pHGF对人中性粒细胞吞噬功能的影响

我们曾报道，从乳猪肝脏提取的促肝细胞生长因子（ｐＨＧＦ）能增强小鼠中性粒细胞的吞噬功能，本文进而用吞噬杀菌试验、化学发光技术和促凝血活性试验证明，ｐＨＧＦ也能显著增强人中性粒细胞的吞噬和杀菌功能。     

全血化学光Ⅰ.介绍一种测定全血中粒细胞吞噬功能和调理活性的简便和快速的方法

本文介绍用鲁米诺依赖性全血化学发光方法(Luminol-dependent Chemilumins-eenee of whole blood,简称CL)来同时测定粒细胞吞噬功能和血清调理活性。前者由全血CL峰值和样品中粒细胞计数来判定,后者由加入未经调理的CL诱导物(见下)后CL峰出现的时间来判定。既往用CL分析吞噬细胞功能常使用纯化的粒细胞,需血量大,且制备过程对粒细胞功能又有很多     

中性粒细胞化学发光测定的实验研究

<正> 我们用国产FG83-1型化学发光测试仪,建立了分离的中性粒细胞(PMN)化学发光(CL)测定法,并对影响CL测定的因素作了探讨。 方法:参照Boyum法,从外周血分离出PMN。取PMN悬液1ml、加佴米诺溶液0.2ml混匀后移入样品室,测定木底,然后加入经正常人混合血清     

人工制备流行性出血热病毒免疫复合物对人中性粒细胞功能的影响

工制备流行性出血热病毒免疫复合物(EHFV-IC),用分离之正常人中性粒细胞(NPL)作吞噬试验,双标记免疫荧光染色表明,NPL吞噬免疫复合物百分率可达100％。体外将IC与正常人NPL共同孵育20min后,用化学发光法(CL)检测NPL功能,发现NPL对调理的酵母多糖(OZ)的应答能力明显下降,并与所加IC的浓度呈负相关。说明EHFV-IC可被NPL吞噬,同时造成NPL功能受损。     

定量测定中性粒细胞吞噬功能及其初步临床应用

<正> 定量测定细胞吞噬功能,对于研究患者的免疫功能及指导临床治疗具有意义。以往大多采用斑蝥发疮、粒细胞吞菌及硝基蓝四氮唑(NBT)等方法观察细胞吞噬功能。但这些方法通常只能定性且操作较繁琐,有一定局限性。 1980年Pick等报道了一个利用生化比色的简便方法,定量测定细胞吞噬功能。即     

慢性阻塞性肺部疾病患者中性粒细胞吞噬和杀灭白色念珠菌功能的初步观察

嗜中性粒细胞是具有高度吞噬功能的细胞,它在防御感染中起着重要作用,能参与非特异性免疫应答。1980年我院曾建立了一个快速、简易、价廉的方法,以测定中性粒细胞吞噬和杀灭白色念珠菌的     

脐血和母血嗜中性粒细胞吞噬功能的观察

<正> 本文对50例脐血和母血进行了嗜中性粒细胞吞噬力的检测,结果脐血和母血分别为76.8±20％和85.7±12％,两者经统计学处理P<0.01,脐血的吞噬力明显低于母血。另对32例脐血标本中加入20％母血清,结果脐血的吞噬力从83.2％上升到91.3％,达母血吞噬力水平。本文还对结果进行了扼要的讨论,现报告如下。 一、试剂:(1)白色葡萄球菌悬液;(2)固定剂:甲醇丙酮(1:1),(3)姬姆萨和瑞氏染液;(4)肝素25～50u/ml。 二、方法:取全血4滴于小葫芦瓶中,然后加     

小鼠巨噬细胞吞噬功能检测方法的改进

<正> 在徐淑云介绍的用新鲜鸡红细胞(CRBC)作为吞噬物,半体内法测定小鼠巨噬细胞(Mφ)吞噬功能的基础上,我们进行了改进。其一将CRBC进行醛化。取阿氏液抗凝的新鲜CRBC用生理盐水离心洗2次,1ml压积的CRBC中加入24ml生理盐水、0.1ml 25％戊二醛,吹打均匀,室温下摇动30分钟,用生理盐水洗3次,4℃可存1年,使用时配成1％浓度;其二省去Mφ与吞噬物在玻片上于37℃温育使之贴壁的步骤,而将1％醛化CRBC注入小鼠腹腔,待作用1小时后,处死小鼠、用洁净玻片在暴露的腹     

慢性重症乙型肝炎患者血糖和中性粒细胞吞噬及细胞内杀菌功能检测的临床意义

慢性重症乙型肝炎常伴有细胞免疫抑制、造成患者免疫功能的降低[1],而中性粒细胞又是细胞免疫的重要组成部分。为了进一步地探讨慢性乙型肝炎的免疫功能水平,本文对患者进行了血糖及中性粒细胞吞噬和细胞内杀菌功能的测定,现将结果报告如下。     

中性粒细胞的化学发光试验

中性粒细胞的活化所产生的活性氧,溶酶体酶等在组织损伤中起重要作用。本文观察某些激动剂对中性粒细胞(PMN)化学发光的作用及山茛菪碱(654-2)等药物对其影响。 按以前报道的方法,由大鼠腹腔取得PMN,用Hank's液调制成5×10~6/ml细胞悬液备     

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。     

Role of serotype-specific polysaccharide in the resistance of Streptococcus mutans to phagocytosis by human polymorphonuclear leukocytes

To clarify the role of cell surface components of Streptococcus mutans in resistance to phagocytosis by human polymorphonuclear leukocytes (PMNs), several isogenic mutants of S. mutans defective in cell surface components were studied with a luminol-enhanced chemiluminescence (CL) assay, a killing assay, and a transmission electron microscope. The CL responses of human PMNs to mutant Xc11 defective in a major cell surface antigen, PAc, and mutant Xc16 defective in two surface glucosyltransferases (GTF-I and GTF-SI) were the same as the response to the wild-type strain, Xc. In contrast, mutant Xc24R, which was defective in serotype c-specific polysaccharide, induced a markedly higher CL response than the other strains. The killing assay showed that human PMNs killed more Xc24R than the parent strain and the other mutants. The transmission electron microscopic observation indicated that Xc24R cells were more internalized by human PMNs than the parental strain Xc. These results may be reflected by the fact that strain Xc24R was more phagocytosed than strain Xc. The CL response of human PMNs to a mutant defective in polysaccharide serotype e or f was similar to the response to Xc24R. Furthermore, mutants defective in serotype-specific polysaccharide were markedly more hydrophobic than the wild-type strains and the other mutants, suggesting that the hydrophilic nature of polysaccharides may protect the bacterium from phagocytosis. We conclude that the serotype-specific polysaccharide, but not the cell surface proteins on the cell surface of S. mutans, may play an important role in the resistance to phagocytosis.     

Role of the capsular polysaccharide-like serotype-specific antigen in resistance of Actinobacillus actinomycetemcomitans to phagocytosis by human polymorphonuclear leukocytes.

Serotype b-specific polysaccharide antigen (SPA) of Actinobacillus actinomycetemcomitans Y4 consists of D-fucose and L-rhamnose. To clarify the role of SPA in phagocytosis of the organism by human polymorphonuclear leukocytes (PMNs), monoclonal antibodies (MAbs) against SPA and SPA-defective mutants, which were constructed by inserting the transposon Tn916 into strain Y4, were used in a chemiluminescence (CL) assay and a phagocytic killing assay. The CL responses of human PMNs to strain Y4 were very low, and the organism was not killed by PMNs. In contrast, SPA-defective mutants induced strong CL responses. The addition of immunoglobulin G MAbs against Y4 SPA enhanced significantly both the CL responses to strain Y4 and the killing of the organism in the presence of complement. The CL responses to SPA-defective mutants were little affected by the addition of these MAbs. We conclude that SPA of A. actinomycetemcomitans plays an important role in the resistance to host defenses by PMNs.     

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.     

Priming effect of fibronectin on respiratory burst of human neutrophils induced by formyl peptides and platelet-activating factor

Fibronectin (FN), a glycoprotein present in the plasma and the extracellular matrix, has been shown to enhance adherence-related functions of polymorphonuclear leukocytes (PMNs). In this study we investigated the effects of FN on the activation of human PMNs in suspension by soluble stimuli, as determined by the generation of Superoxide radicals (respiratory burst). FN (up to 100g/ml) did not directly stimulate the PMN respiratory burst assessed using a sensitive assay, luminol-dependent chemiluminescence (CL). Low FN concentrations (Up to 25gl ml) caused a dose-dependent enhancement of the CL induced by two chemoattractants,N-formyl-methionyl-leucyl-phenylalanine (FMLP) and platelet-activating factor (Paf), and also by phorbol myristate acetate (PMA), a known protein kinase C activator. Higher FN concentrations were less effective. The potentiation involved both initial rate and total CL responses and was more active on extracellular than intracellular generation of oxygen radicals. FN potentiation persisted after cell washing and was abolished by treatment of FN with trypsin. Measurement of the respiratory burst using the cytochrome c reduction assay confirmed that FN enhanced both the initial rate and total amount of Superoxide anion generated by FMLP-stimulated PMNs. These data indicate that FN facilitates the respiratory burst of chemoattractant-stimulated PMNs and suggest that FN can prepare PMNs in suspension for amplified biological functions induced by soluble inflammatory stimuli.     

Saliva inhibits the chemiluminescence response, phagocytosis, and killing of Staphylococcus epidermidis by polymorphonuclear leukocytes.

Saliva inhibited several functional properties of polymorphonuclear leukocytes (PMNs) from murine peritoneal exudate, namely, luminol-mediated chemiluminescence (CL) induced by either Staphylococcus epidermidis or formylmethionyl-leucyl-phenylalanine (FMLP), phagocytosis, and killing of bacteria in vitro. The concentration of saliva in the reaction mixture that caused a complete inhibition of the CL response of PMNs to both S. epidermidis and FMLP was 25%. However, there was no catalase or superoxide dismutase activity in saliva that could influence the CL response of PMNs. The production of superoxide by PMNs stimulated with S. epidermidis was assayed in the presence or absence of saliva by inhibition of the reduction of cytochrome c by superoxide dismutase. In the presence of 50% saliva, O2- generation by PMNs was only 7.3% of that observed in the absence of saliva. After gel filtration of salivary material through Sephadex G-25 or Sephacryl S-200, several fractions were obtained that inhibited the CL response of PMNs to either FMLP or S. epidermidis or to both. Two inhibitory fractions were analyzed. One contained immunoglobulin A, and the other contained a peptide which was composed of 14 different amino acids. The two fractions of high molecular weight included in the first protein peak of Sephacryl S-200 gel filtration were able to inhibit the CL response to S. epidermidis and to inhibit phagocytic activity, while fractions of low molecular weight (under 12,500 Mr) inhibited the CL response to FMLP and to S. epidermidis but did not inhibit phagocytic activity.     

Modulation of human polymorphonuclear leukocyte function by the flavonoid silybin

The effect in vitro of the naturally occurring flavonoid silybin on human polymorphonuclear leukocyte (PMN) functions has been studied. Preincubation of PMNs for 10 min at 37 degrees C with silybin inhibited, in a dose-dependent way, the luminol-enhanced chemiluminescence (CL) generated by stimulated cells without affecting the non-enhanced CL or superoxide anion production evaluated by the cytochrome C reduction assay. No significant effect of silybin on PMN phagocytic or chemotactic activities were found. Silybin did not absorb light at the wavelength of luminol-enhanced CL and was not toxic to PMNs at the concentrations used. Catalase, a scavenger of H2O2, inhibited luminol-enhanced CL to a similar degree as silybin; moreover, when incubated together with PMNs, silybin and catalase did not produce an additive inhibition of CL. On the contrary, the simultaneous addition of silybin and sodium azide, an inhibitor of myeloperoxidase, further increased inhibition over that seen with azide alone. These results suggest that inhibition of H2O2 may be the mechanism by which silybin inhibits the luminol-enhanced CL generated by stimulated PMNs. Such results indicate a possible anti-inflammatory activity for silybin even if their clinical relevance remains to be elucidated.     

Antioxidant Properties of Lunularia Cruciata (Bryophyta) Extract

The effects of Lunularia cruciata (L.) Dum (Bryophyta) acetonic extract was studied in vitro by means of luminol-dependent chemiluminescence (CL) emission from human peripheral whole blood phagocytes and isolated polymorphonuclear leukocytes (PMNs). L. crudata adult thalli underwent extraction with acetone. CL emission was evaluated in an automated luminometer, measuring the oxygen free-radical production by phagocytes incubated with opsonized zymosan (OZ) or phorbol myristate acetate (PMA), in absence or in presence of various concentrations of L. crudata extract. The CL results indicated that L. crudata induced significant changes in light emission from whole blood phagocytes, as well as isolated PMNs. Its inhibitory activity was more evident when resting isolated PMNs were studied. When the cells were activated, the greatest inhibitory effect was observed with PMA. The L, crudata activity could be caused by several compounds, such as flavonoids and or sesquiterpenes, present in the acetonic extract.     

Lucigenin-dependent chemiluminescence as a new assay for NAD(P)H-oxidase activity in particulate fractions of human polymorphonuclear leukocytes

The enzyme responsible for the respiratory burst in human neutrophils is an oxidase that catalyzes the reduction of oxygen to superoxide anion (O-2). Superoxide anion production may be measured by chemiluminescence (CL) in the presence of lucigenin (10,10'-dimethyl-9,9'- biacridinium dinitrate). We established an assay of the oxidase, by measuring the CL of particulate fractions of PMN in the presence of lucigenin . This CL required the addition of NAD(P)H and was very low in fractions of resting cells. In particulate fractions of PMNs stimulated with PMA selectively, the NADPH-dependent CL was found to be increased. CL was linear with protein concentrations up to 100 micrograms and was shown to be at least 10 times more sensitive for the detection of O-2 than the assay based on the spectrophotometric determination of superoxide mediated cytochrome c reduction. CL was abolished by inactivating the enzyme at 56 degrees C.     

Modulation of the inflammatory response by products released from human polymorphonuclear leukocytes during phagocytosis

During phagocytosis of latex particles human polymorphonuclear leukocytes (PMNs) release a product that generates chemotactic activity from fresh human serum. Release of this product is maximal at 20–30 min of phagocytosis. It is present in resting PMNs, may be recovered from granule fractions, and functions at physiologic pH. Gel-filtration chromatography of the activated serum indicates that C5a accounts for most of the chemotactic activity generated. Studies utilizing preparations of C5, EDTA, magnesium-EGTA, CS-deficient serum, and serum heated for 20 min at 50°C demonstrate that generation of C5a results from activation of the complement system as well as from direct cleavage of C5. Activation of the complement system by this PMN-derived serum activator appears to proceed through both the alternate and classical pathways. In addition to this serum activator, an inactivator of C5a chemotactic activity is also released by the PMNs under certain conditions of phagocytosis. These studies suggest that phagocytizing PMNs have secretory functions that contribute to the localization and amplification of inflammatory responses.