Adsorptive Granulocyte/Monocyte Apheresis for the Maintenance of Remission in Patients with Ulcerative Colitis: A Prospective Randomized, Double Blind, Sham-Controlled Clinical Trial

Background/Aims

Weekly granulocyte/monocyte adsorption (GMA) to deplete elevated and activated leucocytes should serve as a non-pharmacological intervention to induce remission in patients with ulcerative colitis (UC). This trial assessed the efficacy of monthly GMA as a maintenance therapy to suppress UC relapse.

Methods

Thirty-three corticosteroid refractory patients with active UC received 10 weekly GMA sessions as a remission induction therapy. They were then randomized to receive one GMA session every 4 weeks (True, n=11), extracorporeal circulation without the GMA column every 4 weeks (Sham, n=11), or no additional intervention (Control, n=11). The primary endpoint was the rate of avoiding relapse (AR) over 48 weeks.

Results

At week 48, the AR rates in the True, Sham, and Control groups were 40.0%, 9.1%, and 18.2%, respectively. All patients were steroid-free, but no statistically significant difference was seen among the three arms. However, in patients who could taper their prednisolone dose to <20 mg/day during the remission induction therapy, the AR in the True group was better than in the Sham (p<0.03) or Control (p<0.05) groups.

Conclusions

Monthly GMA may potentially prevent UC relapse in patients who have achieved remission through weekly GMA, especially in patients on <20 mg/day PSL at the start of the maintenance therapy.     

Successful Treatment of Three Cases of Generalized Pustular Psoriasis With Granulocyte and Monocyte Adsorption Apheresis

Generalized pustular psoriasis (GPP) is a rare form of psoriasis characterized by the presence of variable numbers of sterile pustules appearing in erythematous and scaly lesions, which are associated with moderate to severe constitutional symptoms. It can be life‐threatening especially in the elderly; therefore, medical care must be performed in rapid succession of treatment especially in refractory cases. We have performed granulocyte and monocyte adsorption apheresis (GCAP) on three GPP cases associated with several systemic and laboratory findings. As a result, the edema, erythema and numbers of sterile pustules on the skin lesions were reduced dramatically in all three patients after the first sessions of GCAP therapy. The sizes of the psoriatic lesions were reduced in all three patients following a weekly GCAP treatment for 5 consecutive weeks. Psoriasis area and severity index on discharge had improved in all three patients. No serious adverse effects were observed for up to at least 8 months after treatment. We therefore considered GCAP as one effective alternative to currently existing therapies, especially for recalcitrant cases of GPP.     

重组人粒细胞集落刺激因子对兔颈动脉粥样硬化球囊损伤后再内皮化和内膜增生的影响

Objeelive To investigate the effect of rhG-CSF on mobilizing bone marrow-MSCs, reendothelialization and intima hyperplasia in carotid artery of rabbits post balloon catheter injury, nethods Rabbits were treated with rhG-CSF (25 μkg, twice daily, i. p, n =35) or saline (n =32) for 5 days, then, carotid arteries of rabbits were injured by balloon catheter. The number of peripheral MSCs was detected with FACS. The morphology of injuried artery was examined with hematoxylin and eosin stain, PCNA was determined with immunohistochemistry. Results (1) Number of peripheral MSCs was similar at baseline and significantly increased at 24 hours and peaked at 7 days and remained increased till 14 days post rhG-CSF. (2) Significant endothelial cell deletion was evidenced in the control group, while scatter endothelial cells was observed in the rhG-CSF group at 1 week post injury. Two weeks after injury, new endothelial area was significantly higher in rhG-CSF group compared to control group. At 4 weeks post injury, endothelial connection was evidenced and regularly displayed in rhG-CSF treated group. (3) PCNA-positive cells in the tunica intima were significantly lower in rhG-CSF treated rabbits at 7, 14 and 28 days compared that in control rabbits (all P ＜ 0.01). Conclusion rhG-CSF could mobilize the bone marrow-MSCs and promote re-endothelialization and attenuate intima hyperplasia post balloon catheter injury in carotid arteries of rabbits.     

Antiinflammatory steroids inhibit granulocyte/macrophage colony-stimulating factor production by human lung tissue

Granulocyte/macrophage colony-stimulating factor (GM-CSF) is an important hematopoietic growth factor which has been shown to induce proliferation and activation of inflammatory cells, and may play a role in allergic diseases and experimental allergic reactions. Since little is known about the involvement of cytokines in allergic inflammation in the lung, we investigated whether human lung fragments produce GM-CSF in vitro. The present studies demonstrate that human lung fragments produce GM-CSF in vitro and that glucocorticoids are potent inhibitors of this cytokine production. Human lung was cut into fragments, rinsed, and cultured in 60-mm tissue culture plates containing 50 mg of tissue in RPMI 1640 with antibiotics in the presence or absence of a variety of steroids for 18 h. Lung fragments were rinsed and then incubated for an additional 4 h. Supernatants were harvested and analyzed for GM-CSF activity using the GM-CSF/interleukin (IL)-3 responsive M-07e human luekemic cell line. Steroids alone had no effect on M-07e proliferation. Human lung fragments produced 32.1 ± 11.8 ng of GM-CSF equivalents per gram wet weight of tissue during the 4 h incubation (mean ± S.E.M., n = 5, range 9.2–74.2). While specific antisera against human GM-CSF neutralized 96.8 ± 2.8% (n = 5) of the activity, anti-IL-3 antibody had no effect, suggesting most or all of this activity was GM-CSF. Treatment of lung fragments in vitro for 18 h with hydrocortisone (HC) inhibited the production of GM-CSF dose-dependently. Maximal inhibition of GM-CSF production was 72.8 ± 4.0% at a concentration of 10^–6 m hydrocortisone (n = 5), and the molar concentration of HC that inhibited of GM-CSF production by lung tissue by 50% (IC₅₀) was approximately 4.5 × 10^–7 m. Kinetic studies revealed that a 6 h preincubation with the drug was required for 50% inhibition of GM-CSF production. HC and other glucocorticoids, at a concentration of 0.1 µm, demonstrated significant inhibition of GM-CSF release. Based on the rank order of potency of several glucocorticoids, and the fact that nonglucocorticoid steroids including testosterone and -estradiol (0.1 µm) had no effect, we suggest that this is a specific receptor-mediated effect. We conclude that human lung produces GM-CSF in vitro and that antiinflammatory steroids are potent and effective inhibitors of the production of this cytokine. This may contribute to the therapeutic efficacy of these drugs in pulmonary diseases. Offprint requests to: R. P. Schleimer     

粒细胞集落刺激因子动员骨髓内皮祖细胞促进损伤血管内皮修复

目的探讨大鼠颈动脉球囊损伤后粒细胞集落刺激因子动员对损伤血管内膜增殖和内皮修复的影响及其相关机制。方法36只雄性Wistar大鼠随机分为假手术组(n=12)、损伤 细胞因子组(n=12)和损伤 安慰剂组(n=12)。损伤 细胞因子组在球囊损伤后即刻腹腔注射粒细胞集落刺激因子[100μg/(kg.d)]直至术后2周,损伤 安慰剂组给予等量生理盐水注射,术后2周取血管段HE染色计算内膜/中膜面积比,免疫组织化学观察并计算增殖细胞核抗原阳性细胞比例,通过伊文氏蓝染色计算内皮修复率,血管匀浆检测一氧化氮释放量,用流式细胞术检测外周血CD34 VEGFR-2 双阳性细胞比例。结果与假手术组比较,损伤 安慰剂组血管内膜增殖明显,有较多增殖细胞核抗原阳性细胞,内皮修复不全,血管匀浆中一氧化氮释放量减少;与损伤 安慰剂组相比,损伤 细胞因子组内膜/中膜面积比和增殖细胞核抗原阳性细胞比例降低(分别为1.04±0.05比1.67±0.07和21.3%±5.1%比34.4%±6.1%,P<0.05),内皮修复率和一氧化氮释放量增加(分别为70.1%±4.6%比53.2%±3.8%和71.3±12.9μmol/L比56.7±10.8μmol/L,P<0.05);损伤 细胞因子组外周血CD34 VEGFR-2 双阳性细胞比例较损伤 安慰剂组明显上升(0.954%±0.076%比0.379%±0.052%,P<0.05)。结论粒细胞集落刺激因子明显增强血管损伤后内皮修复并减轻新生内膜增生,其机制可能涉及骨髓动员增加外周血内皮祖细胞比例。     

乙型肝炎患者血清粒细胞-巨噬细胞集落刺激因子检测的临床意义

目的：探讨血清粒细胞—巨噬细胞集落刺激因子(GM-CSF)对乙型肝炎患者的临床意义。方法：采用放射免疫法检测各型乙型肝炎和肝炎后肝硬化患者与健康受试者血清GM-CSF浓度，常规检测外用血白细胞(WBC)计数，聚合酶链反应(PCR)法检测外用血乙肝病毒基因组(HBV DNA)含量。结果：与正常人相比，慢性肝炎、肝炎后肝硬化患者血清GM-CSF水平、WBC水平下降(P＜0．05)；重型肝炎患者血清GM-CSF水平显著下降(P＜0．01)，WBC水平显著升高(P＜0．01)。HBV DNA含量在急性肝炎、肝炎后肝硬化、慢性重型肝炎、慢性肝炎中依次升高。血清GM-CSF浓度基本与WBC的计数量变化一致。结论：乙型肝炎患者血清GM-CSF水平对WBC水平的影响强于乙肝病毒，且有抗病毒作用。     

rhG-CSF和rhSCF联合动员骨髓干细胞治疗大鼠急性心肌梗死

目的:观察联合应用重组人粒细胞集落刺激因子(rhG-CSF)和干细胞因子(rhSCF)对大鼠急性心肌梗死(AMI)心室功能、梗死面积及心室重塑的影响。方法:采用异丙基肾上腺素法将32只雄性SD大鼠制为AMI模型,3h后按皮下注射药物类别随机分为4组,A组为rhG-CSF与rhSCF合用组,B组为rhG-CSF组,C组为 rhSCF组,D组为对照组。每组8只,各组随机分为2个亚组,分别于14d和28d观察心室功能后处死,取出心脏,称重比较心室重塑差异,HE染色,体视学方法观察梗死面积大小及药物治疗作用。结果:(1)给药14d及28d后,A组心功能优于B、C、 D组(P<0．05);左心室质量／体质量(g/G)小于C、D组(P<0．05);梗死面积／左心室总面积(s／S)小于B、C、D组(P<0．05)。(2)A组28 d亚组较14 d亚组心功能提高, 梗死面积减小(P<0．05);心室重塑改善不明显(P>0．05)。B、C、D组的亚组间各指标无明显差异。结论:rhG-CSF和rhSCF合用对AMI大鼠缺血损伤心肌的保护和再生作用优于单用,可明显改善AMI大鼠的心室功能。     

Developmental regulation of granulocyte-macrophage colony-stimulating factor production during human monocyte-to-macrophage maturation

Summary Cells of the macrophage lineage are a major source of various cytokines and hematopoietic growth factors. With regard to the growth factors acting on cells of their own lineage, macrophage colony-stimulating factor (M-CSF) has been proven to be secreted by monocytes (MO) and macrophages (MAC), whereas the production of granulocyte-macrophage colony-stimulating factor (GM-CSF) by human MO/MAC is under debate. Here we report that in elutriation-purified MO, as well as in MAC derived from cultured MO, GM-CSF m-RNA was regularly induced by LPS. In MO the GM-CSF message was still detectable 18h after stimulation under serum-free conditions, but in contrast was already lost at this time point in MAC. Secreted GM-CSF protein was detected in the culture medium using a sandwich ELISA. Furthermore, a factor-dependent cell line (M-07) was used for a biological assay. Here, a neutralizing anti GM-CSF antibody specifically blocked the proliferation-inducing activity of MO/MAC supernatants. Whereas only small amounts of GM-CSF were detected in MO, its secretion increased severalfold upon MO-to-MAC differentation in vitro. A similar increase upon in vitro maturation of MO was observed for the production of granulocyte colony-stimulating factor. The highest amounts of GM-CSF (up to 2.8 ng/10⁶ cells) were produced by MAC that had been derived from MO cultured under serum-free conditions in the presence of 0.5 mg/ml albumin as the only medium supplement.This work was supported by theDeutsche Forschungsgemeinschaft (AN 111).     

The determinants of donor safety and product quality in optimization of apheresis granulocyte harvest: An experience from a tertiary care oncology centre in India

Allogeneic granulocyte transfusions play a substantial role in treatment of lifethreatening neutropenia-associated infections in patients undergoing intensive chemotherapy and hematopoietic stem cell transplant. Granulocyte harvest by apheresis is considered a safe and effective method to obtain adequate therapeutic granulocyte dosage for clinical effectiveness. This study described the experiences associated with apheresis granulocyte harvest procedures in our tertiary care haemato-oncology centre. We have analysed the incidence of adverse events (AEs) with associated potential risk factors contributing to donor safety and improvement in product quality. Retrospective data of 131 healthy allogeneic donors who underwent granulocyte harvest from May 2016 to July 2020 were analyzed. AEs were observed in overall 29 procedures (22.13%), including 14.50% citrate reactions, 7.6% venous access-related reactions, and 1.52% vasovagal reactions. Older age (p = 0.012) and higher body mass index (p = 0.015) in donors were significant variables found associated with a higher incidence of AEs. There was no significant impact of AEs on granulocyte product yield (p = 0.41) with a median collection yield of 1.73 × 10 10 cells/ unit. In multivariate analysis, post-mobilization parameters like total leukocyte counts (p = 0.036), absolute neutrophil counts (p = 0.042), and platelet counts (p = 0.006) showed a positive correlation with higher product yield. All the donors successfully donated and tolerated granulocyte colony stimulating factor plus dexamethasone mobilization and granulocyte apheresis harvest without any serious AEs. Our study shows that optimal technical and procedural modifications during apheresis granulocyte harvest procedures can overcome the associated potential risks by providing donor safety and improving product quality.