Therapy with stem cells in inflammatory bowel disease

Inflammatory bowel disease(IBD) affects a part of the young population and has a strong impact upon quality of life. The underlying etiology is not known, and the existing treatments are not curative. Furthermore, a significant percentage of patients are refractory to therapy. In recent years there have been great advances in our knowledge of stem cells and their therapeutic applications. In this context, autologous hematopoietic stem cell transplantation(HSCT) has been used in application to severe refractory Crohn’s disease(CD), with encouraging results. Allogenic HSCT would correct the genetic defects of the immune system, but is currently not accepted for the treatment of IBD because of its considerable risks. Mesenchymal stem cells(MSCs) have immune regulatory and regenerative properties, and low immunogenicity(both autologous and allogenic MSCs). Based on these properties, MSCs have been used via the systemic route in IBD with promising results, though it is still too soon to draw firm conclusions. Their local administration in perianal CD is the field where most progress has been made in recent years, with encouraging results. The next few years will be decisive for defining the role of such therapy in the management of IBD.     

骨髓干细胞治疗炎症性肠病的研究进展

炎症性肠病（inflammatory bowel disease,IBD）是一种侵及胃肠道的特发的炎症反应,包括溃疡性结肠炎（ulcerative colitis,UC）和克罗恩病（Crohn’s disease,CD）,在中国的发病率逐年上升,其可能与现代生活方式、基因遗传、肠道菌群的免疫耐受缺失、免疫反应激活等因素有关。目前的治疗在于控制活动性炎症和调节免疫紊乱,包括抗炎药物、激素、免疫抑制剂、生物治疗等多种治疗手段,但无一具有长期疗效且副作用颇多。而近年来干细胞再生、营养、免疫调节等方面的研究给IBD的治疗带来了新的希望。本文讨论了骨髓间充质干细胞及造血干细胞在IBD治疗中的新发现,回顾了干细胞在IBD治疗中可能的作用机制。     

间充质干细胞与肺疾病的研究进展

间充质干细胞(mesenchymal stem cells,MSC)是一类具有自我复制和多向分化潜能的成体干细胞,广泛分布于人及动物多种组织和器官中.文献报道MSC在体内能分化为气道及肺组织上皮细胞.本文着重阐述了MSC对肺疾病损伤的修复及其在肺疾病治疗中的应用前景和存在的问题.     

Evaluation of inflammatory activity in Crohn's disease and ulcerative colitis

Crohn's disease and ulcerative colitis evolve with a relapsing and remitting course. Determination of inflammatory state is crucial for the assessment of disease activity and for tailoring therapy. However, no simple diagnostic test for monitoring intestinal inflammation is available. Noninvasive markers give only indirect assessments of disease activity. Histopathological or endoscopical examinations accurately assess inflammatory activity, but they are invasive, time consuming and expensive and therefore are unsuitable for routine use. Imaging procedures are not applicable for ulcerative colitis. The usefulness of ultrasound and Doppler imaging in assessing disease activity is still a matter of discussion for Crohn's disease, and an increased interest in computed tomography enterograph (CTE) has been seen, mainly because it can delineate the extent and severity of bowel wall inflammation, besides detecting extraluminal findings. Until now, the available data concerning the accuracy of magnetic resonance enterography in detecting disease activity is less than CTE. Due to this, clinical activity indices are still commonly used for both diseases.     

间充质干细胞与支气管哮喘

支气管哮喘是一种气道慢性炎症性疾患,同时也是一种免疫紊乱的变态反应性疾病,其发病机制复杂.间充质干细胞(mesenchymal stem cells,MSCs)是来源于中胚层的具有高度自我更新能力和多向分化潜能的多能干细胞,并且具有强大的免疫调节功能.MSCs这些独特性能将赋予它们在支气管哮喘冶疗中的潜在价值.     

自体造血干细胞移植治疗难治性炎症性肠病10例回顾与随访

目的 回顾自体造血干细胞移植(HSCT)治疗10例炎症性肠病(IBD)患者的疗效及安全性.方法 2004年1月至2006年8月,采用HSCT治疗9例糖皮质激素及免疫抑制剂治疗无效的克罗恩病(CD)患者及1例Truelove临床重型溃疡性结肠炎(UC)患者.CD患者中2例CD活动指数(CDAD大于450(严重型),6例CDAI为150～450(活动型).1例UC患者为全结肠炎型.经环磷酰胺(CTX)及粒细胞刺激因子动员后,采集患者外周血于细胞,行CD34+细胞分选并置液氮保存,2周后行CTX及抗淋巴细胞球蛋白预处理,将解冻的干细胞回输.结果 HSCT后3和12个月时分别有5例和1例患者CDAI<150(完全缓解).2例术后CDAI分值下降但未达缓解标准;完全缓解者症状消失,血液检查指标正常,体重明显增加(5～20 kg).平均随访16.1个月,复发4例,除1例病情严重外,余均较术前病情减轻;5例患者获长期缓解.1例UC患者术后10个月无复发症状,血液检查指标均正常,但肠镜复查示无明显改善.与HSCT治疗相关的并发症有干细胞回输期间与白细胞减少相关的发热及感染、术后脱发,1例乙型肝炎病毒(HBV)携带者术后转氨酶水平及HBV-DNA滴度增高.结论 HSCT可改善其他方法治疗无效的难治性IBD,近期疗效满意,部分患者可获较长期的缓解,治疗期间无严重不良反应,但不能阻止病情复发,且不能改善患者肠道病理改变.     

骨髓间充质干细胞与肺间质纤维化

间质性肺疾病（ILD）是由各种不同原因引起的一组疾病,其以肺间质纤维化为共同病理改变,是导致肺功能进行性丧失和呼吸衰竭的主要原因。特发性肺纤维化（IPF）是最常见的一种ILD,其对目前采用的药物治疗几乎无反应,预后差。目前对这类疾病发病机制的研究认为,无论是否有炎症反应,肺纤维化形成都是肺损伤与修复反复失衡的结果。间充质干细胞（MSC）是广泛存在于人体各组织中的一种成体干细胞,具有多向分化潜能。在受损肺,骨髓来源的MSC（BMd-MSC）可以分化为上皮细胞、血管内皮细胞和成纤维细胞。采用MSC疗法有可能是治疗肺纤维化最理想的方法之一。     

Proteome profiling of spinal cord and dorsal root ganglia in rats with trinitrobenzene sulfonic acid-induced colitis

AIM: To investigate proteomic changes in spinal cord and dorsal root ganglia (DRG) of rats with trinitrobenzene sulfonic acid (TNBS)-induced colitis.METHODS: The colonic myeloperoxidase (MPO) activity and tumor necrosis factor-α (TNF-α) level were determined. A two-dimensional electrophoresis (2-DE)-based proteomic technique was used to profile the global protein expression changes in the DRG and spinal cord of the rats with acute colitis induced by intra-colonic injection of TNBS.RESULTS: TNBS group showed significantly elevated colonic MPO activity and increased TNF-α level. The proteins derived from lumbosacral enlargement of the spinal cord and DRG were resolved by 2-DE; and 26 and 19 proteins that displayed significantly different expression levels in the DRG and spinal cord were identified respectively. Altered proteins were found to be involved in a number of biological functions, such as inflammation/immunity, cell signaling, redox regulation, sulfate transport and cellular metabolism. The overexpression of the protein similar to potassium channel tetramerisation domain containing protein 12 (Kctd 12) and low expression of proteasome subunit α type-1 (psma) were validated by Western blotting analysis.CONCLUSION: TNBS-induced colitis has a profound impact on protein profiling in the nervous system. This result helps understand the neurological pathogenesis of inflammatory bowel disease.     

间充质干细胞治疗糖皮质激素耐药性慢性移植物抗宿主病临床疗效观察

目的 探讨间充质干细胞(MSC)治疗糖皮质激素(激素)耐药性慢性移植物抗宿主病(cGVHD)的疗效及作用机制.方法 12例激素耐药性cGVHD患者在原有免疫抑制剂基础上联合MSC治疗.12例患者共接受24例次MSC输注,17例次MSC来自第二方,7例次来源于第三方,其中1例输注1次、10例输注2次、1例输注3次.首次输注细胞量1.0(0.4～2.1)×106/kg,第2次输注1.2(0.8～1.9)×106/kg,末次输注细胞量为1.1×106/kg.5例接受2例不同供者MSC输注.并分别于MSC治疗前、治疗后4周应用流式细胞仪检测患者外周血CD+3、CD+4、CD+8、CD+19、CD+4CD+25、FOXF3+、FOXP3+CD+4及FOXP3+CD+25淋巴细胞亚群比例.结果 12例患者接受MSC输注后3例获得完全缓解(CR)、6例获得部分缓解(PR),总有效率(CR+PR)为75%(9/12).3例CB患者在MSc治疗471(372～731)d后停用免疫抑制剂,随访129(100～292)d cGVHD无复发;6例PR患者在MSC治疗64(60～79)d后开始进入免疫抑制剂减量阶段,目前除1例继发舌瘤致死外,其他患者应用小剂量免疫抑制剂维持治疗随访中位生存时间1152(795～1914)d病情仍稳定并存活.3例无效患者中,1例疾病稳定患者目前仍需联合三种免疫抑制剂治疗,另2例疾病进展患者分别于输注MSC8个月及10个月后死于白血病复发及肺脏cGVHD.输注MSC后12例患者CD4/CD8比例及调节性T细胞均显著升高,B细胞无明显改变.结论 第二方或第三方来源的Msc对于治疗激素耐药性cGVHD具有一定疗效,其作用机制可能与提高CD4/CD8比例及诱导调节性T细胞形成有关.     

肠道干细胞与炎症性肠病的关系

炎症性肠病(IBD)包括克罗恩病(CD)和溃疡性结肠炎(UC),是一类病因和发病机制尚不十分明确的肠道慢性非特异性疾病。近年来其发病率呈上升趋势,但到目前为止还没有找到彻底治愈的方法。肠道干细胞是成体干细胞的一种,主要分布在肠道的隐窝,其在修复损伤的黏膜中发挥重要作用。本文就肠道干细胞与IBD的关系作一概述。     

Mesenchymal stem cells and autoimmune diseases

Mesenchymal stem cell (MSC) immunosuppressive properties offer a potentially attractive therapeutic modality for autoimmune diseases. MSC inhibit virtually all types of immune responses in vitro and prevent the induction of disease in several experimental models of autoimmunity. However, the processes involved in the pathogenesis of human diseases are more complicated and treatment cannot be administered before disease induction. In autoimmune diseases persistent antigenic stimulation recruits endogenous MSC to the site of lesion that contribute to the fibrotic evolution. Therefore, administering MSC to a chronic inflammatory disorder may not be desirable. In fact, MSC are not constitutively immunosuppressive but require a 'licensing' step provided by molecules of acute phase inflammation, like IFNγ and TNF-α, or toll-like receptor (TLR) ligands. Conversely, different cytokines and/or the stimulation of selective TLR make MSC to become immunostimulatory. Therefore, dissecting the inflammatory environment in autoimmune diseases will identify the best conditions amenable to successful MSC therapy.     

Suppression of inflammatory cytokine secretion by granulocyte/monocyte adsorptive apheresis in active ulcerative colitis

To elucidate the molecular mechanisms involved in the therapeutic effects of granulocyte/monocyte adsorption apheresis, changes were investigated in the cytokine responses of peripheral blood mononuclear cells (PBMC) before and after granulocyte/monocyte adsorptive apheresis in ulcerative colitis (UC) patients. Four patients with active UC were enrolled. All patients responded to granulocyte/monocyte adsorptive apheresis. A total of 20 sessions of four patients were analyzed. Peripheral blood mononuclear cells were isolated from peripheral venous blood within 5 min before and after each session of granulocyte/monocyte adsorptive apheresis. The cells were stimulated with interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha for 24 h, and the secreted IL-8 and IL-6 levels were determined by enzyme-linked immunosorbent assay (ELISA). IL-1beta-induced IL-8 and IL-6 secretion was significantly decreased after granulocyte/monocyte adsorptive apheresis. TNF-alpha-induced IL-8 secretion was also significantly decreased after apheresis, but there was no significant difference in TNF-alpha-induced IL-6 secretion (P = 0.052). In conclusion, granulocyte/monocyte adsorptive apheresis down-regulates the IL-1beta- and TNF-alpha-induced inflammatory responses in PBMC. The induction of hyporesponsiveness to pro-inflammatory cytokines may be an important factor mediating the clinical effects of granulocyte/macrophage adsorptive apheresis in UC patients.     

肠道蠕虫与炎症性肠病的治疗

炎症性肠病(IBD)是一种病因不明,反复发作性的肠道特异性疾病。随着遗传因素、环境因素与免疫反应异常等发病机制方面研究的不断进展,IBD的治疗发生了重大的变化,其中直接改善肠道微环境逐渐受到重视。在动物模型和人体寄生虫感染研究中,有丰富的资料支持寄生虫感染有免疫调节作用,近来这个概念已经用来治疗IBD,不断有肠道蠕虫治疗IBD的报道。现就肠道蠕虫治疗IBD的研究作一综述。     

Mesenchymal stromal cell-based therapy: Regulatory and translational aspects in gastroenterology

The past decade has witnessed an outstanding scientific production focused towards the possible clinical applications of mesenchymal stromal cells (MSCs) in autoimmune and chronic inflammatory diseases. This raised the need of novel standards to adequately address quality, efficacy and safety issues of this advanced therapy. The development of a streamlined regulation is currently hampered by the complexity of analyzing dynamic biological entities rather than chemicals. Although numerous pieces of evidence show efficacy in reducing intestinal inflammation, some inconsistencies between the mechanisms of action of rodent vs human MSCs suggest caution before assigning translational value to preclinical studies. Preliminary evidence from clinical trials showed efficacy of MSCs in the treatment of fistulizing Crohn’s disease (CD), and preparations of heterologous MSCs for CD treatment are currently tested in ongoing clinical trials. However, safety issues, especially in long-term treatment, still require solid clinical data. In this regard, standardized guidelines for appropriate dosing and methods of infusion could enhance the likelihood to predict more accurately the number of responders and the duration of remission periods. In addition, elucidating MSC mechanisms of action could lead to novel and more reliable formulations such as those derived from the MSCs themselves (e.g., supernatants).     

Inflammatory bowel disease： Moving toward a stem cell-based therapy

The incidence and prevalence of Crohn＇s disease （CD） and ulcerative colitis （UC）, the two major forms of inflammatory bowel diseases （IBD）, are rising in western countries. The modern hygienic lifestyle is probably at the root of a disease where, in genetically susceptible hosts, the intestinal commensal flora triggers dysregulated immune and inflammatory responses. Current therapies ranging from anti-inflammatory drugs to immunosuppressive regimens, remain inadequate. Advances in our understanding of the cell populations involved in the pathogenetic processes and recent findings on the regenerative, trophic and immunoregulatory potential of stem cells open new paths in IBD therapy. Hematopoietic and mesenchymal stem cells are catalyzing the attention of IBD investigators. This review highlights the pivotal fi ndings for stem cell-based approaches to IBD therapy and collects the encouraging results coming in from clinical trials.     

干细胞疗法治疗炎症性肠病的应用

炎症性肠病是一种慢性炎症性疾病,主要包括克罗恩病和溃疡性结肠炎,其发病原因尚不清楚。干细胞疗法在治疗炎症性肠病方面可能是非常有价值的,近年来,干细胞疗法的研究也更加突出而显著,本文就干细胞疗法治疗炎症性肠病的应用作一概述。     

间充质干细胞与糖尿病慢性并发症

间充质干细胞（MSCs）具有多向分化能力，由于MSCs的分化细胞类型与糖尿病慢性并发症损伤的细胞类型吻合，近年来越来越多的研究针对MSCs在糖尿病慢眭并发症发病机制中的作用。糖尿病时高血糖可能通过影响MSCs的细胞分化类型而导致糖尿病多种并发症的发生。从MSCs多向分化潜能的角度解释糖尿病慢性并发症的发病机制为其治疗提供了新的思路，具有重要的临床意义。     

Use of infliximab in the prevention and delay of colectomy in severe steroid dependant and refractory ulcerative colitis

AIM： To determine if infliximab can prevent or delay surgery in refractory ulcerative colitis （UC）. METHODS： UC patients who failed to have their disease controlled with conventional therapies and were to undergo colectomy if infliximab failed to induce a clinical improvement were reviewed. Patients were primarily treated with a single 5 mg/kg infliximab dose. The Colitis Activity Index （CAI） was used to determine response and remission. Data of 8 wk response and colectomy rates at 6 mo and 12 mo were collected. RESULTS： Fifteen patients were included, 7 with UC unresponsive or intolerant to Ⅳ hydrocortisone, and 8 with active disease despite oral steroids （all but one with therapeutic dosage and duration of immunomodulation）. All the Ⅳ hydrocortisone-resistant/intolerant patients had been on azathioprine/6-MP 〈 8 wk. At 8 wk, infliximab induced a response in 86.7% （13/15） with 40% in remission （6/15）. Within 6 mo of treatment 26.7% （4/15） had undergone colectomy and surgery was avoided in 46.6% （7/15） at 12 mo. The colectomy rate at 12 mo in those on immunomodulatory therapy 〈 8 wk at time of infliximab was 12.5% （1/8） compared with 100% （7/7） in patients who were on long-term maintenance immunomodulators （P 〈 0.02）. CONCLUSION： Infliximab prevented colectomy due to active disease in immunomodulatory-na？ve, refractory UC patients comparable to the use of Cyclosporine. In patients, however, on effective dosage and duration of immunomodulation at time of infliximab therapy colectomy was not avoided.     

Therapeutic and prophylactic thalidomide in TNBS-induced colitis: Synergistic effects on TNF-α, IL-12 and VEGF production

AIM: To evaluated the therapeutic and prophylactic effect of thalidomide on 2, 4, 6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. Thalidomide has been reported to downregulate the expression of tumor necrosis factor α (TNF-α), IL-12, and vascular endothelial growth factor (VEGF), hallmarks of intestinal inflammation in Crohn's disease (CD).METHODS: Male Wistar rats were divided in five groups of ten animals each. Four groups received a rectal infusion of TNBS in ethanol. The first group was sacrificed 7 d after colitis induction. The second and third groups received either thalidomide or placebo by gavage and were sacrificed at 14 d. The fourth group received thalidomide 6 h before TNBS administration, and was sacrificed 7 d after induction. The fifth group acted as the control group and colitis was not induced. Histological inflammatory scores of the colon were performed and lamina propria CD4+ T cells, macrophages, and VEGF+ cells were detected by immunohistochemistry. TNF-α and IL-12 were quantified in the supernatant of organ cultures by ELISA.RESULTS: Significant reduction in the inflammatory score and in the percentage of VEGF+ cells was observed in the group treated with thalidomide compared with animals not treated with thalidomide. Both TNF-α and IL-12 levels were significantly reduced among TNBS induced colitis animals treated with thalidomide compared with animals that did not receive thalidomide.TNF-α levels were also significantly reduced among the animals receiving thalidomide prophylaxis compared with untreated animals with TNBS-induced colitis. Intestinal levels of TNF-α and IL-12 were significantly correlated with the inflammatory score and the number of VEGF+ cells.CONCLUSION: Thalidomide significantly attenuates TNBS-induced colitis by inhibiting the intestinal production of TNF-α, IL-12, and VEGF. This effect may support the use of thalidomide as an alternate approach in selected patients with CD.