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Masanobu Tsuda,
Weici Zhang,
Guo-Xiang Yang,
Koichi Tsuneyama,
Yugo Ando,
Kazuhito Kawata,
Ogyi Park,
Patrick S C Leung,
Ross L Coppel,
Aftab A Ansari,
William M Ridgway,
Bin Gao,
Zhe-Xiong Lian,
Richard Flavell,
Xiao-Song He,
M Eric Gershwin
[show abstract]
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ABSTRACT: Mice with a dominant-negative transforming growth factor b receptor restricted to T cells (dnTGFbRII mice) develop an inflammatory biliary ductular disease that strongly resembles human primary biliary cirrhosis (PBC). Furthermore, deletion of the gene encoding interleukin (IL)-12p40 resulted in a strain (IL-12p40 2/2 dnTGFbRII) with dramatically reduced autoim-mune cholangitis. To further investigate the role of the IL-12 cytokine family in dnTGFbRII autoimmune biliary disease, we deleted the gene encoding the IL-12p35 subunit from dnTGFbRII mice, resulting in an IL-12p35 2/2 dnTGFbRII strain which is deficient in two members of the IL-12 family, IL-12 and IL-35. In contrast to IL-12p40 2/2 mice, the IL-12p35 2/2 mice developed liver inflammation and bile duct damage with similar severity but delayed onset as the parental dnTGFbRII mice. The p35 2/2 mice also demonstrated a distinct cytokine profile characterized by a shift from a T-helper 1 (Th1) to a Th17 response. Strikingly, liver fibrosis was frequently observed in IL-12p35 2/2 mice. In conclusion, IL-12p35 2/2 dnTGFbRII mice, histologically and immunologically, reflect key features of PBC, providing a useful generic model to understand the immunopathology of human PBC. (HEPATOLOGY 2013;57:806-816)
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Yugo Ando,
Guo-Xiang Yang, Masanobu Tsuda,
Kazuhito Kawata,
Weici Zhang,
Takahiko Nakajima,
Koichi Tsuneyama,
Patrick Leung,
Zhe-Xiong Lian,
Kazuichi Okazaki,
William M Ridgway,
Gary L Norman,
Aftab A Ansari,
Xiao-Song He,
Ross L Coppel,
M Eric Gershwin
[show abstract]
[hide abstract]
ABSTRACT: Dominant negative form of transforming growth factor beta receptor type II (dnTGFbRII) mice, expressing a dominant negative form of TGFb receptor II under control of the CD4 promoter, develop autoimmune colitis and cholangitis. Deficiency in interleukin (IL)-12p40 lead to a marked diminution of inflammation in both the colon and the liver. To distinguish whether IL-12p40 mediates protection by the IL-12 or IL-23 pathways, we generated an IL-23p19 2/2 dnTGFbRII strain deficient in IL-23, but not in IL-12; mice were longitudinally followed for changes in the natural history of disease and immune responses. Interestingly, IL-23p19 2/2 mice demonstrate dramatic improvement in their colitis, but no changes in biliary pathology; mice also manifest reduced T-helper (Th)17 cell populations and unchanged IFN-c levels. We submit that the IL-12/Th1 pathway is essential for biliary disease pathogenesis, whereas the IL-23/Th17 pathway mediates coli-tis. To further assess the mechanism of the IL-23-mediated protection from colitis, we gen-erated an IL-17A 2/2 dnTGFbRII strain deficient in IL-17, a major effector cytokine produced by IL-23-dependent Th17 cells. Deletion of the IL-17A gene did not affect the severity of either cholangitis or colitis, suggesting that the IL-23/Th17 pathway contrib-utes to colon disease in an IL-17-independent manner. These results affirm that the IL-12/ Th1 pathway is critical to biliary pathology in dnTGFbRII mice, whereas colitis is caused by a direct effect of IL-23. (HEPATOLOGY 2012;56:1418-1426) M
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[show abstract]
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ABSTRACT: C-type lectin receptors are pattern recognition receptors that are critical for autoimmunity and the immune response. Mincle is a C-type lectin receptor expressed by a variety of antigen presenting cells including macrophages, neutrophils, dendritic cells and B cells; a variety of stimuli including stress are known to induce the expression of Mincle. Mincle is an FcRγ-associated activation receptor that senses damaged cells and upon ligation induces activated macrophages to produce inflammatory cytokines. Recently, while several studies have reported that Mincle plays an important role in macrophage responses to fungal infection its function on B cells remains to be defined. In efforts to elucidate the function of Mincle expressed by B cells, we studied the expression of Mincle on subsets of B cells and analyzed cytokines and synthesized immunoglobulin upon ligation of Mincle. The expression of Mincle on CD27-CD19(+) naïve B cells is significantly higher than CD27 + CD19(+) memory B cells. The stimulation of TLR9 ligand induced Mincle expression on B cells. Furthermore, co-stimulation of TLR9 and Mincle ligand reduced IgG and IgA production from B cells without a significant change in the inflammatory cytokines TNF-α, IL-6, IL-8 and IL-10. Our data identifies Mincle as a potentially critical player in human B cell responses.
Journal of Autoimmunity 06/2012; · 7.37 Impact Factor
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Masanobu Tsuda,
Weici Zhang,
Guo-Xiang Yang,
Koichi Tsuneyama,
Yugo Ando,
Kazuhito Kawata,
Ogyi Park,
Patrick S C Leung,
Ross L Coppel,
Aftab A Ansari,
William M Ridgway,
Bin Gao,
Zhe-Xiong Lian,
Richard Flavell,
Xiao-Song He,
M Eric Gershwin
[show abstract]
[hide abstract]
ABSTRACT: We have previously reported that mice with a dominant negative transforming growth factor β receptor restricted to T cells (dnTGFβRII mice) develop an inflammatory biliary ductular disease that strongly resembles human primary biliary cirrhosis (PBC). Furthermore, deletion of the gene encoding interleukin (IL)-12p40 resulted in a strain (IL-12p40(-/-) dnTGFβRII) with dramatically reduced autoimmune cholangitis. To further investigate the role of the IL-12 cytokine family in dnTGFβRII autoimmune biliary disease, we deleted the gene encoding the IL-12p35 subunit from dnTGFβRII mice, resulting in an IL-12p35(-/-) dnTGFβRII strain which is deficient in two members of the IL-12 family, IL-12 and IL-35. In contrast to IL-12p40(-/-) mice, the IL-12p35(-/-) mice developed liver inflammation and bile duct damage with similar severity but delayed onset as the parental dnTGFβRII mice. The p35(-/-) mice also demonstrated a distinct cytokine profile characterized by a shift from a Th1 to a Th17 response. Strikingly, liver fibrosis was frequently observed in IL-12p35(-/-) mice. In conclusion, IL-12p35(-/-) dnTGFβRII mice, histologically and immunologically, reflect key features of PBC, providing a useful generic model to understand the immunopathology of human PBC. (HEPATOLOGY 2012.).
Hepatology 05/2012; · 11.66 Impact Factor
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Yugo Ando,
Guo-Xiang Yang, Masanobu Tsuda,
Kazuhito Kawata,
Weici Zhang,
Takahiko Nakajima,
Koichi Tsuneyama,
Patrick Leung,
Zhe-Xiong Lian,
Kazuichi Okazaki,
William M Ridgway,
Gary L Norman,
Aftab A Ansari,
Xiao-Song He,
Ross L Coppel,
M Eric Gershwin
[show abstract]
[hide abstract]
ABSTRACT: Dominant negative form of transforming growth factor beta receptor type II (dnTGFβRII) mice, expressing a dominant negative form of TGFβ receptor II under control of the CD4 promoter, develop autoimmune colitis and cholangitis. Deficiency in interleukin (IL)-12p40 lead to a marked diminution of inflammation in both the colon and the liver. To distinguish whether IL-12p40 mediates protection by the IL-12 or IL-23 pathways, we generated an IL-23p19(-/-) dnTGFβRII strain deficient in IL-23, but not in IL-12; mice were longitudinally followed for changes in the natural history of disease and immune responses. Interestingly, IL-23p19(-/-) mice demonstrate dramatic improvement in their colitis, but no changes in biliary pathology; mice also manifest reduced T-helper (Th)17 cell populations and unchanged IFN-γ levels. We submit that the IL-12/Th1 pathway is essential for biliary disease pathogenesis, whereas the IL-23/Th17 pathway mediates colitis. To further assess the mechanism of the IL-23-mediated protection from colitis, we generated an IL-17A(-/-) dnTGFβRII strain deficient in IL-17, a major effector cytokine produced by IL-23-dependent Th17 cells. Deletion of the IL-17A gene did not affect the severity of either cholangitis or colitis, suggesting that the IL-23/Th17 pathway contributes to colon disease in an IL-17-independent manner. These results affirm that the IL-12/Th1 pathway is critical to biliary pathology in dnTGFβRII mice, whereas colitis is caused by a direct effect of IL-23. (HEPATOLOGY 2012).
Hepatology 04/2012; 56(4):1418-26. · 11.66 Impact Factor
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Masanobu Tsuda,
Yuki Moritoki,
Zhe-Xiong Lian,
Weici Zhang,
Katsunori Yoshida,
Kanji Wakabayashi,
Guo-Xiang Yang,
Toshio Nakatani,
John Vierling,
Keith Lindor,
M Eric Gershwin,
Christopher L Bowlus
[show abstract]
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ABSTRACT: The aim of this study was to determine the safety and potential efficacy of B-cell depletion with the anti-CD20 monoclonal antibody rituximab in patients with primary biliary cirrhosis (PBC) and an incomplete response to ursodeoxycholic acid (UDCA). This open-label study enrolled six patients with PBC and incomplete responses to UDCA to be treated with 2 doses of 1000 mg rituximab separated by 2 weeks and followed for 52 weeks. The primary endpoints were safety and changes in B-cell function. Two patients received only 1 dose of rituximab, one due to activation of latent varicella and the other due to a viral upper respiratory infection. Serum levels of total IgG, IgM, and IgA as well as anti-mitochondrial autoantibodies (AMAs) IgA and IgM decreased significantly from baseline by 16 weeks and returned to baseline levels by 36 weeks. Stimulation of B cells with CpG produced significantly less IgM at 52 weeks after treatment compared with B cells at baseline. In addition, transient decreases in memory B-cell and T-cell frequencies and an increase in CD25(high) CD4(+) T cells were observed after treatment. These changes were associated with significant increases in mRNA levels of FoxP3 and transforming growth factor-β (TGF-β) and a decrease in tumor necrosis factor-α (TNF-α) in CD4(+) T cells. Notably, serum alkaline phosphatase levels were significantly reduced up to 36 weeks following rituximab treatment. CONCLUSION: These data suggest that depletion of B cells influences the induction, maintenance, and activation of both B and T cells and provides a potential mechanism for treatment of patients with PBC with an incomplete response to UDCA.
Hepatology 02/2012; 55(2):512-21. · 11.66 Impact Factor
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ABSTRACT: It is important to have a venous line in cardiopulmonary arrest (CPA) patients as an emergency treatment measure in prehospital settings, but establishment of a peripheral venous line is difficult in such patients. This study aimed to investigate the current status of intravenous infusion (IVI) in CPA patients by Emergency Life-Saving Technicians (ELSTs) in Japan. We also considered alternative measures in case IVI was difficult or impossible.
We investigated a nationwide database between 1 January 2005 and 31 December 2008. From a total of 431,968 CPA cases, we calculated the IVI success rate and related parameters.The Bone Injection Gun (BIG) and simulator legs (adult, pediatric, and infant) were used by 100 ELSTs selected for the study to measure the time required and the success rate for intraosseous infusion (IOI).
The number of CPA patients, IVI, adrenaline administration, and the IVI success rate in adult CPA patients increased every year. However, the IVI success rate in pediatric CPA patients did not increase. Although adrenaline administration elevated the ROSC rate, there was no improvement in the 1-month survival rate. The time required for IOI with BIG was not different among the leg models. The success rates of IOI with BIG were 93%, 94%, and 84% (p < 0.05 vs. adult and pediatric) in adult, pediatric, and infant models, respectively.
The rate of success of IVI in adult CPA patients has been increased yearly in Japan. However, as establishing a peripheral venous line in pediatric patients (1-7 years old) by ELSTs is extremely difficult in prehospital settings, there was no increase in the IVI success rate in such patients. As the study findings indicated IOI with BIG was easy and rapid, it may be necessary to consider IOI with BIG as an alternative option in case IVI is difficult or impossible in adult and pediatric patients.
International Journal of Emergency Medicine 01/2012; 5(1):2.
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Weici Zhang, Masanobu Tsuda,
Guo-Xiang Yang,
Koichi Tsuneyama,
Xiao-Song He,
Aftab A Ansari,
William M Ridgway,
Ross L Coppel,
Zhe-Xiong Lian,
Patrick S C Leung,
M Eric Gershwin
[show abstract]
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ABSTRACT: Hepatosplenic T cell lymphoma (HSTCL) is a distinct and lethal subtype of peripheral T cell lymphoma with an aggressive course and poor outcome despite multiagent chemotherapy. Contradictory literature, an unknown etiology, and poor response to treatment highlight the need to define the malignant process and identify molecular targets with potential for successful therapeutic interventions. Herein, we report that mice homozygously expressing a dominant negative TGFβRII (dnTGFβRII) under the control of the CD4 promoter spontaneously develop lymphoma-like T cell infiltration involving both spleen and liver. Splenomegaly, hepatomegaly and liver dysfunction were observed in homozygous dnTGFβRII mice between 10 weeks and 10 months of age associated with a predominant infiltration of CD4(-)CD8(-)TCRβ(+)NK1.1(+) or CD8(+)TCRβ(+)NK1.1(-) T cell subsets. Notch 1 and c-Myc expression at the mRNA levels were significantly increased and positively correlated with the cell number of lymphoid infiltrates in the liver of dnTGFβRII homozygous compared to hemizygous mice. Further, 2×10(4) isolated lymphoma-like cells transplant disease by adoptive cell transfers. Collectively, our data demonstrate that increased copy number of dnTGFβRII is critical for development of lymphoma-like T cell infiltration.
PLoS ONE 01/2012; 7(11):e49413. · 4.09 Impact Factor
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Masanobu Tsuda,
Yoko M Ambrosini,
Weici Zhang,
Guo-Xiang Yang,
Yugo Ando,
Guanghua Rong,
Koichi Tsuneyama,
Kosuke Sumida,
Shinji Shimoda,
Christopher L Bowlus,
Patrick S C Leung,
Xiao-Song He,
Ross L Coppel,
Aftab A Ansari,
Zhe-Xiong Lian,
M Eric Gershwin
[show abstract]
[hide abstract]
ABSTRACT: In primary biliary cirrhosis (PBC), patients develop a multilineage response to a highly restricted peptide of the E2 component of pyruvate dehydrogenase (PDC-E2) involving autoantibody and autoreactive cluster of differentiation (CD)4(+) and CD8(+) T-cell responses. Recent data from murine models have suggested that liver-infiltrating CD8(+) cells play a critical role in biliary destruction in PBC. We hypothesized that chronic antigen stimulation of CD8(+) T cells alters effector memory T cell (T(EM) ) frequency and function similar to that seen with chronic viral infections, including failure to terminally differentiate and relative resistance to apoptosis. We have rigorously phenotyped CD8(+) T-cell subpopulations from 132 subjects, including 76 patients with PBC and 56 controls, and report a higher frequency of T(EM) cells characterized as CD45RO(high) CD57(+) CD8(high), but expressing the gut homing integrin, α4β7, in peripheral blood mononuclear cells of PBC. These CD8(high) T(EM) cells have reduced expression of Annexin V after TCR stimulation. Consistent with a T(EM) phenotype, CD45RO(high) CD57(+) CD8(high) T cells express higher levels of granzyme A, granzyme B, perforin, CCR5 and α4β7, and lower levels of CCR7 and CD28 than other CD8(high) T cells. Furthermore, interleukin (IL)-5 produced by CD8(+) CD57(+) T lymphocytes upon in vitro T-cell receptor stimulation are increased in PBC. Histologically, CD8(+) CD57(+) T cells accumulate around the portal area in PBC. Moreover, CD8(+) CD57(+) T cells respond specifically to the major histocompatibility class I epitope of PDC-E2. CONCLUSION: In conclusion, our data demonstrate that CD45RO(high) CD57(+) CD8(high) T cells are a subset of terminally differentiated cytotoxic T(EM) cells, which could play a critical role in the progressive destruction of biliary epithelial cells.
Hepatology 07/2011; 54(4):1293-302. · 11.66 Impact Factor
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Yuki Moritoki, Masanobu Tsuda,
Koichi Tsuneyama,
Weici Zhang,
Katsunori Yoshida,
Zhe-Xiong Lian,
Guo-Xiang Yang,
William M Ridgway,
Linda S Wicker,
Aftab A Ansari,
M Eric Gershwin
[show abstract]
[hide abstract]
ABSTRACT: There are now several murine models of autoimmune cholangitis that have features both similar and distinct from human PBC. One such model, the NOD.c3c4 mouse, manifests portal cell infiltrates, anti-mitochondrial antibodies but also biliary cysts. The biliary cysts are not a component of PBC and not found in the other murine models. To address the immunopathology in these mice, we generated genetically B cell deficient Igμ(-/-) NOD.c3c4 mice and compared the immunopathology of these animals to control B cell sufficient NOD.c3c4 mice. B cell deficient mice demonstrated decreased number of non-B cells in the liver accompanied by reduced numbers of activated natural killer cells. The degree of granuloma formation and bile duct damage were comparable to NOD.c3c4 mice. In contrast, liver inflammation, biliary cyst formation and salivary gland inflammation was significantly attenuated in these B cell deficient mice. In conclusion, B cells play a critical role in promoting liver inflammation and also contribute to cyst formation as well as salivary gland pathology in autoimmune NOD.c3c4 mice, illustrating a critical role of B cells in modulating specific organ pathology and, in particular, in exacerbating both the biliary disease and the sialadenitis.
Cellular Immunology 01/2011; 268(1):16-23. · 1.97 Impact Factor
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ABSTRACT: IPEX syndrome is a congenital disorder of immune regulation caused by mutations in the FOXP3 gene, which is required for the suppressive function of naturally arising CD4 + CD25 + regulatory T cells. In this case series we evaluated serum samples from 12 patients with IPEX syndrome for the presence of common autoantibodies associated with a broad range of autoimmune disorders. We note that 75% of patients (9/12) had 1 or more autoantibodies, an incidence far above the cumulative rate observed in the general population. The range of autoantibodies differed between patients and there was no predominant autoantibody or pattern of autoantibodies present in this cohort. Surprisingly, one patient had high-titer anti-mitochondrial antibodies (AMA) typically associated with primary biliary cirrhosis (PBC) although the patient had no signs of cholestasis. PBC is a well-characterized autoimmune disease that occurs primarily in women and includes the serological hallmarks of serum AMA and elevated IgM which were both present in this patient. PBC is virtually absent in children with the exception of one reported child with interleukin 2 receptor α (CD25) deficiency which is associated with an IPEX-like regulatory T cell dysfunction. Based on the present data and the available literature we suggest a direct role for CD4 + CD25 + regulatory T cells in restraining B cell autoantibody production and that defects in regulatory T cells may be crucial to the development of PBC.
Journal of Autoimmunity 11/2010; 35(3):265-8. · 7.37 Impact Factor
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Weici Zhang, Masanobu Tsuda,
Guo-Xiang Yang,
Koichi Tsuneyama,
Guanghua Rong,
William M Ridgway,
Aftab A Ansari,
Richard A Flavell,
Ross L Coppel,
Zhe-Xiong Lian,
M Eric Gershwin
[show abstract]
[hide abstract]
ABSTRACT: The role of interleukin-6 (IL-6) in autoimmunity attracts attention because of the clinical usage of monoclonal antibodies to IL-6 receptor (IL-6R), designed to block IL-6 pathways. In autoimmune liver disease, activation of the hepatocyte IL-6/STAT3 (signal transducer and activator of transcription 3) pathway is associated with modulating pathology in acute liver failure, in liver regeneration, and in the murine model of concanavalin A-induced liver inflammation. We have reported that mice expressing a dominant negative form of transforming growth factor beta receptor II (dnTGFbetaRII) under control of the CD4 promoter develop both colitis and autoimmune cholangitis with elevated serum levels of IL-6. Based on this observation, we generated IL-6-deficient mice on a dnTGF-betaRII background (dnTGFbetaRII IL-6(-/-)) and examined for the presence of antimitochondrial antibodies, levels of cytokines, histopathology, and immunohistochemistry of liver and colon tissues. As expected, based on reports of the use of anti-IL-6R in inflammatory bowel disease, dnTGFbetaRII IL-6(-/-) mice manifest a dramatic improvement in their inflammatory bowel disease, including reduced diarrhea and significant reduction in intestinal lymphocytic infiltrates. Importantly, however, autoimmune cholangitis in dnTGFbetaRII IL-6(-/-) mice was significantly exacerbated, including elevated inflammatory cytokines, increased numbers of activated T cells, and worsening hepatic pathology. CONCLUSION: The data from these observations emphasize that there are distinct mechanisms involved in inducing pathology in inflammatory bowel disease compared to autoimmune cholangitis. These data also suggest that patients with inflammatory bowel disease may not be the best candidates for treatment with anti-IL-6R if they have accompanying autoimmune liver disease and emphasize caution for therapeutic use of anti-IL-6R antibody.
Hepatology 07/2010; 52(1):215-22. · 11.66 Impact Factor
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Katsunori Yoshida,
Guo-Xiang Yang,
Weici Zhang, Masanobu Tsuda,
Koichi Tsuneyama,
Yuki Moritoki,
Aftab A Ansari,
Kazuichi Okazaki,
Zhe-Xiong Lian,
Ross L Coppel,
Ian R Mackay,
M Eric Gershwin
[show abstract]
[hide abstract]
ABSTRACT: Our laboratory has reported that mice that express a dominant negative form of transforming growth factor beta receptor restricted to T cells (dnTGFbetaRII) develop an inflammatory biliary ductular disease with elevated serum levels of interleukin (IL)-12p40 and other proinflammatory cytokines and antimitochondrial autoantibodies (AMAs) closely resembling human primary biliary cirrhosis (PBC). We have used this mouse model to address the potential mechanisms of immunomodulation of liver disease by creating two unique genetic strains: IL-12p40 knockout (KO)-dnTGFbetaRII mice and IFN-gamma KO-dnTGFbetaRII mice. The two colonies of genetically modified mice-and, for purposes of controls, the dnTGFbetaRII mice-were monitored for liver immunopathology, AMAs, and intrahepatic cytokine production. Disease expression in the IFN-gamma KO-dnTGFbetaRII mice, including liver immunopathology, were similar to those of dnTGFbetaRII mice, whereas the IL-12p40 KO-dnTGFbetaRII mice had a dramatic reduction in histological autoimmune cholangitis and significant decreases in levels of intrahepatic proinflammatory cytokines, but similar levels of AMAs compared with dnTGFbetaRII controls. CONCLUSION: These data indicate that in this mouse model of PBC, signaling by way of IL-12p40 is an essential requirement for the development of autoimmune cholangitis. The results of these studies will play an important role in identifying pathways and reagents that will selectively inhibit IL-12 signaling for the outlining of future therapeutic strategies for human PBC.
Hepatology 07/2009; 50(5):1494-500. · 11.66 Impact Factor
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ABSTRACT: Immunosuppression after burn injury increases the risk of sepsis and multiple organ failure. We examined changes of immune function in mice after burn injury and investigated the immunostimulatory effect of oligodeoxynucleotides containing CpG motifs.
Male BALB/c mice (8-10 wk old) received a full-thickness burn to 20% of their body surface area, after which the immunological parameters of splenic macrophages were evaluated. To assess the immunostimulatory effect of oligodeoxynucleotide treatment, splenic macrophages harvested from burned mice were incubated with oligodeoxynucleotides. Then cytokine production and major histocompatibility complex class II antigen expression were measured. To assess the in vivo effect of oligodeoxynucleotides, intraperitoneal administration was done on day 4 after burn injury, and class II antigen expression by splenic macrophages was measured 10 d later.
Class II antigen expression and the synthesis of cytokines (interleukin-12, tumor necrosis factor-alpha, interleukin-6, and interleukin-1) by splenic macrophages were significantly reduced after burn injury, while incubation of splenic macrophages from burned mice with oligodeoxynucleotides partially enhanced the production of interleukin-12, tumor necrosis factor-alpha, interleukin-6, and interleukin-1. In addition, intraperitoneal administration of oligodeoxynucleotides enhanced class II antigen expression by splenic macrophages.
The reduction of class II antigen expression and synthesis of cytokines (interleukin-12, tumor necrosis factor-alpha, interleukin-6, and interleukin-1) by splenic macrophages after burn injury was partially reversed by oligodeoxynucleotide treatment. Therefore, immunostimulatory oligodeoxynucleotides may be a potential treatment for post-burn immunosuppression.
Journal of Surgical Research 03/2009; 161(1):111-8. · 2.25 Impact Factor
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ABSTRACT: Adipose tissue-derived stem cells (ADSCs) can be easily obtained from subcutaneous adipose tissue, and ADSCs can be demonstrated to display multilineage developmental plasticity. In this study, using TNBS-induced colitis rats, we show the feasibility of repairing injured intestinal mucosa with adipose tissue-derived stem cells.
The subcutaneous adipose tissue of F344 rats was obtained and digested by collagenase. The digested tissue was cultured in DMEM containing 10% FBS for 1 month. ADSCs were confirmed to differentiate under appropriate conditions into various lineages of cells, including bone, neural cells, adipocytes, and epithelial cells. HGF, VEGF, TGF-beta, and adiponectin in the culture supernatants of ADSCs were determined by ELISA. ADSCs (10(7) cells) were injected into the submucosa of the colon to examine their capacity to repair intestinal mucosa injured by TNBS.
In the experimental colitis model, the injection of ADSCs facilitated colonic mucosal repair and reduced the infiltration of inflammatory cells. High levels of HGF, VEGF, and adiponectin were detected in the culture supernatants of ADSCs. Moreover, injected ADSCs distributed to several layers of the colon, and some of them differentiated into mesodermal lineage cells.
ADSCs can accelerate the regeneration of injured regions in experimental colitis. HGF, VEGF, and adiponectin might be responsible for the regeneration of injured regions in the colon.
Inflammatory Bowel Diseases 07/2008; 14(6):826-38. · 4.86 Impact Factor
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ABSTRACT: The long-term maintenance of hematopoietic stem cells (HSCs) is assessed by serial bone marrow transplantation (BMT), in which HSCs are injected intravenously. Recently, we have found that intra-bone marrow (IBM)-BMT can efficiently reconstitute the hematopoietic system with cells of donor origin, in contrast to conventional intravenous (i.v.) BMT. In the present study, we have compared the long-term maintenance of HSCs using multiple rounds of serial i.v.-BMT and IBM-BMT. The frequencies of donor-derived progenitor cells (Lin(-)/c-kit(+) cells) and more primitive progenitors (Lin(-)/c-kit(+)/CD34(+)/Sca-1(+) cells) were higher in the tertiary recipients by serial IBM-BMT than in those that had received bone marrow cells by serial i.v.-BMT. Furthermore, neither donor-derived progenitor cells nor mature hematolymphoid cells were detected in approximately 25% of the tertiary recipients after serial i.v.-BMT, indicating that progenitor cells can be efficiently maintained by IBM-BMT but not by i.v.-BMT. Finally, we confirmed that the recipients treated with the primary IBM-BMT (without carrying out serial BMT) showed a significantly higher survival rate than those treated with i.v.-BMT. These findings clearly show that IBM-BMT efficiently promotes the longterm maintenance of donor-derived hematopoiesis.
Stem Cells and Development 05/2008; 17(2):291-302. · 4.46 Impact Factor
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Kequan Guo,
Muneo Inaba,
Ming Li,
Jun An,
Wenhao Cui,
Changye Song,
Jianfeng Wang,
Yunze Cui,
Yutaku Sakaguchi, Masanobu Tsuda,
Mariko Omae,
Yugo Ando,
Qing Li,
Xiaoli Wang,
Wei Feng,
Susumu Ikehara
[show abstract]
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ABSTRACT: Donor-specific central tolerance in cardiac allograft can be induced by hematopoietic chimerism via conventional intravenous bone marrow transplantation (IV-BMT). However, there are problems with IV-BMT, such as the risk of graft failure and of the toxicity from conditioning regimens.
A new method for heart transplantation is presented. This method consists of administration of fludarabine phosphate (50 mg/kg) and fractionated low-dose irradiation (3.5 Gyx2 or 4.0 Gyx2), followed by intrabone marrow injection of whole bone marrow cells (IBM-BMT) plus heterotopic heart transplantation.
Cardiac allografts with IBM-BMT were accepted and survived long-term (>10 months) showing neither acute rejection nor chronic rejection including cardiac allograft vasculopathy by such conditioning regimens. In contrast, cardiac allografts with conventional IV-BMT were rejected within 1 month after the treatment with irradiation of 3.5 Gyx2 or within 3 months after the treatment with irradiation of 4.0 Gyx2. Macrochimerism (>70%) was favorably established and stably maintained by IBM-BMT but not IV-BMT. Low levels of transient mixed chimerism (<7%) were induced by IV-BMT with fludarabine plus 4.0 Gyx2, but the chimerism was lost within 1 month after the treatment.
These findings indicate that IBM-BMT is a feasible strategy for the induction of persistent donor-specific tolerance, enables the use of reduced radiation doses as conditioning regimens, and obviates the need for immunosuppressants.
Transplantation 01/2008; 85(1):93-101. · 4.00 Impact Factor
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Muneo Inaba,
Yasushi Adachi,
Hiroko Hisha,
Naoki Hosaka,
Masahiko Maki,
Yusuke Ueda,
Yasushi Koike,
Takeshi Miyake,
Junichi Fukui,
Yunze Cui, [......],
Akihisa Nishida,
Yugo Ando,
Kequan Guo,
Changye Song,
Wenhao Cui,
Wei Feng,
Junko Katou,
Katuyuki Sado,
Shuji Nakamura,
Susumu Ikehara
[show abstract]
[hide abstract]
ABSTRACT: The collection of bone marrow cells (BMCs) using a perfusion method has been advantageous not only because of the low contamination of BMCs with T cells from the peripheral blood but also the enrichment of stromal cells, which support hemopoiesis. Before the application of this new method to humans, its safety needed to be confirmed using cynomolgus monkeys. We therefore performed the perfusion method on more than 100 cynomolgus monkeys using the long bones (such as the humerus and femur) and also the iliac bones (for human application); in the more than 150 trials to date, there have been no accidental deaths. Furthermore, the technical safety of a new method for the intra-bone marrow (IBM) injection of BMCs (termed IBM-bone marrow transplantation) has also been confirmed using 30 monkeys. Disclosure of potential conflicts of interest is found at the end of this article.
Stem Cells 09/2007; 25(8):2098-103. · 7.78 Impact Factor
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Hiromi Mukaide,
Yasushi Adachi,
Naoko Koike-Kiriyama,
Yasuhiro Suzuki,
Keizo Minamino,
Masayoshi Iwasaki, Masanobu Tsuda,
Keiji Nakano,
Yasushi Koike,
Akio Shigematsu,
Yasuo Kamiyama,
Susumu Ikehara
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ABSTRACT: We have previously shown that the combination of allogeneic intra-bone marrow-bone marrow transplantation (IBM-BMT) and donor lymphocyte infusion (DLI) using CD4+ cell-depleted spleen cells is effective in suppressing tumor growth, but that this does not induce graft-versus-host disease (GVHD) in mice. In this report, we show that formalin-fixed tumor cell-pulsed dendritic cells (FFTCP DCs) have an additive effect with IBM-BMT plus DLI on the suppression of tumor growth, but that the DCs do not augment GVHD. BALB/c mice, which had been subcutaneously inoculated with Meth A (BALB/c-derived fibrosarcoma), were irradiated at a low dose (5 Gy) and were transplanted with bone marrow cells (BMCs) from C57BL/6 (B6) mice into the bone marrow cavity (IBM-BMT). Simultaneously, the mice were intravenously injected with spleen cells from B6 mice, and subcutaneously injected with FFTCP DCs derived from the bone marrow (BM) of B6 mice. At the point of the induction of DCs from BMCs, formalin-fixed Meth A cells were added into the culture. The mice treated with the combination of FFTCP DCs, IBM-BMT and DLI using CD4+ cell-depleted spleen cells showed smaller tumor sizes and longer survival than the mice treated with IBM-BMT plus FFTCP DCs or IBM-BMT plus DLI using CD4+ cell-depleted spleen cells. These results suggest that the combination of FFTCP DCs, IBM-BMT plus DLI using CD4+ cell-depleted spleen cells has potent anti-tumor effects without showing GVHD.
International Journal of Oncology 07/2007; 30(6):1309-15. · 2.40 Impact Factor
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ABSTRACT: A P6 substrain of the senescence accelerated mouse (SAMP6) spontaneously develops osteoporosis early in life. These mice show the clinical signs of osteoporosis, such as elevated levels of urinary deoxypyridinoline (Dpd), decreased bone mineral density (BMD), and a significant loss of trabecular and cortical bone thickness at 12 months of age. Here, we describe the transfer of osteoporosis to a normal strain by the injection of bone marrow cells from SAMP6 donors directly into the bone marrow cavity (intra-bone marrow-bone marrow transplantation [IBM-BMT]). More than 1 month after IBM-BMT, hematolymphoid cells were completely reconstituted by donor-derived cells, and bone marrow stromal cells that could differentiate into osteocytes were also found to be of donor origin. In addition, the recipient C57BL/6 mouse showed the features of osteoporosis in the trabecular bone. Decreases in BMD and increases in urinary Dpd were also observed. When the message levels of cytokines (interleukin [IL]-11, IL-6, receptor activator of NF-kappaB ligand [RANKL], osteoprotegerin, macrophage-colony-stimulating factor, and insulin-like growth factor-1) were examined by reverse transcription-polymerase chain reaction (RT-PCR) and real-time RT-PCR analysis, IL-6 and IL-11 were reduced to a level similar to that in SAMP6 mice, whereas that of RANKL was increased. These findings indicate that not only the hemopoietic system but also the bone marrow microenvironment are reconstituted as a result of IBM-BMT, and suggest that the development of senile osteoporosis might be attributable to "stem cell disorders." Disclosure of potential conflicts of interest is found at the end of this article.
Stem Cells 07/2007; 25(6):1356-63. · 7.78 Impact Factor
