CD4~+Foxp3~+T细胞亚群在系统性红斑狼疮中的免疫抑制功能研究

采用五色流式细胞术检测系统性红斑狼疮(systemic lupus erythematosus,SLE)患者和健康对照组的CD4+Foxp3+T细胞及其细胞亚群;以CFSE染色法分析SLE患者和健康对照组CD4+Foxp3+T细胞各亚群的免疫抑制功能。结果发现SLE组CD4+Foxp3+T细胞(占CD4+T细胞的比例)显著高于对照组(12.03%±1.523%vs 6.410%±0.4353%,t=3.790,P<0.01),但其CD4+T细胞的增殖却比对照组活跃(PI:4.052%±0.4004%vs 2.528%±0.2322%,t=3.293,P<0.05)。根据Foxp3和CD45RA表达强度的差异,可将CD4+Foxp3+T细胞清晰地分成3个亚群:CD45RA+Foxp3low(Ⅰ区)细胞、CD45RA-Foxp3high(Ⅱ区)细胞和CD45RA-Foxp3low(Ⅲ区)细胞,SLE组的Ⅰ区和Ⅲ区细胞数量显著增加(分别为t=2.123,P<0.05;t=3.462,P<0.01),而Ⅱ区细胞与对照组之间差异无统计学意义(0.6056%±0.1879%vs0.6571%±0.1764%,t=0.1999,P>0.05)。SLE患者和健康对照者的Ⅰ区和Ⅱ区细胞在体外均具有免疫抑制功能,而Ⅲ区细胞不具有该功能。SLE患者Ⅰ区细胞的免疫抑制能力弱于对照组(t=2.994,P<0.05),而Ⅱ区细胞和Ⅲ区细胞与对照组之间差异均无统计学意义。提示,SLE患者CD4+Foxp3+T细胞比例升高而免疫抑制功能下降,可能是两大原因造成了这种数量和功能的不一致性:①SLE患者CD4+Foxp3+T细胞数量增加主要由于Ⅲ区细胞增多所致,而这区细胞不具有免疫抑制功能;②在SLE患者,具有免疫抑制功能的Ⅰ区细胞的抑制能力明显减弱。     

特发性血小板减少性紫癜患儿外周血T细胞PKC活性变化的研究

为了探讨ITP患儿外周血T细胞蛋白激酶C(PKC)的活性变化及其与T细胞活化和血小板减少程度之间的关系,无菌采集35例ITP患儿及30例正常儿童外周血,采用T细胞分离富集柱法分离纯化T细胞,分别用非同位素标记法检测T细胞PKC的活性变化,用流式细胞仪检测T细胞活化标志FasL蛋白的表达,血细胞计数仪计数血小板的减少程度。结果ITP患儿T细胞PKC的总活性与正常儿童相比明显增强[(0.97±0.21)nmol/ml·min和(0.55±0.13)nmol/ml·min,x±s,P<0.05],T细胞活化标志FasL蛋白表达与正常儿童比较显著升高(CD4+TFasL:32.7%±3.4%和14.7%±4.2%;CD8+TFasL:17.3%±9.7%和11.6%±8.5%,x±s,P<0.05),并且T细胞PKC的活性变化与CD4+TFasL、CD8+TFasL的表达均为显著正相关(r1=0.68,r2=0.53,P<0.05),与血小板计数成显著负相关(r=-0.75,P<0.05)。上述研究结果表明ITP患儿PKC活性增强可能引起T细胞的活化,活性T细胞增多可导致患儿血小板大量损伤,提示PKC信号转导在ITP的免疫病理机制中发挥重要作用。     

抗内皮细胞抗体和血小板生成素测定在ITP与SLE鉴别中的意义

目的：探讨抗内皮细胞抗体(AECA)和血小板生成素(TPO)测定在鉴别特发性血小板减少性紫癜（ITP）和系统性红斑狼疮（SLE）中的临床意义。 方法: 用ELISA法分别测定76例ITP患者、41例SLE患者及50例正常人血清中的AECA和TPO水平。 结果: SLE组、ITP组患者血清AECA水平明显高于正常对照组（P<0.01）；SLE组患者血清AECA水平显著高于ITP组（P<0.01）；ITP组患者血清TPO水平与正常对照组无显著差异(P>0.05)，而SLE组血清TPO水平显著高于ITP组患者和正常对照组(P<0.01)。 结论: 血清AECA和TPO的测定在鉴别诊断ITP和SLE中有显著的临床意义。     

SLE血小板减少与巨核系祖细胞内在缺陷关系的探讨

本文应用改良的血浆凝块培养技术和ABC染色法研究SLE病人血小板减少的发病机理。实验结果表明,正常人外周血所形成的CFU—MK为21.2±4.7;血小板正常的SLE病人的CFU—MK为19.8±3.2;而血小板减少的SLE病人CFU—MK为8.5±3.1,明显低于正常对照(P<0.05)。我们的实验结果提示了SLE病人血小板减少是由于巨核系祖细胞(progenitor)或在CFU—MK水平上存在内在缺陷(intrinsic defect)。     

SLE患者血清对骨髓间充质干细胞衰老的作用及机制研究

目的:探讨系统性红斑狼疮(SLE)患者血清对间充质干细胞(MSC)衰老的作用及机制。方法:收集性别、年龄匹配6例健康对照者及6例SLE患者血清,灭活补体后-80℃保存备用;分离培养3例健康对照及3例SLE患者骨髓MSC,传代至第3~4代备用;以含10%SLE或对照血清的DMEM/F12模拟机体全身血液微环境,培养不同来源骨髓MSC;SA-β-gal染色并计数阳性细胞比例;Real time PCR法检测MSC p53、p21基因水平;Western blot法分别检测p53、p21蛋白及核蛋白NF-κB(p65)表达水平;CCK8测细胞增殖。结果:SLE患者血清增加健康人骨髓MSC的β-半乳糖苷酶阳性比例[(26.6±2.1)%vs(11.8±1.9)%,P<0.05],对SLE患者骨髓MSC的促进作用更为显著[(58.8±2.8)%vs.(31.2±2.2)%,P<0.05];SLE患者血清可同时上调健康对照及SLE患者骨髓MSC的p53及p21水平;SLE患者血清既能够抑制健康骨髓MSC增殖[(0.7±0.1)vs(1.1±0.1),P<0.01],又能抑制SLE患者骨髓MSC增殖能力[(0.5±0.2)vs(0.8±0.3),P<0.05]。此外,SLE患者血清还可显著上调两种来源骨髓MSC核内NF-κB(p65)的蛋白水平。结论:SLE患者血清能够促进骨髓MSC衰老,该过程可能与其胞内NF-κB信号通路的活化有关。     

淋巴瘤患者外周血TCRVα24+Vβ11+自然杀伤T细胞体外活化特性研究

目的： 研究淋巴瘤患者外周血TCRVα24+Vβ11+自然杀伤T（NKT）细胞的数量以及体外活化后的功能状态，与正常人外周血NKT细胞的数量及功能状态进行比较。方法: 制备30例淋巴瘤患者和30例年龄及性别匹配的正常对照外周血单个核细胞（PBMNCs），以流式细胞术（FACS）检测TCRVα24+Vβ11+NKT细胞数量，以α-半乳糖神经酰胺（α-Galcer）及白细胞介素-2（IL-2）从PBMNCs中扩增活化NKT细胞，采用细胞内细胞因子流式细胞术检测手段，测定NKT细胞中胞内白细胞介素-4（IL-4）、干扰素-γ（IFN-γ）、肿瘤坏死因子-α（TNF-α）阳性细胞的比例。结果: 淋巴瘤患者与正常对照PBMNCs中TCRVα24+Vβ11+NKT细胞的细胞比率分别为0.17%±0.10%、0.28%±0.18%(P<0.05)。PBMNCs培养体系中加入α-Galcer及IL-2，将NKT细胞扩增活化7d后，淋巴瘤患者与正常对照的NKT细胞的扩增倍数分别为101.37±44.61、129.66±56.31(P<0.05)。扩增活化后，淋巴瘤患者与正常对照的NKT细胞中胞内细胞因子IFN-γ阳性细胞的比例分别为41.96%±15.06%、52.48%±18.85%(P<0.05)；TNF-α阳性细胞的比例分别为46.30%±16.03%、71.37%±17.28%(P<0.05)；IL-4阳性细胞的比例分别为36.19%±11.74%、33.12%±12.95%(P>0.05)。不同病理分型及分期的淋巴瘤患者之间上述各指标无显著差异。结论: 淋巴瘤患者外周血TCRVα24+Vβ11+NKT细胞数量较正常对照明显减少，经α-Galcer扩增活化后扩增倍数较正常对照降低，分泌细胞因子IFN-γ、TNF-α的功能较正常降低，此数量及功能的降低与淋巴瘤的分型及分期无关。但其仍保持有对α-Galcer刺激后的扩增活化反应能力。     

乳香提取物对免疫低下实验小鼠T细胞功能的影响

研究乳香提取物提高小鼠免疫功能的机理与途径,探索乳香提取物对增强免疫功能的应用潜能。采用环磷酰胺制备免疫低下小鼠模型,以乳香提取物经灌胃给予免疫低下小鼠,连续10d。灌胃结束后取小鼠外周血采用荧光抗体染色并经流式细胞仪检测小鼠T细胞数量变化;取小鼠脾脏单个核细胞加抗CD3和CD28刺激并经3 H-TdR掺入试验,观察小鼠淋巴细胞增殖能力;取小鼠骨髓前体细胞与GM-CSF共培养,3 H-TdR掺入试验观察小鼠骨髓前体细胞增殖能力;ELISA方法检测小鼠血清中细胞因子。结果表明,给免疫低下模型小鼠口服乳香提取物后可有效增加小鼠总T细胞及CD4+T细胞数量:非服用组T细胞为12%±3%,服用组T细胞为28%±4%,两组相比具有显著差异(P<0.01);而CD4+T细胞从5%±2%(非服用组)增加至22%±3%(服用组),两组相比具有显著性统计学差异(P<0.01)。接受乳香提取物处理小鼠的T细胞对抗CD3和CD28刺激的增殖反应明显增高(P<0.05);IFN-γ、TNF-α、IL-12和IL-4水平上调;骨髓粒系细胞受GM-CSF刺激后增殖上调。因此,乳香提取物具有明显上调小鼠T细胞和Th1和Th2细胞数量和功能,其作用可能与刺激骨髓前体细胞增殖有关。     

rhTPO对巨核系的定向分化效应

目的 :探讨重组人促血小板生成素 (rhTPO)对外周造血干细胞中巨核系祖细胞的定向诱导分化能力。方法 :经化疗及G CSF动员后分离的外周造血干细胞 ,进行液体和集落培养 ,研究rhTPO单独及与IL 3、IL 6、SCF的协同作用。结果 :液体培养后 ,TPO组单个核细胞数 (MNC)扩增了 ( 1.73± 0 .49)倍 ,IL 3 IL 6 SCF组MNC比种植时增加 ( 4.2 0± 1.14)倍 ,IL 3 IL 6 SCF TPO组MNC增加了 ( 4.5 3± 1.2 7)倍。单独应用TPO及TPO与IL 3、IL 6、SCF合用产生高比例的CD41a 细胞 ,TPO组培养前CD41a 细胞为 11.70 %± 5 .2 3% ,培养后CD41a 细胞为 19.17%± 6 .2 6 % ( P <0 .0 5 )。CD41a 细胞数在TPO组扩增了 ( 3.5 2± 1.18)倍 ,IL 3 IL 6 SCF组扩增了 ( 5 .32± 1.79)倍 ,IL 3 IL 6 SCF TPO组扩增了 ( 6 .94± 2 .19)倍。结论 :TPO可以定向诱导巨核系细胞的分化 ,IL 3、IL 6、SCF可以协同TPO的作用 ,这在造血调控研究 ,体外定向扩增外周血干细胞 ,促进外周造血干细胞移植患者血小板的恢复具有应用前景     

类风湿关节炎患者外周血和滑液中CD4~+CD25~+调节性T细胞的水平变化

了解具有抑制功能的CD4+CD25+调节性T细胞(Treg)在类风湿关节炎(RA)中的水平变化。分离32例RA患者及35例正常对照者外周血和15例RA关节滑液中的单个核细胞,用荧光抗体标记细胞膜表面CD4、CD25分子和细胞内Foxp3转录因子,进行流式细胞分析,同时用RT-PCR方法测定单个核细胞中Foxp3 mRNA水平。实验发现RA外周血中CD4+CD25hT细胞比例(1.90±1.68)与健康人(1.81±1.79)无明显差异,而RA关节滑液中CD4+CD25+和CD4+CD25hT细胞含量却明显增高(14.98±12.52,8.94±9.67,P<0.01)。RA患者外周血单个核细胞中Foxp3+/CD4+T细胞比值(2.35±2.06)较正常人(7.25±3.98)明显降低(P<0.01),RA外周血中Foxp3 mRNA含量较正常人Treg减少,而RA关节液中Foxp3 mRNA含量较RA外周血更为低下(P<0.01)。RA患者存在CD4+CD25+Treg的异常改变,其外周血和关节液中具有抑制作用的Treg含量明显降低提示RA患者Treg数量减少及抑制功能下降可能是RA自身免疫反应亢强不能控制的原因之一。RA关节液中CD4+CD25hT细胞增高考虑与RA炎症反应造成T细胞过度活化有关。     

脐带血巨核祖细胞的体外扩增

目的联合血小板生成素(TPO)、白细胞介素-11(IL-11)和肝素用脐带血CD34 细胞定向扩增巨核祖细胞。方法采用免疫磁珠法(MACS)分选CD34 细胞,用TPO、IL-11和肝素定向扩增巨核祖细胞,巨核祖细胞集落分析(CFU-MK)测定巨核祖细胞扩增倍数,流式细胞术检测巨核祖细胞分化过程中不同细胞组群(CD34 、CD41a 、CD61 、CD34 CD41a 和CD41a CD61 )的变化,免疫组织化学染色(CD41a)和透射电镜观察巨核细胞形态及趟微结构,血小板体外活化实验及非肥胖性糖尿病／严重联合免疫缺陷鼠异种体内移植实验评价扩增的巨核祖细胞功能。     

Suspects in the tale of lupus-associated thrombocytopenia

Immunologically mediated thrombocytopenia is a frequent clinical manifestation in patients with systemic lupus erythematosus (SLE). Autoantibodies targeting platelet membrane glucoproteins have a central role in peripheral platelet destruction. Autoantibodies against thrombopoietin are also present in about one‐third of patients, but their pathogenetic role is obscure. Thirty‐eight serum samples from SLE patients were tested for anti‐platelet antibodies, anti‐thrombopoietin antibodies and levels of circulating thrombopoietin. Bone marrow histology was also assessed. Thirty‐nine per cent of sera displayed anti‐thrombopoietin antibodies and 29% had circulating anti‐platelet antibodies. Anti‐thrombopoietin antibodies were associated with lower thrombopoietin concentrations, and lower mean platelet values in long‐term follow‐up. Anti‐platelet antibodies were present in about 40% of thrombocytopenic and non‐thrombocytopenic individuals but were absent in patients who had recovered from thrombocytopenia, supporting their pathogenetic role. Both autoantibodies were absent in control sera from patients with rheumatoid arthritis and primary Sjögren’s syndrome. Decreased bone marrow cellularity, normal or low number of hypolobulated, pyknotic megakaryocytes and stromal alterations were prominent findings in thrombocytopenic SLE patients, suggesting a defect in megakaryopoiesis. These findings were not evident in specimens from patients with idiopathic thrombocytopenic purpura who had increased megakaryocytes, normal cellularity and absence of stromal alterations. In conclusion, peripheral destruction due to platelet autoantibodies, anti‐thrombopoetin antibodies, lower effective circulating thrombopoetin and impaired compensatory response due to bone marrow damage interact in SLE and thrombocytopenia ensues.     

CD41+/CD45+ cells without acetylcholinesterase activity are immature and a major megakaryocytic population in murine bone marrow

Murine megakaryocytes (MKs) are defined by CD41/CD61 expression and acetylcholinesterase (AChE) activity; however, their stages of differentiation in bone marrow (BM) have not been fully elucidated. In murine lineage-negative (Lin(-))/CD45(+) BM cells, we found CD41(+) MKs without AChE activity (AChE(-)) except for CD41(++) MKs with AChE activity (AChE(+)), in which CD61 expression was similar to their CD41 level. Lin(-)/CD41(+)/CD45(+)/AChE(-) MKs could differentiate into AChE(+), with an accompanying increase in CD41/CD61 during in vitro culture. Both proplatelet formation (PPF) and platelet (PLT) production for Lin(-)/CD41(+)/CD45(+)/AChE(-) MKs were observed later than for Lin(-)/CD41(++)/CD45(+)/AChE(+) MKs, whereas MK progenitors were scarcely detected in both subpopulations. GeneChip and semiquantitative polymerase chain reaction analyses revealed that the Lin(-)/CD41(+)/CD45(+)/AChE(-) MKs are assigned at the stage between the progenitor and PPF preparation phases in respect to the many MK/PLT-specific gene expressions, including beta1-tubulin. In normal mice, the number of Lin(-)/CD41(+)/CD45(+)/AChE(-) MKs was 100 times higher than that of AChE(+) MKs in BM. When MK destruction and consequent thrombocytopenia were caused by an antitumor agent, mitomycin-C, Lin(-)/CD41(+)/CD45(+)/AChE(-) MKs led to an increase in AChE(+) MKs and subsequent PLT recovery with interleukin-11 administration. It was concluded that MKs in murine BM at least in part consist of immature Lin(-)/CD41(+)/CD45(+)/AChE(-) MKs and more differentiated Lin(-)/CD41(++)/CD45(+)/AChE(+) MKs. Immature Lin(-)/CD41(+)/CD45(+)/AChE(-) MKs are a major MK population compared with AChE(+) MKs in BM and play an important role in rapid PLT recovery in vivo.     

类风关滑膜浸润T细胞特性与致病机制研究

为研究类风湿关节炎时关节滑膜浸润性T细胞生物学特性与致病机制 ,对 10例RA患者滑膜液中淋巴细胞的免疫表型、细胞因子分泌格局与趋化因子受体表达进行了分析。用双色荧光标记法分别测定滑膜液中和外周血淋巴细胞表型与趋化因子受体表达。用ELISA方法检测滑膜液与外周血中IFN γ、IL 10、IL 4与IL 12的含量。结果是滑膜液中的CD4 + T淋巴细胞为 4 0 0 %± 11% ,CD8+ T细胞为 34 0 %± 6 % ,CD4 + 与CD8+ T细胞的比值为 1 2 ,显著低于外周血中CD4 /CD8的比值。滑膜液中CD3和CD2 5双阳性的活化T细胞占 16 %± 6 0 %。趋化因子受体CCR5表达较低 ,与外周血无明显差异。但CX CR3表达水平较高 ,为 16 %± 4 0 % ,远远高于外周血 (仅为 0 5 %± 0 3% )。IFN γ在滑膜液中含量很高 ,达 (36 6 7± 4 3 2 )pg/ml,而外周血中含量仅为 (2 0 1± 3 2 )pg/ml。IL 4含量未能测得 (<15pg/ml ) ,与外周血相似。IL 12含量为 (4 19 9±89 2 )pg/ml,远高于外周血中的含量 (6 5 32± 34 2 )pg/ml。IL 10含量为 (187 7± 34 5 )pg/ml,高于外周血中的含量 (85±12 7)pg/ml。在所测细胞因子中 ,关节滑膜液中IFN γ和IL 12的含量与外周血相比具有显著的统计学差异。表明RA关节滑膜液中有相当数量的T细胞浸润。这些T细胞     

SLE患者Th1/Th2平衡状态及与外周血淋巴细胞CD28及CTLA-4分子表达的关系

目的:了解SLE患者Th1/Th2平衡状态以及共刺激分子CD28/CTLA-4与Th1/Th2平衡状态的关系。方法:研究对象为18例SLE患者(活跃期12例、缓解期6例)。对照组14例,为健康体检者。外周血单个核细胞(PBMCs)经梯度密度离心法分离后置于含PMA(5μg/L)及ionomycin(500μg/L)培养液中培养72 h。采用ELISA方法检测培养的PBMCs上清液中IFN-γ及IL-10的含量。应用流式细胞技术检测培养的淋巴细胞CD28及CTLA-4分子的表达。结果:活跃期SLE患者培养的PBMCs分泌IL-10的量(351.29 ng/L±153.31 ng/L)较对照组(254.48 ng/L±120.69 ng/L)有一定程度的升高,但差异无显著(P0.05),IFN-γ的分泌量(25.76 ng/L±16.09 ng/L)明显低于对照组(50.71 ng/L±27.92 ng/L,P0.05),IL-10/IFN-γ比值(18.74±13.77)明显高于对照组(6.66±4.95,P0.05)。培养前、后SLE患者CD3+及CD8+T细胞CD28分子表达量与对照组比较均无显著差异。培养前活跃期SLE患者CD3+T细胞CTLA-4分子表达量(0.79%+0.37%)较对照组(1.31%+0.61%)明显降低(P0.05)。培养后SLE患者CD3+T细胞及CD8+T细胞CTLA-4分子表达量仍低于对照组,但差异无显著(P0.05)。活跃期SLE患者培养的PBMCs中CD3+T细胞CTLA-4分子的表达量与上清液中IFN-γ含量呈明显的直线正相关关系(r=0.681,P0.05)、与上清液中IL-10及IL-10/IFN-γ比值呈明显的直线负相关关系(r=-0.624,P0.05;r=-0.738,P0.01)。结论:SLE患者存在Th1/Th2平衡向Th2方向偏移,即Th2优势状态。CTLA-4分子可能通过抑制CD28的信号转导参与Th2优势状态的形成。     

Total and biologically active CD154 in patients with SLE

CD154, a member of the tumor necrosis factor receptor family, is involved in several biological responses. In the sera of systemic lupus erythematosus (SLE) patients, the levels of sCD154 have been shown to be increased, however, few reports have dealt with the biologically active tetramer. Here, we assessed the biological activity of the serum CD154 tetramer using bioassays for BC activation and production nitrite or peroxide. The patients showed a markedly increased total sCD154 serum concentration (12.5 ± 8.2 vs. 3.9 ± 1.2 ng/ml; p < 0.001). ba-sCD154 was significantly increased in non-treated patients (7.4 ± 3.4 ng/ml, n = 22; p < 0.001) and patients with the highest SLE disease activity index (SLEDAI) scores (5.3 ± 2.9 ng/ml, n = 8), but not in stable patients (1.3 ± 1.2 ng/ml, n = 30) whose values were similar to normal healthy donors (NHD; 0.8 ± 0.2 ng/ml). Patients with SLEDAI above 8 that recovered after successful treatment displayed significantly decreased levels of ba-sCD154. We conclude that the bioassay is a useful tool discriminating active and stable SLE, as well as non-treated patients.     

用CD133免疫磁珠分离脐血内皮祖细胞的实验研究

目的：从脐血中分离、培养血管内皮祖细胞,研究内皮祖细胞的生长特性和诱导分化条件。 方法： 应用MACS磁球抗体标记法纯化脐血中的CD133+细胞，通过流式细胞仪、免疫细胞化学、免疫荧光等技术及形态学（光镜、电镜）观察研究内皮祖细胞；将细胞接种于添加（或未添加）VEGF、bFGF、干细胞因子（SCF）的含20％胎牛血清（FBS）的IMDM培养基中,观察内皮祖细胞的生长特性。 结果： 分离新鲜脐血所得CD133阳性细胞占单个核细胞的(1.41±1.14)%,经流式细胞仪鉴定CD133+细胞纯度为75%-85%；将分离细胞接种于纤维连接蛋白包被的24孔板内，培养1-2 h即有细胞贴壁，7-10 d可见贴壁细胞呈铺路石样排列；14 d后细胞出现小圆形、梭形等多样性变化，可见毛细血管管腔样结构，电镜观察可见胞浆内典型的Weibel-Palade小体；在VEGF、bFGF、SCF存在条件下,检测贴壁细胞培养14 d后细胞表面抗原表达情况：与培养开始时相比，祖细胞标志CD133和CD34阳性率呈明显下降趋势，分别由(77.0±3.3)%和(93.1±4.7)%降至(1.6±2.2)%和(37.4±4.9)%，P<0.05，内皮细胞特异性标志Flk-1表达明显增加，由(22.3±3.3)%增至(94.3±4.1)%，P<0.05，同时vWF抗原呈强阳性表达，阳性率为(77.9±3.3)%。 结论： 根据细胞表面特异性分子标志（CD133+/CD34+/Flk-1+）可以从脐血中分离出EPCs，EPCs可在体外一定的诱导因子作用下，培养7-10 d分化为成熟内皮细胞。     

Structural changes in the megakaryocytes of patients infected with the human immune deficiency virus (HIV-1).

Although immune mechanisms are known to be partially responsible for the thrombocytopenia of patients infected with HIV-1, an understanding of the mechanism underlying this disorder is incomplete. A casual observation that bone marrow biopsies of HIV-infected individuals seem to exhibit an unusually large number of denuded megakaryocyte nuclei (DN-MK) prompted a study comparing MK of 20 HIV-seropositive individuals with those of 10 patients with HIV-negative idiopathic thrombocytopenic purpura and 10 hematologically normal subjects. In normal marrows the number of DN-MK average 2.1 +/- 0.5 SE per 10 low power field. In patients with ITP the average number was 6.5 +/- 1.4 SEM, whereas HIV-ITP marrows had an average of 42.5 +/- 3.7 SEM. Electron microscopy of AIDS megakaryocytes exhibited ballooning of the peripheral zone to an extent not seen by us in any other myelodysplastic syndromes. These observations support the concept that the pathophysiology affecting MK/platelets in HIV-infection should not be equated with the destructive process underlying other immune thrombocytopenias.     

Deficiency of red cell bound CD55 and CD59 in patients with systemic lupus erythematosus

CD55 and CD59 are glycosylphosphatidylinositol-anchored proteins with complement inhibitory properties. Autoimmune hemolytic anemia (AIHA) has been associated with antiphospholipid antibodies (APLA). The aim of this study was to evaluate the presence of APLA and its possible correlation with diminished CD55 and CD59 in red blood cells from patients with primary AIHA or secondary to systemic lupus erythematosus (SLE). Flow cytometric analyses were performed on CD55 and CD59 stained erythrocytes from 24 patients (primary AIHA, n=8; AIHA plus SLE, n=11; and SLE without AIHA, n=5) and 20 healthy controls. Antibodies to several phospholipids were detected in the sera by ELISA. Most patients with AIHA plus SLE and few with primary AIHA showed deficiency of either or both CD55 and CD59 expression and was not associated to the presence of APLA, while SLE patients exhibited a normal expression of these molecules. Although our findings showed CD55 and CD59 deficiency in primary or secondary AIHA, it appears that this defect plays a facilitator rather than a triggering role for the hemolytic process. Additionally, a role of anti-phospholipid antibodies as causative of this acquired defect is questionable.     

Complement activating cryoglobulins in the nephritis of systemic lupus erythematosus

Complement activation in vitro by cryoglobulins isolated from the sera of 28 patients with systemic lupus erythematosus (SLE) was examined by incubating the cryoglobulin with normal human serum and performing crossed-immunoelectrophoresis of the mixture to detect C3 conversion. Eighteen of the 28 SLE cryoglobulins activated complement; eight by the classical pathway, four by the alternative pathway exclusively, and six by both pathways. In contrast only two out of 20 cryoglobulins isolated from the sera of normal subjects activated complement and both did so by the classical pathway. Twenty-three of the 28 SLE sera activated complement and complement activating cryoglobulins were isolated from 15 of these 23 sera. The parent sera of cryoglobulins activating complement had lower C4 and C3 concentrations than sera whose cryoglobulins did not split complement but these differences were not significant. The ability of SLE cryoglobulins to activate complement in vitro suggests that these immune complexes activate complement in vivo and thus may contribute to tissue damage in this disease. The activation of both classical and alternative complement pathways is in keeping with other evidence that both pathways are involved in SLE.     

系统性红斑狼疮患者血清IV型胶原和层粘连蛋白测定的意义

为探讨血清IV型胶原 (IV C)和层粘连蛋白 (LN)在系统性红斑狼疮 (SLE)患者中的临床意义 ;我们采用放射免疫法(RIA)对 34例SLE和 6 3例正常人的血清IV C和LN含量进行了测定 ,并进行治疗前后对比以及血清IV C和LN水平与其它临床指标间的直线相关关系分析。结果发现SLE组血清IV C和LN均较对照组显著升高 (P <0 0 0 0 1) ;治疗后随病情缓解较治疗前明显降低 (分别为P <0 0 0 1;0 0 0 5 )。血清LN与血清白蛋白呈显著负相关 (P <0 0 0 1) ,与 2 4h尿蛋白定量、血清免疫球蛋白M和补体C3水平之间呈明显的正相关 (P <0 0 5 ) ;血清IV C与血沉呈显著正相关 (P <0 0 5 )。上述结果提示血清IV C和LN水平在SLE患者中普遍升高 ,是评估SLE患者病情活动的重要指标