CD4+调节性T细胞在再生障碍性贫血免疫发病中的作用

目的探讨CD4 调节性T细胞(Tregul atory,Treg)在再生障碍性贫血(aplasticanemia,AA)免疫发病机制中的作用。方法对2005年7月至2006年3月苏州大学附属第一医院23例发病期AA患者、10例恢复期AA患者及15名正常对照者测定骨髓中CD4 CD25 Treg、CD4 CTLA-4 Treg、CD4 PD-1 Treg、CD3 CD8-IL-10 Treg、CD3 CD8-TGF-β1 Treg、CD3 CD8-IL-4 Treg变化,分析其与免疫启动因素CD28及免疫效应因素干扰素-γ(IFN-γ)的关系。结果AA发病期、恢复期及正常对照组CD4 CD25 Treg数比较差异无显著性意义;AA发病期CD4 CTLA-4 Treg表达较对照组明显下降,PD-1与对照组比较差异无显著性意义;AA恢复期CD4 CT-LA-4 Treg及PD-1表达均较发病期明显升高,与对照组相当;AA恢复期患者IFN-γ较发病期明显下降(P=0.021),与正常对照组相当(P=0.402),IL-4、TGF-β及IL-10与AA发病组及对照组比较差异均无显著性意义;AA发病期患者骨髓CD3 CD4 T淋巴细胞膜表面CD28表达较正常对照组明显增加;AA恢复期患者的CD28表达率较发病期显著降低,亦低于正常对照组(P=0.048)。AA发病期CD4 CTLA-4 Treg明显下降,其余Treg无显著变化;AA恢复期CD4 CTLA-4 Treg、CD4 PD-1 Treg升高显著。CD28/CTLA-4、CD28/PD-1发病期与对照组比,均显著升高;AA恢复期为明显低于发病期,与对照组相当。IFN-γ /IL-4 、IFN-γ /TGF-β 及IFN-γ /IL-10 发病期较对照组均显著升高;AA恢复期明显低于发病期,而与对照组相当。结论在免疫应答起始阶段或效应阶段,AA正性调控共刺激因子表达增加,而负性调控共刺激因子表达减少或无变化,使免疫平衡向持续增强偏移,调节性因素增高利于造血恢复;对于增强的免疫应答,AA的CD4 Treg呈下降趋势,可能与发病有关。     

再生障碍性贫血患者外周血T淋巴细胞亚群及HLA-DR抗原的变化及临床意义

目的：探讨再生障碍性贫血（再障）患者治疗前、后外周血T淋巴细胞亚群及HI。A-DR抗原的变化及其临床意义。方法：采用流式细胞术检测28例再障患者治疗前、后外周血T淋巴细胞亚群及HLA-DR抗原的变化，并与健康对照组进行比较。结果：①再障患者治疗前CD4^＋、CD4^＋／CD8^＋明显低于健康对照组，CD8^＋、HLA—DR^＋则明显升高；重型再障组与普通型再障组比较，CD8^＋、HLA-DR^＋明显升高，CD3^＋、CD4^＋、CD4^＋／CD8^＋无统计学意义。②再障患者治疗后与治疗前相比，CD4^＋、CD4^＋／CD8^＋明显升高，CD8^＋、HLA-DR^＋明显降低。③再障患者治疗有效者与无效者比较，CD4^＋、CD4^＋／CD8^＋明显升高，CD8^＋、HLA-DR^＋明显降低。结论：再障患者存在T淋巴细胞亚群的失调及T淋巴细胞的异常活化，检测外周血T淋巴细胞亚群及HLA-DR抗原有助于再障患者的病情监测、疗效评价及预后判断。     

重型再生障碍性贫血NSCT移植前后T细胞亚群与造血负调控因子的研究

目的检测行非清髓性造血干细胞移植（NSCT）前后重型再生障碍性贫血（SAA）患者的T细胞亚群和造血负调控因子水平,分析二者异常与疗效的相关性,评价NSCT在重型SAA发病机制中的作用及意义。方法用流式细胞术检测30例重型SAA患者（SAA组）及16例正常者（对照组）的外周血T细胞亚群、IFN-γ及IL-2。结果SAA组76．6％T细胞亚群异常,IFN-γIL-2较对照组明显升高（P〈0．01）;治疗后,T细胞亚群异常及IL-2、IFN-γ升高者比无T细胞亚群异常及IL-2、IFN-γ正常或低下者的临床有效率高;部分患者治疗1a后T细胞亚群及血浆细胞因子恢复正常。结论异常升高及活化的免疫效应细胞对骨髓造血细胞直接或间接损伤,可能是导致SAA的重要机制;NSCT能影响SAA患者T细胞亚群及血浆细胞因子水平。     

抗T细胞单抗治疗再生障碍性贫血12例疗效观察

再生障碍性贫血 (简称再障 )是一种因骨髓造血功能衰竭而引起的以全血细胞减少为特征的造血系统疾病。近年来 ,国内已陆续报道抗 T淋巴细胞单克隆抗体 (简称抗 T细胞单抗 )治疗再障 ,并取得了较好的疗效。我院应用抗 T细胞单抗 ( CD3 ,由山东医科大学附属医院血液病研究室提供 )治疗再障 1 2例 ,临床效果满意 ,现报告如下。1　资料与方法1 .1 　 一般资料本组 1 2例 ,男 8例 ,女 4例 ,年龄 1 7～ 63岁。病程 1个月～ 1 0年。重型再障 型 ( SAA- ) 2例 , 型 ( SAA- ) 1例 ,慢性再障 ( CAA) 8例 ,纯红再障 ( PRCA) 1例。 1 2例患者…     

抗人CD3T淋巴细胞单克隆抗体治疗重型再生障碍性贫血的初步观察

目的　探讨特异性免疫抑制剂鼠抗人CD3 T淋巴细胞单克隆抗体 (CD3 单抗 )对重型再生障碍性贫血 (SAA)的临床疗效。方法　 13例SAA患者中位数年龄 2 2岁 ,既往未经特殊治疗者4例 ,既往治疗无效的 9例中 ,加环孢素≥ 3个月者 4例 ;给药方法 :CD3 单抗 5mg静脉滴注 ,日 1次 ,连用 10d为一疗程。结果　共随访 3～ 15个月 ,骨髓象有 8例明显好转 ;外周血WBC平均升高 1 5 9× 10 9/L ,中性粒细胞升高 0 72× 10 9/L ,Hb升高 4 0 g/L ,血小板升高 4 7× 10 9/L(Р值均 <0 0 1) ;其中 2例基本治愈 ,2例达缓解 ,7例明显进步 ,2例无效 ;T淋巴细胞亚群的变化 :CD4/CD8比值由1 12± 0 34上升至 1 4 2± 0 4 3、HLA DR的表达率由 ( 2 9 2± 13 3) %下降至 ( 15 2± 5 6 ) % (Р值均<0 0 1) ;体外培养患者外周血单个核细胞分泌下列淋巴因子含量的中位数值 (U/ml) :肿瘤坏死因子α、干扰素γ与白细胞介素 2分别由 2 6 7、784和 92降至 15 2、5 70和 5 1(Р值均 <0 0 1)。不良反应 :单抗治疗期间 ,全部病例均有发热 ,4例出现胸闷、呼吸困难 ,但无 1例死亡。结论　与其他常用的免疫抑制剂相比 ,CD3 单抗对SAA的临床疗效更快、有效率可能更高 ,且安全性较好 ;关于该单抗的长期疗效、远期不良作用 ,有待于积累更多     

T细胞大颗粒淋巴细胞白血病伴纯红细胞再生障碍性贫血1例

大颗粒淋巴细胞(LGL)是淋巴细胞的一种亚群。正常情况下,LGL占外周血单个核细胞的10%～15%。根据功能和免疫表型,LGL可分为:①CD3+LGL,代表活化的细胞毒T淋巴细胞;②CD3-LGL,代表自然杀伤细胞。而大颗粒淋巴细胞白血病(large granular lymphocytic leukemia,LGLL)就是起源于这两类细胞的克隆性疾病。临床分为T细胞型LGLL和NK细胞型LGLL,无论T细胞或NK细胞亚型临床上均可表现为惰     

再生障碍性血患者白细胞介素－2与T细胞亚群的检测及意义探讨

本文检测了１７例再障患者外周血淋巴细胞经植物血凝素（ＰＨＡ）刺激后白细胞介素２（ＩＬ－２）的活性水平及Ｔ细胞亚群。结果：再障患者白细胞介素２的活性水平明显高于正常对照（Ｐ＜０．０１），Ｔ＿８明显增高，Ｔ＿４／Ｔ＿８比值倒置，ＩＬ－２活性水平与Ｔ＿８成正相关。表明再障患者Ｔ淋巴细胞及淋巴因子网络失衡，与再障的发病有关。     

Preliminary study correlating CX3CL1/CX3CR1 expression with gastric carcinoma and gastric carcinoma perineural invasion

AIM:To study the relationship between the CX3CL1chemokine,its receptor CX3CR1,and gastric carcinoma/gastric carcinoma perineural invasion(PNI).METHODS:Thirty cases of gastric carcinoma were surgically resected(radical resection or palliative resection)between February 2012 and July 2012.Tumour and tumour-adjacent tissues were evaluated for the presence of CX3CL1(ELISA)and CX3CR1(immunohistochemistry and Western blotting)in an effort to analyse the relationship between CX3CL1/CX3CR1 and gastric carcinoma/gastric carcinoma PNI.RESULTS:Of these 30 cases,14 were PNI-positive(46.7%).No significant differences in CX3CL and CX3CR1 expression in tumour-adjacent tissues were found between the PNI positive and negative groups.Expression levels of CX3CL and CX3CR1 in tumour tissues were significantly higher than those in adjacent tissues(P<0.01),and were significantly higher in tumour tissues from the PNI-positive group compared to the PNI-negative group(P<0.01).CONCLUSION:CX3CL1/CX3CR1 expression may be associated with the occurrence and development of gastric carcinoma as well as gastric carcinoma PNI.     

缺血性脑血管病患者趋化因子受体CX3CR1基因V2491多态性的研究

目的 探讨缺血性脑血管病(ICVD)患者趋化因子受体CX3CR1基因V2491的多态性及其频率.方法 采用聚合酶链反应和限制性片段长度多态性方法检测ICVD患者(脑梗死、腔隙性脑梗死、短暂性脑缺血发作)及健康对照者的CX3CR1基因V2491的多态性及频率.结果 对照组CX3CR1基因V2491只有ⅤⅤ和Ⅵ基因型,ICVD组有ⅤⅤ、Ⅵ和Ⅱ3种基因型;ICVD组Ⅰ等位基因频率明显高于对照组(P<0.01);CX3CR1基因型及Ⅰ等位基因频率在不同类型ICVD患者之间无统计学差异.结论 CX3CR1基因V2491多态性可能与ICVD有关.     

Polymorphisms of Fractalkine receptor CX3CR1 and plasma levels of its ligand CX3CL1 in colorectal cancer patients

Background and aims The chemokine Fractalkine/CX3CL1, which is expressed by epithelial cells within normal colorectal mucosa and in colorectal cancer (CRC), is thought to have a crucial role in colorectal mucosal immunity by recruiting leucocytes via the receptor CX3CR1. The purpose of this study was to investigate two single-nucleotide polymorphisms of the Fractalkine receptor/CX3CR1 gene, V249I and T280M, in CRC to find out whether they occur more often in patients with CRC than in non-CRC individuals. In the search for tumour markers, we also intended to determine whether plasma levels of Fractalkine were correlated with parameters such as Dukes’ stage, tumour localisation, gender and age in CRC patients. Materials and methods Genomic deoxyribonucleic acid from 223 CRC patients and 229 controls was amplified by polymerase chain reaction, and the polymorphisms were detected by the restriction fragment length polymorphism analysis. Fractalkine/CX3CL1 was analysed in plasma from 62 CRC patients and 78 controls using enzyme-linked immunosorbent assay. Results The variant V249I was significantly different in genotype and allelic distribution between CRC patients and control subjects, P = 0.028 and P = 0.048, respectively. We also found that individuals with the I249 allele in homozygote state were less frequent in the CRC group (3.1%) compared with controls (9.2%; P = 0.008). No significant difference was observed regarding Fractalkine/CX3CL1 levels in plasma between patients and the control group. Conclusion Our results suggest that the lack of the allele I249 of the CX3CR1 gene may play a partial or minor role in CRC and that plasma Fractalkine/CX3CL1 does not seem to be a useful tumour marker that reflects the disease outcome of CRC.     

CX3CR1基因多态性与冠心病的相关性

目的 研究Fractalkine受体CX3CR1基因多态性(249V/I和280T/M)与冠心病的相关性.方法 应用聚合酶链反应限制片长多态性方法对139例冠心病患者和90例对照者的CX3CR1基因多态性进行分析,比较CX3CR1基因多态性在两组之间的差异性.结果 等位基因I249在对照组中的分布频率明显高于冠心病组(P<0.05);冠心病组280T/M基因型和等位基因频率分布与对照组比较无显著性差异(P>0.05).结论 Fractalkine受体CX3CR1等位基因I249变异可能与冠心病的发病危险性下降有关,CX3CR1基因多态性与中国南方汉族人群冠心病的发生存在相关性.     

硫化氢对动脉粥样硬化的影响及其与 CX3CR1 的关系

目的在高脂饮食ApoE基因敲除小鼠动脉粥样硬化模型中,探讨硫化氢对动脉粥样硬化斑块的影响及其与斑块内趋化因子受体CX3CR1的关系。方法10周龄、雄性纯合子ApoE基因敲除小鼠予以高脂饮食喂养,在高脂饮食喂养第4、8、12、24周时处死小鼠并留取血浆和主动脉,通过化学比色法和Western Blot技术检测血浆硫化氢水平和主动脉胱硫醚-γ-裂解酶（CSE）表达情况。一部分小鼠在高脂饮食4或12周后开始每天予以硫化氢供体药物NaHS（1mg-kg^-1,i．P．）或生理盐水。高脂饮食24周后,通过超声生物显微镜成像技术评估小鼠主动脉及其主要分支内的动脉粥样硬化情况,随后处死小鼠并留取主动脉,通过H＆E染色和免疫组化技术进一步观察小鼠头臂干动脉粥样斑块的病变情况及CX3CR1的表达水平。结果在动脉粥样硬化斑块形成早期,即出现了CSE表达水平的显著降低,随着斑块的进展,CSE的表达水平进一步下调和CSE活性明显下降,最终导致血浆硫化氢水平的显著降低。动脉粥样硬化早期或中晚期予以NariS均可显著延缓动脉粥样硬化斑块的形成和进展,但是NaHS早期干预,其抗动脉粥样化的益处显著优于中晚期干预。NariS的抗动脉粥样硬化益处可能与其抑制斑块内CX3CR1的表达有关。结论动脉粥样硬化过程中存在着内源性硫化氢代谢紊乱,予以NaHS干预可抑制斑块内CX3CR1的表达和延缓动脉粥样硬化的进展,早期NaHS干预的疗效显著优于中晚期干预。     

Fractalkine/CX3CL1 in rheumatoid arthritis

Fractalkine is a CX3C chemokine that exists in both membrane-bound and soluble forms. Interaction between fractalkine and its unique receptor (CX3CR1) induces cell adhesion, chemotaxis, crawling, “accessory cell” activity, and survival. The serum level of fractalkine is elevated in patients with rheumatoid arthritis (RA) and is correlated with disease activity. Peripheral blood CD16⁺ monocytes and a subset of T cells express CX3CR1, while fractalkine is expressed on fibroblast-like synoviocytes and endothelial cells in the synovial tissue of patients with RA. Fractalkine expression is enhanced by tumor necrosis factor-α and interferon-γ, and it promotes the migration of monocytes, T cells, and osteoclast precursors into RA synovial tissue. Fractalkine also induces the production of inflammatory mediators by macrophages, T cells, and fibroblast-like synoviocytes. Moreover, fractalkine promotes angiogenesis and osteoclastogenesis. In an animal model of RA, arthritis was improved by the abrogation of fractalkine. Recently, a clinical trial of an anti-fractalkine monoclonal antibody for the treatment of RA commenced in Japan. We review the multiple roles of fractalkine in the pathogenesis of RA and its potential as a therapeutic target for this disease.     

Intestinal microbes direct CX3CR1+ cells to balance intestinal immunity

ABSTRACT

Intestinal damage driven by unrestricted immune responses against the intestinal microbiota can lead to the development of inflammatory diseases including inflammatory bowel disease. How such breakdown in tolerance occurs alongside the mechanisms to reinforce homeostasis with the microbiota are a focus of many studies. Our recent work demonstrates coordinated interactions between intact microbiota and CX₃CR1 expressing intestinal antigen presenting cells (APCs) that limits T helper 1 cell responses and promotes differentiation of regulatory T cells (Treg) against intestinal antigens including pathogens, soluble proteins and the microbiota itself. We find a microbial attachment to intestinal epithelial cells is necessary to support these anti-inflammatory immune functions. In this addendum, we discuss how our findings enhance understanding of microbiota-directed homeostatic functions of the intestinal immune system and implications of modulating this interaction in ameliorating inflammatory disease.     

High CX3CR1 expression predicts poor prognosis in paediatric acute myeloid leukaemia undergoing hyperleukocytosis

Introduction

Paediatric AML patients with hyperleukocytosis have a poor prognosis and higher early mortality. Therefore, more studies are needed to explore relevant prognostic indicators and develop effective prevention strategies for this type of childhood AML.

Methods

All original data were obtained from the TARGET database. First, we explored meaningful differentially expressed genes (DEGs) between the hyperleukocytosis group and the non-hyperleukocytosis group. Next, we screened and identified valuable target genes using univariate Cox regression, Cytoscape software, and Kaplan–Meier survival curves. Finally, the coexpressed genes, functional networks, and immune-related activities associated with the target gene were deeply analysed by the GeneMANIA, LinkedOmics, GEPIA2021, TISIDB, and GSCA databases.

Results

We selected 1229 DEGs between the hyperleukocytosis group and the non-hyperleukocytosis group in paediatric AML patients. Among them, 495 DEGs were significantly linked with the overall survival of paediatric AML patients. Further, we discovered that CX3CR1 was a promising target gene. Meanwhile, we identified CX3CR1 as an independent prognostic predictor. Besides, we showed that CX3CR1 had strong physical interactions with CX3CL1. Additionally, functional network analysis suggested that CX3CR1 and its coexpressed genes modulated immune response pathways. Subsequent analysis found that immune cells with a high median value of CX3CR1 were monocytes, resting NK cells and CD8 T cells. Finally, we observed that CX3CR1 expression correlated with infiltrating levels of immune cells and immune signatures.

Conclusion

Elevated CX3CR1 expression may be an adverse prognostic indicator in paediatric AML patients undergoing hyperleukocytosis. Moreover, CX3CR1 may serve as an immunotherapeutic target for AML with hyperleukocytosis in children.     

Lymphocyte subsets in patients with aplastic anemia

Lymphocyte subsets were enumerated in a group of 31 patients with aplastic anemia. Abnormal numbers of immunoregulatory T-cells were found in some patients: 26% of them showed a reversed helper/suppressor ratio. Seven of 18 patients showed significantly decreased proliferation in response to PWM; this hyporesponsiveness was present in 75% of patients with a reversed helper/suppressor ratio and in 10% of those with a normal helper/suppressor ratio (R = 0.66, P = 0.008). Eight of 18 patients showed suppressor activity over PWM-induced allogeneic cell proliferation. This suppressive activity did not correlate with T-cell phenotype. Of the patients with a low number of T-cells, 73% had responded to treatment, whereas of those patients with a normal number of T-cells, 26% had responded (P = 0.016). The results are consistent with abnormal immune response in selected patients with aplastic anemia, and suggest a possible influence of T-cells on disease process.