Pathological role of CD44 on NKT cells in carbon tetrachloride-mediated liver injury

Aim: CD44 has a variety of functions in immune regulation and signal transduction. Although CD44 is involved in the induction of several inflammatory diseases, it remains unknown whether CD44-targeting therapies are useful for liver diseases. Here, we examined whether CD44 blockade is effective in a chemical-induced liver injury model. Methods: We injected CD44 knock out (KO) or wild type mice with carbon tetrachloride (CCl(4)) and examined the difference of liver injury by immunological or histological analysis. Results: Although CD44KO mice exhibited suppressed liver injury at 6 h after CCl(4) injection with decreased inflammatory cell numbers and cytokine production, these mice showed severe liver injury at 24 h. We found that NKT cells played an important role in liver injury with increased infiltration of theliver after migration, which was independent of the CD44 pathway. In CD44NKT double-KO mice, liver injury was suppressed with reduced cytokine production and macrophage infiltration compared with CD44KO mice. Furthermore, MIP-2 derived from NKT cells or tumor necrosis factor alpha from macrophages contributed to exacerbation of the liver injury, since neutralization of MIP-2 provided significant protection against liver injury in CD44KO mice. Finally, we found that CD44KO mice exhibited excessive liver fibrosis compared with wild-type mice after repeated CCl(4) injections. Conclusion: We found that CD44 has unique characteristics for inflammatory liver diseases associated with NKT cell infiltration and activation. Furthermore, CD44-targeting therapies may need to be viewed with caution for liver diseases due to the actions of the liver immune system.     

Role of CD44 in CTL-induced acute liver injury in hepatitis B virus transgenic mice

Background  There are many uncertain points regarding leukocyte movement in the liver, especially interaction between liver sinus endothelial cells (LSECs) and cytotoxic T lymphocytes (CTLs). We examined the role of CD44 in these interactions using the hepatitis model. Methods  CTLs were administered to hepatitis B virus transgenic mice (HBVTg) mice and HBVTg × CD44 knockout (KO) mice, and alanine aminotransferase activity (ALT), number of intrahepatic leukocytes, cytokines, and chemokine mRNA level were examined. To determine the number and distribution of CTLs in the liver, 5,6-carboxyfluoroscein succinimidyl ester (CFSE)-labeled CTLs was administered to HBVTg mice with or without CD44. Results  Serum ALT activity increased after 12 h, although it had declined to 4 h in the CD44KO × HBVTg mice after CTL injection. Similarly, the levels of tumor necrosis factor (TNF)-α, interferon (IFN)-γ, and macrophage inflammatory protein (MIP)-2 mRNAs were reduced in 4 h, although the levels were increased after 12 h in the CD44KO × HBVTg mice. The number of apoptotic hepatocytes increased intentionally at 24 h in the CD44KO × HBVTg livers, and this was thought to result from the lower activity of initial nuclear factor kappa B (NF-κB). Although the number of CTLs was lower at 4 h in the CD44KO × HBVTg livers, the difference of intercellular adhesion molecule (ICAM)-1 and CD86 expression on LSECs was not detected. Conclusions  CD44 exerts and important effect on LSECs for CTL migration into the hepatocytes. However, because the CD44-deficient state exacerbated hepatic injury, attention is necessary for hepatitis treatment as CD44 target therapy.     

CD44V6和CD44S表达与膀胱癌转移的相关性研究

采用免疫组化ＳＡＢＣ法对８１例膀胱移行细胞癌患者癌组织中细胞粘附分子ＣＤ４４ｖ６和ＣＤ４４ｓ的表达进行检测。结果显示,ＣＤ４４ｖ６的阳性表达与膀胱癌临床分期、病理分级有显著关系,伴淋巴结转移组的ＣＤ４４ｖ６阳性表达率显著低于未转移组。     

Expression of CD44 in rat liver allografts during rejection

As CD44 is believed to be a homing receptor involved in lymphoid trafficking and inflammatory responses, it is expected to be closely linked to transplant rejection. In this study, the expression of CD44 during liver transplant rejection was compared with the expression of lymphocyte-function associated antigen-1 (LFA-1) and intercellular adhesion molecule-1 (ICAM-1), which play an essential role in cell interactions and the initiation of immune responses. Male Brown Norway (BN) and Lewis (LEW) rats were used as donors and recipients, respectively. Orthotopic liver transplantation (OLTX) was done using the cuff technique of Kamada and Calne. Animals were killed on days 3, 5, and 7 after OLTX, and a piece of tissue from each of the liver grafts was obtained. Immunohistochemical staining was used to investigate the expression of CD44, ICAM-1, and LFA-1. CD44 was strongly expressed in portal areas of the rejected liver, and LFA-1 and ICAM-1 were expressed mainly on sinusoids and hepatocytes. These findings indicate that CD44 is closely involved in lymphocyte infiltration, which is dominant in portal areas, and that lymphocyte infiltration during the rejection process may involve a homing mechanism.     

Immunohistochemical Study of Hyaluronate Receptor (CD44) in Alcoholic Liver Disease

Background: It has been suggested that the elevation of serum hyaluronate (HA) levels in liver diseases may be due to increased synthesis of HA by hepatic stellate cells or decreased degradation by sinusoidal endothelial cells. The increase in serum HA levels in patients with cirrhosis is thought to be a response to a reduction in HA receptors (CD44) in hepatic sinusoidal endothelial cells. To learn more about how alcohol affects the number and distribution of HA receptors of hepatic sinusoidal endothelial cells, we immunohistochemically studied CD44 levels in liver biopsy obtained from patients with alcoholic liver disease (ALD patients) and also from patients with nonalcoholic liver disease (non-ALD); ALD patients were evaluated when they were currently drinking and again after they became abstinent. Normal liver tissue obtained from three autopsy cases served as a control. Methods: Liver biopsy specimens were obtained from 18 ALD patients and 12 non-ALD patients. In ALD patients, liver biopsy was performed twice within 3 days and 4 to 8 weeks after abstinence when serum levels of aspartate aminotransferase and alanine aminotransferase became normal. CD44 in liver specimens was stained with anti-CD44 antibody by streptavidine-biotin-peroxidase complex. The intensity of the staining of CD44 in liver tissue was determined by a computer-assisted imaging analyzer. We also measured serum levels of CD44 in both ALD and non-ALD patients. Results: The intensity and the number of CD44 staining increased in both ALD and non-ALD patients compared with those in normal liver, which was negative. The staining intensity of CD44 in liver specimens obtained from patients with ALD who were active in alcohol consumption were significantly higher when compared with patients with ALD after abstinence. Serum levels of CD44 in patients with liver disease increased compared with those of healthy subjects. Conclusions: The results suggest that HA receptors may increase to degrade the increased HA in serum and/or liver.     

结直肠癌组织中CD44V3,v6蛋白的表达意义

目的研究CD44v3,v6蛋白在结直肠癌组织中的表达及其预后意义.方法应用EnvisionTM免疫组化法,回顾性分析121例结直肠癌石蜡组织CD44v3,v6的表达.患者中位随访时间为67.77mo.结果CD44v3,v6在结直肠癌中的阳性率分别为60.3%和57.9%.CD44v3的阳性表达与患者肿瘤部位,淋巴结转移状况、远处转移与否、临床分期密切相关(P＜0.05,Spearman等级相关检验).CD44v6的阳性表达与患者性别、淋巴结转移状况、远处转移与否、临床分期密切相关(P＜0.05,Spearman等级相关检验).Kaplan-Meier生存曲线,Log-rank检验单因素生存分析结果显示,CD44v3阴性、阳性患者的五年生存率分别为81.25%,60.27%,两者之间存在显著性差异(P=0.035);CD44v6阴性、阳性患者的5 a生存率分别为80.39%,60.00%,两者之间亦存在显著性差异(P=0.0299).Cox模型多因素生存分析结果显示,CD44v3表达是影响结直肠癌预后的独立因素.结论CD44v3,v6蛋白与结直肠癌转移、预后相关,CD44v3是影响结直肠癌预后的独立因素.     

CD44v17在胃癌组织中的表达及其与临床生物学特性之间的关系

目的 探讨一种新的CD44变异体（CD44v17）在胃癌组织中的表达及其与临床生物学特性之间的关系.方法根据本实验室在MCF-7／Adr中新发现的1种CD44剪切拼接变异体CD44v17设计特异性引物,应用SYBR Green I荧光染料,采用实时PCR法检测87例胃癌组织及相应癌旁组织CD44v17 Mrna的表达,根据标准曲线计算CD44v17mRNA的表达量,分析CD44v17mRNA表达与胃癌临床生物学特性之间的关系.结果胃癌组织CD44v17 Mrna表达明显高于相应癌旁正常组织（P〈0.05）;肿瘤〉5 cm组CD44v17 Mrna表达与肿瘤≤5 cm组表达比较差异无显著性（P〉0.05）;未分化腺癌组表达明显高于乳头状腺癌及管状腺癌组（P〈0.05）,乳头状腺癌组CD44v17 Mrna表达与管状腺癌组比较差异无显著性（P〉0.05）;肿瘤累及浆膜及浆膜外组CD44v17 Mrna表达显著高于侵及黏膜、黏膜下层及肌层组（P〈0.05）,而肿瘤侵及黏膜及黏膜下层组的CD44v17 Mrna表达与侵犯肌层组表达比较差异无显著性（P〉0.05）.淋巴结转移组CD44v17 Mrna表达明显高于淋巴结无转移组（P〈0.05）;有肝脏转移组CD44v17 Mrna表达显著高于无肝脏转移组（P〈0.05）.结论CD44v17 Mrna在胃癌组织中高表达,可能与胃癌的侵袭、转移及预后有关.     

NIV毒素和硒对软骨细胞CD_（44）代谢的影响

目的进一步探讨大骨节病（KBD）的发病机制。方法取4月龄引产胎儿软骨细胞原代分离、培养并随机分为四组。对照组不处理,加硒组加入硒浓度为0.1 mg/L,雪腐镰刀菌烯醇毒素（NIV）组加入NIV毒素使终质量浓度为0.1 mg/L,联合组加硒和NIV毒素,浓度同上。RT-PCR法检测各组CD44 mRNA在软骨细胞中的表达;流式细胞仪检测各组软骨细胞表面CD44蛋白的表达;酶联免疫吸附法检测细胞培养液中可溶性CD44（sCD44）水平。结果 NIV组、联合组软骨细胞CD44 mRNA及蛋白表达水平均明显高于对照组和加硒组（P均〈0.05）。与NIV组比较,联合组CD44表达有所降低,但不明显。NIV组细胞培养液中溶解的sCD44水平最高,其次为联合组,对照组水平最低。结论 NIV毒素和硒能影响软骨细胞CD44的代谢,这可能是造成软骨细胞损伤的机制。     

Protective effect of bicyclol on lipopolysaccharide-induced acute lung injury in mice

Bicyclol is synthesized based on schisandrin, which is one of the main active components of Chinese herb Fructus Schisandrae. The purpose of this study is to investigate whether bicyclol has a beneficial effect on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. Bicyclol was given to mice by gavage for three times. ALI was induced by vena caudalis injection of LPS. The last dose of bicyclol was administrated 1 h before LPS given. Mice in each group were sacrificed at different time point after LPS administration. As revealed by survival study, pretreatment with high doses of bicyclol reduced the mortality of mice from ALI. Bicyclol pretreatment significantly improved LPS-induced lung pathological changes, inhibited myeloperoxidase (MPO) activity, and reduced lung/body and lung wet/dry weight ratios. Bicyclol also inhibited the release of TNF-α, IL-1β and HMGB1, whereas simultaneously increased the expression of IL-10. Furthermore, the phosphorylation level of NF-κB p65 was markedly decreased by bicyclol. Taken together, our study showed that bicyclol improves survival rate and attenuates LPS-induced ALI. The protective mechanism may be due to the inhibition of NF-κB activation and regulation of cytokine secretion.     

结直肠癌组织中ESA和CD44的表达及其临床病理意义

目的:探讨原发性结直肠癌组织中E SA和CD44蛋白表达的关系及临床病理意义.方法:应用免疫组织化学SP法检测10例正常结直肠黏膜组织、13例伴不典型增生的结直肠腺瘤及67例结直肠癌组织中ESA和CD44的表达,并比较两者不同病理参数的相关性.结果:结直肠癌组织中ESA和CD44的表达明显高于其在正常肠黏膜组织和伴不典型增生的结直肠腺瘤中的表达( P<0.05); 在结直肠癌中,CD44的表达与淋巴结转移,分化程度及Dukes分期有关(χ2 = 8.375,14.284,7.948,均P<0.05); ESA与年龄、浸润深度、分化程度及Dukes分期有关(χ2 = 16.918,15.195,10.395,7.681,均P<0.05); ESA的表达与CD44的表达呈正相关( r = 0.614,P = 0.000).结论:ESA与CD44在结直肠癌的发生发展、浸润转移中密切相关,同步检测二者在结直肠癌组织中的表达并综合分析两者之间的关系对评价结直肠癌的侵袭转移能力判断具有一定价值.     

CD44v3和CD44v6在溃疡性结肠炎与其他类型结肠炎中的表达

目的比较CD44v3和CD44v6在溃疡性结肠炎(UC)与对照组(包括感染性结肠炎、阿米巴痢疾、血吸虫性结肠炎及克罗恩病等其他类型结肠炎)中表达的差异.方法免疫组化SP法检测两组患者结肠粘膜CD44v3和CD44v6的表达.结果 UC组患者CD44v3和CD44v6表达阳性率分别为68.8%和56.3%,与对照组比较有显著性增高(P<0.05).UC组与感染性结肠炎、阿米巴痢疾、血吸虫性结肠炎病及克罗恩病分别比较差异仍有显著性(P<0.05).结论 CD44v3和CD44v6的检测有助于鉴别UC和感染性结肠炎等其他类型结肠炎.     

肝癌组织骨桥蛋白及CD44v6的表达与肝癌肝移植术患者预后的意义研究

目的研究骨桥蛋白(OPN)、CD44v6预测肝细胞癌肝移植患者预后效能,为寻求新的受体选择标准提供理论依据。方法收集2002年8月至2006年7月期间在西安交通大学医学院第一附属医院行经典原位肝移植术,且病理确诊为肝细胞癌的病例共30例。应用组化染色法,检测OPN、CD44v6的表达,进行统计分析。结果 30例肝细胞肝癌组织中OPN和CD44v6表达正相关。肝移植术后肿瘤复发转移与符合米兰标准情况显著负相关(P<0.01),与OPN、CD44v6的表达正相关(P<0.05)。在超出米兰标准的患者中,肿瘤复发与OPN、CD44v6阳性表达的相关性更强(P<0.01)。随访期内OPN或CD44v6染色阳性患者的生存率明显低于阴性患者(P=0.001),双阳性患者的生存率最低。对于超米兰标准的肝移植患者更为显著(P=0.004)。结论 OPN、CD44v6可以作为预测肝细胞癌肝移植患者预后的预测标签,联合检测OPN、CD44v6能够更好地反映超米兰标准的肝移植患者预后,检测OPN、CD44v6可以纳入受体选择标准。     

Expression of CD44 variants in osteosarcoma

The standard form of CD44 (CD44H) is a transmembranous glycoprotein, widely distributed on a variety of human lymphoid cells, epithelial cells and tumours. CD44 has many variant forms, which are generated by alternative splicing. In recent years, CD44 has been reported to be related to the degree of tumour differentiation, tumour cell invasion, and metastasis. We investigated 44 tumour specimens in 39 patients with osteosarcoma immunochemically to analyse the expression of CD44 standard (CD44H) and variant exon-encoded gene products (CD44v3, v4, v5, v6, v7, v9, and v10). Furthermore, the relationship between CD44 expression and the clinical outcome of patients with osteosarcoma was analysed. Membrane accentuation and exclusive cytoplasmic reactivity were analysed as separate staining patterns. Tumour cells and some multinucleated giant cells were markedly stained. CD44H, v3, v4, v5, v6, v7, v9, and v10 were expressed in 85%, 49%, 54%, 59%, 46%, 5%, 28%, and 10% of the specimens respectively. The cumulative 5-year metastasis-free survival was 58% in CD44v6-negative cases and 24% in CD44v6-positive cases (P=0.046). However, the cumulative 5-year metastasis-free survival was not significantly different between cases positive and negative for other variants of CD44. Multivariate analysis (Cox proportional-hazard model) with CD44v6 expression (positive or negative), chemotherapy (intensive or non-intensive), tumour site (proximal or distal), and age (at least 30 years or less than 30 years) showed that expression of CD44v6 and chemotherapy were important prognostic factors in patients with osteosarcoma. Overexpression of CD44 isoforms containing variant v6 is correlated with poor prognosis in patients with osteosarcoma. Received: 4 March 1999 / Accepted: 7 June 1999     

明胶酶、CD44v6在结直肠癌中的表达及意义

目的探讨明胶酶在结直肠癌中的表达及其CD44v6在结直肠癌侵袭转移过程中的关系。方法采用明胶酶谱（gel zymography）和S-P免疫组化对50例结直肠癌癌组织和相应正常组织中的明胶酶、CD44v6进行检测。结果明胶酶在结直肠癌中的表达水平显著高于正常组织（P〈0．05）；明胶酶与结直肠癌的Duke’s分期呈同步的正相关（P〈0．05）；明胶酶在CD44v6强阳性组中表达量升高显著（P〈0．05）。结论明胶酶与结直肠癌的侵袭转移性有着密切的关系，不仅在癌细胞的酶解，而且在癌细胞的黏附、运动过程中有可能发挥着重要作用。     

Expresion of cell adhesion molecule CD44 in primary tumors of the liver: an immunohistochemical study

Abstract: CD44, a widely distributed integral membrane protein, has been implicated in tumor invasion and metastatic spread in some human carcinomas and lymphomas. In this study, 35 cases of hepatocellular carcinoma from 32 patients (11 cholangiocarcinomas, 9 hepatic adenomas, and 5 cases of focal nodular hyperplasia, a non-neoplastic lesion) were examined by imunohistochemical methods for expression of CD44. The mouse monoclonal antibody A3D8 was used on formalin-fixed, paraffin-embedded tissue; this antibody does not distinguish between standard CD44 and splice variants. Positive membrane staining was seen in 13 of 35 cases of hepatocellular carcinoma (12 of 32 patients), 8 of 11 cases of cholangiocarcinoma, and 1 of 9 cases of hepatic adenoma. The strongest staining for CD44 was seen in two cases of fibrolamellar carcinoma, but CD44 expression was otherwise not related to degree of tumor differentiation. All five cases of focal nodular hyperplasia were negative for CD44. In non-neoplastic liver, hepatocytes were negative; sinusoidal lining cells and portal lymphocytes were positive; bile ducts and proliferating bile ductules were focally positive in some cases. Anatomic stage at time of presentation was similar in both groups of patients, with most patients presenting with stage III or IV disease. A trend towards slightly longer survival in patients whose hepatocellular carcinomas were CD44 negative was noted. These results show that aberrant CD44 expression is present in a subset of hepatocellular carcinomas and in most cholangiocarcinomas. The relationship between CD44 expression and tumor spread is unclear in this group of tumors, but is unlikely to be a simple association between CD44 expression and metastatic potential.     

CD44蛋白在胃癌中的表达及与临床关系的探讨

目的:探讨CD44蛋白在胃癌中表达及与临床的关系。方法:采用免疫组化S-P方法检测CD44S及CD44变异体V6在54例胃良性病变(BGD)及63例胃癌(GC)中的表达,并比较了阳性表达与肿瘤组织分型及临床预后等相关性。结果:CD44S在浅表性胃炎(CSG)、萎缩性胃炎伴肠上皮化生(CAG/IM)及GC中表达阳性率分别为48.5％、57.1％和52.4％。CD44V6在CSG中无表达,在CAG/IM及GC中表达阳性率为19.1％和53.9％。在肠型胃癌中,CD44V6表达率(73.7％)明显高于弥漫型胃癌(24％)。CD44S阳性表达与组织分型无关。CD44S阳性表达与术后复发率呈正相关,与5年生存率呈负相关。结论:CD44S和CD44V6在胃癌中均有表达,CD44V6表达率与胃癌组织来源有关。CD44S表达率与临床预后有关。     

Interaction between hyaluronan and CD44 in the development of dimethylnitrosamine-induced liver cirrhosis

BACKGROUND: A significant increase in serum hyaluronan (HA) levels has been reported in patients with liver cirrhosis. This mechanism is not yet clear, and receptors for HA have not been characterized. In this study, we examined the expression of both HA and its receptors, CD44 and intercellular adhesion molecule-1 (ICAM-1), in dimethylnitrosamine-induced liver cirrhosis. METHODS AND RESULTS: Using biotinylated HA binding protein, HA was detected in the area of periportal fibrosis and around the sinusoidal wall where hepatic fibrosis was developing. Electron microscopy revealed that HA was localized on Ito cells and sinusoidal endothelial cells (SEC). Conversely, CD44, which was only expressed weakly in normal liver, was present in large amounts in cirrhotic liver. The distribution pattern of CD44 was similar to that of HA, however, CD44 was mainly localized on the infiltrating lymphocytes and Kupffer cells. Moreover, CD44 was detected on part of factor VIII-positive SEC. Intercellular adhesion molecule-1, another receptor for HA, was detected on the surface of hepatocytes and around the sinusoidal wall in cirrhotic liver, but its distribution was not accompanied by expression of HA. With respect to CD44 isoforms, the standard form m-RNA predominated in both normal and cirrhotic liver. Variant pMeta-1 mRNA was detected at low levels. CONCLUSIONS: An interaction between HA and CD44 may play a role in the recruitment of numerous infiltrating cells and HA accumulation in hepatic sinusoids. Together with phenotypic changes in the SEC, these results may lead to a disturbance in the elimination of HA during the progression of liver cirrhosis.     

CD44v6和MMP-9在胎盘原位滋养细胞肿瘤中的表达及相关性

目的探讨CD44v6和基质金属酶-9（MMP-9）在胎盘原位滋养细胞肿瘤（PSTT）中的表达及相关性。方法采用SABC免疫组化法检测30例正常绒毛、15例PSTT的CD44v6和MMP-9表达情况。结果CD44v6在正常胎盘绒毛、良性PSTT、恶性PSTT的阳性表达率分别是3．3％、30．3％、100％,组间比较差异有统计学意义（P〈0．05）;而MMP-9的阳性表达率分别为6．7％、38．5％、100％,组间比较差异有统计学意义（P〈0．05）。结论CD44v6和MMP-9的过表达促进PSTT的浸润转移,二者具有正协同作用,联合检测CD44v6和MMP-9蛋白可作为PSTT的浸润转移及评价预后生物学指标。     

T-2毒素对软骨细胞CD44表达的影响

目的 研究T-2毒素对软骨细胞CD44表达的影响，为阐明T-2毒素损伤软骨组织的机制提供科学依据。方法 利用RT-PCR方法检测CD44 mRNA在软骨细胞中的转录情况；利用免疫组化方法检测软骨细胞表面CD44的表达情况。结果 在T-2毒素作用5d后，毒素组、毒素加硒组CD44 mRNA表达水平均低于对照和对照加硒组。同时免疫组化结果显示，毒素组体外再建软骨组织CD44表达最低，其次为毒素加硒组，对照加硒组最高。结论 T-2毒素对于软骨细胞的CD44表达具有明显的抑制作用。