate-buffered saline with 1% FCS and 0·1% sodium azide, the cells were incubated with biotin-conjugated antirat mouse IgG for 20 min on ice. The cells were then washed in washing buffer, followed by reaction with SA-APC. For the staining of chemokine receptors, the cells were reacted on ice with FITC-conjugated anti-CCR4 or FITC-conjugated anti-CCR5 with a mixture of PE-conjugated anti-CD45RO, PerCP-conjugated anti-CD4 or PerCP-conjugated anti-CD8 for 30 min. For the staining of CXCR3, PBMCs were incubated with a mixture of anti-β7 integrin and anti-CXCR3, then FITC-conjugated antimouse rat IgG was used as the second antibody for CXCR3. The four-colour-stained cells (CCR3; CCR4; CCR5; CXCR3/CD4; CD8/CD45RO/β7 integrin) were analysed using a FACScan flow cytometer (Becton Dickinson, San Jose, CA, USA). The expression of β 7 integrin was classified into β7high or β 7low, according to the intensity.

Statistical analysis

The percentages of chemokine receptors were expressed as interquartile values. Statistical analyses were performed, using the Kruskal–Wallis test for the difference among the groups, followed by the unpaired Mann–Whitney test for comparisons between any two of the groups. Wilcoxon's signed-rank test was used for comparisons between findings before and after treatment. Correlations between chemokine receptor expression and clinical activity index were evaluated by Spearman's correlation coefficient test. Although probabilities <0·05 were considered to be significant, the P-values were interpreted after Bonferroni correction for multiple comparisons among the groups.

RESULTS

Comparison of memory T lymphocyte population among the groups

Percentages of memory (CD45RO+) CD4+ T cells and memory (CD45RO+) CD8+ T cells in PBMCs are shown in Table 2. There was no significant difference in the percentage of memory CD4+ T or memory CD8+ T cells among CD, UC and NC.

Table 2.

Percentage of memory CD4+ and CD8+ T cells in lymphocyte gate

CD4+ CD45RO+ CD4+ CD45RO+ β7high CD8+ CD45RO+ CD8+ CD45RO+β7high
Crohn's disease 15·8–22·9 3·0–5·7 5·6–13·7 1·8–4·2
Ulcerative colitis 14·0–33·0 3·0–5·2 8·4–14·7 1·6–4·6
Normal controls 17·7–22·1 2·7–5·9 5·6–11·7 3·3–5·7
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Results are expressed as percentage positivity in lymphocytes gate (25th to 75th percentile).

Comparison of CCR4 expression among the groups

The percentage of CCR4+ memory CD4+ T (3·3–5·2%versus 1·7–3·3%, P < 0·005) and CCR4+ memory β 7high CD4+ T cells (0·62–1·40%versus 0·38–0·53%, P < 0·01) in PBMCs was significantly higher in CD than in NC. The percentage of CCR4+ memory CD4+ T (0·8–2·6%) and CCR4+ memory β 7high CD4+ T cells (0·30–0·52%) in UC was not significantly different from NC.

CCR4 expression on memory CD4+ T (5·7–12·0%versus 12·0–17·2%) and memory β 7low CD4+ T cells (4·4–10·7%versus 7·8–14·6%) was significantly lower in UC than in NC (P < 0·005) (Fig. 1a). Five UC patients with long-term remission showed a high expression of CCR4 on memory CD4+ T cells. In contrast, CCR4 expression on memory CD4+ T (15·0–24·1%versus 12·0–17·2%, P < 0·001) and memory β 7high CD4+ T cells (14·2–23·0%versus 7·0–12·2%, P < 0·0001) was significantly higher in CD when compared to NC (Fig. 1a,b). When the values were compared between CD and UC, CCR4 expression in CD was significantly higher on memory CD4+ T cells (15·0–24·1%versus 5·7–12·0%, P < 0·0001) (Fig. 1a), memory β 7high CD4+ T cells (14·2–23·0%versus 6·6–11·6%, P < 0·0001) (Fig. 1b) and memory β 7low CD4+ T cells (13·2–22·5%versus 4·4–10·7%, p < 0·0001) than in UC.

Fig. 1.

Fig. 1

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Comparison of CCR4 expression on peripheral blood memory T cells among Crohn's disease (CD), ulcerative colitis (UC) and the controls (NC). CCR4 expression on memory CD4+ T cells is significantly higher in CD than in UC or the controls (a) (*P < 0·0001, ***P < 0·001, respectively), but it is significantly lower in UC than in NC (a) (**P < 0·005). CCR4 expression on β 7high memory CD4+ T cells is significantly higher in CD than in UC or the controls (b) (*P < 0·0001).

CCR4 expression on memory CD8+ T and memory β 7low CD8+ T cells (P < 0·005) was significantly higher in CD than in NC (both P < 0·005) (data not shown). In contrast, CCR4 expression by memory CD8+ T and memory β 7low CD8+ T cells was lower in UC than in NC (both P < 0·05) (data not shown).

Comparison of CCR3, CCR5, and CXCR3 expression among the groups

The number of CCR3-positive cells was small in each group, and comprised less than 1·0% of the CD4+ T cells. CCR5 expression on CD4+ T cells did not differ among the groups (Table 3); however, CXCR3 expression on memory β 7high CD4+ T cells was lower in UC (P < 0·005) than in NC (Table 3). CCR5 expression by memory CD8+ T cells was lower in CD than in NC (P < 0·005) (Table 4). CXCR3 expression on memory CD8+ T and memory β7low CD8+ T cells was significantly lower in CD than in NC (P < 0·005, P < 0·001). CXCR3 expression on memory CD8+ T and memory β 7low CD8+ T cells was also significantly lower in UC than in NC (P < 0·0005).

Table 3.

CCR5 and CXCR3 expression rate on memory CD4+ T cells

CD4+ CD45RO+ CD4+ CD45RO+β7high CD4+ CD45RO+ β7high
CXCR3+ cells CCR5+ cells CXCR3+ cells CCR5+ cells CXCR3+ cells CCR5+ cells
Crohn's disease 41·1–54·1 14·0–31·8 38·5–55·8 19·7–32·9 41·2–56·5 17·6–33·1
Ulcerative colitis 41·4–54·7 19·9–33·3 42·1–51·3 a 25·6–36·6 44·9–55·8 18·7–40·1
Normal controls 45·9–58·6 20·9–33·8 52·8–59·8 a 21·8–38·9 47·8–56·5 20·9–31·3
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Results are presented as 25th−75th percentile. P-values show a significant difference between the two groups as calculated by the unpaired Mann–Whitney test

a

(P < 0·005). P-value < 0·006 is considered to be significant by Bonferroni correction.

Table 4.

CCR5 and CXCR3 expression rate on memory CD8+ T cells

CD8+ CD45RO+ CD8+ CD45RO+β7high CD8+ CD45RO+β7high
CXCR3+ cells CCR5+ cells CXCR3+ cells CCR5+ cells CXCR3+ cells CCR5+ cells
Crohn's disease 33·6–53·1a 46·7–64·5 a 46·0–66·2 58·5–75·7 34·7–45·1c 42·9–60·0
Ulcerative colitis 28·0–57·0b 49·1–74·5 36·1–70·0 60·3–81·7 26·2–53·6b 44·8–70·3
Normal controls 50·0–61·5a,b 62·3–79·7a 56·6–68·9 76·4–84·3 47·9–65·0b,c 55·8–74·4
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Results are presented as 25th −75th percentile. P-values show a difference between the two groups as calculated by the unpaired Mann–Whitney test

a

P < 0·005

b

P < 0·0005

c

P < 0·001

A P-value < 0·006 is considered to be significant by Bonferroni correction.

Correlation between chemokine receptor expression and disease activity

As indicated in Fig. 2, there was a significant positive correlation between CCR4 expression on memory CD4+ T cells and clinical activity in 24 different patients with CD (P < 0·05). In UC, however, no such significant correlation was found between chemokine receptor expression and disease activity.

Fig. 2.

Fig. 2

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Correlation between CCR4 expression on memory CD4+ T cells and Crohn's disease activity index (CDAI) in 24 different CD patients. CCR4 expression on memory CD4+ T cells exhibits a significant positive correlation with CDAI (P < 0·05).

We then assessed possible correlations between the various chemokine receptor expression levels. As shown in Fig. 3, CCR4 expression negatively correlated with CXCR3 expression on memory CD4+ T cells in CD (P < 0·005). However, no significant correlations were found between chemokine receptor expression levels in UC group or in NC.

Fig. 3.

Fig. 3

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Correlation between CCR4 and CXCR3 expression on memory CD4+ T cells in 24 different CD patients. CCR4 expression exhibits a negative correlation with CXCR3 expression which is statistically significant (P < 0·005).

Effect of therapy on CCR4 expression

In CD, CCR4 expression on memory CD4+ T, memory β 7high CD4+ T and memory β 7low CD4+ T cells was decreased significantly after treatment (from 19·0–26·5% to 10·5–21·0%, P < 0·05; from 14·7–24·8% to 7·3–17·0%, P < 0·05; and from 13·2–22·5% to 9·8–16·8%, P < 0·05, respectively) (Fig. 4a,b). In DC, CCR4 expression on neither memory CD4+ T (from 11·6–20·5% to 11·0–22·0%), memory β 7high CD4+ T (from 10·3–17·3% to 8·8–18·9%) nor memory β7low CD4+ T cells (from 10·4–15·0% to 9·5–15·7%) changed significantly after TPN (Fig. 4c,d). In UC, CCR4 expression on all the phenotypes remained unchanged (data not shown).

Fig. 4.

Fig. 4

Fig. 4

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Changes in CCR4 expression on memory CD4+ T cells before and after TPN in CD (a, b) and in disease controls (c, d). In CD, CCR4 expression on memory CD4+ T (a) and β7high memory CD4+ T cells (b) is decreased significantly after treatment (P < 0·05). Such decreases in CCR4 expression were not found in disease controls (c, d).

There were no significant changes in CCR5 or CXCR3 expression on memory CD4+ T cells during therapy in either the CD or UC groups. Similarly, there were no significant differences with regard to chemokine receptor expression on CD8+ T cells between before and after treatment in CD or UC.

DISCUSSION

The assessment of PBMCs partly gives rise to an understanding of the immunopathogenesis of IBD [19–22]. We confirmed in the present investigation that CCR4 expression on memory CD4+ T cells is increased, that it is related to the severity of the disease, and that it decreases after treatment in CD. Moreover, we were able to show that in CD, CCR4 expression has a negative correlation with CXCR3 expression on memory CD4+ T cells. On the other hand, CCR4 expression on memory CD4+ and CD8+ T cells was decreased in UC, regardless of the severity of the disease or the medications given. However, caution should be paid to the interpretation of our results because the behaviour of PBMCs is not equivalent to that in the topical inflamed mucosa of IBD [20–22].

It has been shown recently in vitro that different chemokine receptors are expressed preferentially in polarized Th1 and Th2 cell lines. Prior investigations have indicated that CCR5 and CXCR3 are expressed predominantly on Th1 cells [10,23], and CCR3 and CCR4 on Th2 cells [10–12]. Among Th2 cells, the expression of CCR4 has been shown to be much higher than that of CCR3. While CCR4 was up-regulated in our patients with CD, CCR3 expression was extremely low in all the groups examined. It has been recently shown that CCR4+ CD4+ cells had Th2 profiles in healthy subjects and in patients with atopic dermatitis [10,11,25]. It thus seems likely that CCR4 expressing memory CD4+ T cells functioned as Th2 cells in our patients with IBD. From this point of view, our results indicate the possibility that a Th2 response may occur in PBMCs of CD patients in response to the severity of the disease.

Previous reports have suggested that CD is characterized by a Th1 response within the mucosa, whereas a mucosal Th2 response is evident in both UC and early CD lesions [2]. The possible enhancement of a Th2 response in peripheral blood T cells in CD may be a counterpart of the early intestinal lesions in CD or a consequence of immunosuppression of Th1 cytokines, which predominate within the intestinal lesions [1,2]. However, because CCR4 expression has not been proven to be restricted to Th2 cells, the up-regulation of CCR4 in CD may not be attributable to an enhanced Th2 response alone.

Because β 7 has been shown to be expressed on mitogen-stimulated T cells and, more recently, on cytotoxic T lymphocytes involved in renal allograft rejection as a heterodimer with CD103 [26], we investigated whether memory integrin β 7high CD4+ T cells could be intestinal homing lymphocytes. CCR4 expression on memory integrin β 7high CD4+ T cells was found to be higher in CD than UC or NC. These findings suggest that CCR4 expression on memory CD4+ T intestinal homing lymphocytes is up-regulated in CD. However, a previous study suggested that only a small number of CCR4-expressing cells were intestinal homing cells [27]. In addition, infiltrating lymphocytes in the mucosal lesions of CD were shown to be negative for CCR4 [28]. Although the percentage of CCR4+ memory β 7high CD4+ T cells was increased in the PBMCs of our CD patients, the absolute number was in fact small. The small number of the cells, even in PBMCs, may explain the absence of the cells in the mucosal lesions shown in previous investigations [28]. Another explanation may be that the expression of CCR4 on intestinal T lymphocytes attenuates after infiltration into the intestinal wall via receptor internalization. Further investigations into CCR4 expression on intestinal homing T cells are necessary to clarify this issue.

In a recent report, increases in MDC and TARC mRNA expression were demonstrated in the inflamed mucosa of active CD. MDC protein has been shown to be produced by isolated PBMC or LPMC from patients with CD [29]. So far, CCR4 has been shown to be the only receptor for MDC and TARC [30], which are produced predominantly by antigen-presenting cells (APC), such as mature dendritic cells [31]. Therefore, enhanced CCR4 expression in peripheral memory T cells in CD, especially in those with positive integrin β 7high, seems to be related to a rise in expression of its ligands in systemic circulation and intestine.

There are other possible functions of CCR4-expressing memory CD4+ T cells. A recent study identified professional regulatory T cells among CD4+ CD25+ T lymphocytes from human peripheral blood T cells [32,33], and such T cells have been shown to express CCR4 and CCR8 [34]. These T cells are non-proliferating and they suppress conventional CD4+ T cells in a contact- and dose-dependent fashion without antigen-specificity. Another study discussed CCR4+ CD4+ T cells which preferentially express CD25 in patients with atopic dermatitis [11]. These studies imply that CCR4+ memory CD4+ T cells are candidates for ‘professional regulatory T cells’in vivo, and that these cells possibly attenuate T cell activation.

In contrast to CD, CCR4 expression on CD4+ and CD8+ cells was decreased in UC, and it remained low even after treatment. Kurashima et al. [35] reported that steroids reduced CCR4 expression on PBMCs in patients with bronchial asthma. In our study, 21 of 30 UC patients were receiving prednisolone at the time of assessment. This may have resulted in a decrease in CCR4 expression on CD4+ memory T cells. The effect of prednisolone on CCR4 expression is further suggested by the fact that CCR4 expression on memory CD4+ T cells was not decreased in those patients with UC who were not administered prednisolone over a prolonged period.

In contrast to western countries, TPN is an accepted strategy for patients with active CD in Japan [36]. While the expression of CCR4 on memory CD4+ T cells correlated well with disease activity in CD, it decreased further significantly after TPN without any changes in the expression of the other chemokine receptors. In addition, such a decrease in CCR4 expression was not found in DC during TPN. It thus seems likely that the decrease of CCR4 seems to be a consequence of the improvement of CD, rather than that of TPN itself. Because the precise mechanism of the decrease is not yet understood, further investigations are necessary.

Recently, chemokine expression in the inflamed tissue of CD and UC has been investigated vigorously. An investigation of CCL5/RANTES, CCL2/monocyte chemoattractant protein MCP-1 and CCL6/MCP-3 expression in colonic biopsy specimens from patients with CD and UC revealed that these chemokines were abundant in vascular endothelial cells, intraepithelial lymphocytes or lamina propria mucosa in both diseases [37]. Uguccinoni et al. [38] provided immunohistochemical evidence for the enhanced expression of IFN-γ-inducible protein-10 (IP-10), IL-8, MCP-1 and MCP-3 in colonic biopsy specimens from patients with UC. IP-10 receptor and CXCR3-expressing cells have also been shown to be increased in the colonic mucosa of UC [4] and the submucosa of CD [39]. CCR5 is the receptor for MIP-1α, MIP-1β and RANTES, while CXCR3 is the receptor for IP-10 and Mig [4]. In our investigation, the expression of CCR5 and CXCR3 on CD4+ and CD8+ peripheral blood T cells was somewhat decreased in CD and UC.

The mechanisms and kinetics of down-regulation of CCR5 and CXCR3 expression on memory T cells are unclear. In this study, CCR4 expression showed a negative correlation with CXCR3 expression on memory CD4+ T cells in CD. Down-regulation of CXCR3-expressing memory T cells on peripheral memory CD4 T cells may imply the counter-regulation of CCR4-expressing T cells at the systemic level. In a gene-targeted experimental animal model of IBD, Andres et al. [40] demonstrated a reduction in colony lesions during dextran sodium sulphate-mediated chronic colitis in CCR2 and CCR5-deficient mice. CCR5 and CXCR3 in patients with IBD may be down regulated to prevent an excess of lymphocyte recruitment to the bowel wall. In such circumstances, the expression of CCR5, which depends partly on IL-2 [23], may be inhibited by IL-12 through induction of β-chemokine [41]. Changes in the cytokine profiles in CD and UC may contribute to the reduction of CCR5 and CXCR3 expression on memory T cells.

In summary, our results indicate that CCR4+ memory CD4+ T cells are up-regulated and CCR5, CXCR3+ memory T cells are down-regulated in CD, presumably through a modification of Th1/Th2 balance. In addition, down-regulation of CCR5 and CXCR3 expression of memory T cells may be the characteristic chemokine receptor profile of CD and UC. Although the correlation of chemokine receptor expression with the Th1/Th2 balance and the relationship between peripheral and topical lymphocytes need to be investigated further, these chemokine receptors may be potential therapeutic targets for the treatment of IBD.

Acknowledgments

The English language used in this manuscript was revised by Miss K. Miller (Royal English Language Centre, Fukuoka, Japan).

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