A randomized, placebo controlled trial of an antisense oligodeoxynucleotide to intercellular adhesion molecule-1 in the treatment of severe rheumatoid arthritis

OBJECTIVE: To determine the safety of an antisense oligodeoxynucleotide to intercellular adhesion molecule-1 (ICAM-1) (ISIS 2302), administered in an intensive 4 week regimen with dose escalation; and to provide preliminary evidence for efficacy in rheumatoid arthritis (RA). METHODS: Patients with active RA were enrolled in a 6 month, double blind, placebo controlled, dual center, dose escalation (0.5, 1, and 2 mg/kg) study. Subjects received a total of 13 intravenous ISIS 2302 infusions, given on alternate days for 2 weeks and then 3 times a week for another 2 weeks. Doses of corticosteroids (< or = 10 mg/day) and disease modifying antirheumatic drugs (stable > or = 3 months) remained constant throughout the study. The primary efficacy endpoint was the Day 26 Paulus index, with secondary evaluations at Months 2-6. RESULTS: A total of 43 patients were enrolled with 11, 10, 3, and 19 patients receiving placebo or 0.5, 1, or 2 mg/kg of ISIS 2302, respectively. There were no differences between groups after randomization and the mean baseline swollen joint count was 22.5. Pharmacokinetic studies revealed a T(1/2) of 63 min and first-order kinetics with slight dose dependency, suggesting a saturable clearance process, although no accumulation was noted with repeat dosing. The Paulus 20% responses at Day 26 were 20%, 0%, and 5% for patients treated with ISIS 2302 (0.5, 1, 2 mg/kg, respectively) and 36% with placebo. For Months 2-6, the average intent-to-treat Paulus 20% responses were 21.2% for ISIS 2302 and 12.6% for placebo. Only ISIS 2302 treated subjects (19%) achieved Paulus 50% responses. ISIS 2302 was well tolerated. An expected and transient mean activated partial thromboplastin time increase of roughly 7 s was observed at the highest dose (2 mg/kg), as were small and clinically insignificant increases in serum C3a levels. T/B cell immunophenotyping, recall antigen skin testing, and serum immunoglobulin levels revealed no significant immunosuppressive effects. CONCLUSION: This study shows that 13 ISIS 2302 infusions over 4 weeks are well tolerated in patients with active RA. Although significant efficacy was not evident at the primary endpoint (1 month), the study lacked sufficient power to draw any formal conclusions. We tested a 4-fold drug concentration range, which led to a lower area under the curve range than was therapeutic in a subsequent Crohn's disease trial. Any further evaluation of this well tolerated ICAM-1 antisense agent should therefore be conducted at higher dosing.     

Colorectal cancer extracellular acidosis decreases immune cell killing and is partially ameliorated by pH-modulating agents that modify tumor cell cytokine profiles

Tumor cells upregulate myriad proteins that are important for pH regulation, resulting in the acidification of the extracellular tumor microenvironment (TME). Abnormal pH is known to dampen immune function, resulting in a worsened anti-tumor immune response. Understanding how extrinsic alterations in pH modulate the interactions between immune cells and tumors cells will help elucidate opportunities for new therapeutic approaches. We observed that pH impacts the function of immune cells, both natural killer (NK) and T cells, which is relevant in the context of a highly acidic TME. Decreased NK and T cell activity was correlated with decreasing pH in a co-culture immune cell-mediated tumor cell-killing assay. The addition of pH-modulating drugs cariporide, lansoprazole, and acetazolamide to the co-culture assay was able to partially mitigate this dampened immune cell function. Treatment of colorectal cancer (CRC) cells with NHE1 inhibitor cariporide increased CRC cell-secreted cytokines involved in immune cell recruitment and activation and decreased cytokines involved in epithelial-mesenchymal transition (EMT). Cariporide treatment also decreased CRC cell shed TRAIL-R2, TRAIL-R3, and PD-L1 which is relevant in the context of immunotherapy. These experiments can help inform future investigations into how the pH of the tumor microenvironment may be extrinsically modulated to improve anti-tumor immune response in solid tumors such as colorectal cancer.     

Ex vivo expansion with stem cell factor and interleukin-11 augments both short-term recovery posttransplant and the ability to serially transplant marrow

The characterization of many cytokines involved in the control of hematopoiesis has led to intense investigation into their potential use in ex vivo culture to expand progenitor numbers. We have established the optimum ex vivo culture conditions that allow substantial amplification of transient engrafting murine stem cells and which, simultaneously, augment the ability to sustain serial bone marrow transplantation (BMT). Short-term incubation of unfractionated BM cells in liquid culture with stem cell factor (SCF) and interleukin-11 (IL- 11) produced a 50-fold amplification of clonogenic multipotential progenitors (CFU-A). Following such ex vivo expansion, substantially fewer cells were required to rescue lethally irradiated mice. When transplanted in cell doses above threshold for engraftment, BM cells expanded ex vivo resulted in significantly more rapid hematopoietic recovery. In a serial transplantation model, unmanipulated BM was only able to consistently sustain secondary BMT recipients, but BM expanded ex vivo has sustained quaternary BMT recipients that remain alive and well more than 140 days after 4th degree BMT. These results show augmentation of both short-term recovery posttransplant and the ability to serially transplant marrow by preincubation in culture with SCF and IL-11.     

Systemic hematologic effects of PEG-rHuMGDF-induced megakaryocyte hyperplasia in mice

PEG-rHuMGDF injected daily in normal mice causes a rapid dose-dependent increase in megakaryocytes and platelets. At the same time that platelet numbers are increased, the mean platelet volume (MPV) and platelet distribution width (PDW) can be either decreased, normal, or increased depending on the dose and time after administration. Thus, PEG-rHuMGDF at a low dose causes decreases in MPV and PDW, MGDF at an intermediate dose causes an initial increase followed by a decrease in MPV and PDW, and PEG-rHuMGDF at higher doses causes an increase in MPV and PDW followed by a gradual normalization of these platelet indices. In addition to the expected thrombocytosis after 7 to 10 days of daily injection of high doses of PEG-rHuMGDF, a transient decrease in peripheral red blood cell numbers and hemoglobin is noted accompanied in the bone marrow by megakaryocytic hyperplasia, myeloid hyperplasia, erythroid and lymphoid hypoplasia, and deposition of a fine network of reticulin fibers. Splenomegaly, an increase in splenic megakaryocytes, and extramedullary hematopoiesis accompany the hematologic changes in the peripheral blood and marrow to complete a spectrum of pathologic features similar to those reported in patients with myelofibrosis and megakaryocyte hyperplasia. However, all the PEG-rHuMGDF-initiated hematopathology including the increase in marrow reticulin is completely and rapidly reversible upon the cessation of administration of PEG-rHuMGDF. Thus, transient hyperplastic proliferation of megakaryocytes does not cause irreversible tissue injury. Furthermore, PEG-rHuMGDF completely ameliorates carboplatin-induced thrombocytopenia at a low-dose that does not cause the hematopathology associated with myelofibrosis.     

一氧化氮对人肝癌细胞SMMC-7721辐射敏感性的增强效应

目的:观察一氧化氮对肿瘤细胞SMMC-7721辐射敏感性的作用效果。方法:实验于2005-06/09在兰州大学生命科学学院和中科院近代物理研究辐射医学实验室进行。处于对数生长期的肝癌细胞SMMC-7721,在用X射线照射前4h,换入含有0.1mmol/L硝普钠(一氧化氮的前体)的培养液,与对照组(不加硝普钠)一起,在200cGy/min的剂量率下,分别照射0,1,2,4,6,8Gy,换为正常培养液培养。用集落形成法计算细胞的存活率,用吖啶橙/溴乙啶双染法检测细胞的死亡情况,用流式细胞仪检测细胞周期。结果:①存活曲线细胞存活率随照射剂量增加而减少,硝普钠组细胞的克隆形成率低于对照组(2Gy时,P<0.01)。②细胞死亡百分率(坏死细胞与凋亡细胞总数/总细胞数):与照射剂量呈正相关,硝普钠组高于对照组(P<0.05)。对照组从(9.95±3.53)%(0Gy)逐渐升至(58.74±3.46)%(6Gy),而硝普钠组则从(18.53±12.02)%(0Gy)迅速升至(61.57±9.53)%(2Gy)。③细胞周期检测结果:对照组细胞经过X射线照射后,出现了G2/M期阻滞[从0Gy时(12.50±5.76)%逐渐增加到8Gy(40.36±2.74)%],而硝普钠组细胞在低剂量时主要表现为G0/G1期阻滞[0Gy:(16.06±7.19)%;2Gy:(17.93±0.92)%],而G2/M期阻滞仅在高剂量时明显[8Gy时为(50.10±3.93)%,P<0.05]。结论:经硝普钠产生的一氧化氮,通过与X射线协同作用,减少了肝癌细胞SMMC-7721的细胞存活率,促进细胞死亡,阻止细胞被阻滞至G2/M期,是一种有效的辐射增敏剂。     

针刺足三里和悬钟治疗缺血性脑卒中

目的:观察针刺足三里、悬钟2穴对缺血性脑卒中脑血管功能的影响,分析其可能的作用机制,并对临床疗效做出评价。方法:选择2004-11/2006-05湖北中医药高等专科学校附属古城医院针灸科、荆州市第五人民医院中医康复科、荆州市第三人民医院中医科3单位缺血性脑卒中患者合适病例160例,采用查随机数字表的方法,将其随机分为对照组和针刺组,各80例。对照组采用现代医学常规干预方法进行治疗:卧床,保持呼吸道通畅,预防感染,控制颅内压、血压,维持水电解质平衡。针刺组在此基础上加针刺足三里、悬钟2穴,采用慢速捻转进针法针刺,留针20～30min,每隔5min行针1次。1次/d。两组患者治疗30d。并以经颅多普勒检测观察缺血性脑卒中患者治疗前后脑血管舒缩反应能力、脑血流自动调节功能、大脑半球侧枝循环代偿功能的变化,同时以治疗前后神经功能缺损程度为指标评价其临床疗效。结果:160例病例全部进入结果分析。①针刺组与治疗前相比,脑血管舒缩反应能力明显加强,差异有显著性意义(t=2.97,P<0.05),且优于对照组(t=2.45,P<0.05)。②针刺组与治疗前相比,脑血流自动调节能力明显改善,差异有非常显著性意义(t=8.01,P<0.01),且优于对照组(t=7.67,P<0.05)。③针刺组与治疗前相比,大脑半球侧枝循环代偿功能得到加强,差异有显著性意义(t=3.15,P<0.05),且优于对照组(t=5.16,P<0.05)。④针刺组与治疗前相比,神经功能缺损积分明显降低,差异有非常显著性意义(t=4.83,P<0.01),且优于对照组(t=5.43,P<0.05)。结论:针刺足三里、悬钟2穴对缺血性脑卒中患者脑血管舒缩反应能力、脑血流自动调节功能、大脑半球侧枝循环代偿功能有明显改善作用,并能促进神经功能的恢复。     

Predictors of success of intervention programs for persons with fibromyalgia

OBJECTIVE: To determine which sociodemographic, psychological, and behavioral characteristics of persons with fibromyalgia (FM) will predict a positive response to treatment; and to determine if subjects classified according to the Multidimensional Pain Inventory (MPI) responded differently to the interventions. METHODS: One hundred twenty-eight women with FM underwent baseline testing and were randomized into one of 3 intervention groups or a control group. After the 12 week program, the subjects were reexamined on the same pretest measures. Stepwise regression analyses were conducted to determine the variables that could significantly predict the change in the dependent variables. A reliability of change index was calculated to determine the proportion of responders and nonresponders. RESULTS: The stepwise regression revealed significant predictor variables for change for all dependent variables except the Fibromyalgia Impact Questionnaire; however, the percentage of the variance in the change scores explained by the independent variables ranged from 4 to 15%. Results from the reliability of change index indicated that no MPI subgroup responded more than another group on any measure. CONCLUSION: Select sociodemographic and psychosocial variables and type of intervention were not strong predictors of improvement in a variety of measures after a treatment program. The low percentage of explained variance may be due to the heterogeneity of FM. Additionally, the low percentage of responders suggests that current forms of treatment are not effective for a large portion of the FM population.     

Simultaneous measurement of gallbladder emptying with cholescintigraphy and US during infusion of physiologic doses of cholecystokinin: a comparison

Both ultrasonography (US) and cholescintigraphy are used to study gallbladder dynamics. The present study was undertaken to determine whether the two methods provide the same or different information relating to gallbladder emptying. Emptying was simultaneously studied with both methods during infusion of graded physiologic doses of cholecystokinin (CCK) in six healthy subjects. Infusion of stepwise increasing doses of CCK, ranging from 0.03 to 0.5 Ivy dog units per kilogram of body weight per hour (IDU/kg.h), induced significant dose-related increases in plasma CCK, decreases in gallbladder volume assessed with US, and gallbladder emptying assessed with cholescintigraphy. The threshold dose for inducing significant gallbladder emptying was 0.13 IDU/kg.h, as determined with both techniques, indicating similar detection limits. There was a highly significant correlation between decreases in gallbladder volume and decreases in radioactive counts over the gallbladder region, with a tendency toward greater gallbladder responses at sonography during the early phase of gallbladder contraction and toward greater responses at cholescintigraphy during the later phase of gallbladder contraction. It is concluded that these methods can be used interchangeably for the quantitation of gallbladder emptying.     

Spondyloarthritis Research Consortium of Canada magnetic resonance imaging index for assessment of spinal inflammation in ankylosing spondylitis

OBJECTIVE: To develop a feasible magnetic resonance imaging (MRI)-based scoring system for spinal inflammation in patients with spondylarthropathy that requires minimal scan time, does not require contrast enhancement, evaluates the extent of lesions in 3 dimensional planes, and limits the number of vertebral levels that are scored because MRI demonstrates characteristic inflammatory lesions in the spine of patients with ankylosing spondylitis (AS) prior to the development of typical features on plain radiographic. METHODS: Our scoring method was based entirely on the assessment of increased signal denoting bone marrow edema on T2-weighted STIR sequences. Blinded MRI films were assessed in random order at 2 sites by 3 blinded readers at each of the 2 sites (the Universities of Alberta and Toronto). Intra- and interreader reliability was assessed by intraclass correlation coefficient. The 24-week response of patients with AS randomized to infliximab:placebo (8:3) was assessed by effect size and standardized response mean. RESULTS: An initial analysis of all discovertebral units (DVUs) in the spine of 11 patients demonstrated a mean of 3.2 (95% confidence interval 3.2, 5.2) affected units, while limiting the scoring to a maximum of 6 units captured most of the affected units. We scanned 11 patients with AS with clinically active disease and 20 additional patients randomized to a 24-week trial of either infliximab or placebo. Intraobserver reproducibility for the 6-DVU STIR score ranged from 0.93 to 0.98 (P < 0.0001). Interobserver reproducibility of scores by readers from both sites was 0.79 (P < 0.0001) for status score and 0.82 (P < 0.0001) for change score. Analysis of pretreatment and posttreatment scores for all 20 patients randomized to infliximab/placebo showed a large degree of responsiveness (standardized response mean = 0.87). Reproducibility and responsiveness were only slightly improved by using contrast enhancement with gadolinium diethylenetriaminepentaacetic acid. CONCLUSION: The Spondyloarthritis Research Consortium of Canada MRI index is a feasible, reproducible, and responsive index for measuring spinal inflammation in AS.