慢性乙型肝炎患者外周血T细胞亚群和共刺激分子的表达及意义

探讨慢性乙型肝炎患者外周血T细胞亚群及共刺激分子表达的临床价值。采用流式细胞术检测了 6 8例慢性乙型肝炎患者和 5 0例正常人外周血T细胞亚群及共刺激分子的表达。慢性乙型肝炎中、重型患者CD 4 细胞及CD4/CD8细胞比值明显低于正常组。三组患者CD2 8、CD 8CD 2 8表达水平均显著低于正常组 ,而CD80 、CD 8CD-2 8表达水平均显著高于正常组。CD2 8、CD80 、CD 8CD 2 8、CD 8CD-2 8的表达及CD4/CD8细胞比值在各型患者间均有明显差异 ,而CD3 、CD8表达水平无明显变化。T细胞亚群及共刺激分子的检测 ,对指导乙肝的临床治疗、判断预后有一定意义     

急性冠状动脉综合征患者B7∶CD28/CTLA4共刺激分子的表达

目的:探讨急性冠状动脉综合征(ACS)患者外周血中单核细胞共刺激分子CD28、细胞毒T淋巴细胞抗原(CTLA)4、CD80(B7-1)的变化,探讨这些分子在发病中的意义。方法:采用直接荧光标记流式细胞仪测定23例ACS患者(ACS组)和31例稳定型心绞痛(SA)患者(SA组)入院时外周血CD4 ,CD8 T淋巴细胞CD28、CTLA4、B7-1分子的表达,同时选健康人群15例作为对照(对照组)。结果:与对照组相比,SA组及ACS组发病时CD4 ,CD8 T淋巴细胞表面共刺激分子CD28均显著升高(均P<0.01);SA组与ACS组比较差异无统计学意义。与对照组相比,SA组CD4 ,CD8 T淋巴细胞表面CTLA4表达均显著升高(P<0.01);而ACS组CD4 ,CD8 T淋巴细胞表面CTLA4表达均显著下降(P<0.01)。各组B7-1的表达差异无统计学意义。结论:①共刺激分子B7-1:CD28/CTLA4途径参与了冠心病的发病过程;②ACS的强烈的炎症反应与CT-LA4的低表达有关。     

肾移植排斥反应外周血CD28、CD40共刺激通路相关分子的表达

目的探讨肾移植受者外周血T细胞CD28和CD40共刺激通路相关分子的表达及其与排斥反应的关系。方法采用流式细胞仪检测了42例肾移植受者外周血CD28、CTLA4、CD40及CD40L分子在CD4     

Graves病甲状腺共刺激分子GL50-ICOS的表达及其免疫病理意义

目的 研究共刺激分子GL50-ICOS在Graves病(GD)甲状腺组织中的表达及其免疫病理意义.方法运用细胞培养、流式细胞术、RT-PCR、Western印迹和免疫组化技术,检测GD甲状腺组织和原代培养的甲状腺滤泡细胞(TFC)共刺激分子GL50和ICOS的表达.结果 (1)与正常同龄对照相比,CD4+CD28-T细胞群体在GD患者外周血中显著升高,其表面ICOS表达上调.(2)RT-PCR显示,GD患者甲状腺组织中有ICOS mRNA表达,与对照非毒性甲状腺肿(NTG)组相比具有统计学差异(P＜0.01).(3)Western印迹显示,GL50蛋白在10例GD患者组织中全部表达,较对照组差异有统计学意义(P＜0.01).(4)与对照甲状腺腺瘤组相比,GL50在20例GD患者组织切片中全部检出,而对照组无阳性表达(P＜0.01).(5)炎性细胞因子刺激体外原代培养的甲状腺滤泡细胞表面上调表达GL50(P＜0.05).结论共刺激分子GL50-ICOS在Graves病甲状腺组织异常表达.     

负性共刺激分子介导免疫下调与免疫逃逸的关系

T细胞充分活化需要识别双信号系统,即主要组织相容性复合体-抗原肽提供的第一信号和共刺激分子提供的第二信号.共刺激分子在调节T细胞的免疫应答中起关键作用,其中负性共刺激分子介导的免疫下调在病毒、细菌、肿瘤细胞等的免疫逃逸中发挥重要作用,也可能与寄生虫慢性感染的免疫逃逸有关.该文就负性共刺激分子介导免疫下调与免疫逃逸关系的...     

T淋巴细胞共刺激分子在支气管哮喘患者发病中的作用

研究T淋巴细胞表面共刺激分子CD154、CD152 介导的信号途径,对支气管哮喘(简称哮喘)患者外周血T淋巴细胞产生白细胞介素13(IL 13)及B淋巴细胞分泌IgE抗体的影响,以明确这些共刺激信号途径在哮喘发生、发展过程中的作用特点。对象与方法　从我院门诊及住院患者中筛选出对蒿草过敏者15例,男8例,女7例,平均年龄(40±16 )岁,均符合中华医学会呼吸病学分会哮喘学组制定的诊断标准[1] ,酶联免疫吸附试验(ELISA)检测出血清中蒿草抗原的特异性IgE高出健康对照组2倍以上。健康对照组16名,男8名,女8名,平均年龄(38±18)岁,为本院健康自愿者。…     

负性共刺激分子介导免疫下调与免疫逃逸的关系

T细胞充分活化需要识别双信号系统,即主要组织相容性复合体-抗原肽提供的第一信号和共刺激分子提供的第二信号.共刺激分子在调节T细胞的免疫应答中起关键作用,其中负性共刺激分子介导的免疫下调在病毒、细菌、肿瘤细胞等的免疫逃逸中发挥重要作用,也可能与寄生虫慢性感染的免疫逃逸有关.该文就负性共刺激分子介导免疫下调与免疫逃逸关系的研究进展作一综述.     

ITP患者外周血淋巴细胞共刺激分子表达及意义

目的　研究共刺激分子在特发性血小板减少性紫癜 (ITP)外周血淋巴细胞的表达 ,并探讨其发病机制。方法　应用流式细胞术检测 2 8例 ITP患者及 15例正常对照者的外周血淋巴细胞 CD80 、CD2 8、CD86 表达 ,用酶联免疫吸附法检测血小板相关抗体 (PAIg G)水平 ;并与患者的临床资料进行相关性分析。结果　 1ITP组外周血淋巴细胞 CD2 8表达率降低 ,CD80 表达率略增加 ,与对照组均无统计学差异 ;CD86 表达率明显高于对照组 (P<0 .0 1)。 2 ITP组 2 1例 PAIg G水平升高 ,均值为 (197.39± 6 7.81) ng/ 10 7PA。 3ITP组外周血淋巴细胞 CD86 表达率与其巨核细胞数值呈正相关 (P<0 .0 5 )。结论　 ITP患者外周血淋巴细胞共刺激分子 CD2 8、CD80 表达无缺陷 ;CD86 表达明显增加。表明 CD86 可能参与 ITP的发病机制 ,应用抗 CD86 单克隆抗体方法可能治疗 ITP。     

免疫治疗对支气管哮喘小鼠树突细胞共刺激分子的影响

目的应用卵白蛋白(OVA)建立特异性免疫治疗小鼠模型,探讨特异性免疫治疗对支气管哮喘(简称哮喘)小鼠树突细胞(DC)表面分子CD80、CD86表达的影响。方法120只BALB/c小鼠按随机数字表法分为哮喘模型组(A组)40只:用0.1%OVA 10μg连续腹腔注射(共70μg)及1%OVA雾化吸入(共300 mg);免疫治疗模型组(B组)40只:用与A组同剂量的OVA致敏和激发,同时连续尾根部皮下注射OVA 1 mg;对照组(C组)40只:以磷酸盐缓冲液代替OVA,其余同A组。留取各组肺组织切片,经苏木精-伊红(HE)染色观察炎症反应;用酶联免疫吸附测定(ELISA)法检测血清OVA特异性免疫球蛋白IgE(sIgE)及脾脏T淋巴细胞中白细胞介素2(IL-2)和IL-4的分泌。分离各组小鼠脾脏DC,用流式细胞仪检测其表面CD80、CD86分子表达。分离正常小鼠脾脏T淋巴细胞,与上述各组DC共培养;用ELISA法检测T淋巴细胞IL-4、IL-5的分泌量;用3H-胸腺嘧啶核苷(3H-TdR)掺入法检测其增殖反应。结果(1)B组小鼠肺组织中支气管及血管周围以大量淋巴细胞和嗜酸粒细胞为主的炎性细胞浸润明显轻于A组,但并未完全消失;A组血清sIgE吸光度(A)值为712±129,B组为124±59,C组为20±13,A、C组间比较差异有统计学意义(P<0.05),B、C组比较差异无统计学意义(P>0.05)。B组T淋巴细胞分泌IL-2、IL-4水平分别为(8±3)、(8.4±4.3)pg/m l,A组分别为(22±8)、(32.4±12.1)pg/m l,C组分别为(6±4)、(5.1±1.1)pg/m l,A、B两组比较差异有统计学意义(P<0.05),B、C组间比较差异无统计学意义(P>0.05);(2)B组DC表面CD86、CD80阳性表达率分别为58.23%、95.63%,A组分别为77.59%、96.98%,C组分别为77.37%、77.84%;(3)与B组DC共培养的正常小鼠T淋巴细胞体外经OVA刺激后,IL-4、IL-5水平分别为(10.8±2.3)、(18.8±3.8)pg/m l,A组分别为(17.3±4.7)、(35.7±7.9)pg/m l,C组分别为(5.7±2.7)、(11.0±2.2)pg/m l,A、B两组比较差异有统计学意义(P<0.05),B、C组间比较差异无统计学意义(P>0.05);B组DC与正常小鼠T淋巴细胞共培养时,刺激指数(SI)为3.8±0.7,A组为11.5±3.2,C组为5.8±1.5,A、B组间比较差异有统计学意义(P<0.05);B、C组间比较差异无统计学意义(P>0.05)。结论建立了OVA特异性免疫治疗小鼠模型;DC表面CD86分子表达的下调可能是OVA特异性免疫治疗诱导T淋巴细胞功能丧失的机制之一。     

类风湿关节炎患者外周血B淋巴细胞共刺激分子的表达及其意义

目的 初步探讨类风湿关节炎(RA)患者免疫功能紊乱的机制。方法 采用流式细胞仪检测RA患者外周血B淋巴细胞共刺激分子CD80、CD86、CD40的表达并用ELISA检测RA患者血清和关节滑膜液中Th1细胞分泌的细胞因子白细胞介素(IL)-2、干扰素-γ和Th2细胞分泌的细胞因子IL-6、IL-10的水平。结果 RA患者外周血B淋巴细胞CD86表达比正常对照组明显下降(P＜0．01),B淋巴细胞CD40表达比正常对照组明显增多(P＜0．05),而B淋巴细胞CD80表达与正常对照组之间差异无显著性(P＞0．05),同时RA患者血清及滑膜液中IL-2、干扰素-γ的水平比正常对照组明显升高(P＜0．01或P＜0．05),而IL-6、IL-10的水平降低(P＜0．01或P＜0．05)。结论 B淋巴细胞共刺激分子CD86、CD40的异常表达可能与RA患者Th1／Th2细胞分泌的细胞因子失衡密切相关,这将为临床治疗RA提供新的思路。     

树突细胞表面共刺激分子在小鼠哮喘模型中的作用及其机制的研究

目的　探讨树突细胞 (DC)表面共刺激分子在哮喘发病中的作用及其机制。方法BALB/c小鼠 12 0只 ,分为 3组 (每组 4 0只 ) :哮喘组 [用卵蛋白 (OVA)致敏和激发建立小鼠哮喘模型 ]、磷酸盐缓冲液 (PBS)对照组 (以PBS替代OVA致敏和激发 )、健康对照组。对 3组小鼠取肺组织做病理观察 ,行支气管肺泡灌洗液 (BALF)计数细胞并分类 ,ELISA测血清特异性IgE(sIgE)及脾脏T淋巴细胞白细胞介素 (IL) 4和IL 5水平。分离、培养脾脏DC ,用流式细胞仪 (FACS)测CD11c,鉴定表型。用FACS分析哮喘小鼠DC表面共刺激分子CD80 、CD86表达的变化 ;并将哮喘小鼠的DC与健康小鼠的T淋巴细胞共同培养 ,ELISA测培养上清IL 4和IL 5水平。结果　哮喘小鼠肺组织表现为以嗜酸性粒细胞、淋巴细胞浸润为主的炎症改变 ,BALF中嗜酸性粒细胞计数显著增加 (P <0 0 1) ,血清sIgE水平显著升高 (P <0 0 1) ,脾脏T淋巴细胞IL 4和IL 5的水平亦显著升高 (P <0 0 1)。与PBS对照组比较 ,哮喘小鼠CD80 表达上调 ,CD86无明显变化。哮喘小鼠DC能刺激健康小鼠T淋巴细胞IL 4和IL 5水平升高 (P <0 0 1)。结论　DC可能通过上调CD80 在哮喘Th2类免疫反应的维持和放大中发挥作用。     

他克莫司对类风湿关节炎关节滑膜液淋巴细胞协同刺激分子的作用

目的研究他克莫司对类风湿关节炎（RA）患者关节滑膜液淋巴细胞协同刺激分子的作用．并初步探讨其免疫抑制的机制。方法分离培养RA关节滑膜液淋巴细胞，经100nmol/L的他克莫司处理后．用流式细胞仪检测T淋巴细胞协同刺激分子CD28、CD154（CD40L）和B淋巴细胞协同刺激分子CD80、CD86、CD40的表达。同时检测T、B淋巴细胞活化标志CD69、CD25、HLA—DR和CD71的表达．用酶联免疫吸附法（ELISA）检测淋巴细胞培养上清Th1细胞分泌的细胞因子白细胞介素（IL）-2、干扰素（IFN）-γ和Th2细胞分泌的细胞因子IL-6、IL-10的水平，并设不加他克莫司处理的为对照组。结果经他克莫司处理后。T淋巴细胞的CD154（CD40L）和B淋巴细胞的CD86表达阳性率低于对照组（P〈0．05），而T淋巴细胞的CD28和B淋巴细胞的CD80、CD40表达阳性率与对照组相比差异无统计学意义；T、B淋巴细胞的HLA—DR表达阳性率明显低于对照组（P〈0．01），而CD69、CD25、CD71表达阳性率与对照组相比差异无统计学意义；淋巴细胞培养上清Th1细胞分泌的细胞因子IL-2、IFN-γ和Th2细胞分泌的细胞因子IL-6、IL-10的水平与对照组相比均显著下降（P〈0．01）。结论他克莫司能明显抑制RA关节滑膜液淋巴细胞的活化，降低Th1细胞因子和Th2细胞因子的分泌水平，这种作用可能是通过下调淋巴细胞的协同刺激分子的表达而实现。     

Expression of the Fas antigen on primary human leukemia cells

Summary The antigen defined by the monoclonal antibody anti-Fas can mediate apoptosis, is associated with the receptor for tumor necrosis factor, and is expressed on a limited number of human tissues. In this study we analyzed the expression of Fas on primary human leukemic cells and on mononuclear cells from other hematologic disorders. A total of 95 samples of blood or bone marrow were studied by indirect immunofluorescence. These samples included the normal controls, 47 cases of acute myelogenous leukemia (AML), 11 cases of acute lymphoblastic leukemia (ALL), 21 cases of leukemic lymphoma, seven cases of chronic myelogenous leukemia (CML), five cases of plasma cell leukemia or multiple myeloma, and five cases of myelodysplastic or myeloproliferative syndromes. Normal controls were negative without exception. Among AML, 13/47 cases (28%) were positive; among ALL, 1/11 cases (9%) was positive; among leukemic lymphomas, 3/21 cases (14%) were positive. In a case of plasma cell leukemia which strongly expressed the Fas antigen, we demonstrated that the antibody mediates cell lysis, which was synergistically enhanced by the addition of rabbit complement. In patients with AML, Fas positivity had no obvious clinical relevance. Taken together, our results show that approximately 30% of cases of AML and occasionally other leukemias express the Fas antigens, whereas normal controls are negative in our test system. These findings may be useful in the treatment of refractory leukemias or may permit the purging of autologous transplants.Presented in part at the 34th annual meeting of the American Society of Hematology, Anaheim, CA, December 1992     

Expression of class II major histocompatibility complex antigens on pancreatic B cells in the NOD mouse

Summary To elucidate the role of class II major histocompatibility complex antigen expression on pancreatic B cells in the development of diabetes in the non-obese diabetic (NOD) mouse, indirect immunofluorescence was employed for I-A staining on Bouin-fixed pancreas sections of NOD mice (I-A of which was reported as d), B10.GD (I-A, d), BALB/c (I-A, d) and C3H/He (I-A, k). I-A positive islets were observed in all NOD mice examined. Positive reaction was detected in islets both with and without lymphocytic infiltrations. Double staining with anti-insulin, glucagon, somatostatin or pan creatic polypeptide antibodies revealed that I-A positive cells corresponded with insulin cells, while other types of pancreatic islet cells were virtually negative for I-A. Weaker staining was seen in islets of B10.GD and, to a lesser extent, in those of BALB/c mice. C3H/He mouse islet cells showed no I-A expression. These results demonstrated the expression of I-A antigens on pancreatic B cells in the NOD mouse.     

Decreased expression of B7 costimulatory molecules and major histocompatibility complex class-I in human hepatocellular carcinoma

BACKGROUND AND AIM: We analyzed the expression of antigen-processing and antigen-presenting molecules in surgically resected fresh samples of human hepatocellular carcinoma (HCC) tissue to elucidate a mechanism of immune escape. We also examined the expression of interleukin (IL)-10 protein, which might act to downregulate expression of antigen-processing and antigen-presenting molecules. METHODS: Twenty-eight HCC samples obtained by surgical resection were analyzed for the expression of beta2-microglobulin, heat-shock protein (HSP)-70, human leukocyte antigen (HLA) class-I, CD80 (B7-1), CD86 (B7-2) and IL-10 by immunostaining. RESULTS: Beta2-microglobulin and HSP-70 were preserved in all samples. In contrast, the expression of HLA class-I molecules was significantly reduced according to lowering in the histological grading of tumor differentiation (P = 0.024). Furthermore, B7-1 and B7-2 expression was reduced in tumor cells compared with corresponding areas of liver tissue without malignant involvement irrespective of the histological grading of tumors (21% and 36%, respectively). Although IL-10 protein was expressed in 54% of HCC, no relationship between the expression of IL-10 and downregulation of B7-1, B7-2, and HLA class-I was evident. CONCLUSION: These findings suggest the potential role of B7 co-stimulatory molecules and HLA class-I molecules in facilitating HCC escape from immune surveillance without the involvement of IL-10.     

Rhinovirus infection induces major histocompatibility complex class I and costimulatory molecule upregulation on respiratory epithelial cells

Human respiratory epithelial cells may act as antigen-presenting cells during respiratory viral infections. In addition to major histocompatibility complex (MHC) molecules, antigen presentation requires participation of costimulatory molecules. Here the authors investigated class I and class II antigens and B7-1 and B7-2 costimulatory molecule expression in human A549 pulmonary epithelial cells and primary bronchial epithelial cells (HBECs) at baseline and after rhinovirus infection. Constitutive expression of MHC class I and B7-1 molecules was observed on both cell types. MHC class I molecules were up-regulated by rhinovirus infection, while B7-1 was up-regulated only on A549 cells. B7-2 molecules were constitutively expressed at a low level and were up-regulated by rhinovirus only on HBECs. Rhinovirus induction of antigen-presenting molecule expression on A549 cells was accompanied by cellular activation in terms of induction of release of the chemokines RANTES and Groalpha. These data show that respiratory epithelium expresses full antigen-presentation machinery and that rhinovirus infection up-regulates this expression.     

High expression of costimulatory molecules correlates with low relapse-free survival probability in acute myeloid leukemia (AML)

Costimulatory molecules such as lymphocyte function-associated antigen (LFA)-1 (CD11a), LFA-3 (CD58), intercellular adhesion molecule (ICAM)-1 (CD54), neuronal cell adhesion molecule (NCAM) (CD56), B7-1 (CD80), or B7-2 (CD86) are important regulatory elements in healthy immunological cascades, but their role in acute myeloid leukemia (AML) has only been rarely investigated. We studied their expression on mononuclear bone marrow (BM) cells from 105 patients with AML at initial diagnosis and evaluated their prognostic significance. Fluorescence-activated cell sorter (FACS) analyses were performed using antibodies directly conjugated with fluorescein. A BM sample was considered positive if more than 20% of the cells in the blast containing gate expressed the respective marker. The surface expression of CD11a (27 of 29 cases positive with an average of 71% positive blasts; 27⁺/29, 71%), CD54 (23⁺/33, 37%), CD56 (24⁺/93, 20%), CD58 (29⁺/29, 95%), CD80 (13⁺/28, 30%), and CD86 (19⁺/29, 39%) was measured. The expression of these markers in different French-American-British (FAB) classification types (M0–M5) was heterogeneous, except for CD56, which showed a higher proportion of positive cells in monocytic subtypes of AML. In addition, cases with a poor risk karyotype as well as patients succumbing to early death after double induction therapy according to the AML Cooperative Group (CG) protocol were characterized by a high expression of CD56. Relapse-free survival analyses demonstrated that patients with more than 8% CD56⁺ cells in the BM relapsed significantly sooner. CD54 was preferentially expressed in AML M4_eo and in addition in favorable cytogenetic risk groups and in cases that had responded to AML-CG therapy. Only very high proportions (>60%) of CD54⁺ cells were associated with a lower probability for relapse-free survival. CD80 and CD86 expressions were similar in all FAB types. Patients who had responded to AML-CG therapy showed higher CD80 proportions and lower CD86 proportions compared to the nonresponder group. Whereas cases with more than 15% CD80⁺ cells had a significantly lower probability for relapse-free survival, only cases with more than 65% CD86⁺ were characterized by a significantly lower probability for relapse-free survival. Expression profiles of CD11a and CD58 were not associated with specific FAB types or prognostically relevant groups. We can conclude: (1) Expression of costimulatory molecules in AML is very variable. This reflects the great diversity of immunophenotypes in AML. (2) CD56 is mainly expressed in monocytic subtypes of AML. CD56⁺ subtypes of AML seem to be a separate entity with a worse prognosis independent of the karyotype. (3) High expression of some costimulatory molecules correlates with a worse prognosis concerning relapse-free survival times.     

慢性肾炎患者外周血共刺激分子CD28和CD137水平变化研究

目的　研究慢性肾炎患者外周血共刺激分子CD2 8和CD1 37的表达特点及其在慢性肾炎免疫病理机制中的作用。方法　采用免疫荧光标记和流式细胞仪分析 ,对 5 2例慢性肾炎患者外周血共刺激分子CD2 8、CD1 37和T淋巴细胞亚群的表达进行检测。结果　慢性肾炎患者T细胞亚群明显失衡 ,表现为CD4减少 ,CD8增加 ,CD4 CD8比值显著降低。共刺激分子CD2 8表达显著低于正常对照组 (P <0 0 1) ,且CD+4 CD+2 8T细胞和CD+8CD+2 8T细胞均显著减少 (P <0 0 1)。共刺激分子CD1 37表达显著高于正常对照组 (P <0 0 1)。结论　慢性肾炎患者外周血T细胞亚群失衡和T细胞活化所必需的共刺激分子CD2 8、CD1 37异常表达 ,可能在慢性肾炎发生和病变进展中起着重要作用