Augmented expression of secondary lymphoid tissue
chemokine and EBI1 ligand chemokine in Crohn’s disease
D Kawashima, N Oshitani, Y Jinno, K Watanabe, S Nakamura, K Higuchi, T Arakawa
See end of article for
Dr N Oshitani, Department
Osaka City University
Graduate School of
Medicine, 1-4-3, Asahi-
machi, Abeno-ku, Osaka
545-8585, Japan; nobu@
Accepted for publication
3 February 2005
J Clin Pathol 2005;58:1057–1063. doi: 10.1136/jcp.2004.024828
Background: A dominant T helper type 1 (Th1) immune response is thought to be involved in Crohn’s
disease (CD). SLC/CCL21 and ELC/CCL19, chemokines that regulate T cell homing and promote
recirculating T and dendritic cell (DC) interactions, help control antigen specific T cell responses.
Aims: To investigate the Th1 response and SLC and ELC in CD pathogenesis.
Methods: Surgically resected intestine and mesenteric lymph nodes (MLNs) from controls and patients with
CD and ulcerative colitis (UC) were investigated. CD3, CD83, HECA452, VEGFR3, SLC, ELC, and CCR7
expression was studied immunohistochemically. CCR7 mRNA was quantified using real time RT-PCR.
Results: ELC was almost undetectable in intestinal samples. SLC was found sporadically in lymphoid
follicles, lymphoid aggregate venules, and lymphatic vessels. In MLNs, SLC was highly expressed in high
endothelial venules (HEVs), lymphatic vessels, and stromal DCs, predominantly in T cell areas. ELC was
highly expressed in mature DCs. There were significantly more SLC positive HEVs and ELC positive mature
DCs, important components of T cell areas, in CD. SLC, ELC, and CCR7 mRNA was significantly higher in
CD MLNs compared with UC. CD MLNs had increased expression of SLC and ELC, mainly in HEVs,
mature DCs, and lymphatic vessels, inducing T cell hyperplasia. CCR7 mRNA was increased in T cell
Conclusion: The dominant Th1 immune response is facilitated by interaction of SLC positive HEVs/
lymphatic vessels, ELC positive mature DCs, and CCR7 positive T cells in hyperplastic T cell areas. In CD,
memory T cells and mature DCs may home to MLN.
and large intestine. Longitudinal ulcers and a cobblestone
appearance are characteristic findings of Crohn’s disease, and
a pathognomonic feature of the disease is a histological
finding of non-caseating epithelioid cell granuloma. It has
been suggested that immunological dysregulation is involved
in the pathogenesis of Crohn’s disease,1 2and recent studies
revealed that T helper type 1 (Th1) skewing may affect the
immune response of patients with Crohn’s disease.3–9In
addition, macrophage dysfunction may play a role in the
aetiology of Crohn’s disease because macrophages in patients
with this disease phagocytose and process ingested antigens
in the intestine and lymph nodes, resulting in granuloma
formation.3Antigen presenting cells such as macrophages
and dendritic cells are key cells that influence the Th1 or Th2
differentiation of T cells. We have reported the expression of
costimulatory molecules on macrophage lineages in Crohn’s
disease intestine that function as antigen presenting cells to
elicit Th1 dominant T cell immunity.5 10
rohn’s disease is an intractable intestinal disease of
unknown aetiology that can involve the entire digestive
tract from mouth to anus, but particularly the ileum
‘‘Recent studies revealed that T helper type 1 skewing may
affect the immune response of patients with Crohn’s
Chemokines, categorised as CC and CXC subgroups
according to their molecular structure, are not only chemoat-
tractant to various white blood cells but are involved in the
Th1 and Th2 differentiation of T cells.11–13
lymphoid tissue chemokine (SLC), known as CCL21, and
EBI1 ligand chemokine (ELC), known as CCL19, are CC
chemokines derived from the p12 gene on chromosome 9,
which recognise chemokine receptor 7 (CCR7) as their
cellular receptor. SLC was established from the plt/plt mouse,
which has immature peripheral lymph organs.14SLC is found
in the high endothelial venules (HEVs) of lymphoid tissue
and lymph vessels, which are involved in the homing of T
cells and dendritic cells.15ELC was first identified as a
functional ligand of CCR7.16ELC is found in mature dendritic
cells and plays an important role in T cell interactions.17 18
We investigated intestinal and mesenteric lymph node
localisation and expression of SLC and ELC in controls, in
patients with ulcerative colitis (UC), and in patients with
Crohn’s disease. We also measured the expression of SLC,
ELC, and CCR7 mRNA to clarify the role of these chemokines
in the pathogenesis of Crohn’s disease.
MATERIALS AND METHODS
Surgical specimens taken from the lower intestine and
mesenteric lymph nodes were obtained from 22 patients
with Crohn’s disease (seven ileitis, 13 ileocolitis, and two
colitis), 18 patients with UC (16 total colitis type and two left
sided colitis type), and 10 controls (table 1). An approxi-
mately 5 65 62 mm segment was cut from the intestine,
and whole mesenteric lymph nodes were fixed in periodate/
lysine/2% paraformaldehyde at 4˚C for six hours, then
continuously incubated in a 10%, 15%, and 20% sucrose
gradient in phosphate buffered saline (PBS) at 4˚C for six
hours each. Specimens were embedded in OCT compound
(Miles Sankyo, Tokyo, Japan) in dry ice/acetone, and stored
at 280˚C until use. Informed consent was obtained from all
Abbreviations: ELC, EBI1 ligand chemokine; GALT, gut associated
lymphoid tissue; HEV, high endothelial venule; IL, interleukin; PBS,
phosphate buffered saline; PBS-T, phosphate buffered saline Tween;
PCR, polymerase chain reaction; RT, reverse transcription; SLC,
secondary lymphoid tissue chemokine; Th1/2, T helper type 1/2
be the primary sites of T cell recruitment to the inflamed
intestine and may perpetuate the inflammatory reaction by
activating memory T cells in patients with Crohn’s disease.
Dendritic cells in peripheral tissues begin maturation after
phagocytosing antigens, followed by CCR6 downregulation
and CCR7 upregulation, and are recruited to the lymph nodes
by a reciprocal reaction with SLC, which is produced by
lymphatic vessels.15 17In addition, CCR7 is expressed on naive
T cells and Th1 T cells.33Increased expression of SLC in
lymphatic vessels and HEVs triggers the recruitment of CCR7
positive dendritic cells and T cells, which leads to T cell zone
hyperplasia in Crohn’s disease lymph nodes. Dendritic cells
recruited in lymph nodes produce CCR7 to interact with
CCR7 positive T cells and expand the Th1 immune response
in Crohn’s disease. Suppression of SLC and ELC in Crohn’s
disease GALT by antibodies or drugs is a possible future
treatment for patients with Crohn’s disease.
In conclusion, differential expression of SLC, ELC, and
CCR7 in the mesenteric lymph nodes of patients with Crohn’s
disease characterises abnormal antigen processing and Th1
skewing as a pathogenetic mechanism in Crohn’s disease.
Mesenteric lymph nodes play an important role in T cell and
dendritic cell recruitment in patients with this disease.
D Kawashima, N Oshitani, Y Jinno, K Watanabe, S Nakamura,
K Higuchi, T Arakawa, Department of Gastroenterology, Osaka City
University Graduate School of Medicine, 1-4-3, Asahi-machi, Abeno-
ku, Osaka 545-8585, Japan
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