异基因造血干细胞移植后合并慢性移植物抗宿主病患者T细胞免疫状态的研究

目的 研究异基因造血干细胞移植后合并慢性移植物抗宿主病（cGVHD）时的T细胞免疫状态。方法 应用RT-PCR扩增14例白血病异基因造血干细胞移植后合并cGVHD患者及5名正常供者的外周血T细胞受体（TCR）BV24个家族的基因序列，并通过基因扫描的方法判断TCRBV家族的克隆表达情况、CDR3克隆性质，并计算BV家族的利用率。结果 在移植后6～19个月，所有患者TCRBV家族的利用仍处于不均一状态，检测到部分BV家族缺失，部分家族出现寡克隆、单克隆T细胞增殖。在24个BV家族的表达为6／24～15／24，并且均出现1～5个单克隆家族表达。单克隆及双克隆表达分布在不同的BV家族，未发现共用的BV家族。结论 异基因造血干细胞移植后合并cGVHD患者的T细胞免疫存在缺陷。在不同的TCRBV家族出现与cGVHD相关的克隆增殖的T细胞克隆。     

Evolving Therapeutic Options for Chronic Graft-versus-Host Disease

Despite improvements in prevention and treatment of acute graft-versus-host disease (GVHD), chronic GVHD (cGVHD) remains a significant contributor to morbidity and mortality of allogeneic transplant patients. Chronic GVHD remains a leading cause of late complications posttransplant and is impacted by donor-, patient-, and transplant-related (hematopoietic cell transplant [HCT]) factors. Advances in the biological understanding of cGVHD have provided opportunities to improve clinical interventions for prevention and treatment. Expansion of posttransplantation cyclophosphamide beyond haploidentical HCTs has transformed alternative donor, matched, and mismatch GVHD outcomes and is currently being investigated in two upcoming clinical trials network prophylaxis studies. Although corticosteroids remain the cornerstone therapy, several clinical trials are prospectively investigating the utility of using novel agents in combination with corticosteroids as upfront therapy to mitigate prolonged steroid exposure. Several treatment options for patients with steroid-refractory cGVHD are currently being investigated, and advances have resulted in ibrutinib becoming the first cGVHD agent approved by the U.S. Food and Drug Administration. We review recent advances in understanding of cGVHD pathophysiology and new approaches for the prevention and treatment of cGVHD.     

Progress in the management of chronic GVHD insights into novel therapies to treat and manage GVHD lasting longer than 12 Months

Chronic graft-versus-host disease (cGVHD) is a major cause of poor outcomes after hematopoietic stem cell transplantation (HCT). An increased understanding of the pathobiology of cGVHD has led to the development of novel therapies. This review summarized the underlying pathogenesis of cGVHD and has provided considerations for integrating new agents into practice.     

慢性移植物抗宿主病狼疮肾炎模型的建立和淋巴细胞功能的变化

目的:探讨慢性移植物抗宿主病(cGVHI))狼疮小鼠肾炎模型方法的建立及其T、B淋巴细胞功能的变化,为进一步探讨狼疮肾炎发病机制奠定基础.方法:(1)将亲代BALB/C小鼠淋巴细胞经静脉途径输入(BALB/C×C57 BL/6)F1代小鼠体内诱导狼疮小鼠模型;(2)测定血清中抗核抗体(ANA)及肾脏病理以检测模型是否成功;(3)用ConA刺激脾和胸腺T细胞,采用MTT法检测成熟与未成熟T细胞对于丝裂原增殖转化能力.结果:(1)模型组血清中ANA均为阳性,而对照组均为阴性;(2)模型小鼠的肾脏有免疫复合物沉积,肾脏有明显的病理损害;(3)模型组成熟T细胞(脾脏细胞)和未成熟T细胞(胸腺细胞)经ConA刺激后增殖能力较对照组减弱.结论:运用cGVHD方法诱导的狼疮肾炎模型,小鼠发病迅速,病理及临床表现与人狼疮肾炎相似,并且存在T、B淋巴细胞功能异常,为应用此模型研究狼疮肾炎提供了理论依据.     

Quantitative ultrasound detects bone impairment after bone marrow transplantation in children and adolescents affected by hematological diseases

PURPOSE: Bone marrow transplantation (BMT) recipients are at risk of bone mass impairment and skeletal morbidity. We investigated bone status with quantitative ultrasound (QUS) technique in children and adolescents with hematological diseases before and after BMT. METHODS: Phalangeal QUS measures for amplitude-dependent speed of sound (AD-SoS) and bone transmission time (BTT) were obtained in 144 hematological patients (81M, 63F; 11.6+/-5.2 years); forty two were evaluated before BMT and 102 after allogeneic or autologous BMT. Bone parameters were expressed as Z-scores based on age-sex-matched normal controls. RESULTS: Mean BTT Z-score was reduced in subjects after BMT compared to patients before BMT (M, -0.35+/-1.04 vs. 0.70+/-1.11, P<0.001; F, -0.60+/-1.23 vs. 0.23+/-1.17, P<0.05). Females and males with hormone deficiencies showed reduced BTT Z-scores when compared with subjects without hormone defects (M, -0.52+/-1.0 vs. 0.05+/-1.17, P<0.05; F, -0.50+/-1.27 vs. -0.19+/-1.26; P=0.06). AD-SoS and BTT Z-scores were reduced in 15 subjects with fractures and/or avascular osteonecrosis compared to patients without bone events (-1.52+/-1.7 vs. -0.41+/-1.32 and -0.85+/-1.19 vs. -0.10+/-1.18; both Ps<0.05). Bone event cumulative incidence was 4 times greater in subjects who suffered from chronic GVHD. CONCLUSIONS: Assessment of phalangeal QUS in young BMT survivors points towards impairment of bone status and endocrine dysfunction and chronic GVHD as risk factors of adverse bone events.     

含对硝基苯丙氨酸的B淋巴细胞刺激因子疫苗对狼疮肾炎模型小鼠的保护作用

考察含对硝基苯丙氨酸的B淋巴细胞刺激因子(BAFF)治疗BAFF过表达的自身免疫性疾病的效果。利用本研究前期构建的工程菌,表达纯化了B淋巴细胞刺激因子的可溶型突变体(smBAFF)以及在其65位定点引入了对硝基苯丙氨酸的改构体(pNO₂Phe⁶⁵smBAFF)。通过考察pNO₂Phe⁶⁵smBAFF体外促小鼠淋巴细胞增殖活性、免疫原性以及所诱导的抗血清对天然BAFF活性的抑制作用,评价了其用于治疗BAFF过表达的自身免疫性疾病的可行性;同时采用了cGVHD(graft-versus-host disease)诱导的狼疮肾炎小鼠模型评价了pNO₂Phe⁶⁵smBAFF的药理活性。结果表明:定点引入了对硝基苯丙氨酸的pNO₂Phe⁶⁵smBAFF不具有促进小鼠淋巴细胞增殖的能力;由于对硝基苯丙氨酸的引入显著增强了蛋白的免疫原性,诱导机体产生了可以抑制天然BAFF活性的交叉抗体;在cGVHD诱导的狼疮肾炎小鼠模型中,pNO₂Phe⁶⁵smBAFF可显著减轻疾病症状。定点引入了对硝基苯丙氨酸的pNO₂Phe⁶⁵smBAFF可以作为治疗BAFF过表达的自身免疫性疾病的候选分子。     

一种实用性狼疮样肾炎动物模型--鼠慢性移植物抗宿主病的制备

目的建立一种狼疮样肾炎动物模型，为人类狼疮性肾炎（ＬＮ）实验和临床研究提供基础。方法取雌性亲代ＤＢＡ／２的淋巴细胞分４次经静脉注射雌性子代Ｆ１代杂交鼠（Ｃ５７ＢＬ６× ＤＢＡ２）。注射时间为０，３，７，１０ ｄ。每次注射淋巴细胞数为５０× １０６。结果尿白蛋白于第４次注射后２周开始升高，第１２周达高峰；血抗ｄｓ－ＤＮＡ水平于第４次注射后２周已明显升高，至第４周达高峰，各时点与对照组差异明显。普通光镜（ＰＡＳ、ＰＡＳＭ）示肾小球基底膜增厚，系膜轻度增生，小管间质炎症细胞浸润；免疫荧光示ＩｇＧ沿基底膜呈线状或颗粒状沉积，部分小管基底膜亦可见其沉积；电镜显示膜性肾炎改变，基底膜增宽，内有电子致密物沉积，上皮细胞足交融合。结论本实验所建立狼疮样肾炎动物模型，主要为膜性肾炎。     

Severe COVID‐19 in a patient with chronic graft‐versus‐host disease after hematopoietic stem cell transplant successfully treated with ruxolitinib

Graft‐versus‐host disease (GVHD) is a common complication of hematopoietic stem cell transplant, which is known to be mediated by cytotoxic T‐cell effectors and dysregulated inflammatory cytokines. Similarly, the lung injury observed in severe COVID‐19 cases appears to be related to a massive production of pro‐inflammatory cytokines. The selective JAK1/2 inhibitor ruxolitinib has shown promising results in the context of GVHD, and different trials are currently underway in patients with severe COVID‐19; nevertheless, no clinical observation of safety or efficacy of treatment with ruxolitinib in this context has been published yet. We describe a first case of severe COVID‐19 developed after hematopoietic stem cell transplantation in a patient with a concomitant chronic GVHD (cGVHD), in which a treatment with ruxolitinib was administered with good tolerance and positive outcome.     

The oral manifestations of chronic graft-versus-host disease (cGVHD) in paediatric allogeneic bone marrow transplant recipients

The oral manifestations of chronic graft-versus-host disease (cGVHD) in eight allogeneic bone marrow transplant (BMT) paediatric recipients were studied clinically, and lip biopsies were performed in seven of them. A prominent lichenoid reaction was observed in four patients, two with accompanying ulceration. Superficial mucoceles were present in three children. Clinically obvious xerostomia was seen in seven patients. Lip biopsies were positive and correlated with the clinical manifestations. Both clinical and histological findings confirmed the diagnosis of cGVHD. In three additional children, with systemic manifestations indicating cGVHD, the oral mucosa was clinically and histologically normal, and the systemic manifestations were, thus, attributed to drug reactions. The above findings indicate the high value of oral examination in diagnosing or confirming paediatric cGVHD. Superficial mucoceles, reported for the first time in paediatric recipients, seem to be important in the early diagnosis of cGVHD.     

Evaluation of prognostic factors among patients with chronic graft-versus-host disease

Background

Chronic graft-versus-host disease (cGVHD) is a major complication after allogeneic stem cell transplantation with an adverse effect on both mortality and morbidity. In 2005, the National Institute of Health proposed new criteria for diagnosis and classification of chronic graft-versus-host disease for clinical trials. New sub-categories were recognized such as late onset acute graft-versus-host disease and overlap syndrome.

Design and Methods

We evaluated the prognostic impact of the new sub-categories as well as the clinical scoring system proposed by the National Institute of Health in a retrospective, multicenter study of 820 patients undergoing allogeneic stem cell transplantation between 2000 and 2006 at 3 different institutions. Patients were retrospectively categorized according to the National Institute of Health criteria from patients’ medical histories.

Results

As far as the new sub-categories are concerned, in univariate analysis diagnosis of overlap syndrome adversely affected the outcome. Also, the number of organs involved for a cut-off value of 4 significantly influenced both cGVHD related mortality and survival. In multivariate analysis, in addition to NIH score, platelet count and performance score at the time of cGVHD diagnosis, plus gut involvement, significantly influenced outcome. These 3 variables allowed us to develop a simple score system which identifies 4 subgroups of patients with 84%, 64%, 43% and 0% overall survival at five years after cGVHD diagnosis (score 0: HR=15.96 (95% CI: 6.85–37.17), P<0.001; score 1: HR=5.47 (95% CI: 2.6–11.5), P<0.001; score 2: HR=2.8 (95% CI: 1.32–5.93), P=0.007).

Conclusions

In summary, we have identified a powerful and simple tool to discriminate different subgroups of patients in terms of chronic graft-versus-host disease related mortality and survival.