再生障碍性贫血患儿血清TNF-α、IFN-γ和T细胞亚群检测的临床意义

目的：探讨再生障碍性贫血患儿血清TNF-α、IFN-γ和T细胞亚群的变化及意义。方法：应用放免法、酶联免疫双抗体夹心法和单克隆抗体检测法对33例再生障碍性贫血患儿进行血清TNF-α、IFN-γ和T细胞亚群检测,并以35名正常健康儿童作对照。结果：再生障碍性贫血患儿血清TNF-α、IFN-γ水平均非常显著地高于正常儿组（P〈0.01）,而CD3、CD4、CD4/CD8比值又非常显著地低于正常儿组（P〈0.01）。结论：再生障碍性贫血患儿血清TNF-α、IFN-γ和T细胞亚群的变化可作为病情和预后判断的重要检测指标。     

急性多发性神经根神经炎外周血T淋巴细胞亚群的研究

一般认为,急性多发性神经根神经炎与感染及免疫调节紊乱有关。为探讨T淋巴细胞在发病中的作用,我们用抗人T淋巴细胞单克隆抗体OKT3,OKT4和OKT8检测了23例该病患者外周血T细胞亚群。     

再生障碍性贫血患者外周血T细胞亚群和T细胞受体表达水平及其意义

采用流式细胞仪及间接免疫荧光法检测30例再生障碍性贫血患者外周血中T细胞亚群及T细胞表面受体表达水平,并与健康对照组相比,结果表明:70％再障患者存在CD4/CD8比例倒置及CD8~+％异常增高;50％再障患者外周血γδ-T细胞亚群及其在T淋巴细胞总体中所占比例均显著增高;而αβT细胞亚群及TirA~+细胞百分率与正常对照组相比无显著性差异。提示:半数以上再障患者外周血中存在异常增多的γδT细胞及Ts细胞亚群。并可通过其直接或间接作用抑制造血,从而导致再障的发生。     

普通变量对单克隆抗体测定T细胞亚群的影响

用异种抗血清和单抗已阐明,T细胞功能仅限于带某些分化抗原的T细胞亚群.已证明疾病变化与体内T_H/T_S细胞比例的变化有关.因此定量分析淋巴细胞亚群已成为临床诊断的手段.而普通变量对淋巴细胞亚群的影响尚未详细研究.作者测定了两性T细胞亚群的正常值,研究了一个时期内T细胞亚群的个体变量,并研究了年龄、身体锻练、饮食、取血时间、血样保存期限及冰冻分离淋巴细胞等对亚群的影响.为说明这些问题,作者使用了普通单抗如OKT3、OKT4、OKT8及OKIal(主要鉴定B细胞)     

小儿单纯性肾病T淋巴细胞亚群变化的初步观察

本文通过OKT系统检测34例单纯性肾病的周围血中T淋巴细胞亚群的变化,并对7例肾病活动期和缓解期进行了动态观察,结果发现肾病活动期存在着明显的T细胞亚群紊乱,T_3~ 、T_4~ 细胞减少,T_8~ 细胞增多,T_4、/T_8比值低于正常健康儿童对照组,经过激素和     

人T细胞亚群研究进展

根据分化抗原不同,通常可以将人外周血淋巴细胞分成OKT4~+和OKT8~+两个亚群(简称T4~+和T8~+)。一般认为T4~+亚群具有辅助和诱导功能(T_H/T_i),T8~+亚群具有抑制和细胞毒功能(T_S/TC)。近年来,由于单克隆抗体技术的发展,已获得了一些新的人T细胞单克隆抗体(下称     

5′-核苷酸酶活性比色分析

血清及细胞膜上的5′—核苷酸酶活性定量测定法简单,快速、敏感,一般实验室可测,血清中此酶的定量测定能用于肝脏疾病的诊断。因淋巴细胞膜上结合有此酶,所以可用于白细胞亚群的临床检测及培养的淋巴细胞的测定,也能用于白血病及恶性淋巴瘤的研究。据报导,原发性低丙种球蛋白血症(CVK);几种联合性免疫缺陷病;T急性淋巴细胞性白血病、B慢性淋巴性白血病及其他B细胞疾病(例如:B急性淋巴性白血     

人脂肪间充质干细胞与再生障碍性贫血患者T淋巴细胞的体外共培养

背景:目前已有临床应用造血干细胞联合间充质干细胞输注治疗再生障碍性贫血的报道。目的:观察人脂肪间充质干细胞对再生障碍性贫血T淋巴细胞分泌功能的调节作用。方法:从健康人脂肪中提取并培养人脂肪间充质干细胞,从再生障碍性贫血患者外周血中分离T淋巴细胞,将两者共培养后,以健康志愿者的T淋巴细胞单独培养及与人脂肪间充质干细胞共培养做对照。ELISA法检测上清白细胞介素2、白细胞介素4、白细胞介素10和γ-干扰素的水平,Realtime RCR和Western blot测定T-bet和GATA-3的表达。结果与结论:共培养7 d后,间充质干细胞+再生障碍性贫血T淋巴细胞组Th1类细胞因子γ-干扰素、白细胞介素2水平明显低于再生障碍性贫血T淋巴细胞组(P0.05),Th2类细胞因子白细胞介素4、白细胞介素10水平明显高于再生障碍性贫血T淋巴细胞组(P0.05);间充质干细胞+再生障碍性贫血T淋巴细胞组T-bet mRNA及蛋白表达水平均低于再生障碍性贫血T淋巴细胞组,GATA-3 mRNA及蛋白水平均高于再生障碍性贫血T淋巴细胞组。提示人脂肪间充质干细胞在体外对再生障碍性贫血患者T淋巴细胞具有免疫调节作用,其机制可能是通过调节转录因子T-bet和GATA-3的表达从而抑制Th细胞向Th1细胞方向分化。     

传染性单核细胞增多症患儿外周血淋巴细胞亚群的分布及意义

探讨传染性单核细胞增多症(infectious mononucleosis,IM)患儿外周血中淋巴细胞亚群的分布及临床意义。采用流式细胞术检测131例IM患儿和50例体检健康儿童外周血中淋巴细胞亚群的表达,131例IM患儿中有26例患儿收集到对应的丙种球蛋白(intravenous immunoglobulin,IVIG)治疗后全血,流式细胞术检测淋巴细胞亚群在IM患儿IVIG治疗前后分布比例的差异。结果显示,与健康对照组相比,IM患儿CD3~+T细胞和CD8+T细胞的表达水平明显增高(均P<0.000 1),CD4~+T细胞、NK细胞、B淋巴细胞及CD4~+/CD8+T细胞比值水平明显降低(均P<0.000 1)。IM患儿急性期CD4~+、CD8~+T细胞及CD4~+/CD8~+T细胞比值升高及降低水平与患儿外周血异型淋巴细胞升高的百分率呈现正相关趋势。与IVIG治疗前相比,IM患儿IVIG治疗后外周血CD3~+T细胞和CD8~+T细胞的表达水平有所减低,但仍高于健康对照组(均P<0.01),治疗后CD4~+T细胞、NK细胞、B淋巴细胞及CD4~+/CD8~+T细胞比值水平增高,但其中治疗后CD4~+T细胞、B淋巴细胞及CD4~+/CD8~+T细胞比值水平仍显著低于健康对照组(均P<0.01)。综上所述,检测淋巴细胞亚群对IM患儿的细胞免疫状况评估、辅助诊断和指导治疗具有重要的临床意义。     

川崎病患儿精细免疫分型分析及其临床意义

目的分析急性期川崎病(KD)患儿淋巴细胞各亚群的数量变化,探讨对临床诊治的意义。方法前瞻性分析重庆医科大学附属儿童医院收治的20例急性期KD患儿的临床资料以及外周血淋巴细胞各亚群(CD3~+、CD4~+及各亚群、CD8~+及各亚群、TCRαβ~+DNT、TCRγδ~+、CD4/CD8、CD19~+及各亚群、CD16~+CD56)的数量,并选取20例健康儿童为对照组,比较2组间的变化并分析临床指标与淋巴细胞各亚群的关系。结果 20例患儿的临床表现均符合典型KD的诊断标准,与对照组相比,KD组的CD8 naive、TCRαβ~+DNT、CD4/CD8、CD19~+及各亚群水平均明显升高(P0.05),而CD4 EM(CD4~+CD45RA-effector memory T cells)、CD4 Temra(CD4~+CD45RA~+effector memory T cells)、CD8~+T、CD8 CM(CD8~+CD45RA-central memory T cells)、CD8 Temra(CD8~+CD45RA~+effector memory T cells)、CD16~+CD56水平明显降低(P0.05)。有冠状动脉损害的KD患儿与未出现冠状动脉损害的KD患儿相比,TCRγδ~+T细胞水平明显升高(P0.05),CD4 Temra细胞水平有下降趋势。合并呼吸道或消化道系统并发症的KD患儿与未合并呼吸道或消化道系统并发症的KD患儿相比,CD3~+T、CD8 CM、CD8 EM、CD8 Temra细胞水平明显下降(P0.05),Transitional B细胞水平明显升高(P0.05)。结论急性期KD患儿的T、B淋巴细胞均处于异常活化的状态,其中TCRγδ~+T和CD4 Temra细胞与川崎病合并冠状动脉损害相关,CD3~+T、CD8 CM、CD8 EM、CD8 Temra和Transitional B细胞与川崎病合并呼吸道或消化道系统并发症相关。     

溶组织阿米巴提出液中的T诱导/辅助细胞有丝分裂原

溶组织阿米巴提出液(E.H.e)中含有对 T 细胞的有丝分裂原。为了进一步研究 E.h.e对 T 细胞亚群的有丝分裂作用,将纯化的 T4~+及 T8~+亚群细胞分别与 E.h.e 温育,以~3H-TdR掺入为指标,观察各细胞亚群对 E.h.e 的增殖反应,结果发现仅 T4~+细胞对 E.h.e 有增殖反应,说明 E.h.e 中含有的有丝分裂原分子具有对 T4~+亚群特异性。实验还证明,E.h.e 刺激 T4~+细胞增殖是通过 IL2来实现的。     

Heterogeneity of concanavalin A-induced suppressor T cells in man defined with monoclonal antibodies.

Human peripheral blood T lymphocytes were enriched for OKT4+ or OKT8+ subpopulations using complement mediated lysis with OKT8 or OKT4 monoclonal antibodies. These subpopulations and unfractionated T cells were separately stimulated with concanavalin A (Con A) for a period of 48 hr and were then examined for their suppressive influence on proliferative response of autologous T cells to phytohaemagglutinin (PHA) or allogeneic non-T cells. Con A-activated unfractionated T cells, OKT4+ and OKT8+ T cell subsets markedly suppressed both these responses. Both OKT4+ and OKT8+ T cell subsets when enriched following Con A-activation of unfractionated T cells also caused significant suppression of proliferative responses of autologous T cells to PHA and allogeneic non-T cells in mixed lymphocyte cultures. The suppressive influence of Con A-activated T subsets was abolished by irradiation (2,000 rad) of activated cells. These studies indicate that Con A-induced suppressor T cells are heterogeneous. Precursors of Con A-induced suppressor T cells appear to reside in both OKT4+ and OKT8+ T cell populations.     

Lymphocyte subpopulations during acute and convalescence phases of malaria

Lymphocytes of normal healthy persons were separated from blood by Ficoll-Hypaque gradient centrifugation and iron-magnet application. peripheral blood lymphocytes (PBL) were stained by various dye-labeled monoclonal antibodies. Cells positive for specific surface markers were enumerated by a fluorescence activated cell sorter (FACS) and fluorescence microscope (FM). The results revealed that the percentages of cells positive with one monoclonal antibody counted by these two techniques were similar while the percentages of cells with double staining were higher when counted by FACS than by FM. Lymphocyte subpopulations of 18 patients infected with Plasmodium falciparum during acute and convalescence period were studied. Lymphocytopenia occurred during the acute infection while total white blood cell counts were normal. PBL of the patients were stained with OKT3, OKT4, OKT8, Leu-11 and a combination of Leu-7, Leu-1 monoclonal antibodies. The absolute numbers of all lymphocyte subpopulations were decreased during the acute infection while T8 positive cells were decreased in both percentage and absolute number. Thus T4:T8 ratio (1.7:1) became higher than normal (1.3:1) at this period. During convalescence phase, absolute numbers and percentages of Leu-7+, Leu-1+ and perhaps Leu-7+, Leu-11- cells which had low NK cell activity were significantly higher than during acute illness. The finding might explain why the NK cell activity was low during the convalescence period.     

T cell subpopulations in CLL: methods of T cell enrichment artificially alter proportions of OKT4 and OKT8 positive cells

There is growing speculation about the meaning of reported imbalances in subpopulations of T lymphocytes in patients with chronic lymphocytic leukaemia (CLL). This study compared two techniques for producing T cell enriched subpopulations from both patients with CLL and normal individuals and the effects of these techniques on relative proportions of OKT4 and OKT8 positive cells. A sheep red cell rosetting technique resulted in significantly larger OKT8 and smaller OKT4 positive populations than did nylon wool column elution. Similar results were obtained in normal individuals. The nylon wool column elution technique produced less distortion of unfractionated OKT4/OKT8 ratios in normals than did the rosetting technique. Studies of T lymphocyte subpopulations should be interpreted with great caution as the methods used to study them can influence the results.     

肿瘤病人外周血T细胞及其亚群和NK细胞活性变化的研究

一般认为,T细胞和NK细胞可能是构成机体肿瘤免疫功能的重要因素。肿瘤病人外周血T细胞及其亚群和NK细胞活性的变化,国内外均有报道。但对同一肿瘤病人T细胞及其亚群和NK细胞活性变化的对比研究报道得较少。本文用OKT系列的单克隆抗体检测T细胞及其亚群,用~(51)Cr-释放试验检测NK细胞活性,对同一肿瘤病人外     

Immunoregulatory T cell subpopulations in patients with scleroderma using monoclonal antibodies.

Twenty-eight patients with scleroderma were compared with 22 healthy age-matched subjects. Monoclonal antibodies were used to detect the whole T cell population (OKT3), T helper cells (OKT4), and T suppressor/cytotoxic cells (OKT8) by indirect immunofluorescence on isolated peripheral blood mononuclear cells. A subset of scleroderma patients (i.e. 30% or eight of 28 patients) exhibited an elevated ratio of OKT4/OKT8 cells which could be accounted for, mainly by a reduction in OKT8 cells compared with controls. The scleroderma patients with an elevated OKT4/OKT8 ratio tended to be younger, have a shorter disease duration and more extensive skin involvement than patients with a normal OKT4/OKT8 ratio. There was no correlation with the presence of autoantibodies, drug therapy, or HLA-DR type. In order to further determine whether this imbalance in immunoregulatory cell subpopulations was specific for scleroderma, we further studied 16 patients with psoriatic arthritis but without manifest autoimmunity and delineated a similar subset of patients with an elevated OKT4/OKT8 cell ratio (i.e. 38% or six of 16 patients). The results demonstrate similar immunoregulatory T cell imbalances in patients with scleroderma and psoriatic arthritis. These findings suggest that numerical imbalances in lymphocyte subpopulations may not be specific for autoimmune disorders.     

Utilization of monoclonal antibodies and immuno-scanning electron microscopy for the positive identification of human leukemic cells

Monoclonal antibodies generated against normal and leukemic human leukocytes were tested for their differential reactivity with leukemia and lymphoma cell lines as well as with circulating lymphoid and myeloid leukemic cells by means of immuno-scanning electron microscopy (immuno-SEM). Anti-T (OKT3), anti-mu-chain, anti-CALLA (J5), anti-BA-1, anti-BA-2, and anti-nonlymphoid (Mol) monoclonal antibodies were covalently conjugated to polystyrene latex microspheres (immunolatex), using a two-step glutaraldehyde reaction, and subsequently incubated with the various cell types. Cultured B-type Burkitt lymphoma cells (Daudi) and chronic lymphocytic leukemia (CLL) cells displayed extensive labeling with monoclonal anti-mu, anti-B1, and anti-BA-1 immunolatex conjugates, while cultured malignant T cells (HD-Mar) showed positive labeling with OKT3 immunolatex alone. Cultured myelomonocytic cells (GDM-1) and cells obtained from patients with acute myeloblastic (AML) and monoblastic leukemia (AMoL) labeled only with anti-Mol immunolatex, while cultured promyelocytic cells (HL-60) displayed far less labeling with this conjugate. Common-type acute lymphoblastic leukemia (C/ALL) cells were labeled predominantly with the J5 (anti-CALLA) and anti-BA-2 immunolatex conjugates. Evidence is presented indicating that immuno-SEM employing monoclonal antibodies is a reproducible technique which may be used in the study of leukocyte maturation and may provide additional information in the classification of poorly differentiated leukemias.     

Imbalance of T cell subpopulations in patients with chronic lymphocytic leukaemia of the B cell type.

T cell enriched mononuclear cells from the peripheral blood of 20 patients with histologically and immunologically defined chronic lymphocytic leukaemia of the B cell type (B-CLL) and 20 healthy individuals of various ages were investigated with T cell-specific monoclonal antibodies (OKT 4 and OKT 8) with regard to their subpopulation distribution. In B-CLL, a significant increase of lymphocytes reacting with OKT 8 could be demonstrated. Whereas there was a ratio of OKT 4 to OKT 8 of 1 X 72 in the control group, an OKT 4 to OKT 8 ratio of 0 X 67 was found in the B-CLL as a whole. With increasing clinical stage in accordance with the Rai scheme (Rai et al., 1975), a further displacement of this ratio in favour of OKT 8 positive cells was found. These results clearly show that, in peripheral blood of patients with B-CLL, an abnormal distribution pattern of circulating T cell subpopulations is present and that this also has prognostic relevance.     

Establishment of a human T-acute lymphoblastic leukemia cell line with a (16;20) chromosome translocation

A new T-cell line, Loucy, was established from the peripheral blood of a patient with T-cell acute lymphoblastic leukemia (T-ALL). The surface marker analysis of the cell line is OKT3+, OKT4+, THB4+, J5 +/-, OKT6-, TdT-, and HLA-DR-, indicating stage IV in T-cell lineage. Karyotype analysis revealed 45,X,5q-,t(16;20)(p12;q13). The translocation between chromosomes 16 and 20 has not been previously detected in ALL. This cell line may be of value in evaluating the role of t(16;20) in the etiology of T-ALL.     

Studies on immunoregulatory mechanisms in acute and chronic hepatitis B

Patients with acute hepatitis B and HBV-induced chronic hepatitis as well as normal control persons participated in the study. Hepatitis patients of both groups have decreased OKT4+/OKT8+T cell ratios due to an percental increase of OKT8+T cells in peripheral blood compared to the data of controls. Lymphocyte cultures of chronic hepatitis patients show reduced DNA synthesis after stimulation by allogeneic non-T cells, PHA, Con A and PWM. PWM-induced immunoglobulin secretion by B cells, determined by means of a reverse haemolytic plaque assay (RHPA) and a solid phase ELISA, showed comparable results in hepatitis B patients and controls. The AMLR, which is thought to reflect an autologous immunoregulatory phenomenon, is slightly impaired in cultures of hepatitis B patients in comparison to controls. Con A-induced suppressor cell activity on T cell reactions is decreased in hepatitis, whereas suppressor cell activity on B cell activation is within the same range as in cultures of controls. It is concluded from these data, that suppressor cell activity on T cell function is impaired in hepatitis B, whereas B cell functions and suppressor cell activity on B cell function are in the normal range. The results with the functional assays and the finding of increased proportions of OKT8+T cells in hepatitis B are considered to reflect properties of different T cell subpopulations, responsible for different immunoregulatory functions.