人淋巴细胞Fc段受体流式细胞仪测定

本文研究了正常人外周血淋巴细胞(PBL)表面IgGFc段受体(FcrR)和IgMFc段受体(FeHR)。细胞荧光标记与常规免疫荧光法不同,以Avidin-Biotin放大体系为荧光载体标记细胞,使对Fc段受体的测定更灵敏和特异。取48名健康成人静脉血,制备单个核细胞。细胞和AggIgG-Biotin(生物素化聚合IgG),IgM-Biotin(生物素化IgM)及Avidin-FITC(结合异硫氰酸荧光黄的Avidin)分二步孵育,用荧光标记淋巴细胞表面FcrR与FcμR。流式细胞仪分析结果显示,正常人FcrR+PBL (X±SD)为36.63±9.07％,FcμR+PBL为12,76±5.39％。我们用自己建立的Fc段受体(FcR)流式细胞仪分析法,对国人淋巴细胞表面FcrR与FcμR的研究,为探索FcR对免疫系统的调节作用,及一些免疫性疾病的发病机理奠定了一定基础。     

应用四色荧光标记流式细胞术检测外周血淋巴细胞亚群的变化

目的 探讨四色荧光标记流式细胞术(Flow cytometry,FCM)检测鼻咽癌患者服用云芝丹参胶囊后外周血淋巴细胞亚群:辅助性T淋巴细胞(CD4 ),抑制性和细胞毒T淋巴细胞(CD8 ),B淋巴细胞(CD19 ),NK(CD16 CD56 ) 细胞的绝对数和百分率等的变化.方法 收集鼻咽癌患者27例,设立中药组和安慰剂对照组,采用四色荧光标记流式细胞术对服用云芝丹参胶囊后的鼻咽癌患者外周抗凝全血的淋巴细胞亚群CD4 细胞、CD8 细胞、B细胞、NK细胞进行绝对计数和相对计数,并对两组结果进行比较分析.结果 四色荧光标记流式细胞术结果显示:接受放射治疗的鼻咽癌患者在服用云芝丹参胶囊16周后,外周血T淋巴细胞的绝对数和百分率,以及Ts、Th细胞绝对数的下降均明显低于安慰剂组(P＜0.05);中药组和安慰剂组NK细胞百分率的变化均显著增高(P＜0.05); 但两组NK细胞绝对值的变化不存在明显差异(P＞0.05).结论 云芝丹参能明显减轻放射治疗对鼻咽癌患者的淋巴毒性.     

单链抗体经流式细胞仪检测细胞表面受体

用流式细胞仪检测两个自然杀伤细胞抑制受体的单链抗体（ＮＫＡＴ２－ｓｃＦｖ；ＮＫＡＴ４－ｓｃＦｖ），识别细胞表面受体分子的能力。单链抗体是吸附固相的抗原筛选一噬菌体抗体库而获得。结果显示，含ＮＫＡＴ２－ｓｃＦｖ的细菌上清及胞质提取物均可识别到表达在细胞表面的ＮＫＡＴ２受体分子，但含ＮＫＡＴ４－ｓｃＦｖ的细菌上清及胞质提取物均未成功     

流式细胞仪检测巨细胞病毒感染

巨细胞病毒(CMV)是巨细胞包涵体病的病原体, 是疱疹病毒家族的DNA病毒, 具有典型的疱疹病毒结构.人群中CMV感染非常广泛, 60%-90%的成人已有CMV抗体.     

流式细胞仪国产鞘液的研制

流式细胞仪(ＦＣＭ)已成为当代分析细胞学领域检测细胞分子特征的主要工具之一,在医学、生物学和肿瘤学等学科中得到广泛应用〔１,２〕。ＦＣＭ所用的鞘液是维持仪器正常运转的主要消耗试剂,其主要作用是经荧光染色的单细胞悬液和鞘液同时进入流动室,在气体压力的作用下,形成包绕细胞悬液的壳液流,使细胞依次排列成单行,一个跟一个地以均等的速度通过检测区,而得到精确测量〔３〕。与仪器配套的Ｃｏｕｌｔｅｒ鞘液消耗量大(从开机到停机一直连续使用),需用外汇进口,不仅价格较高,而且周期较长。为解决这一问题,我们经过两年半的反复试验,研…     

流式细胞术测定淋巴细胞的分裂

为确立流式细胞仪检测淋巴细胞的分裂的新方法 ,利用CFSE标记新鲜分离外周及胸腺淋巴细胞后 ,加入PMA、ConA、抗TCR抗体等培养 3d ,染色进行FACS分析。结果 ,诱导T细胞活化分裂的 3种刺激剂 ,以PMA刺激作用最强 ,ConA次之 ,抗TCR抗体最弱。表明CFSE标记细胞 ,流式细胞仪分析法不仅可以检测单细胞水平上细胞的分裂 ,还可根据荧光强度判断细胞的分裂次数。     

流式细胞分选仪研究进展与展望

流式细胞技术一直是生物细胞研究的重要手段,近年来流式细胞分选仪在细胞富集、筛选后再利用等方面的应用使得对能够满足高通量、高细胞信息提取、高细胞活性要求的流式细胞分选仪的需求增加。本文阐述基于光学探测方法的流式细胞分选仪结构、工作原理及其发展现状出发;提出在光学流式细胞分选仪研究领域出现无标记化和微流控芯片化两种新的发展趋势。在两种发展趋势下,无标记成像方法、微流控片上分选系统和片上压电致动器集成分选等新兴技术越来越多地应用到光学流式细胞分选仪的研发设计中,本文从技术特性、性能优势、应用前景等方面对这些技术的研究进展以及面临的挑战进行了总结和展望,供流式细胞分选技术的使用者及研究者参考。     

卵巢上皮性肿瘤细胞DNA含量流式细胞仪分析

卵巢上皮性肿瘤细胞ＤＮＡ含量流式细胞仪分析傅兴生，徐燕兰，魏宝秀，潘凌云我们采用流式细胞仪分析４４例用石蜡包埋的卵巢上皮性肿瘤的细胞核ＤＮＡ含量、细胞周期各时相分布与临床关系。一、材料与方法我们选择１９７２～１９８８年之间的４８例卵巢上皮性肿瘤、其中...     

Two Distinct Fc Receptors for IgG on Human Peripheral T Lymphocytes

The proportion of human peripheral T lymphocytes forming rosettes with IgG-coated ox erythrocytes (ORBC) is increased after controlled hypotonic treatment. This increment may be as high as 40% of total T cells, depending on the lymphocyte donor. Such treatment is shown not to result in selective cell loss. Induced rosetting is mediated by a receptor specific for the Fc portion of human IgG (Fc gamma R). Inhibition of induced Fc gamma R activity is equally well accomplished by monomeric and by aggregated IgG of defined size. This is in contrast to the Fc gamma R detected before hypotonic treatment, which is not significantly inhibited by monomeric IgG. Capping studies established the structural independence of these two types of Fc gamma R in the lymphocyte membrane by virtue of selective cross-linking of either receptor while leaving the respective counterpart unaffected. The biochemical basis of the hypotonic effect is not yet resolved. However, the data presented suggest that hypotonicity results in removal of Fc gamma R-bound cytophilic IgG. Operationally, we propose the term induced Fc gamma R (Fc gamma R-I) for the here-described new type of receptor with high affinity for monomeric IgG.Fc gamma R that are directly assayable without hypotonic induction and not inhibited by monomeric IgG are termed free Fc gamma R (Fc gamma R-F).     

白血病、肾病综合征的外周血淋巴细胞糖皮质激素受体与糖皮质激素依赖性关系的临床意义

用放射配体结合分析检测了急性白血病和肾病综合征（肾综）外周淋巴细胞的糖皮质激素受体（ＧＲ）的含量。结果提示：白血病和肾综多为糖皮质激素依赖性。急性淋巴细胞性白血病ＧＲ含量明显增高，对激素治疗的反应性较高；成人和小儿肾综的ＧＲ含量均高于正常人（Ｐ＜０．０５）。小儿肾综的ＧＲ含量大于正常值者，对激素治疗的反应敏感比率为７０．５９％；小于正常值者则为５０％，且易复发，这表明ＧＲ含量高的患者激素疗效好，反之疗效低。本方法简便，用于观察激素疗效较灵敏，可多次取材重复检测，具有一定临床价值。     

Induction of Ige Receptors on Human Lymphocytes

A mouse myeloma IgE protein was cross-linked with dimethylsuberimidate to induce polymer ization and incubated with normal human peripheral blood T lymphocytes in tissue culture. After 24 hours at 37°C approximately 30% of the cells formed rosettes with IgE sensitized red cells as compared with much lower levels in the controls. A variety of controls established that the cellular receptors involved in rosette formation were specific for IgE. Monomeric mouse IgE failed to induce a response.     

Transferrin Receptors on Mitogen-Stimulated Human Thymus-Derived Lymphocytes

The appearance of transferrin receptors on mitogen-stimulated human thymus-derived (T) lymphocytes was studied. When indirect immunofluorescence with immunoadsorbent-purified antitransferrin antibodies was used, approximately 10% of resting T cells were stained. This proportion increased to 50-80% of the cells 3-4 days after stimulation with the mitogenic lectins concanavalin A (Con A) and leucoagglutinin (La) from Phaseolus vulgaris. Almost all blast cells (greater than or equal to 90%) were positive. Cell binding experiments with 125I-labelled transferrin indicated the presence of 1-5 x 10(5) transferrin receptor molecules/cell with high avidity for transferrin (K = 2 - 12 x 10(8) l/mol). Analysis by sodium dodecyl sulphate polyacrylamide gel electrophoresis and autoradiography of cell lysates containing 125I-labelled T-cell surface components revealed two surface peptides (90 kdaltons and 80 kdaltons, reducing conditions), which selectively bound to insolubilized antitransferrin antibodies. The 90-kdalton peptide also bound to insolubilized transferrin. The 80-kdalton peptide is most probably transferrin and the 90-kdalton peptide the transferrin receptor. Unreduced transferrin receptor had a molecular weight of 180 kdalton. It is probably a glycoprotein, since it reacted with wheat germ agglutinin, La, and probably also Con A. The properties of the lymphocyte transferrin receptor are similar to those described for transferrin receptors on various in-vitro-grown transformed cells. This speaks in favour of a common receptor present on all proliferating human cells.     

原发性癫痫患者糖皮质激素和糖皮质激素受体改变的研究

用放射配体结合分析，测定了４１例（男１６例，女２５例）原发性癫痫患者外周血白细胞糖皮质激素受体（ｇｌｕｃｏｃｏｒｔｉｃｏｉｄ ｒｅｃｅｐｔｏｒ ＧＲ），同时用放射免疫分析测定了患者血浆皮质醇（Ｆ），即糖皮质激素（ｇｌｕｃｏｃｒｔｉｃｏｉｄＧＣ），并与正常对照组比较。结果表明，无论年龄、性别及服用何种抗痫药物，原发性癫痫患者的ＧＣ都明显高于对照组（Ｐ〈０．０１）；ＧＲ都明显低于对照组，（Ｐ〈０．０     

Reactivity of Antiserum to Human Brain with Peripheral Blood Lymphocytes

Antisera to human brain (AHBS) and human thymocytes (AHTS) were produced in rabbits and selectively absorbed to render them specific for T cells. After absorption AHBS, but not AHTS, lost most of its cytotoxic activity against T cells. Absorbed AHBS bound up to 95% of peripheral blood T lymphocytes as detected by indirect immunofluorescence and inhibited up to 46% of the lytic activity of AHTS; however, it was incapable of inhibiting the E-rosette formation of T lymphocytes. All 10 samples of human peripheral blood lymphocytes, pretreated with AHBS, were significantly suppressed in their response to antigens, but fewer samples were affected in their response to mitogens and to allogeneic stimulation, indicating diversity in the nature of the receptors involved in the cellular responses.     

Receptors for the Third Complement Component on a Proportion of Large Granular Lymphocytes from Human Peripheral Blood

Large granular lymphocytes (LGL) are nonadherent cells with cytoplasmic azurophilic granules, avid receptors for the Fc portion of IgG, and a paranuclear localization of alpha-naphthyl acid esterase or acid phosphatase. LGL constitute the bulk of TG cells (cells with receptors for sheep erythrocytes and for IgG molecules) and null cells (non-T, non-B cells). In the present study we demonstrate that 20-33% of the circulating human LGL express receptors for the third complement component (C3R). When TG cell or null cell fractions from normal individuals or non-T cells from a patient with infantile agammaglobulinaemia (which contained almost exclusively LGL) were rosetted with erythrocytes coated with antibody and complement, a variable number of C3R-bearing cells were detected. Such cells were isolated and analysed further; the great majority of them displayed the cytochemical and ultrastructural features of LGL.     

Decreased Expression of HLA-DR Antigens on Peripheral Blood B Lymphocytes during Glucocorticoid Treatment

In 18 patients the expression of HLA-A, B, C and -DR antigens on peripheral blood T and B lymphocytes (PBTL and PBBL) was assessed using the lymphocytotoxic microtechnique before and during glucocorticoid administration. During steroid treatment we found a significant reduction in the reactivity of PBBL to allogeneic specific anti HLA-DR antisera as well as a xenogeneic multispecific anti HLA-DR antiserum, corresponding to approx. 1–2 dilution steps. In contrast, the reactivity of both PBBL and PBTL to allogeneic anti HLA-A, B, C antisera and anti β₂-microglobulin was unaffected during the treatment.
Membrane marker studies of the isolated PBBL and PBTL showed no differences in the relative distribution of mononuclear cell subpopulations before and during treatment, which seems to exclude extravascular redistribution of particular HLA-DR positive lymphocyte or monocyte subsets during steroid administration as an explanation for the finding.
Incubation of isolated normal PBBL with dexamethasone under various conditions did not affect the reactivity of these cells to anti HLA-DR antisera. Furthermore, preliminary experiments gave no indication of decreased rate of HLA-DR antigen synthesis in the presence of dexamethasone in vitro.
Thus, we conclude that glucocorticoid administration in vivo decreases the expression of HLA-DR antigens on PBBL, possibly in an indirect way, the mechanism of which is still unknown.     

Association between HLA Determinants and Complement Receptors on Human Lymphocytes

The relationship between the major histocompatibility complex (MHC) and functional receptors on the surface of human lymphocytes was studied. HLA antisera were tested for their effect on the formation of E-, EA-, and EAC'-rosettes by human peripheral blood lymphocytes (PBL). Antisera to various HLA specificities inhibited the formation of EAC'-rosettes, but had no effect on the formation of E-rosettes. The formation of EA-rosettes was inhibited by HLA antisera only in part among the individuals tested. Anti β₂-microglobulin serum resembled HLA antisera in its effect on the formation of the various rosettes.
HLA determinants and complement receptors are different entities on the cell surface since elimination of complement receptors by trypsin treatment does not seem to affect the expression of HLA antigens on the cell surface. It is suggested that EAC'receptors are located close to HLA determinants.     

Carbohydrate Metabolism in PHA Stimulated Human Peripheral Blood Lymphocytes

Human peripheral blood lymphocytes (PBL) stimulated in vitro by phytohemoagglutinin (PHA) manifest augmented glycolysis and oxidation of glucose-1-¹⁴C, indicating an increased utilization of the pentose pathway. Lactic acid production, as index of increased glycolysis, follows the same kinetic of thymidine incorporation and can he easily quantitated by an enzymatic assay.