柘树根多糖的体外免疫增强作用

枯树[Cudrania tricuspidata(Carr)Bur]属桑科植物,在《本草纲目》等资料中记载可治疗关节疼痛、黄疸等。研究了提取的三种柘树根多糖直接及协同有丝分裂原ConA或LPS对小鼠脾淋巴细胞增殖的影响和三种多糖对腹腔巨噬细胞(Mφ)吞噬功能的影响。     

IL—12,IL—7,IL—2体外协同诱导人PBMC增殖和PBMC杀瘤活性的？…

研究了重组ＩＬ－１２、ＩＬ－７、ＩＬ－２协同对健康人外周血单个核细胞（ＰＢＭＣ）的体外增殖和杀瘤活性的影响。结果表明，高浓度ＩＬ－２与低浓度ＩＬ－２协同可引起ＰＢＭＣ明显增殖，若在该培养体系中加入低浓度的ＩＬ－２，可使这种增殖效应显著增强。如果加大ＩＬ－１２剂量时会产生更强的细胞毒活性，并呈现剂量依赖关系。在此培养条件下的培养上清中亦检出了较高水平的ＩＦＮ－γ。对培养３￣１４ｄ的协同刺激细胞表型分     

IL-12、IL-7、IL-2体外协同诱导人PBMC增殖和PBMC杀瘤活性的实验研究

研究了重组ＩＬ１２、ＩＬ７、ＩＬ２协同对健康人外周血单个核细胞（ＰＢＭＣ）的体外增殖和杀瘤活性的影响。结果表明，高浓度ＩＬ２与低浓度ＩＬ２协同可引起ＰＢＭＣ明显增殖，若在该培养体系中加入低浓度的ＩＬ２，可使这种增殖效应显著增强。如果加大ＩＬ１２剂量时会产生更强的细胞毒活性，并呈现剂量依赖关系。在此培养条件下的培养上清中亦检出了较高水平的ＩＦＮγ。对培养３～１４ｄ的协同刺激细胞表型分析显示，ＣＤ５６抗原阳性细胞百分率呈递增趋势，提示ＣＤ５６＋的ＮＫ细胞构成了产生增殖和杀伤效应的主要细胞群体。     

二 干扰素的诱生和作用机制

50年前Issacs和Lindenmann在研究病毒干扰时发现了干扰素。半个世纪以来，干扰素研究始终是现代分子生物学的一个前沿领域。对此，我们可以从几个方面来认识。首先，干扰素是第一个被发现的细胞因子，也是研究得最清楚的细胞因子之一。干扰素及其他细胞因子是高等动物体内的重要生物调节分子，有关干扰素及细胞因子的研究，长期是细胞与分子生物学的热点。     

女贞子多糖免疫增强作用的体外实验研究

应用３Ｈ－ＴｄＲ掺入法检测女贞子多糖（ＬＬＳ）体外对小鼠免疫细胞增殖的影响。实验结果显示：１５６￣１２５０ｕｇ／ｍｌ的女贞子多糖对正常小鼠脾淋巴细胞、Ｂａｌｂ／ｃ裸鼠脾淋巴细胞（Ｂ细胞）及通过尼龙毛柱的脾细胞（Ｔ细胞）均有直接的刺激增殖作用，量效关系明显；巨噬细胞在脾淋巴细胞增殖反应中起一定的辅助作用。     

拉米夫定对PBMC及血清内HBV-DNA的阴转和细胞因子的诱导作用

目的探讨拉米夫定对慢性乙肝患者外周血单个核细胞(PBMC)及血清内HBV-DNA的阴转效果和对细胞因子的影响。方法将81例慢性乙肝患者分为A(47例)、B(34例)两组,分别采用拉米夫定和常规治疗,于治疗前后分别检测患者PBMC、血清中HBV-DNA和细胞因子水平。结果拉米夫定治疗36周后,慢性乙肝患者PBMC内和血清中HBV-DNA阴转率分别为55.32%(26/47)和61.70%(29/47),常规治疗组PBMC内和血清中HBV-DNA阴转率分别为26.47%(9/34)和32.35%(11/34),两者相比差异有统计学意义(P<0.01)。拉米夫定治疗24、36周后,慢性乙肝患者血清HBeAg的阴转率分别为46.81%(22/47)和68.09%(32/47),与常规治疗组相比,差异有统计学意义(P<0.05和P<0.01)。采用拉米夫定和常规治疗后,丙氨酸转氨酶(ALT)、门冬氨酸转氨酸(AST)、ALT/AST水平分别为(30.1±9.6)U/ml、(32.3±10.7)U/ml、0.9±0.1和(48.4±10.7)U/ml、(44.7±11.0)U/ml、1.1±0.2,差异有统计学意义(P<0.01)。慢性乙肝患者IL-6、IL-8、TNF-α表达水平较高,拉米夫定可显著下调IL-6、IL-8、TNF-α至(250.5±33.3)pg/ml、(153.4±22.2)pg/ml、(232.6±21.2)pg/ml,与常规治疗组相比差异有统计学意义(P<0.01)。结论拉米夫定对PBMC及血清内HBV-DNA均具有较好的治疗效果,其阴转率明显高于常规治疗法。拉米夫定比常规药物更具有减轻局部炎性细胞因子浸润、分泌和促进肝细胞功能恢复的作用。     

内毒素诱生的细胞因子及其作用研究进展

革兰阴性菌感染是通过其菌壁脂多糖（ＬＰＳ，内毒素０成分而起作用。ＬＰＳ可诱生多种细胞因子，后者介导内毒素对机体的损伤，引起相应病理变化，并可导致内毒素性休克。细胞因子还能通过调节急性期蛋白的合成参与机体的防御机制。     

细胞因子的复合诱生及初步纯化

采用PHA和LPS对人胎脾脏单个核细胞进行细胞因子复合诱生,结果显示上清中同时含有淋巴因子IL-2、IL-6和单核因子IL-1、IL-6、TNF,而且比单刺激组含量有所提高。进一步采用超滤或盐析技术进行初步纯化,纯化倍数分别为650～730倍和64～78倍,收率分别为74.4～82.6％和36.00～40.5％,继而证明复合细胞因子在低剂量IL-2(100μ/ml)时即可诱导出LAK活性,仅为单纯基因重组IL-2用量的1/10。     

“生精胶囊”对大鼠的生精作用的研究

本文用病理学的计数方法研究了“生精胶囊”对大鼠的生精作用。结果表明“生精胶囊”能明显增加睾丸曲细精管内生殖细胞数目、睾丸内精子数目、曲细精管数量及其直径和曲细精管内细胞层数,增加精子活率而对初级精母细胞直径及精子活动力无明显影响。为临床该药治疗“男性不育少精症”提供了科学依据。     

LFA-1/ICAM-1单抗对ConA诱导脾淋巴细胞增殖及分泌细胞因子的影响

通过研究ＬＦＡ－１／ＩＣＡＭ－１单抗对ＣｏｎＡ诱导的脾淋巴细胞活化增殖及其分泌细胞因子的影响　，探　讨了ＬＦＡ－１／ＩＣＡＭ－１分子在Ｔ细胞活化过程中所起的共刺激作用。结果表明，单独应用ＬＦＡ－１α链单抗（Ｍ１７／４．４．１１．９）或ＩＣＡＭ－１单抗（ＹＮ１／１．７．４）均不能引起脾淋巴细胞的增殖，但在加入ＣｏｎＡ诱导脾淋巴细胞增殖反应的最初８小时内加入ＬＦＡ－１单抗可以剂量依赖性地抑制ＣｏｎＡ诱导的脾淋巴细胞增殖反应及脾淋巴细胞分泌ＩＬ－２、ＩＰＮ－γ，而加入ＩＣＡＭ－１单抗却无此效应。说明ＬＰＡ－１在ＣｏｎＡ诱导的Ｔ细胞活化过程中起着重要的共刺激作用，ＬＰＡ－１通过与除ＩＣＡＭ－１以外的其它配体分子（如ＩＣＡＭ－３）相互识别，提供Ｔ细胞活化所必需的共刺激信号。     

IL-12体外促进慢性乙型肝炎患者PBMC IFN-γ的产生

目的：探讨体外IL-12对慢性乙型病毒性肝炎患者细胞免疫功能的影响，为临床治疗慢性乙型病毒性肝炎提供理论依据。方法：慢性乙型肝炎患者PBMCs与HBsAg、HBsAg＋IL-12、HBcAg或HBcAg＋IL-12孵育后，利用酶联免疫吸附实验（ELISA）检测细胞培养上清中IFN-γ的含量；用酶联免疫斑点实验（ELISPOT）在单个细胞水平上检测PBMCs中IFN-γ产生细胞的频率；用流式细胞记数仪（FACS）检测PBMCs中IFN-γ^＋的细胞亚群。结果：当乙肝患者PBMCs与HBsAg或HBcAg孵育后，只有少数病例发生反应，产生低水平的IFN-γ。当IL-12加入细胞培养后，能显著地增加HBsAg或HBcAg诱导IFN-γ的产生；分泌IFN-γ的细胞频率显著增加；IFN-γ^＋的细胞主要包括CD8^＋T细胞和非T细胞。结论：IL-12体外能增强慢性乙型肝炎患者的细胞免疫应答。     

Study on Anti-Tumor Effect of Total Glycosides from Radix Paeoniae Rubra in S180 Tumor-Bearing Mice

The objective of the paper was to study the anti-tumor effect of total glycosides from Radix paeoniae rubra in S180 tumor-bearing mice, and to preliminarily explore its mechanism of action. Mice were made into S180 solid tumor model, grouped and administered with the extracts; tumor inhibition rate was measured by harvesting the tumors, and serum IL-2 and IL-4 levels were measured by taking blood samples. Total glycosides of Radix paeoniae rubra significantly inhibited the growth of tumor cells in tumor-bearing organisms, enhanced the cytotoxic activity of NK cells, and increased the serum IL-2 and IL-4 levels. Total glycosides of Radix paeoniae rubra have some anti-tumor effect in vivo, which might have been accomplished through the regulation of the immune system.     

Differential effects of Radix Paeoniae Rubra (Chishao) on cytokine and chemokine expression inducible by mycobacteria

Background

Interstitial pulmonary fibrosis is characterized by an altered cellular composition of the alveolar region with excessive deposition of collagen. Lung inflammation is also common in pulmonary fibrosis. This study aims to test the inhibition of 5-lipooxygenase (5-LOX) by boswellic acid (BA) extract in an experimental model of pulmonary fibrosis using bleomycin (BL).

Methods

Boswellic acid extract (1 g/kg) was force-fed to rats seven days prior to administration of BL or gamma irradiation or both. BL (0.15 U/rat) in 25 μl of 0.9% normal saline (NS) or 0.9% NS alone was administered intratracheally. Rats were exposed to two fractionated doses of gamma irradiation (0.5 Gy/dose/week) with a gamma cell-40 (Cesium-137 irradiation units, Canada) during the last two weeks of the experiment. BA was administered during BL or irradiation treatment or both. After the animals were sacrificed, bronchoalveolar lavage was performed; lungs were weighed and processed separately for biochemical and histological studies.

Results

In rats treated with BL, levels of transforming growth factor-β1 (TGF-β1) and tumor necrosis factor-α (TNF-α) were significantly elevated (P = 0.05 and P = 0.005). Hydroxyproline was highly and extensively expressed. Immunoreactive compounds were abundantly expressed, represented in the levels of macrophages infiltrate, accumulation of eosinophils and neutrophils in the lung as well as the aggregation of fibroblasts in the fibrotic area. The levels of lipoxygenase enzyme activity were significantly increased (P = 0.005). Antioxidant activities measured in BL-treated rats deteriorated, coupled with the elevation of both levels of plasma lipid peroxide (LP) content and bronchoalveolar lavage lactate dehydrogenase activity. BA-treated rats had reduced number of macrophages, (P = 0.01), neutrophils in bronchoalveolar lavage (P = 0.01) and protein (P = 0.0001). Moreover, the hydroxyproline content was significantly lowered in BA-treated rats (P = 0.005). BA extract inhibited the TGF-ß induced fibrosis (P = 0.01) and 5-LOX activity levels (P = 0.005). Histologically, BA reduced the number of infiltrating cells, ameliorated the destruction of lung architecture and attenuated lung fibrosis.

Conclusion

BA attenuates the BL-induced injury response in rats, such as collagen accumulation, airway dysfunction and injury. This study suggests that the blocking of 5-LOX may prevent the progression of fibrosis.     

Differential effects of Radix Paeoniae Rubra (Chishao) on cytokine and chemokine expression inducible by mycobacteria

Background  

Upon initial infection with mycobacteria, macrophages secrete multiple cytokines and chemokines, including interleukin-6 (IL-6), IL-8 and tumor necrosis factor-α (TNF-α), to mediate host immune responses against the pathogen. Mycobacteria also induce the production of IL-10 via PKR activation in primary human monocytes and macrophages. As an anti-inflammatory cytokine, over-expression of IL-10 may contribute to mycobacterial evasion of the host immunity. Radix Paeoniae Rubra (RPR, Chishao), a Chinese medicinal herb with potentials of anti-inflammatory, hepatoprotective and neuroprotective effects, is used to treat tuberculosis. This study investigates the immunoregulatory effects of RPR on primary human blood macrophages (PBMac) during mycobacterial infection.     

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。     

脱氧雪腐镰刀菌烯醇对人外周血单个核细胞TAP-1表达的抑制作用

目的: 探讨脱氧雪腐镰刀菌烯醇 (DON)对人外周血单个核细胞 (PBMC)中TAP- 1表达的影响。方法: 采用流式细胞仪(FCM)和半定量RT- PCR方法, 从蛋白和mRNA水平上分析不同浓度的DON对体外培养的人PBMC中TAP- 1分子表达的影响及量 效关系。结果: FCM定量检测表明, 用不同浓度的DON处理, 均可一定程度地抑制人PBMC中TAP -1蛋白的表达, 且二者呈显著的负相关 (r= -0. 865,P<0. 01)。半定量RT -PCR检测显示, 不同浓度的DON处理均可抑制人PBMC中TAP -1mRNA的表达。结论: DON可剂量依赖地抑制体外培养的人PBMC中TAP- 1蛋白和其mRNA的表达, 对阐明食管癌高发区被DON污染的粮食与食管癌发生的关系具有重要的意义。     

黄芪注射液对体外血管新生的影响

背景：血管新生受生长因子的影响,黄芪可与血管内皮生长因子具有协同促血管新生的功效,但机制尚不明确。 目的：通过与单一血管内皮生长因子干预比较,观察中药黄芪对体外血管新生的作用机制。 方法：将黄芪注射液及血管内皮生长因子作用于SD大鼠胸主动脉内皮细胞,应用细胞增殖、细胞迁移、小管形成实验观察黄芪注射液对体外血管新生的促进作用,用Western blot实验检测血管内皮生长因子的表达。 结果与结论：黄芪能明显促进大鼠胸主动脉内皮细胞的增殖(P < 0.01),迁移的细胞数增加(P < 0.01),胸主动脉内皮细胞的小管形成数增加(P < 0.01),并明显促进内皮细胞血管内皮生长因子的表达(P < 0.01)。说明黄芪能明显促进内皮细胞增殖、细胞迁移、小管形成,具有显著的促进体外血管新生作用,其机制可能是通过增加血管内皮生长因子的表达而实现的。     

Sumatriptan increases the proliferation of peripheral blood mononuclear cells from HIV-infected individuals and healthy blood donors in vitro

HIV infection is characterized by the loss of CD4+ T cells as well as the loss of T-cell function, leading to severe immunodeficiency. The proliferative capacity of T cells measured in vitro as responses to antigens and mitogens is severely reduced during HIV infection. An increased level of the intracellular second messenger adenosine 3',5'-cyclic monophosphate (cAMP) has been shown to cause impaired proliferative capacity of peripheral blood mononuclear cells (PBMC) from HIV-infected individuals in vitro. Sumatriptan, a 5HT1d receptor agonist, inhibits the activity of adenylyl cyclases, the enzymes responsible for regulation of the intracellular levels of cAMP. In a preliminary study sumatriptan increased the proliferative responses of PBMC to a polyclonal activator in vitro in 9 of 10 HIV-seropositive individuals (p=0.007), and in 7 of 9 healthy blood donors (p=0.05). This was probably due to a decrease in the intracellular level of cyclic AMP.     

Increased inflammatory responsiveness of peripheral blood mononuclear cells (PBMCs) to in vitro NOD2 ligand stimulation in patients with ankylosing spondylitis

Abstract

Background: Ankylosing spondylitis (AS) is a common debilitating rheumatic disease in which the innate immune components especially the Interleukin (IL)-23/IL-17 axis related genes play important role in its pathogenesis. Nucleotide binding oligomerization domain-containing protein (NOD)2, as an innate receptor, is critical for IL-23 production in cells. Therefore, we aimed to stimulate NOD2 signaling and study its effects on cytokine production in peripheral blood mononuclear cells (PBMC) of these patients.

Methods: PBMCs from 18 patients with active AS and 18 healthy individuals were separated by Ficoll-Hypaque density gradient centrifugation and cultured in the presence of muramyl dipeptide (MDP), as NOD2 ligand. Quantitative expression analysis of NOD1, NOD2, RIPK2, SLC15A4, NLRP1, NLRP3, IL23A, IL17A, IL1B, and TNFA genes was performed using Real-time polymerase chain reaction (PCR). Finally, protein changes of IL23A and IL17A expression were validated using enzyme linked immunosorbent assay (ELISA).

Results: Apart from NOD1 that tend to be downregulated in the controls, all the selected genes showed overexpression in response to MDP in cells from the studied groups. Except RIPK2, all the genes had higher expression changes upon MDP stimulation in the AS population. Overexpression of IL23A and IL17A were confirmed at protein levels using ELISA. The strong positive correlation between NLRP3 and NOD2 was decreased after stimulation but new correlations between NLRP3 and IL1B, RIPK2 and SLC15A4 were observed after treatment.

Conclusions: This study indicated that AS PBMCs were hyper-responsive to MDP stimulation. This observation implies an important role of NOD2 in the pathogenesis of inflammatory diseases including AS.