CD44 Deficiency Attenuates Chronic Murine Ileitis

doi:10.1053/j.gastro.2008.08.053

Gastroenterology

Volume 135, Issue 6, December 2008, Pages 1993-2002

https://doi.org/10.1053/j.gastro.2008.08.053 Get rights and content

Background & Aims

Lymphocyte recruitment to sites of inflammation requires the sequential engagement of adhesion molecules and chemokine receptors. In the current studies we analyzed the role of CD44 for the development of chronic small-intestinal inflammatory infiltrates in vivo.

Methods

By using a tumor necrosis factor (TNF)-driven model of chronic ileitis (ie, B6.129P-TNF^{ΔAU-rich element [ARE]}) that recapitulates many features of Crohn's disease, we noticed dynamic changes in the expression and functional state of CD44 and its ligand hyaluronan via enzyme-linked immunosorbent assay, real-time reverse-transcription polymerase chain reaction, immunohistochemistry, and flow cytometry. In addition, we assessed the role of lymphocyte populations during induction of ileitis through adoptive transfer studies, and generated CD44-deficient TNFΔARE mice to assess the role of CD44 for development of ileitis.

Results

Soluble hyaluronan levels and expression of hyaluronan synthase-1 were increased in TNFΔARE mice. This coincided with increased expression of CD44 (including variant 7) and reactivity towards hyaluronan on CD4⁺ T cells. CD44 was spatially colocalized with the gut-homing integrin α₄β₇, spatially linking lymphocyte rolling with arrest. These cells had an effector phenotype because they lacked L-selectin and a higher proportion in diseased mice produced TNF and interleukin-2 compared with wild-type littermates. Lastly, CD4⁺ but not CD8⁺ T cells conferred ileitis to RAG^−/− recipients and deficiency of one or both alleles of the CD44 gene resulted in attenuation of the severity of ileitis in TNFΔARE mice.

Conclusions

Our findings support an important role of CD44 expressed by CD4⁺ and CD8⁺ for development of ileitis mediated by TNF overproduction.

Section snippets

Mice

The B6.129S-Tnf^tm2Gkl/Jarn strain (MGI: 3720980) was generated by continuous backcrosses between TNF^ΔARE mice on mixed genetic background (ie, C57BL6 and 129S6)⁴ to C57BL6/J mice as previously described.⁷ Mice were kept under specific pathogen-free conditions. All progeny were either heterozygous (TNF^ΔARE/+, TNFΔARE) or carried no mutated alleles, wild-type (WT). The latter were used as the noninflamed controls. CD44-, integrin β₇-, and L-selectin–deficient mice on the C57BL6/J background were

Increased Expression of Hyaluronan and Hyaluronan Synthase-1 in TNFΔARE Mice

We compared the levels of hyaluronan in serum, the expression of hyaluronan synthases, and the reactivity of hyaluronan binding protein within the terminal ileum. Levels of soluble hyaluronan were increased significantly in 30-week-old TNFΔARE mice with ileitis compared with WT mice (822 ± 121 vs 410 ± 51 ng/mL; P < .01; Figure 1A).

To try to identify the enzyme(s) responsible for increased hyaluronan levels, we assessed the expression of hyaluronan synthases 1–3 (HAS-1, -2, and -3) (Figure 1B)

Discussion

A prior study in the TNFΔARE mouse model of CD showed that the CD8/CD44^high T-cell population played an important role in disease development.⁶ The current study extends those observations to additionally show that CD4/CD44⁺ effectors also are important players. These CD4⁺ T cells have an effector surface phenotype, show enhanced reactivity to HA, produced TNF and interleukin-2, and were required for the induction of ileitis in RAG−/− mice. We additionally show that the synthesis of the CD44

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The role of HA in intestinal inflammation may function through its receptor CD44. In a TNF-driven mouse model of chronic ileitis, soluble HA levels and expression of Has1 were increased coincided with increased expression of CD44 on CD4 + T cells, and deficiency in CD44 resulted in attenuated the severity of ileitis of the mice [245]. Interestingly, reports revealed that platelet-derived HYAL2 is able to cleave HA into fragments that stimulate mononuclear leukocytes in the immediate microenvironment to produce proinflammatory cytokines [41].
Accumulation and turnover of extracellular matrix is a hallmark of tissue injury, repair and remodeling in human diseases. Hyaluronan is a major component of the extracellular matrix and plays an important role in regulating tissue injury and repair, and controlling disease outcomes. The function of hyaluronan depends on its size, location, and interactions with binding partners. While fragmented hyaluronan stimulates the expression of an array of genes by a variety of cell types regulating inflammatory responses and tissue repair, cell surface hyaluronan provides protection against tissue damage from the environment and promotes regeneration and repair. The interactions of hyaluronan and its binding proteins participate in the pathogenesis of many human diseases. Thus, targeting hyaluronan and its interactions with cells and proteins may provide new approaches to developing therapeutics for inflammatory and fibrosing diseases. This review focuses on the role of hyaluronan in biological and pathological processes, and as a potential therapeutic target in human diseases.
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: The authors disclose the following: Supported by US Public Health Service/National Institutes of Health grants DK067254 and DK073280 (J. R.-N.) and by the University of Virginia Silvio Conte Digestive Health Research Center (grant DK56703).

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Basic—Alimentary TractCD44 Deficiency Attenuates Chronic Murine Ileitis

Background & Aims

Methods

Results

Conclusions

Section snippets

Mice

Increased Expression of Hyaluronan and Hyaluronan Synthase-1 in TNFΔARE Mice

Discussion

Gastroenterology

Immunity

Immunity

Gastroenterology

Gastroenterology

Gastroenterology

J Immunol Methods

J Immunol Methods

Am J Pathol

Biochim Biophys Acta

Exp Gerontol

J Biol Chem

Blood

J Biol Chem

Basic—Alimentary Tract
CD44 Deficiency Attenuates Chronic Murine Ileitis