用CL法和微量MPO法观察人参皂甙对人PMN吞噬杀菌作用

采用化学发光（ＣＬ）法和髓过氧化物酶（ＭＰＯ）微量测定法检测了人参茎叶皂甙对人多形核白细胞（ＰＭＮ）吞噬金黄色葡萄球菌的影响。实验结果表明，人参茎叶皂甙作用过的ＰＭＮ－ＣＬ反应（ＣＬ３０和Ｐｅａｋ）和ＭＰＯ释放水平均增强，对ＣＬ３０与ＣＦＵ、ＭＰＯ与ＣＦＵ行直线相关分析均呈密切相关（ｒ＝０．９５８，ｐ＜０．００１；ｒ＝０．８７５，ｐ＜０．０１）。本实验提示，ＰＭＮ－ＣＬ法和ＭＰＯ微量测定法是研究药物对ＰＭＮ吞噬杀菌功能影响的客观、定量和快速的方法。     

用CL法和微量MPO法观察人参皂甙对人PMN吞噬杀菌…

采用化学发光（ＣＬ）法和髓过氧化物酶（ＭＰＯ）微量测定法检测了人参茎叶皂甙对人多形核白细胞（ＰＭＮ）吞噬金黄色葡萄球菌的影响。实验结果表明，人参茎叶皂甙作用过的ＰＭＮ－ＣＬ反应（ＣＬ３０和Ｐｅａｋ）和ＭＰＯ释放水平均增强，对ＣＬ３０与ＣＦＵ、ＭＰＯ与ＣＦＵ行直线相关分析均呈密切相关（ｒ＝０．９５８，ｐ＜０．００１；ｒ＝０．８７５，ｐ＜０．０１）。本实验提示，ＰＭＮ－ＣＬ法和ＭＰＯ微量测定法是研究药     

多形核白细胞在内毒素休克时肝组织损伤中的作用

家兔输注内毒素０．３ｍｇ／ｋｇ复制内毒素休克模型。输注４ｈｒ以后ＰＭＮ吞噬发光和Ｏ＾－２生成呈持续升高（Ｐ＜０．０５～０．０１），同时激活的ＰＭＮ释放氧自由基损伤肝细胞，引起肝细胞内ＭＤＡ含量和培养上清ＬＤＨ活性升高（Ｐ＞０．０５～０．０１）。体外内毒素与ＰＭＮ共孵，ＰＭＮ吞噬发光和Ｏ＾－２生成呈先升高后回降的变化，经内毒素在体外激活的ＰＭＮ也能明显引起肝细胞内ＭＤＡ含量和上清ＬＤＨ活性升高（Ｐ＜     

锌在蚕噬细胞免疫功能中的作用

本文通过对断奶３天的Ｗｉｓｔｅｒ大鼠进行缺锌喂养，观察其在生长６周后体重，饲料效价、外周血白细胞（ＰＭＮ）及化学发光值（ＣＬ１）、腹腔吞噬细胞（Ｍφ）吞噬率、吞噬指数及化学发光值（ＣＬ２）的变化。发现缺锌大重增长及饲料效价均明显低于自由喂养组（Ｐ＜０．０１），外周血白细胞数明显低于自由喂养组（Ｐ＜０．０１）；腹腔吞噬细胞（Ｍφ）吞噬指数和吞噬率明显低于自由喂养组（Ｐ＜０．０１）；化学发光值ＣＬ１和     

绿脓杆菌外毒素A对人PMN功能的影响

为了证明绿脓杆菌外毒素Ａ（ＰＥＡ）对人中性粒细胞（ＰＭＮ）的破坏以及对ｐＭＮ功能的影响，本文报道用不同剂量的ｐＥＡ和作用时间作用于ＰＭＮ，并用抗ｐＥＡ抗体进行中和保护，分别测定完存的ＰＭＮ数目、超氧（Ｏ２－）水平、特殊颗粒（ＳＧ）数目、胞内杀菌力（ＩＣＢＡ）和吞噬百分数。结果表明，ＰＥＡ４０～１００μｇ／ｍｌ作用１～３ｈ，使ＰＭＮ数目、Ｏ２－水平、ＳＧ数目和ＩＣＢＡ测值明显下降，作用２ｈ以后使其吞噬百分数也明显下降；抗ｐＥＡ抗体能阻止上述指标测值的下降（ｐ＜０．０５）。上述结果证实：ＰＥＡ是造成ＰＭＮ杀菌力和完存ＰＭＮ数目下降的直接原因。     

P物质对人多形核白细胞功能的影响

目的研究Ｐ物质（ＳＰ）对人多形核白细胞（ＰＭＮ）有关方面作用及可能的意义。方法应用硝基四氮唑蓝（ＮＢＴ）还原法，荧光法和Ｇｒｉｅｓｓ反应等测定不同浓度ＳＰ单独或在细菌衍生肽类似物ＦＭＬＰ甲酯存在下对ＰＭＮ产生超氧阴离子（Ｏ－２，）过氧化氢（Ｈ２Ｏ２），一氧化氮（ＮＯ）和ＰＭＮ膜上一功能酶中性内肽酶（ＮＥＰ）活性及膜流动性的影响。结果ＳＰ（≥１０－５ｍｏｌ／Ｌ）可剌激ＰＭＮ显著产生Ｏ－２，Ｈ２Ｏ２和ＮＯ，后者被Ｌ－单甲基精氨酸（ＮＭＭＡ）所抑制；ＳＰ（１０－８～１０－４ｍｏｌ／Ｌ）能显著增加ＦＭＬＰ甲酯剌激ＰＭＮ产生Ｈ２Ｏ２，并显浓度递增依赖性；ＳＰ与ＦＭＬＰ甲酯协同可下调ＮＥＰ活性；ＳＰ能提高ＰＭＮ膜流动性。结论ＳＰ可通过ＰＭＮ介导调节炎症反应，对ＰＭＮ的杀菌功能有增强作用，ＳＰ对ＰＭＮ的这些影响可能是神经系统参与炎症和免疫调节的途径之一。     

锌在吞噬细胞免疫功能中的作用

本文通过对新奶３天的Ｗｉｓｔｅｒ大鼠进行缺锌喂养，观察其在生长６周后体重、饲料效价、外周血白细胞（ＰＭＮ）及化学发光值（ＣＬ１）、腹腔吞噬细胞吞噬率、吞噬指数及化学发光值（ＣＬ２）；的变化。发现缺锌大鼠体重增长及饲料效价均明显低于自由喂养组（ｐ＜０．０１），外周血白细胞数明显低于自由喂养组（Ｐ＜０．０１）；腹腔吞噬细胞吞噬指数和吞噬率明显低于自由喂养组（Ｐ＜０．０１）；比学发光值ＣＬ１和ＣＬ２均明显低于自由喂养组（Ｐ＜０．０１）。提示缺锌不仅影响大鼠的正常生长，且影响大鼠的吞噬功能和灭菌能力，由此降低大鼠的免疫功能。     

NIDDM患者中性粒细胞膜流动性的改变及其影响因素

目的检测６５例非胰岛素依赖型糖尿病（ＮＩＤＤＭ）患者中性粒细胞（ＰＭＮ）膜流动性（ＭＦ）的改变,探讨这种改变与年龄、病程、ＢＭＩ、ＦＢＧ、ＰＢＧ、ＴＧ、ＴＣ、ＷＢＣ、ＲＢＣ、ＰＬＴ、ＧＰＩ的相关关系。方法６５例无酮症及感染的ＮＩＤＤＭ患者,３０例性别、年龄、ＢＭＩ相匹配的正常人为对照,以ＤＰＨ为探针测定其ＰＭＮ的荧光偏振值（ρ）,并计算相应的细胞膜流动性、膜脂各向异性及微粘度,对ＰＭＮ ρ值的影响因素进行分析。结果１ＮＩＤＤＭ患者的ＰＭＮｐ值显著高于对照组（Ｐ＜０．０１）,ＰＭＮ　ＭＦ明显低于对照组（Ｐ<０．０５）,ＰＭＮ膜脂各向异性及微粘度均显著升高（Ｐ<０．０１）。 ２ ＰＭＮ ＭＦ与ＦＢＧ、ＰＢＧ呈负相关（ｒ＝－０．３４１７,ｒ＝－０．３２３７,Ｐ<０．０５）,与ＧＰＩ相关性显著（ｒ＝－０．３９１３,Ｐ<０．０５）,与年龄、病程、ＢＭＩ、ＷＢＣ、ＲＢＣ、ＰＬＴ、ＴＧ、ＴＣ无相无性。结论ＮＩＤＤＭ患者ＰＭＮ的ＭＦ下降,显示ＤＭ患者ＰＭＮ膜的物理性质有明显改变;ＰＭＮ ＭＦ下降与血糖升高、血清蛋白糖化程度增加有关。     

抗氧化剂PDTC抑制氧化的低密度脂蛋白诱导的人肾小球系膜细胞NF_(-k)B活化

目的 研究氧化的低密度脂蛋白（Ｏｘ－ＬＤＬ）对体外培养的人肾小球系膜细胞（ＨＭＣ）核因子－ＫＢ（ＮＦ－ＫＢ）活化的影响，以及抗氧化剂毗咯二硫氨基甲酸酯（Ｐ ＤＴＣ）对ＮＦ－ＫＢ活化的抑制作用，探讨ＯＸ－ＬＤＬ介导肾损害的基因调控机制及抗氧化剂ＰＤＴＣ防治脂质肾损害的可能性。方法 将Ｏｘ－ＬＤＬ或ＰＤＴＣ与ＨＭＣ共培养后，提取细胞核蛋白进行凝胶迁移率变动分析（ＥＭＳＡ）检测ＮＦ－ＫＢ的活化，用细胞ＥＬＩＳＡ法检测细胞内ＩＫＢα蛋白含量的变化，反映ＩＫＢα的降解及免疫组化染色检测细胞内的Ｐ６５向核转位。结果 正常对照组未见ＮＦ－ＫＢ活化，当用不同浓度（１０、２５、５０及１００ｍｇ／Ｌ）的Ｏｘ－ＬＤＬ，刺激肾小球系膜细胞 ｌｈ后，均可引起细胞 ＮＦ－ＫＢ活化及 ＩＫＢα降解。与对只组相卜较差异显著（ｐ＜０．０５），以５０ｍｇ／Ｌ 的ＯＸ－ＬＤＬ刺激ＨＭＣ１ｈ，ＮＦ－ＫＢ活化及 ＩＫＢα降解最明显。ＮＦ－ＫＢ活化的同时，伴有Ｐ６５由胞浆向胞核的转位。１００μｍｏｌ／Ｌ ＰＤＴＣ，能明显抑制 ＮＦ－ＫＢ的活化、ＩＫＢα降解（ｐ＜０．０１）及 Ｐ６５的核转位。结论Ｏｘ－ＬＤＬ。能诱导ＨＭＣ的ＩＫＢαａ降解、Ｐ６５的核转位，最终使Ｎ     

败血症新生儿外周血中性粒细胞膜流动性变化的观察

为了探讨新生儿败血症时外周血中性粒细胞（ＰＭＮ） 功能异常的膜分子机制， 应用ＤＰＨ 荧光探针标记ＰＭＮ膜， 荧光偏振法测定２８ 例败血症新生儿、１９ 例健康新生儿ＰＭＮ膜流动性。结果： 败血症新生儿ＰＭＮ 膜流动性显著降低， 与对照组比较有极显著差异（ Ｐ＜ ０－０１ ）； 相关分析结果表明， ＰＭＮ 膜微粘度（η）与吞噬、杀菌率呈负相关， 相关系数（γ） 分别为 ０－３２１、 ０－３２６。研究结果提示， ＰＭＮ膜流动性降低是造成败血症新生儿ＰＭＮ 吞噬、杀菌功能降低导致易发感染的重要原因。     

Phagocytosis of virulent and avirulent leptospires by guinea-pig and human polymorphonuclear leukocytes in vitro

Chemiluminescence (CL) and electron microscopy were used to study the phagocytosis of both virulent and avirulent strains of Leptospira interrogans serovar copenhageni by guinea-pig and human polymorphonuclear leukocytes (PMN). A significant CL response was observed when guinea-pig PMN were incubated with virulent leptospires in the presence but not in the absence of specific immune serum. This response was markedly enhanced by the addition of guinea-pig complement. Phagocytosis was confirmed by the observation of intracellular leptospires in guinea-pig PMN by electron microscopy. The phagocytosis of avirulent leptospires by guinea-pig PMN and of both virulent and avirulent leptospires by human PMN required the presence of both specific immune serum and complement. Thus the ability of leptospires to resist phagocytosis by PMN in the absence of immune serum does not appear to be a major determinant of virulence.     

In vitro effect of asbestos fibers on polymorphonuclear leukocyte function

Incubation of chrysotile and amphibole asbestos fibers with normal human peripheral blood polymorphonuclear leukocytes (PMN) resulted in a significant stimulation of PMN metabolic activity and generation of toxic oxygen by-products as measured by chemiluminescence (CL). Although all asbestos fibers tested were cytotoxic to PMN, cytotoxicity and CL varied disproportionately with fiber type. Anthophyllite asbestos produced the greatest PMN cytotoxicity. It also depressed PMN phagocytosis of latex beads the most and induced the greatest PMN CL response of the fiber types examined. We postulate that asbestos-induced release of toxic oxygen by-products from PMN which have infiltrated into the pulmonary alveoli may contribute to disease pathogenesis in asbestosis.     

Leukocyte spreading behavior on vascular biomaterial surfaces: consequences of chemoattractant stimulation

Chemoattractant-induced phenomena of polarity and migration of polymorphonuclear leukocytes (PMN) are believed to play a key physiological role in controlling bacterial infections on implantable vascular biomaterials. Our study targeted the spreading behavior of human PMN adherent to expanded polytetrafluoroethylene (ePTFE), pretreated with various plasma proteins, in response to the chemoattractant, N-formyl-methionyl-leucyl-phenylalanine (fMLP). To this end, a novel imaging configuration was developed to allow in situ reconstructive analysis of PMN 3-D morphology on opaque ePTFE surfaces, using optical sectioning confocal microscopy. Following fMLP stimulation, PMN morphological polarity was enhanced on all substrates studied except fibrinogen treated ePTFE. 3-D PMN morphometry revealed that in the absence of fMLP, overall cell spreading was minimized on albumin-treated ePTFE and maximized on fibrinogen and immunoglobulin-G-treated ePTFE. Following fMLP stimulation, overall PMN spreading increased markedly on untreated and albumin-coated ePTFE, while it stayed invariant on IgG and plasma treated ePTFE, and decreased on fibrinogen-treated ePTFE. Spatial analysis of PMN spreading following fMLP stimulation revealed enhanced PMN attachment on untreated and albumin treated ePTFE and diminished attachment on fibrinogen and plasma treated ePTFE. Thus, chemoattractant stimulation altered a wide range of PMN spreading attributes on ePTFE, including morphological polarity, substrate attachment, and 3-D membrane spreading, in a substrate dependent manner. These chemoattractant-induced spreading responses may also have important consequences for PMN phagocytosis. We report that fMLP stimulation led to enhanced unopsonized particulate phagocytosis on untreated and albumin treated ePTFE, but caused no discernible change in phagocytosis on other protein substrates. Thus, chemoattractant modulation of PMN spreading on ePTFE is highly substrate-regulated, and manifests in concerted effects on PMN phagocytosis.     

猪苓及猪苓多糖对BBN诱导的膀胱癌大鼠胸腺、脾指数及CD86表达的影响

目的观察猪苓及猪苓多糖(PPS)对BBN诱导膀胱癌过程中对大鼠胸腺、脾指数和膀胱组织免疫细胞浸润及CD86表达的影响,从而探讨其抗肿瘤作用中的免疫学机理。方法用N-丁基-N-4-羟丁基亚硝胺(BBN)和糖精水诱导建立Fisher344大鼠膀胱肿瘤模型,实验分6组即对照组、模型组、PPS组(130 mg.kg-1)及猪苓低、中、高剂量组(剂量分别为50、250、500 mg.kg-1)。用称重法测定大鼠的体质量和胸腺、脾脏质量,计算胸腺指数和脾指数;苏木素-伊红(HE)染色观察各组间质淋巴细胞浸润情况,免疫组化法观察各组大鼠膀胱组织CD86的表达情况。结果模型组大鼠胸腺指数显著降低(P<0.05);PPS组的胸腺指数明显高于模型组(P<0.05),猪苓各组脾指数与模型组比较差异无统计学意义。PPS组、猪苓低剂量和中剂量组淋巴细胞浸润较明显,可见淋巴滤泡形成;PPS组与模型组比较有显著性差异(P<0.05)。模型组膀胱肿瘤组织中CD86弱表达或几乎不表达;PPS组膀胱癌上皮CD86表达明显增强,且癌旁上皮CD86有表达。结论猪苓和PPS可通过影响膀胱癌模型大鼠胸腺、脾指数和膀胱组织及癌旁组织淋巴细胞浸润及CD86表达,PPS的作用更显著,猪苓和PPS参与了抑制肿瘤的发生发展过程。     

The interaction of ruminant IgG with receptor type II for IgG on human phagocytes

The interaction of ruminant IgG with human phagocytes was assessed using Fc receptor (FcR)-mediated ingestion and the triggering of a respiratory burst as effector functions indicative of receptor-specific interaction. In monomeric form, ruminant IgG was three to five orders of magnitude less potent than homologous IgG in inhibiting FcR-specific phagocytosis by monocytes. However, when attached to tanned sheep erythrocytes (Es-T), ruminant IgG was opsonic, as it promoted enhanced phagocytosis of Es-T, comparable to ingestion of rabbit IgG-coated Es. This phagocytosis was inhibitable by high concentrations of human IgG in the fluid phase. Moreover, Es-T precoated with ferritin could be opsonized to a similar degree by anti-ferritin IgG from rabbit and cow. However, only bovine IgG1, but not IgG2, was opsonic. Bovine and goat IgG of some, but not other, suppliers were inactive. Similar results were obtained by measuring the respiratory burst triggered by heat-aggregated IgG, using a luminol-enhanced chemiluminescence assay. Thus, human IgG and ruminant IgG stimulated monocytes and, to a lesser extent, polymorphonuclear leucocytes (PMN), to generate CL. Depending on the manufacturer, some preparations of bovine and goat IgG were inactive, and bovine IgG2 failed to induce CL. These findings prove that certain ruminant IgG preparations, including bovine IgG1 interacting weakly with homologous PMN and monocytes, do interact with human PMN, monocytes and macrophages in a FcR-specific manner when offered in complexed form. Inhibition studies suggest that bovine IgG1 interacts mainly with human FcR type II. In contrast, bovine IgG2, regarded as cytophilic for homologous PMN, fails to interact with human PMN, monocytes and macrophages.     

Activation of human polymorphonuclear leucocytes by particulate zymosan is related to both its major carbohydrate components: glucan and mannan.

Unopsonized particulate zymosan and its major carbohydrate component glucan were phagocytosed under serum-free conditions by adherent polymorphonuclear leucocytes (PMN) in a dose- and time-dependent manner. Preincubation of PMN monolayers with mannan did not cause a reduction in the phagocytosis of either particle. The phagocytic response was inhibited by preincubation of the cells with trypsin at a concentration that did not inhibit the phagocytosis of sheep erythrocytes coated with IgG or of latex particles. Homology of the recognition mechanisms for glucan and zymosan was confirmed when cells cultured on fixed glucan or on fixed zymosan failed to ingest either particle to more than 40% of control phagocytosis. Similarly, zymosan and glucan activated PMN in suspension, in a dose- and time-dependent manner, to generate reactive oxygen species which were measured as luminol-dependent chemiluminescence (CL). There was, however, a four-fold greater CL response to zymosan. Preincubation of PMN with mannan resulted in a significantly decreased CL response to zymosan, while the response to glucan was unaffected. The CL response was also sensitive to a range of concentrations of trypsin. In contrast, two other complex polysaccharide particles (barley-derived beta-glucan and algae-derived laminarin) were not phagocytosed by PMN, nor did they cause the generation of CL, despite the fact that they possessed the capacity, in common with zymosan and glucan, to activate the alternative pathway of complement. The identification of a trypsin-sensitive recognition mechanism on the surface of human PMN for unopsonized zymosan and glucan represents a response not hitherto characterized. Furthermore, our data indicate that the phagocytosis of unopsonized zymosan by human PMN is dependent primarily on its glucan content, but that its capacity to activate the respiratory burst may involve mannan and the recruitment of a second cell surface recognition mechanism.     

The selective augmentation by recombinant human tumour necrosis factor-alpha of neutrophil responses to pathogenic Escherichia coli.

Endotoxin release may amplify the neutrophil (PMN) responses to bacterial infection through the release of monocyte-derived tumour necrosis factor (TNF). The present study was designed to assess the effect of recombinant human TNF-alpha (rhTNF-alpha) on the in vitro response of human PMN to two defined strains of pathogenic Escherichia coli. In the absence of rhTNF-alpha, a P-fimbriate strain caused significant release of the PMN secondary granule marker vitamin B12-binding protein (B12 BP), and a low level of release of leukotriene B4 (LTB4). Type 1-fimbriate strain 504, however, stimulated the release of the primary granule marker myeloperoxidase (MPO) and PMN chemiluminescence (CL), in addition to B12 BP and LTB4 release. Following rhTNF-alpha (10(-9) M) pretreatment, the release of LTB4 by PMN stimulated with the P-fimbriate strain was synergistically augmented, while B12 BP and MPO release were additively increased. In contrast, rhTNF-alpha did not significantly affect any of the responses by the type 1-fimbriate strain. These results suggest selectivity in the priming of PMN by rhTNF-alpha and confirm the independence of PMN responses to phagocytic stimuli.     

Effects of human interferon preparations on neutrophil function

We have studied effects of two partially purified human leukocyte (alpha) interferon (IFN) preparations (PIF-A and PIF-B) and a highly purified fibroblast (beta) IFN on the functional activity of normal human neutrophils (PMNs). In vitro, PIF-B conferred a significant and dose-dependent enhancement of chemiluminescence (CL) induced both by phagocytosis and a soluble stimulus, f-Met-Leu-Phe, and decreased killing of Staph. aureus. In contrast, PIF-A caused only a slight inhibition of bactericidal activity and had no effects on CL. beta-IFN had no effects on either bactericidal activity or CL. Migration under agarose was decreased with all of the IFN but phagocytosis and release of enzymes was not affected. PMNs from seven patients treated with PIF-A for multiple myeloma exhibited increased CL responses but no other PMN functions were affected. The findings that human IFN preparations affect PMN functions indicate that high-dose IFN therapy of immunocompromised patients should be carefully evaluated for the possibility of increased infectious complications.