Effects of heparin on growth factor-induced mitogenic and proliferative responses of rat pulmonary a

目的：探讨肝素是否能抑制生长因子诱导的大鼠肺动脉平滑肌细胞（ＰＡＳＭＣ）分裂和增殖．方法：应用含１０％ＦＢＳ的Ｍ１９９培养液培养大鼠ＰＡＳＭＣ．细胞分裂及细胞增殖分别用［ｍｅｔｈｙｌ３Ｈ］ＴｄＲ和细胞计数监测．结果：ＦＢＳ（１０％），以及ＦＢＳ（１％）与ＰＤＧＦ（５０μｇ·Ｌ－１），ＦＧＦ（５０μｇ·Ｌ－１），或ＩＬ１α（１００ｎｇ·Ｌ－１）联合应用均能增加大鼠ＰＡＳＭＣ分裂．肝素（１００ｍｇ·Ｌ－１）抑制１０％ＦＢＳ诱导的大鼠ＰＡＳＭＣ增殖（２８％±６％）和胸腺嘧啶摄取反应（２７％±７％），抑制ＦＢＳ（１％）与ＰＤＧＦ（５０μｇ·Ｌ－１），ＦＧＦ（５０μｇ·Ｌ－１），或ＩＬ１α（１００ｎｇ·Ｌ－１）联用诱导的大鼠ＰＡＳＭＣ增殖（２５％±６％，２７％±７％，２０％±４％），以及胸腺嘧啶摄取反应（２３％±７％，２６％±６％，２０％±６％）．结论：肝素抑制生长因子诱导的大鼠ＰＡＳＭＣ的分裂与增殖．     

肝素对生长因子诱导的大鼠肺动脉平滑肌细胞分裂和增殖的影响

目的：探讨肝素是否能抑制生长因子诱导的大鼠肺动脉平滑肌细胞（ＰＡＳＭＣ）分裂和增殖。方法：应用含１０％ＦＢＳ的Ｍ－１９９培养液培养大鼠ＰＡＳＭＣ。细胞分裂及细胞增殖分别用［ｍｅｔｈｙｌ－＾３Ｈ］ＴｄＲ和细胞计数监测。结果：ＦＢＳ（１０％），以及ＦＢＳ（１％）与ＰＤＧＦ（５０μｇ·Ｌ＾－１），ＦＧＦ（５０μｇ·Ｌ＾－１），或ＩＬ－１α（１００ｎｇ·Ｌ＾－１）联合应用均能增加大鼠ＰＡＳＭＣ分裂。肝素（     

系统性红斑狼疮患者外周血淋巴细胞增殖功能及其产生白细胞 …

目的 研究系统性红斑狼疮（ＳＬＥ）患才吉淋巴细胞（ＰＢＬ）增殖反应及其产生白细胞介素２（ＩＬ－２）的情况。方法 淋巴细胞增殖法及ＩＬ－２检测法,结果 ＳＬＥ患者ＰＢＬ的增殖反应明显高于正常人,但ＳＬＥ患者ＰＢＬ产生ＩＬ－２的能力明显低于正常人。结论 ＳＬＥ患者ＰＢＬ增殖反应提高和其产生ＩＬ－２能力降低可能与Ｔ细胞两亚群ＴＨ１／ＴＨ２的失衡有关。     

Tumor necrosis factor mediated release of platelet-derived growth factor from bovine cerebral microvascular endothelial cells

研究ＴＮＦ介导的ＢＣＭＥＣ释放ＰＤＧＦ及药物对ＢＣＭＳＭＣ增殖的保护作用．方法：体外培养ＢＣＭＥＣ和ＢＣＭＳＭＣ，用结晶紫染色法观察ＴＮＦ引起的ＢＣＭＥＣ释放ＰＤＧＦ继而促进ＢＣＭＳＭＣ增殖作用．结果：ＴＮＦ不能促进无血清培养的ＢＣＭＳＭＣ增殖，但ＴＮＦ（５－２０μｇ·Ｌ－１）剂量依赖地促进ＢＣＭＥＣ释放ＰＤＧＦ．ＴＮＦ（２０μｇ·Ｌ－１）促进ＢＣＭＳＭＣ增殖的百分率为３４±４％．药物Ｉｍｐｅｒａｔｏｒｉｎ，ｉｓｏｉｍｐｅｒａｔｏｒｉｎ和ＰＭＤＰ不影响ＴＮＦ引起ＢＣＭＥＣ释放ＰＤＧＦ，但能剂量依赖地（１－１００μｍｏｌ·Ｌ－１）抑制ＰＤＧＦ促ＢＣＭＳＭＣ增殖的作用．结论：ＴＮＦ促进ＢＣＭＥＣ释放ＰＤＧＦ继而引起ＢＣＭＳＭＣ的增殖．药物能抑制ＰＤＧＦ引起的ＢＣＭＳＭＣ增殖．     

Basic fibroblast growth factor enhanced LAK cell cytotoxicities against human bladder neoplasm cells 1

目的：探讨白细胞介素Ⅱ（ＩＬ２）和碱性成纤维细胞生长因子（ｂＦＧＦ）对膀胱癌患者淋巴因子激活的杀伤细胞（ＬＡＫ细胞）的作用．方法：用细胞计数观察不同浓度ｂＦＧＦ对ＬＡＫ细胞增殖的影响．以膀胱癌细胞系ＥＪ及新鲜分离患者自体肿瘤细胞（ＢＴＣ）为靶细胞,用ＭＴＴ法测定ＬＡＫ细胞对膀胱癌细胞的细胞毒作用．结果：虽然外周血单核细胞（ＰＢＭＣ）的增殖可被ｂＦＧＦ５μｇ·Ｌ－１所抑制,ＩＬ２所诱导的ＬＡＫ细胞的增殖却不受ｂＦＧＦ的影响,ｂＦＧＦ明显加强ＬＡＫ对ＥＪ细胞和ＢＴＣ的细胞毒作用．结论：虽然ｂＦＧＦ抑制ＰＢＭＣ的增殖,但ｂＦＧＦ又增强膀胱癌患者ＬＡＫ细胞对肿瘤细胞的细胞毒性．     

雷藤氯内酯醇对正常人及类风湿关节炎患者周围血单个核细胞及滑膜细胞产生免疫球蛋白的影响

研究了１０例正常人和６例类风湿关节炎（ＲＡ）患者外周血单个核细胞（ＰＢＭＣ）及３例ＲＡ患者消化的单个滑膜细胞（ＤＳＳＣ），经雷公藤的有效单体雷藤氯内酯醇（Ｔ４）处理２ｈ对其分泌免疫球蛋白（Ｉｇ）的影响。结果显示：Ｔ４在浓度为５～３５ｎｇ·ｍｌ－１对正常人ＰＢＭＣ分泌Ｉｇ有明显抑制作用，呈剂量依赖趋势。在２５ｎｇ·ｍｌ－１浓度下，Ｔ４对ＲＡ患者ＰＢＭＣ及ＤＳＳＣ分泌Ｉｇ均有明显抑制作用。     

IL-2-PE40对免疫活性T细胞的影响

目的研究ＩＬ－２－ＰＥ４０对免疫活性Ｔ细胞的影响。方法采用ＣｏｎＡ刺激的淋巴细胞增殖试验、混合淋巴细胞培养（ＭＬＣ）及细胞毒试验。结果ＩＬ－２－ＰＥ４０对ＣｏｎＡ诱导的小鼠脾细胞有十分强的细胞毒性，能选择性地抑制ＭＬＣ中抗原活化的Ｔ细胞活性，保留未活化Ｔ细胞对ＣｏｎＡ诱导的丝裂原反应，在培养ｄ３加入ＩＬ－２－ＰＥ４０比培养开始时加入对ＭＬＣ抑制作用强。结论ＩＬ－２－ＰＥ４０能够高度选择性抑制免疫活性Ｔ细胞，是ＩＬ－２Ｒ靶向治疗中具有潜力的一种免疫抑制剂。     

Detection of DNA damage in peripheral lymphocytes by 7 compounds using comet assay

目的：检测过氧化氢（Ｈ２Ｏ２）、甲磺酸乙酯（ＥＭＳ）、丝裂霉素Ｃ（ＭＭＣ）、二甲基亚硝胺（ＤＭＮＡ）、苯并（ａ）芘（ＢａＰ）、２氨基芴（２ＡＦ）和环磷酰胺（ＣＰ）诱发小鼠、大鼠及人外周血淋巴细胞ＤＮＡ单链断裂．方法：体外单细胞微量凝胶碱性电泳试验（慧星试验）．结果：除ＥＭＳ０９７ｍｍｏｌ·Ｌ－１在小鼠淋巴细胞，ＭＭＣ３０μｍｏｌ·Ｌ－１在小鼠、人淋巴细胞中呈阴性外，其余均为阳性．最低可检测浓度分别为Ｈ２Ｏ２１μｍｏｌ·Ｌ－１，ＥＭＳ０４８ｍｍｏｌ·Ｌ－１，ＢａＰ５０μｍｏｌ·Ｌ－１，ＣＰ２０ｍｍｏｌ·Ｌ－１，ＭＭＣ１０μｍｏｌ·Ｌ－１，ＤＭＮＡ２７３ｍｍｏｌ·Ｌ－１，２ＡＦ６２５μｍｏｌ·Ｌ－１．ＣＰ、ＢａＰ、２ＡＦ需经Ｓ９Ｍｉｘ代谢活化才显示毒性．结论：彗星试验检测出ＭＭＣ诱导大鼠，ＥＭＳ诱导大鼠和人，以及Ｈ２Ｏ２、ＤＭＮＡ、ＢａＰ、ＣＰ和２ＡＦ诱导小鼠、大鼠和人外周血淋巴细胞ＤＮＡ单链断裂损伤．     

褪黑素对类风湿关节炎病人外周血淋巴细胞增殖反应的影响及其G蛋白-cAMP信号转导机制的研究

目的研究褪黑素（ＭＴ）对类风湿关节炎（ＲＡ）病人外周血淋巴细胞（ＰＢＬ）增殖反应的影响及其Ｇ蛋白 ｃＡＭＰ信号转导机制。方法淋巴细胞增殖法、ｃＡＭＰ放免测定及Ｇ蛋白功能分析。结果ＲＡ病人ＰＢＬ增殖能力低于正常人１０倍左右，ＭＴ在１×１０－８～１×１０－６ｍｏｌ·Ｌ－１能明显促进ＲＡ病人ＰＢＬ的增殖反应；ＲＡ病人ＰＢＬ的ｃＡＭＰ水平明显高于正常人，ＭＴ（１×１０－６ｍｏｌ·Ｌ－１）可部分使其降低，且ＭＴ的这一作用能被百日咳毒素（ＰＴ）取消。结论ＭＴ可促进ＲＡ病人ＰＢＬ的增殖反应，ＰＴ敏感的Ｇｉ蛋白 ｃＡＭＰ信号转导机制参与了ＭＴ上调ＰＢＬ的作用     

已酮可可碱与蛋白激酶C抑制剂对佛波醇酯诱导脑微血管内皮细胞？…

目的：研究佛波醇酯（ＰＭＡ）地大鼠脑微血管内皮细胞（ＲＢＭＥＣ）表达细胞间粘附分子－１（ＩＣＡＭ－１）及ＰＫＣ抑制剂Ｈ７与已酮可可碱（ＰＴＸ）的抑制作用。方法：采用ＥＬＩＳＡ方法测定培养ＲＢＭＥＣ表达ＩＣＡＭ－１。结果：ＰＭＡ在１０－１００．Ｌ＾－１范围内剂量依赖性地诱导ＲＢＭＥＣ表达ＩＣＡＭ－１;在４－１６ｈ范围内时间依赖性诱导ＲＢＭＥＣ表达ＩＣＡＭ－１。Ｈ７和ＰＴＸ分别在５－５０μｍｏｌ．Ｌ＾     

The role of monocytes and T cells in 1,25-dihydroxyvitamin D3 mediated inhibition of B cell function in vitro

1,25-Dihydroxyvitamin D3 (1,25-(OH)2D3) inhibits immunoglobulin production by human mononuclear cells (MNC) in vitro. The present study was undertaken to evaluate the role of T cells and monocytes in 1,25-(OH)2D3 induced suppression of B cell functions. The synthetic vitamin D3 analogue MC 903 was examined in parallel. 1,25-(OH)2D3 and MC 903 showed a dose-related inhibition of IgM, IgG and IgA plaque-forming cells in poke-weed mitogen (PWM) activated cultures of MNC. This effect was most likely mediated through impairment of T cell and monocyte functions. First, the inhibitory effect was seen after PWM stimulation, but not after Epstein-Barr virus stimulation which activates B cells independently of T cells and monocytes. Second, 1,25-(OH)2D3 was not effective in T cell and monocyte-depleted cultures. Third, the effect of 1,25-(OH)2D3 on PWM driven MNC was reversed by addition of the recombinant monokines: interleukin (IL)-1 beta, tumour necrosis factor alpha (rTNF alpha), rIL-6, as well as the lymphokines: lymphotoxin (rLT) and rIL-2. This is consistent with the finding that 1,25-(OH)2D3 also inhibited IL-1 alpha, TNF alpha and LT production in these cultures. The assumption that B cells are not directly affected by 1,25-(OH)2D3 was further supported by the fact that 24 h of culture with 10(-8) M 1,25-(OH)2D3 failed to reduce immunoglobulin production by in vivo activated B cells.     

Effects of FK506 and other immunosuppressive anti-rheumatic agents on T cell activation mediated IL-6 and IgM production in vitro

The objective of this study was to investigate the therapeutic potential of FK506 and other immunosuppressive agents for the treatment of rheumatoid arthritis (RA), focusing on the effects on in vitro IL-6 production and IL-6-mediated immune response. We employed an in vitro model producing IL-6 via T cell activation in human PBMC, based on the hypothesis that T cells play a central role in the pathogenesis of RA. FK506 potently inhibited IL-6 production from PBMC stimulated with anti-CD3 and anti-CD28 monoclonal antibody (anti-CD3/CD28). Cyclosporin A (CsA) also inhibited the anti-CD3/CD28 induced IL-6 production but was about 100 times less potent than FK506. Dexamethasone (DEX) inhibited both anti-CD3/CD28 and LPS induced IL-6 production at almost the same concentration. Methotrexate (MTX) did not affect cytokine production. Anti-CD3/CD28 stimulated PBMC culture supernatants were found to enhance IgM production in SKW6.4 cells. The effects of anti-CD3/CD28 stimulated culture supernatants in the presence of agents on IgM production in SKW6.4 cells were investigated. FK506 and CsA led to suppression of IgM production induced by culture supernatants probably via inhibition of IgM inducible cytokine production from PBMC. DEX profoundly enhanced IgM production, although IL-6 production from PBMC was strongly inhibited by the agent. MTX decreased IgM production although it has no inhibitory effect on IL-6 production. The present study suggests that FK506 is the most effective among the four agents for the suppression of IL-6 production and IL-6-mediated autoantibody production in T cell activation related autoimmune diseases such as RA.     

Screening of new derivatives of arylheteroalkanecarboxylic acid on the immune system

The screening of 13 original compounds from the group of derivatives of arylheteroalkanecarboxylic acid on immunity were performed. The compounds exhibit strong myelostimulating/myelosuppressive property, increased or decreased influence on the: PFC (IgM and IgG), DTH at the sheep erythrocytes in CBF1 in vivo. In contrast, in vitro the compounds had no effect or inhibited the spontaneous, ConA or PWM induced proliferation of the splenocytes from normal mice. The problems of the universal methods of the screening of immunoactive properties of compounds are discussed.     

Effects of sulindac on in vitro immunologic function and on prostaglandin production by human peripheral blood mononuclear leukocytes

Prostaglandins (PG) appear to regulate immune-mediated inflammation, but the mechanisms involved remain unclear. Several families of nonsteroidal antiinflammatory drugs (NSAID) have been observed to inhibit PG synthesis. Among these drugs, sulindac sulfide is a potent inhibitor of PG production while its parent pro-drug, sulindac sulfoxide (sulindac), lacks PG synthesis inhibitory activity in cell-free systems. We have studied the effects of sulindac sulfoxide on the blastogenic response of human peripheral blood mononuclear cells (PBMC) stimulated by exposure to alloantigens and mitogens in vitro. Sulindac inhibited proliferation of activated PBMC in a dose-dependent manner but had little effect on the proliferation of unstimulated cells. The inhibition of mitogen-induced blastogenesis correlated with both the uptake of radiolabeled drug and the inhibition of in vitro production of PG (PGE and PGF) by mitogen-activated PBMC. These data indicate a functional relationship between PG synthesis and immune cell activation which may also apply to PBMC activated in vivo by autoimmune disease. Metabolism of sulindac sulfoxide by PBMC in vitro produced too little sulindac sulfide to adequately explain the inhibition of PG production. These data suggest that immunomodulation by sulindac may be due to a direct inhibition of cellular activation. Thus, it is proposed that decreased PG production may be a result rather than the cause of the hypoproliferative response.     

Effect of ozone on immunoglobulin production by human B cells in vitro.

Animal studies indicate that ozone (O3) inhalation results in reduced ability to generate a humoral response to soluble and particulate antigens. In this study, human lymphocytes have been exposed to O3 in vitro (1.0, 0.5, and 0.1 ppm/2 h) and then evaluated for the ability of B cells to produce immunoglobulin G (IgG) in response to the T-cell-dependent stimulus pokeweed mitogen (PWM), and to the T-cell-independent stimulus Staphylococcus aureus Cowan I strain (SAC). Suppression of IgG production was found with O3-exposed PWM-stimulated lymphocytes, while no effect of O3 was seen with SAC-stimulated cells, suggesting that T cells, but not B cells, were sensitive to O3. However, exposing either cell type alone to O3 indicated that both T cells and B cells were affected by the pollutant. The O3-exposed B cells produced less IgG in response to PWM but produced more IgG in response to SAC. On the other hand, O3-exposed T cells were suppressive in both PWM and SAC responses. Since the differentiation of B cells into plasma cells is regulated by complex interactions of cytokines secreted by T cells and antigen-presenting cells, possible O3-induced alterations in secretion of some of these regulatory lymphokines (IL-2, IL-4, IL-6, and IFN-gamma) were investigated in lymphocyte cultures stimulated with PWM. A decrease in IL-2 production was found, while in contrast, IL-6 production was significantly increased. IFN-gamma secretion was not altered, and IL-4 levels were below the limits of detectability. These results suggest that O3-induced changes in IgG production may be mediated by altered production by T cells of important immunoregulatory molecules, in addition to any direct effects of O3 on the IgG-producing cells themselves.     

Immunological alterations induced by polyamine derivatives on murine splenocytes and human mononuclear cells

Three polyamine derivatives assigned as bis-naphthalimidopropyl putrescine (BNIPPut), spermidine (BNIPSpd) and spermine (BNIPSpm) were studied to determine their effects on the proliferation of murine splenocytes and human peripheral blood mononuclear cells (PBMC) induced by the mitogens, Con A, LPS and PHA. All compounds showed a dose dependent inhibitory effect on mouse and human T cell proliferation induced by the mitogens, with BNIPPut exhibiting the most potent antiproliferative activity, followed by BNIPSpd and by BNIPSpm, respectively (Put > Spd > Spm), when considering human T cells. This suppressive activity also affects the capacity of mouse spleen cells to produce Th1 cytokines, namely IL-2 and INF-gamma after in vitro stimulation with Con A. The polyamine-induced inhibition also occurred in the case of LPS-stimulated B cells with a marked decrease of CD69 expression by these cells. Furthermore, the ability for these polyamine derivatives to induce apoptosis on Con A-stimulated splenocytes could be related to their antiproliferative activity.     

FK506 potently inhibits T cell activation induced TNF-alpha and IL-1beta production in vitro by human peripheral blood mononuclear cells

The aim of this study was to elucidate the in vitro inhibitory potency of FK506 on production of the inflammatory cytokines, tumour necrosis factor (TNF)-alpha and interleukin (IL)-1beta, with a view to assessing this immunosuppressive agent as a potential anti-rheumatic drug. We employed an in vitro model which produces TNF-alpha and IL-1beta through T cell activation. Human peripheral blood mononuclear cells (PBMC) were cultured with immobilized anti-CD3/CD28 monoclonal antibody in this model. FK506 inhibited anti-CD3/CD28 induced TNF-alpha and IL-1beta production at concentrations less than 1 ng ml(-1). Flow cytometric analysis of intracellular TNF-alpha and IL-1beta positive cells showed that FK506 potently suppresses inflammatory cytokine production from CD14+ monocytes as well as from T cells. Cyclosporin A (CsA) and dexamethasone (DEX) also inhibited the anti-CD3/CD28 induced cytokine production, but were less potent than FK506. FK506 and CsA, but not DEX, specifically inhibited anti-CD3/CD28 induced inflammatory cytokine production without affecting the lipopolysaccaride (LPS) induced effect. Methotrexate (MTX) was completely inactive for suppressing cytokine production under either condition. Anti-CD3/CD28 stimulated PBMC culture supernatants were found to enhance the expression of adhesion molecules in human vascular endothelial cells. FK506, CsA and DEX led to the suppression of adhesion molecule expression probably by inhibiting cytokine production from PBMC. The inhibitory potency of agents on TNF-alpha and IL-1beta production was compared with cytotoxicity and FK506 was not cytotoxic at concentrations several orders of magnitude greater than those required for cytokine inhibition. These results strongly suggest that FK506 may be most effective to specifically prevent T cell activation mediated inflammatory cytokine production in a clinical setting.     

Effect of tacrolimus derivatives on immunosuppression

In this study, the effects of the FK506-mPEG on immune cell activity, skin grafting rejection, and Freund’s complete adjuvant arthritis were investigated. The proliferation of T cells was inhibited with increase with the FK506 and FK506-mPEG concentrations. FK506 and FK506-mPEG at concentrations between 0.01 nM and 1000 nM had very similar effects on the proliferation of the T cells. On the other hand, in the case of the proliferation of T cells by calcium ionophore A23187 (1 μM), when the FK506-mPEG concentration was increased from 0.01 to 1000 mM, the proliferation was decreased from 90.8 to 40.3%. This was 1.8-fold higher than that of paramethoxyamphetamine (PMA). The inhibitory effect of FK506-mPEG on mast cell proliferation was higher than that of FK506. When B cells were cultured for 7 days in basal medium with no pokeweed mitogen (PWM), the IgG production was 156.2 ng/mL. On the other hand, in the case of the same treatment with 0.25% of PWM, it was 876.4 ng/mL. This is about 5.6-fold higher than with no PWM. These results show that FK506-mPEG may be practically applicable as a prodrug for the immunosuppressant FK506.     

重组人CTLA4Ig融合蛋白体外对人T淋巴细胞功能的影响

目的　研究重组人CTIA4Ig对人T淋巴细胞功能的影响。方法　采用初次混合淋巴细胞培养、再次混合淋巴细胞培养及细胞毒实验的方法。结果　CTLA4Ig在浓度高于3mg·L-1时能有效地抑制初次混合淋巴细胞反应 (P <0 0 5 ) ,其在作用 2 4h内不能发挥有效的抑制作用 ,72h能达最大抑制效率 ;在再次混合淋巴细胞反应实验中CT LA4Ig作用后的T淋巴细胞对同一供者的外周血单核细胞的刺激表现出特异性无反应 ,而对无关供者的外周血单核细胞的刺激能产生正常的应答反应。结论　CTLA4Ig在体外能抑制T细胞的增殖 ,并且能诱导T细胞产生抗原特异性无反应