Activity of acid phosphatase in resting and mitogen-stimulated human peripheral blood lymphocytes

Activity of acid phosphatase was found to be higher in T than in non-T cells of human peripheral blood lymphocytes. The T cell mitogen PHA induced an increase in the magnitude of acid phosphatase activity, while the B cell mitogen LPS was not able to produce any effect on acid phosphatase activity. Acid phosphatase activity in lymphocytes correlated with their spontaneous proliferative activity.     

Inhibiting effects of serotonin antagonists on the proliferation of mercuric chloride stimulated human peripheral blood T lymphocytes.

The serotonin antagonists ketanserin and methiotepine were tested for a modulating effect on the proliferative response of human peripheral blood T lymphocytes to mercuric chloride. This response was inhibited by ketanserin at 8 x 10(-5) mol/l and by methiotepine at 8.0 x 10(-6) mol/l. There were no additive effects at these concentrations of antagonists and at 10(-5) mol/l of serotonin. Low concentrations of ketanserin eliminated the inhibiting effect of serotonin on mercuric chloride induced proliferation of T lymphocytes. It thus seems as if the inhibiting effect of serotonin on T lymphocytes is mediated by 5-HT1c or 5-HT2 receptors, while the mechanism for the intrinsic inhibiting effect of the antagonists at present is unknown.     

脐血来源间充质干细胞可抑制异体T细胞的反应

目的 研究脐血间充质干细胞(HUCB-MSCs)对异体T细胞的抑制作用.方法 体外培养HUCB-MSCs,流式细胞术测表面标记;取正常人外周血,免疫磁珠分离CＤ３+T细胞,将分离的CD3+T与HUCB-MSCs 1:1混合培养5d,PHA刺激或不刺激,采用3H-TdR掺入法观察T细胞增殖,ELISA方法检测细胞因子,流...     

Targeting of peripheral blood T lymphocytes

Conclusions In summary, human single-chain gene transduced T cells were shown: to express the scFv on their surface, to recognize their relevant ligand (tumor-associated antigen) on tumor target cells, to produce cytokines and, to lyze tumor cells. In our earlier review on retargeting T lymphocyte specificity [11], we concluded that a number of questions needed to be answered: (1) Is triggering of cytolysis by CTL or lymphokine production most important to generate anti-cancer effects? Both are important, especially to eliminate bystander cells which do not express tumor-associated antigen [35, 75, 83, 106]. (2) Can targeted CTL traffick and home to the tumor site? Yes, they can. (3) Does humanization of mouse mAbs reduce HAMA responses? Our preliminary experiences suggest that this is the case (unpublished data).Significant progress has therefore been made in the laboratory and in clinical tests, and will continue to be made. We are now preparing for the clinical phase I/H testing of in vivo anti-tumor activity of T lymphocytes retargeted by transfer of chimeric receptor genes encoding Ab-type specificity.     

I-region-associated determinants: expression on mitogen-stimulated lymphocytes and detection by cytotoxic T cells

We have studied the expression of Ia-antigens, controlled by genes in the I-region of the H-2 complex, on phytohemagglutinin (PHA)-stimulated lymph node cells and on lipopolysaccharide (LPS)-stimulated spleen cells, and have compared these two types of cell populations as targets for killer cells in the cell mediated lympholysis (CML) assay. PHA targets are almost completely insensitive to complement-mediated lysis by anti-Ia sera while the majority of LPS targets are killed. T cell-mediated lysis against I-region determinants was also detected, and these determinants, in contrast to H-2K and H-2D CML determinants, seem to be much more strongly expressed on LPS-stimulated cells. No differences in the kinetics of the response to K- or I-region CML determinants were observed. Lysis by killer cells can also be obtained against incompatibilities which do not give rise to strong skin graft rejection and against determinants which are most probably controlled by genes outside the K-D interval.     

Indomethacin enhances the proliferation of mitogen-stimulated T lymphocytes of homosexual males with persistent generalized lymphadenopathy

The possibility that cyclooxygenase products of arachidonic acid might contribute to the defective T lymphocyte function of homosexual men with the reactive lymph node syndrome was examinedin vitro. T lymphocyte proliferation, assessed by the uptake of [³H]thymidine after the addition of phytohemagglutin, was 72,870±66,816 counts per minute (mean±SD) for eight patients and 119,589 ± 64,913 counts per minute for 30 controls (P<0.05, Student'st test). Treatment with the cyclooxygenase inhibitor indomethacin increased the phytohemagglutin-induced proliferation of the T lymphocytes from five of eight patients, but none of 12 healthy homosexual and heterosexual control subjects. The production of prostaglandin E₂ by T lymphocytes from six patients was 16.1±10.5 pg/5×10⁶ cells/hr, as contrasted with that of 4.9±1.3 and 4.3±2.1 pg/5×10⁶ cells for four healthy homosexual and six healthy heterosexual control subjects, respectively (P<0.01, Student'st test). The production of prostaglandin E₂ by the patients' monocytes was normal. Abnormalities of the cyclooxygenase pathway of T lymphocytes of patients with the reactive lymph node syndrome may reflect an immuno-regulatory defect, which predisposes to infections and may evolve into the more severe abnormalities of the acquired immune deficiency syndrome.     

多种活化因子共刺激对人外周血单个核细胞体外增殖和表型的影响

目的:观察多种活化因子共刺激后对人外周血淋巴细胞增殖和表型的影响。方法:用淋巴细胞分离液分离人外周血单个核细胞(PBMC),根据加入刺激因子(CD3 mAb、CD28 mAb、IFN-γ、IL-1α、IL-2、IL-15和IL-21)种类和方法不同将实验分为7组。用全自动五分类血液分析仪计数不同细胞因子诱导培养的PBMC增殖力、流式细胞术测定诱导后共刺激细胞表面CD3,CD4,CD8,CD28,CD16、CD56+CD16,CD3+CD8+,CD3+CD4+,CD3+CD56+,CD45RO等分子的变化、乳酸脱氢酶释放法测定诱导后的共刺激细胞对SGC-7901、SW-1990和SW-116细胞株的杀伤活性。结果:在PB-MC培养体系中加入不同的刺激因子其细胞增殖能力有明显的差异,以含刺激因子CD3、CD28、IFN-γ、IL-2、IL-1α、IL-15和IL-21组增殖倍数最高,在培养第10 d时该组的增殖倍数为255.3 6.3,明显高于仅含CD3、IFN-γ和IL-2培养体系组(166.6 5.5)(P<0.05)。在PBMC培养体系中加入不同的刺激因子其部分细胞表面标志有所差异,在培养体系中无IL-15时CD16+CD56+(NK细胞)细胞和CD3+CD56+细胞比例明显高于其他组;CD45RO+的记忆性T细胞以延迟3 d添加IL-15和IL-21组升高最明显。经不同活化因子刺激培养10 d的PBMC对SGC-7901、SW-1990和SW-1116细胞杀伤活性有明显差异,以延迟3 d添加IL-15和IL-21组最高(分别为76.2%、60.3%和70.6%),明显高于仅含CD3、IFN-γ和IL-2培养体系的细胞组(分别为54.9%、44.6%和50.4%)(P<0.05)。培养的细胞对胃腺癌细胞SGC-7901杀伤活性最强。结论:不同刺激因子活化的PBMC其增殖能力、表面标记和杀伤活性有明显差异,在培养体系中增加相应的刺激因子对细胞定向培养有一定价值。     

Vitamin C and E suppress mitogen-stimulated peripheral blood mononuclear cells in vitro

BACKGROUND/AIMS: The antioxidant properties of vitamin C and E are considered to be important for their anti-inflammatory activity. Recently, antioxidant resveratrol was found to suppress neopterin production and tryptophan degradation in mitogen-treated human peripheral blood mononuclear cells. METHODS: In this study, the effects of vitamin C and E were investigated in unstimulated peripheral blood mononuclear cells and in cells stimulated with the mitogens phytohaemagglutinin and concanavalin A in vitro. RESULTS: The mitogens induced a significant production of neopterin and a degradation of tryptophan. Vitamin C (0.1-10 microM) and E (5-100 microM) suppressed these immunobiological pathways in a dose-dependent way (p < 0.01). CONCLUSION: Neopterin production and tryptophan degradation in monocyte-derived macrophages are both triggered by the pro-inflammatory cytokine interferon-gamma. Thus, their concurrent suppression by vitamin C and E suggests an effect on the formation and release of this cytokine by stimulated T cells. These findings may be related to the general health benefits which are associated with the antioxidant nature of these vitamins.     

CD28复合刺激信号对人外周血T淋巴细胞c-Re1的影响

报道ＣＤ２８信号传导对正常人外周血Ｔ淋巴细胞原癌基因ｃ－Ｒｅ１的影响。用免疫磁珠阴性选择法从外周血单个核细胞中分离出Ｔ淋巴细胞，经ＣＤ２８单克隆抗体及ＰＭＡ复合刺激，提取胞核和胞浆蛋白， 分别用于与ｃ－Ｒｅｌ抗血清或磷酸化酪氨酸的单克隆抗体做免疫沉淀反应和／或ＳＤＳ－ＰＡＧＥ，用ｃ－Ｒｅｌ抗体作为探针。最后用放射自显影技术显示ｃ－Ｒｅｌ的存在和酪氨酸的磷酸化。采用ＣＴＬＬ法测定Ｔ细胞上清液的ＩＬ－２量。研究结果表明：ＣＤ２８的复合刺激促进ｃ－Ｒｅｌ的诱生、核内转移和酪氨酸成酸化，并使活化的Ｔ细胞ＩＬ－２的产生增加。这些结果证明ＣＤ２８依赖的酪氨酸磷酸化的Ｔ细胞途径与ｃ－Ｒｅｌ有直接的相关。     

不同来源的树突状细胞对扩增脐血NKT的影响

目的 比较不同来源的DC对扩增脐血NKT的影响.方法 分别以外周血和脐血为来源诱生DC,在α-Galcer和IL-2的联合刺激下,对脐血中的NKT进行扩增.用流式细胞检测技术分析不同来源DC的表型区别,并对它们扩增的NKT细胞(TCR Va24 TCR Vβ11 )进行鉴定.结果 在14 d的培养时间里,由PB-Dc参与的TCR Vαβ NKT细胞的扩增量占淋巴细胞的(20.8±5.25)%,是原来的(8.22±2.75)×102倍;而由CB-DC参与的TCR Vαβ NKT为(11.26±5.63)%,是原来的(4.66±2.12)×102倍;未加DC组,单用α-Galcer和IL-2,NKT的最终扩增量也达到了(10.84±4.00)%.3组较原来都呈现显著性扩增(P＜0.01),其中以PB-DC参与的NKT扩增组效率最高(P＜0.001).经流式检测,PB来源的DC,其表面CDld表达明显高于CB来源的DC(P＜0.001),而共刺激分子则较脐血DC低.结论 α-Galeer对NKT细胞有特异地扩增功能;NKT的扩增效率与DC细胞之间有着密切的关联,以外周血为来源的DC,其协同刺激脐带血NKT的扩增功能较脐血DC强.     

Accessory cells provide more than one signal for lectin mitogen-stimulated proliferation of human lymphocytes

A culture system was developed in which human peripheral blood mononuclear cells (PBMC) depleted of Ia-expressing cells did not proliferate in response to the lectin mitogen phytohemagglutinin (PHA). These cells were able to respond to mitogen if purified autologous accessory cells were added back to the culture, thus showing an absolute requirement for Ia-expressing accessory cells in mitogen-driven T-cell proliferation. The identity of these accessory cells was shown to be not only monocytes but also Ia-expressing B cells and possibly other unidentified Ia-bearing cell types. Human interleukin-1 (IL-1) in the lectin mitogen-unresponsive culture system was found unable to reconstitute normal human T-cell proliferation. This suggests that those cells with Ia surface antigen acting as accessory cells must deliver more than an IL-1 signal for T-cell proliferation. The phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA) was tested for its ability to replace necessary Ia-expressing accessory cells. TPA was able to replace accessory cells in culture, thus mimicking the Ia-expressing accessory cells and all their delivered signals in the triggering of human lymphocytes.     

Expression of receptors for IgA on mitogen-stimulated human T lymphocytes

This study demonstrates that activation of human peripheral blood mononuclear cells (PBMC) by the T cell mitogens phytohemagglutinin (PHA) and concanavalin A (Con A) induces the expression of receptors for IgA without addition of IgA to the culture medium. Cells bearing receptors for IgA were determined by indirect immunofluorescence using human secretory IgA and fluoresceinated goat anti-human IgA or goat anti-secretory component antibodies. Among freshly isolated PBMC, 4.7 +/- 1.7% of T cells, 12.7 +/- 12.5% of B cells and 14.4 +/- 7.6% of monocytes were found to be IgA receptor positive. In unstimulated PBMC cultures the percentage of IgA receptor-positive cells slightly increased at 48 h and was more elevated after 7 days. In Con A-stimulated cultures 24.3 +/- 18.5% of the cells expressed receptors for IgA after 48 h. Then, the number decreased and rose again thereafter. PHA stimulation induced an increase of smaller magnitude with similar kinetics. Induction of receptor for IgA on activated T cells was demonstrated by double-labelling experiments showing more CD8+ than CD4+ cells with receptors for IgA among Con A-activated PBMC. Furthermore, PHA or Con A stimulation of B cell-depleted PBMC suspensions resulted in a marked increase of cells bearing receptors for IgA. Expression of these receptors was down-regulated by recombinant interferon-gamma (250 units/ml) and by prostaglandin PGE2 (100 nM) both on unstimulated and mitogen-activated PBMC. The receptor for IgA was distinct from the asialoglycoprotein receptor and did not cross-react with the poly-Ig receptor of epithelial cells. It was concluded that, in the absence of inducing exogenous IgA, T cell mitogens trigger the expression of receptors for IgA. Therefore, T cell activation is associated with the down-regulation of receptors for IgM and the increased expression of receptors for IgG, IgA and IgE.     

脐血来源间充质干细胞可抑制异体T细胞的反应

目的研究脐血间充质干细胞(HUCB-MSCs)对异体T细胞的抑制作用。方法体外培养HUCB-MSCs,流式细胞术测表面标记;取正常人外周血,免疫磁珠分离CD3+T细胞,将分离的CD3+T与HUCB-MSCs 1∶1混合培养5 d,PHA刺激或不刺激,采用3H-TdR掺入法观察T细胞增殖,ELISA方法检测细胞因子,流式细胞术观察细胞凋亡。结果 HUCB-MSCs呈纺锤样的细胞形态,不表达CD14、CD34、CD45、HLA-DR,而表达CD29、CD44、HLA-ABC。HUCB-MSCs抑制PHA引起的T细胞增殖(5 230±550 vs 10 500±800 counts/min,P<0.001);HUCB-MSCs还能抑制异体T细胞分泌IFN-γ(510±60 vs 1 580±100 pg/mL,P<0.001)和TNF-α(590±20 vs 1 180±30 pg/mL,P<0.001),上调IL-4(16.3±8.2 vs 4.1±1.8 pg/mL,P<0.001)和IL-10(105±5 vs 17±2 pg/mL,P<0.001)分泌;HUCB-MSCs不诱导T细胞的凋亡。结论 HUCB-...     

The immunosuppressive potency of various steroids on peripheral blood lymphocytes, T cells, NK and K cells

The immunosuppressive effect of natural and synthetic steroids was tested in vitro on phytohemagglutinin (PHA) stimulated T lymphocytes and peripheral blood lymphocytes (PBL), as well as on NK and K cell activity. Three groups of steroids, significantly different in their immunosuppressive activity, were identified. Fluorohydrocortisone, and methylprednisolone were highly potent in suppressing PHA stimulation of T lymphocytes and PBL. Hydrocortisone was of intermediate potency, whereas cortisone, dihydrocortisol, and tetrahydrocortisol were of low potency. T lymphocytes were more sensitive to the suppressive effect of fluorohydrocortisone and methylprednisolone than were PBL cultures. NK and K cell activity was suppressed only by the high potency synthetic steroids and even then the suppression of K cell activity was not significant except at high in vitro steroid concentrations. The present findings support the conception that different lymphocyte subpopulations have different susceptibility to the effect of highly potent steroids. Thus, lymphocyte heterogeneity must be taken into account when the immunosuppressive potencies of different glucocorticoids are studied. Furthermore, the findings in different lymphocyte populations ranked the relative in vitro immunosuppressive potency of glucocorticoids different from the relative anti-inflammatory potencies reported in the literature.     

羊水来源间充质细胞对于外周单核淋巴细胞增殖抑制作用的机制研究

目的鉴定羊水来源间充质细胞(AF-MCs)并探讨其对于外周单核淋巴细胞(PBMC)增殖活性的影响及机制。方法通过选择性消化和单克隆培养分离及培养人源羊水细胞(AFCs),而后通过核型分析细胞来源和流式分析细胞表型鉴定其为间充质细胞(MCs)。另一方面,将鉴定得到的AF-MCs与PBMC共培养,检测其是否具有免疫抑制功能。运用流式分析CD4~+CD25~+Foxp3~+T细胞频率在共培养前后变化,探讨AF-MCs具有免疫抑制功能的机制。结果AF-MCs来自于胚胎发育过程中的脱落细胞,其具有与骨髓间充质干细胞(BM-MSCs)相似的表型,且其能上调CD4~+CD25~+Foxp3~+T细胞的频率而产生免疫抑制功能。结论AF-MCs具有与BM-MSCs类似的表型,并且在免疫调节方面与BM-MSCs类似。这为临床应用间充质干细胞提供了一个新的,安全的种子细胞来源。     

The effects of chromium and copper supplementation on mitogen-stimulated T cell proliferation in hypercholesterolaemic postmenopausal women

The purpose of this study was to analyse effects of chromium and/or copper supplementation on immune function in hypercholesterolaemic postmenopausal women. A 2 x 2 factorial research design was used and 40 subjects were supplemented with 0.394 g lactose, 200 microg Cr, 3.0 mg Cu, or 200 microg Cr and 3.0 mg Cu/d for 12 weeks. A significant interactive effect of Cr and Cu supplementation on lymphocyte proliferation was observed with ConA 50 microg/ml stimulation. After 12 weeks of supplementation, ConA-stimulated (50 microg/ml) lymphocyte proliferation was significantly lower when Cu was added to the Cr supplementation group. Moreover, ConA-stimulated (100 microg/ml) lymphocyte proliferation was significantly lower in the Cu supplementation group compared to the Cr supplementation group after 12 weeks of supplementation. These results suggest that Cu blocks enhancement of lymphocyte proliferation by Cr supplementation and that Cu supplementation has potential suppressive effects on the immune function in these subjects.     

不同浓度泌乳素对葡萄膜炎患者外周血淋巴细胞增殖活化的影响

目的：探讨不同浓度泌乳素（Prolactin，PRL）对葡萄膜炎患者外周血单个核细胞增殖及活化的影响，研究PRL在葡萄膜炎自身免疫中的作用。方法：用化学发光法测定葡萄膜炎患者及正常对照者血清PRL的水平；分离获得葡萄膜炎患者及正常对照者外周血单个核细胞，与S抗原及不同浓度PRL进行培养，108小时后用双标记免疫荧光染色流式细胞检测技术测定CD3^＋CD25^＋细胞的百分数；培养96小时后加入^3H-胸腺嘧啶核苷，12小时后测定每分钟脉冲数；观察在S抗原的刺激下，不同浓度的PRL对葡萄膜炎患者外周血单个核细胞增殖活化的影响。结果：葡萄膜炎患者血清PRL水平与正常人差异无显著意义（t=1．963，P=0．051）。葡萄膜炎患者对S抗原呈阳性反应时，12．5-1000μg/L的PRL有促进葡萄膜炎患者外周血单个核细胞增殖与活化的趋势，在200μg／L的浓度时促进作用最显著（活化t=2．736，P=0．012；增殖t=2．547，P=0．036），1000μg/L时趋势下降。葡萄膜炎患者对S抗原呈阴性反应时，则没有上述作用。结论：在小样本研究中葡萄膜炎患者血清中PRL水平与正常对照者相比差异无显著意义；中等浓度的PRL（200μg／L）能同S-Ag发挥共刺激作用，促进葡萄膜炎患者单个核细胞的增殖与活化，过高浓度（1000μg／L）则失去促进作用。     

T cell derived IgE binding factors inhibit IgE-specific rosettes on basophilic leukaemic cells and lymphocytes

T cell derived IgE-binding factors inhibited the formation of IgE-specific rosettes on both rat basophilic leukaemic cells and lymphocytes. This was demonstrated by IgE-binding factors released from either IgE-pulsed T cells or adherent cells as well as from IgE-binding factor producing hybridomas. The different binding factors inhibited the IgE-rosettes to a different degree when tested on basophils and lymphocytes, most likely due to the higher affinity for IgE of the former cell type. The results indicate that rat basophils and mouse T cells may recognize common epitopes on the Fc portion of IgE.     

Androgens inhibit proliferation of human peripheral blood lymphocytes in vitro

The effect of sex hormones on human lymphocytes was examined in vitro on cell cultures of human peripheral blood mononuclear cells (HPBM). Cells were stimulated using T- and B-cell mitogens, and hormones in physiological (nM) or pharmacological (microM) concentrations were added. Proliferation was determined by measuring the incorporation of tritiated thymidine. It was found that both testosterone and dihydrotestosterone, in physiological concentrations, can attenuate DNA synthesis. The effect was dose dependent in that pharmacological concentrations of both testosterone and dihydrotestosterone caused a strong inhibitory effect on proliferation of in vitro cultured HPBM. However, cell cultures of a few individuals were insensitive to the androgens even at pharmacological concentrations. Also, no difference could be detected in the response between cultured cells of females and males. Although a slight reduction in antibody production was evident in pokeweed mitogen-stimulated cultures, in the presence of both testosterone and dihydrotestosterone, it was, however, not statistically significant.