自身免疫性溶血性贫血患者同种抗体的检测及其临床意义

<正> 自身免疫性溶血性贫血(AIHA)的患者在多次输血免疫刺激后可以产生同种免疫抗体,但此类抗体由于患者体内同时存在的自身抗体的掩盖作用而不易被发现,致使再次输血时可能发生溶血性输血反应。因此,对 AIHA患者进行同种抗体检测具有重要的临床意义。     

CD4^＋CD25^＋调节性T细胞与自身免疫病

目的:CD4 CD25 调节性T细胞是一群具有免疫调节或免疫抑制功能的细胞。越来越多的实验证明,CD4 CD25 调节性T细胞在维持外周免疫耐受中起重要作用,这种T细胞的数量减少或功能缺失可导致自身免疫性疾病的发生。本文就CD4 CD25 调节性T细胞及其在自身免疫性疾病中作用的研究进展做一综述。资料来源:应用计算机检索CNKI、Medline、EMCC数据库和手工检索2006-2007年的相关文献。检索词为"CD4 CD25 T调节性细胞,自身免疫病,免疫耐受,CD4 CD25 regulatory T cell,Treg,autoimmune disease,immune tolerance"。资料选择:检索范围包括临床研究(不限研究对象的年龄、性别、种族)和基础研究,不限体内和体外研究。资料提炼:共收集到相关文献675篇,选择其中33篇英文文献进行重点阅读和分析。资料综合:CD4 CD25 调节性T细胞具有免疫抑制功能,在机体的免疫调节中发挥重要作用。与其免疫调节功能相关的杀伤性T细胞淋巴细胞相关抗原4、CD45RO、糖皮质激素诱导的肿瘤坏死因子受体、淋巴细胞的无能相关基因等细胞表面分子和白细胞介素2、白细胞介素10、白细胞介素4、转化生长因子β等细胞因子的研究不断深入。此外,CD4 CD25 调节性T细胞功能的发挥还与FOXP3的表达密切相关。CD4 CD25 调节性T细胞数量的减少、抑制功能的受损和(或)细胞表面分子表达的缺陷可能导致1型糖尿病、多发性硬化和炎症性肠病等多种自身免疫病的发生。结论:CD4 CD25 调节性T细胞主要通过细胞接触依赖机制和抑制性细胞因子依赖机制发挥免疫抑制效应。其数量的减少、功能的受损和(或)表面分子表达的缺陷与自身免疫病的发生发展密切相关。     

COPD病人外周血CD4^＋CD25^＋调节性T细胞检测及意义

目的检测慢性阻塞性肺疾病（COPD）病人外周血中CD4^＋CD25^＋调节性T细胞（Treg）的频率及血清γ-干扰素（IFN-γ）、白细胞介素4（IL-4）浓度,探讨Treg在COPD发病中的免疫调节作用。方法采用流式细胞仪检测45例COPD急性发作期病人、33例COPD临床缓解期病人以及25例健康人外周血中Treg亚群的频率,ELISA法检测血清中IFNγ-、IL-4浓度。结果 COPD病人急性发作期组IL-4浓度较临床缓解期组和健康对照组显著增高,临床缓解期组低于健康对照组,差异有统计学意义（F=192.86,q=11.63-27.66,P〈0.01）;COPD病人急性发作期组IFN-γ浓度高于临床缓解期组和健康对照组,差异有统计学意义（F=35.53,q=9.72、10.13,P〈0.01）,而临床缓解期组与健康对照组差异无统计学意义（q=1.13,P〉0.05）;临床缓解期组IFNγ-/IL-4比值最高,急性发作期组次之,健康对照组最低,差异有统计学意义（F=230.95,q=7.79-30.10,P〈0.01）;CD4^＋CD25^＋Treg频率在COPD急性发作期组高于临床缓解期组和健康对照组,而临床缓解期组低于健康对照组,差异有显著性（F=146.08,q=8.68-12.72,P〈0.01）。结论 COPD病人存在Th1/Th2动态变化,急性发作期以Th2免疫为主,临床缓解期则以Th1免疫为主;Treg在COPD的发生发展中担负着重要的免疫调节作用。     

自身免疫性溶血性贫血患者外周血T细胞亚群的检测

目的 分析自身免疫性溶血性贫血（AIHA）患者外周血T细胞亚群极化状态,探讨AmA的免疫学发病机制。方法 收集AIHA患者及健康者外周抗凝静脉血,分离纯化淋巴细胞。FITC标记的抗CD3单抗,Cy5标记的抗CD4、CD8单抗,PE标记的CRTH2单抗,以CD3／CD4、CD3／CD8设门作双色流式细胞术,分析AIHA患者T细胞亚群极化状态。结果与正常对照相比,AIHA患者外周血CD4^＋细胞百分率显著升高（P〈0．01）,CD8^＋细胞百分率变化不显著（P〉0．05）,CD4^＋／CD8^＋比例显著升高（P〈0．05）;CD3^＋CD4^＋CRTH2^-T细胞百分率、CD4^＋CRTH2^-T／CD4^＋CRTH2^-T和CD8^＋CRTH2^-T／CD8^＋CRTH2^＋T比例均显著升高（P〈0．01）,而CD3^＋CD4^＋CRTH2^＋T、CD3^＋CD8^＋CRTH2^-T及CD3^＋CD8^＋CRTH2^＋T细胞百分率的变化不显著（P〉0．05）。结论 AIHA患者外周血存在细胞免疫功能失调,T细胞亚群极化状态发生改变,呈明显的Th1／Tc1型细胞优势;Th1／Tc1型细胞介导的细胞免疫可能与AIHA的免疫学发病机制有关。     

自身免疫性甲状腺炎大鼠CD4^＋CD25^＋调节性T细胞变化及意义

目的研究CD4^＋CD25^＋Foxp3^＋调节性T细胞在自身免疫性甲状腺炎（AITD）大鼠外周血中的比例变化,并初步探讨其在疾病进程中的意义。方法选取17例A1TD大鼠,取静脉血,应用流式细胞术及定量PCR的方法,分别在蛋白质和mRNA水平检测Foxp3^＋表达,并与甲状腺炎球蛋白抗体（TgAb）,甲状腺过氧化酶抗体（TPOAb）,甲状腺微粒体抗体（TmAb）和甲状腺组织活检结果作相关分析。取正常大鼠作为对照。结果AITD大鼠CD4^＋CD25^＋调节性T细胞（5．35％±1．27％）显著低于正常对照组（8．02％±2．29％,P〈0．01）,AITD大鼠CD4^＋CD25^＋Foxp3^＋T细胞占CD4‘T细胞比例（3．12％±0．78％）明显低于正常对照组（6．45％±2．12％,P〈0．01）,AITD大鼠Treg与TgAb、TmAb、TPOAb呈负相关（r=-0．291,-0．357,-0．389,P〈0．05,0．05,0．05）,并与小鼠甲状腺组织的炎症程度呈负相关（r=-0．511,P〈0．01）;Foxp3mRNA表达水平与蛋白质表达水平变化相一致。结论AITD大鼠CD4^＋CD25^＋T细胞明显减少,这群调节性T细胞可能参与了AITD的病理进程。     

哮喘患者外周血CD4^＋CD25^＋调节性T细胞的测定及临床意义

目的探讨CD4^＋CD25^＋调节性T细胞在哮喘患者外周血的变化及意义。方法采用流式细胞术检测60例哮喘患者和40例健康对照者外周血中CD4^＋CD25^＋调节性T细胞的表达量,同时测定20例急性发作期哮喘患者的肺功能。结果哮喘患者外周血CD4^＋CD25^＋调节性T细胞占CD4^＋T细胞的百分比明显低于健康对照组,差异有统计学意义（P〈0．01）;急性发作期哮喘患者外周血CD4^＋CD25^＋调节性T细胞明显低于非急性发作期患者,差异也有统计学意义（P〈0．01）。结论CD4^＋CD25^＋调节性T细胞参与了哮喘的发生。     

CD4^＋CD25^＋调节性T细胞与自身免疫性甲状腺炎

调节性T细胞主要在机体免疫系统中发挥负向调节作用,既能抑制不恰当的免疫反应,又能限定免疫应答的范围、程度及作用时间,对效应细胞的增殖、免疫活性的发挥起抑制作用。近年来,CD4^＋CD25^＋调节性T细胞在自身免疫性甲状腺疾病中的作用日益引起人们的关注,动物实验和人体研究发现CD4^＋CD25^＋调节性T细胞数目和功能的异常与自身免疫性甲状腺炎（autoimmune thyroiditis,AIT）的关系密切,可能在AIT的发病机制中发挥关键作用。综述如下。     

消化道肿瘤患者外周血中CD4^＋CD25^＋调节性T细胞的检测

CD4 CD25 调节性T细胞(regulatory T,Treg)是一类维持机体免疫动态平衡的T淋巴细胞亚群,具有免疫无能性和免疫抑制性特点[1-2]。本文对不同的消化道肿瘤、组织类型及治疗前后外周血CD4 CD25 Treg细胞的变化进行观察,以探讨CD4 CD25 Treg细胞的临床意义。1对象与方法1.1研究对象     

自身免疫性溶血性贫血患者的输血治疗

自身免疫性溶血性贫血是贫血的一种类型。是由于某种原因，体内产生了自身抗体，这种抗体与红细胞表面抗原结合或游离于血清中，使红细胞破坏增加的一种贫血。短期内出现严重贫血的患者往往需要输血治疗。鉴于自身免疫性溶血性贫血的免疫血清学特点，输血时会出现配血不合或输血后贫血反而加重的情况，甚至危及患者的生命。因此自身免     

CD4^＋CD25^＋调节性T细胞的生物学研究概述

如同许多其他生理过程一样，免疫也需要兴奋和抑制两种对立统一的调节，才能维持其正常的功能。免疫学的早期更多地关注免疫应答的诱导，后来免疫应答的抑制性调节日益成为免疫学研究的热点之一。尽管胸腺的阴性选择克隆清除了自身反应性T细胞，并建立了机体的自身耐受，但仍然有一部分自身反应性T细胞逃脱了这一过程，并与机体的某些保护性机制建立了一个微妙的平衡，从而形成自身耐受。最初的观点认为，这些机制主要包括T细胞无能以及T细胞忽视。     

T淋巴细胞在自身免疫性溶血性贫血发病机制中的作用

自身免疫性溶血性贫血(AIHA)为常见的自身抗体介导的免疫性红细胞减少症,除了B淋巴细胞活化外,T淋巴细胞在AIHA的发病机制中亦发挥重要作用.根据功能不同,T淋巴细胞分为辅助性T淋巴细胞(Th)、调节性T淋巴细胞(Treg)及细胞毒性T淋巴细胞(CTL).为了进一步了解T淋巴细胞在AIHA发病机制中的作用,笔者现对AIHA中Th与Treg的生物学特性及其分泌的细胞因子的研究进展进行综述,以期了解不同阶段AIHA的免疫学特征,指导临床观察及治疗.     

Thyroid storm and warm autoimmune hemolytic anemia

Graves’ disease is often associated with other autoimmune disorders, including rare associations with autoimmune hemolytic anemia (AIHA). We describe a unique presentation of thyroid storm and warm AIHA diagnosed concurrently in a young female with hyperthyroidism. The patient presented with nausea, vomiting, diarrhea and altered mental status. Laboratory studies revealed hemoglobin 3.9 g/dL, platelets 171 × 10⁹ L^?1, haptoglobin <5 mg/dL, reticulocytosis, and positive direct antiglobulin test (IgG, C3d, warm). Additional workup revealed serum thyroid stimulating hormone (TSH) <0.01 μIU/mL and serum free-T4 (FT4) level 7.8 ng/dL. Our patient was diagnosed with concurrent thyroid storm and warm AIHA. She was started on glucocorticoids to treat both warm AIHA and thyroid storm, as well as antithyroid medications, propranolol and folic acid. Due to profound anemia and hemodynamic instability, the patient was transfused two units of uncrossmatched packed red blood cells slowly and tolerated this well. She was discharged on methimazole as well as a prolonged prednisone taper, and achieved complete resolution of the thyrotoxicosis and anemia at one month. Hyperthyroidism can affect all three blood cell lineages of the hematopoietic system. Anemia can be seen in 10–20% of patients with thyrotoxicosis. Several autoimmune processes can lead to anemia in Graves’ disease, including pernicious anemia, celiac disease, and warm AIHA. This case illustrates a rarely described presentation of a patient with Graves’ disease presenting with concurrent thyroid storm and warm AIHA.     

Finding the elusive and causative autoantibody: An atypical case of autoimmune hemolytic anemia

An isolated IgA‐mediated autoimmune hemolytic anemia can present a diagnostic challenge. When a routine direct antiglobulin test (DAT) is negative but clinical suspicion remains high, further testing with monospecific antisera should be performed. As with IgG‐mediated WAIHA, steroids are first‐line treatment, though splenectomy is often required to achieve a durable treatment response.     

自身免疫性溶血性贫血的诊断和治疗现状

自身免疫性溶血性贫血(AHIA)是由于机体产生抗自身红细胞膜抗原的抗体导致红细胞破坏增加,寿命缩短.据有无病因,分为原发性和继发性,继发者约占50％.据致病抗体最佳活性温度分为温抗体型、冷抗体型和冷温抗体混合型.诊断方面注意查明抗体类型及原发病.治疗方案多为经验性治疗,温抗体型AIHA首选糖皮质激素,有效率可达80％,但易复发,停药后持续缓解率小于20％;二线治疗方案包括脾切除和利妥昔单抗(R).近年来国外研究表明,R有效率高达70％以上,部分学者认为应作为首选二线治疗方案,甚至用于一线治疗.传统治疗如环孢素、达那唑、环磷酰胺、硫唑嘌呤、人免疫球蛋白、吗替麦考酚酯、促红细胞生成素(EPO)、血浆置换、全血置换等均报道有效;新药研究示阿伦单抗、奥法木单抗有效,但均需要大样本前瞻性随机对照试验提供循证学依据.冷抗体型AIHA对糖皮质激素和脾切除疗效差,R应作为一线治疗方案.     

Massive thymic deletion results in systemic autoimmunity through elimination of CD4+ CD25+ T regulatory cells

Incomplete deletion of KRN T cells that recognize the ubiquitously expressed self-antigen glucose-6-phosphate-isomerase (GPI) initiates an anti-GPI autoimmune cascade in K/BxN mice resulting in a humorally mediated arthritis. Transgenic (Tg) expression of a KRN T cell receptor (TCR) agonist under the major histocompatibility complex class II promoter resulted in thymic deletion with loss of anti-GPI T and B cell responses and attenuated arthritis course. However, double Tg mice succumbed to systemic autoimmunity with multiorgan inflammation and autoantibody production. Extensive thymic deletion resulted in lymphopenia and elimination of CD4+ CD25+ regulatory T cells (Tregs), but spared some CD4+ T cells expressing endogenous TCR, which oligoclonally expanded in the periphery. Disease was transferred by these T cells and prevented by cotransfer of CD4+ CD25+ Tregs. Moreover, we extended our findings to another TCR system (anti-hen egg lysozyme [HEL] TCR/HEL mice) where similarly extensive thymic deletion also resulted in disease. Thus, our studies demonstrated that central tolerance can paradoxically result in systemic autoimmunity through differential susceptibility of Tregs and autoreactive T cells to thymic deletion. Therefore, too little or too much negative selection to a self-antigen can result in systemic autoimmunity and disease.     

Vaccination-associated immune hemolytic anemia in two children

BACKGROUND: Two children in whom acute autoimmune hemolytic anemia (AIHA) developed after vaccination were studied. CASE REPORTS: The children were a 20-month-old girl and a 21-month-old boy. The diagnosis of AIHA was made in accordance with established criteria (hemolysis, positive DAT, and lack of other reasons for the hemolysis). Serologic tests were performed according to standard technique. RESULTS: The girl experienced two attacks of hemolysis. The first episode occurred 2 weeks after oral polio vaccination, and the second episode was observed 7 months later, when she received a simultaneous vaccination against mumps, rubella, and measles. The DAT was strongly positive with anti-C3d. No autoantibodies were detectable in either episode. The boy experienced acute hemolysis a few days after a simultaneous revaccination against diphtheria-pertussis-tetanus, Haemophilus influenzae, hepatitis B, and polio. The DAT using anti-IgG was strongly positive, and the DAT performed with anti-C3d was weakly positive. CONCLUSION: Vaccination-induced AIHA resembles those forms of AIHA related to infectious diseases, and it may occur more frequently than has been reported.     

Plasma exchange in autoimmune hemolytic anemia (AIHA)

Plasma exchange therapy in autoimmune hemolytic anemia (AIHA) was used in four patients (two with warm hemolytic anemia and two with cold hemolytic anemia). The size of each plasma exchange approximated 1 plasma volume; three consecutive daily exchanges removed 80–90% of the immunoglobulins—immunoglobulin G (IgG) and immunoglobulin M (IgM)—. complement (C₃, C₄), and reduced antibody titers. Transfusion requirements dramatically decreased after plasma exchange in each case. In two patients, red blood cell (RBC) survival studies were performed to more accurately assess the effect of plasma exchange therapy, since steroid and/or immunosuppressive therapy was given concomitantly. In one case of cold AIHA, homologous ⁵¹Cr-RBC were injected 4 days prior to plasma exchange and repeat injection (same donor) following completion of plasma exchange. The survival curve prior to plasma exchange therapy had a T 1/2 = 7.8 days (r = ?0.988) and after plasma exchange therapy had a T 1/2 = 20.4 days (r = ?0.925). RBC survival studies using homologous ⁵¹Cr-RBC were also performed in a patient with warm AIHA. The survival curve before plasma exchange had a T 1/2 = 2 days (r = ?0.95), and postplasma exchange a T 1/2 = 1.8 days (r = ?0.91). Plasma exchange therapy seems to have a beneficial effect in cold rather than warm autoimmune hemolytic anemia.     

CD4+CD25+Treg细胞和T细胞亚群评估再生障碍性贫血患者免疫状态的对比研究

目的：检测再生障碍性贫血(AA)患者外周血 CD4+ CD25+ Treg 细胞和 T 细胞亚群的表达情况,分析两者与 AA 病情严重程度及免疫抑制剂疗效的关系,为评估 AA 患者病情、选择合理治疗方案提供依据。方法以46例接受免疫抑制剂(单用环孢素 A)治疗的初诊 AA 患者和26例正常健康体检者为研究对象,采用多参数流式细胞术检测其细胞免疫功能,包括 T 细胞亚群、CD4+ CD25+ Treg 细胞和效应性 T 细胞,按照正常对照组水平将患者分组：CD127 low/CD127 high 比值正常型和降低型;CD4+/CD8+比值正常型、倒置型及正置型,分析比较各亚型与病情、疗效之间的关系,并进一步比较 CD4+ CD25+ Treg 细胞与 T 细胞亚群在评估 AA 患者免疫状态中的优劣。结果①CD127 low/CD127 high 比值正常型占28.3%(13/46),降低型占71.7%(33/46);重型 AA(SAA)与非重型 AA(NSAA)两组降低型患者所占比例分别为89.5%(17/19)、59.3%(16/27),差异有统计学意义(χ2=5.022,P <0.05);单用环孢素治疗,总有效率为48.1%(13/27),有效者降低型比例92.3%(12/13),明显高于无效者降低型比例50.0%(7/14)(χ2=3.395,P <0.05)。②CD4+/CD8+比值正常型占39.1%(18/46),倒置型占39.1%(18/46),正置型占21.7%(10/46);SAA 与 NSAA 两组中按 CD4+/CD8+分型,两组间构成比例差异无统计学意义(χ2=0.211,P >0.05);单用环孢素治疗,有效和无效者按 CD4+/CD8+分3种亚型,分别占有效者46.2%(6/13)、46.2%(6/13)、7.7%(1/13),无效者为35.7%(5/14)、28.6%(4/14)、35.7%(5/14),差异无统计学意义(χ2=2.983,P >0.05)。③CD4+ CD25+ Treg和 T 细胞亚群监测 AA 患者免疫状态异常的阳性率分别为71.7%(33/46)和60.9%(28/46),CD4+ CD25+ Treg 监测免疫状态异常的敏感性高于 T 细胞亚群(χ2=4.290,P <0.05)。结论大多数患者存在 CD4+ CD25+ Treg 细胞和(或)T 细胞亚群的异常,与 AA 发病机制的免疫异常密切相关;在监测 AA 患者免疫功能异常、评估病情及免疫抑制剂疗效时,CD4+ CD25+ Treg 细胞比 T 细胞亚群更敏感;CD4+ CD25+ Treg 细胞联合 T 细胞亚群的监测对临床了解AA 病情和发病机制、合理用药、提高诊断和治疗水平有重要价值。     

骨髓间充质干细胞上调再生障碍性贫血患者CD4~+ CD25~+ Foxp3~+调节性T细胞的临床研究

目的:初步探讨再生障碍性贫血(AA)患者外周血CD4+CD25+Foxp3+调节性T细胞(Treg)的表达及临床意义;探讨临床使用异体骨髓间充质干细胞(MSC)治疗对CD4+CD25+Foxp3+Treg表达的影响。方法:(1)流式细胞仪检测13例重型再生障碍性贫血(SAA),17例慢性再生障碍性贫血(CAA)及10例健康对照者外周血CD4+CD25+Foxp3+Treg表达;(2)检测7例AA患者MSC治疗前后外周血CD4+CD25+Foxp3+Treg表达。结果:(1)AA患者外周血CD4+CD25+Foxp3+Treg表达明显低于健康对照者(P<0.01);(2)SAA患者CD4+CD25+Foxp3+Treg表达较CAA者低(P<0.05);(3)MSC治疗后AA患者外周血CD4+CD25+Foxp3+Treg表达升高(P<0.05)。结论:(1)AA患者存在外周血CD4+CD25+Foxp3+Treg表达减低,其表达异常可能参与AA的发病;(2)CD4+CD25+Foxp3+Treg表达在SAA患者更低,提示其表达水平可能可以作为判断AA病情轻重的指标;(3)MSC治疗可上调AA患者CD4+CD25+Foxp3+Treg表达,为MSC纠正AA患者的免疫异常提供了临床依据。