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Arthritis Res Ther. 2014;16(3):R126.

Therapeutic effect of anti-C-X-C motif chemokine 10 (CXCL10) antibody on C protein-induced myositis mouse.

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제1저자 김진현 · 교신저자 송영욱

PMID 24939012 PMC4095607 DOI 10.1186/ar4583 IF 5.4 류마티스내과

Abstract

INTRODUCTION C-X-C motif chemokine 10 (CXCL10) is a chemokine that plays a critical role in the infiltration of T cells in autoimmune diseases and is reported to be expressed in muscle tissue of polymyositis. To determine the therapeutic efficacy of CXCL10 blockade, we investigated the role of CXCL10 and the effect of anti-CXCL10 antibody treatment in C protein-induced myositis (CIM), an animal model of polymyositis.

METHODS CIM was induced with human skeletal muscle C protein fragment in female C57BL/6 mice. Immunohistochemistry of CXCL10 and C-X-C motif chemokine receptor 3 (CXCR3) and measurement of serum CXCL10 were performed. Cell surface markers and interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) in CIM lymph node cells was investigated by flow cytometry. Mice with CIM were treated with anti-CXCL10 antibody or control antibody (anti-RVG1) and the inflammation in muscle tissue was assessed.

RESULTS Immunohistochemistry showed increased expression of CXCL10 and CXCR3 in the inflammatory lesions of muscle in CIM. Especially, CD8+ T cells invading myofiber expressed CXCR3. Serum level of CXCL10 was increased in CIM compared to the level in normal mice (normal mouse, 14.3 ± 5.3 pg/ml vs. CIM, 368.5 ± 135.6 pg/ml, P < 0.001). CXCR3 positivity in CD8+ T cells was increased compared to that of CD4+ T cells in the lymph node cells of CIM (CXCR3+ among CD8+ T cell, 65.9 ± 2.1% vs. CXCR3+ among CD4+ T cell, 23.5 ± 4.7%, P <0.001). Moreover, IFN-γ+ cells were increased among CXCR3+CD8+ T cells compared to CXCR3-CD8+ T cells (CXCR3+CD8+ T cell, 28.0 ± 4.2% vs. CXCR3-CD8+ T cell, 9.5 ± 1.5%, P = 0.016). Migration of lymph node cells was increased in response to CXCL10 (chemotactic index was 1.91 ± 0.45). CIM mice treated with anti-CXCL10 antibody showed a lower inflammation score in muscles than those with anti-RVG1 (median, anti-CXCL10 treatment group, 0.625 vs. anti-RVG1 treatment group, 1.25, P = 0.007).

CONCLUSIONS CXCL10/CXCR3 expression was increased in the inflammation of CIM model and its blockade suppressed inflammation in muscle.

Figures

Figure 1
Figure 1 Expression of CXCL10 in C-protein-induced myositis (CIM). (A) Immunohistochemistry of CXCL9, CXCL10 and CXCL11 in the muscle of CIM (×400). In the inflammatory lesion of CIM, CXCL10 was strongly stained whereas CXCL9 or CXCL11 was weakly stained. (B) Serum level of CXCL10 in normal mice and CIM mice. The level of CXCL10 was measured by ELISA in the sera of normal mice (n = 10) and CIM mice (n = 10) at 3 weeks after induction. The serum level of CIM was more elevated in CIM than in normal mice (normal mouse, 14.3 ± 5.3 pg/ml versus CIM, 368.5 ± 135.6 pg/ml, P <0.001). The horizontal lines indicate the mean.
Figure 2
Figure 2 Expression of CXCR3 in the lymph node and muscle of C-protein-induced myositis (CIM). (A) Immunohistochemistry of CIM muscle tissue and lymph node. CXCR3 was expressed in some cells of the lymph node cells (left, ×200) and inflammatory lesion in the muscle of CIM (right, ×400). (B) Immunostaining of cells invading myofiber in muscle of CIM. Differential interference contrast (DIC) showed myofiber and invasive cells in the middle of myofiber. The CD8 was co-stained with CXCR3 whereas CD4 was not (×1,000). (C) Immunostaining of inflammation in muscle of CIM showed F4/80 co-stained with CXCR3 (×1,000). (D) CXCR3-positive immune cells in CIM lymph node. CIM lymph node cells were analyzed by flow cytometry. CXCR3-positive cells were gated from lymph node cells. CXCR3-positive cells were analyzed by cell-specific markers (CD3 for T cells; B220 for B cells; F4/80 for macrophages).
Figure 3
Figure 3 The IFN-γ and TNF-α positivity by CXCR3 expression in CD8+ T cells. Lymph node cells of C protein-induced myositis (CIM) were stimulated with phorbol 12-myristate 13-acetate, ionomycin and brefeldin A for 4 hours. CD3+CD8+ cells were gated (left) and analyzed according to the presence of CXCR3 and effector molecules. Representative figure of three experiments.
Figure 4
Figure 4 Therapeutic effects of anti-CXCL10 or control antibody treatment in C-protein-induced myositis (CIM). After inducing CIM, mice were treated with anti-CXCL10 antibody or control antibody (anti-RVG1) or were not treated (n = 17 per group). The group treated with anti-CXCL10 showed a lower inflammation score in muscles than those with anti-RVG1 or no treatment. No treatment: no treatment group, anti-CXCL10: anti-CXCL10 treatment group, anti-RVG1: anti-RVG1 treatment group. anti-RVG1, mouse anti-rotavirus IgG1.

그림 출처: PubMed Central 공개 논문 (PMC4095607)

그 밖의 정보
출판 형태
Journal Article, Research Support, Non-U.S. Gov't
저널
Arthritis research & therapy · eISSN 1478-6362
전자 출판
2014-06-17
MeSH
Animals · Antibodies, Blocking / immunology / pharmacology · Carrier Proteins · Chemokine CXCL10 / antagonists & inhibitors / immunology / metabolism · Disease Models, Animal · Female · Flow Cytometry · Humans · Immunohistochemistry · Interferon-gamma / metabolism · Mice, Inbred C57BL · Myositis / blood / chemically induced / drug therapy · Polymyositis / blood / drug therapy · Receptors, CXCR3 / metabolism · Treatment Outcome · Tumor Necrosis Factor-alpha / metabolism · Myosin Binding Protein C
Substances
Antibodies, Blocking · Carrier Proteins · Chemokine CXCL10 · Interferon-gamma · Receptors, CXCR3 · Tumor Necrosis Factor-alpha · Myosin Binding Protein C · Cxcr3 protein, mouse
참고문헌
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