Protective effects of lactoferrin in <Emphasis Type="Italic">Escherichia coli</Emphasis>-induced bacteremia in mice: Relationship to reduced serum TNF alpha level and increased turnover of neutrophils

Objective and Design:Previous studies demonstrated that lactoferrin (LF), given intravenously (i.v.), 24 h before lethal Escherichia coli (E. coli) infection, protects mice against mortality. The aim of this investigation was to determine whether downregulation of serum TNF alpha activity and increase of neutrophil number in the circulation and bone marrow by LF could contribute to the protective action of LF against E. coli-induced sepsis. Materials and subjects:CBA female mice, 10–12 week old, weight 20–22 g, were used. Treatment:Mice were given 10 mg LF i.v. either 2 h or 24 h before i.v. administration of lethal dose of E. coli (5 × 10⁸). Methods:Serum activities of TNF alpha and IL-1 were determined by bioassays 2 h following E. coli or LF injection. The blood and bone marrow smears were stained with Giemsa and May-Grünwald reagents and reviewed histologically. Results:LF given 24 h before E. coli caused a 60% reduction of TNF alpha released into circulation. However, pretreatment of mice with LF 2 h before bacterial challenge resulted in strong (15 fold) increase of TNF alpha serum level. Analysis of bone marrow cell composition revealed a significant increase in neutrophil lineage cell content (myelocytes, bands and mature neutrophils) following 24 h pretreatment with LF (51.8% of the total cell count), versus PBS control (32.7%) and 2 h LF pretreatment (35.8%). The percentage of neutrophils (bands and mature forms) in the peripheral blood rose to 47.4% versus 32% and 32%, respectively. Intravenous administration of LF increased also interleukin 1 (IL-1) concentration in the circulation of noninfected mice. Conclusions:This investigation has added more information regarding the mechanism of the protective action of LF in E. coli-induced bacteremia by revealing the phenomenon of accelerated neutrophil recruitment and down-regulation of E. coli-induced TNF alpha serum level.Received 22 September 2003; returned for revision 31 October 2003; accepted by M. J. Parnham 14 January 2004     

Serum protects against azurophil granule dependent down-regulation of complement receptor type 1 (CR1) on human neutrophils

We have investigated the effect of gradual degranulation on the expression of functional receptors (CR1 and CR3) on human neutrophils. Incubation with increasing concentrations of fMLP (10^–10–10^–7M) translocated CR1 and CR3 to the cell surface in a similar kinetic pattern. When reaching maximal expression of receptors (10^–7 M fMLP), 78 ± 10% and 87 ± 9% of the total pool of CR1 and CR3, respectively, were translocated to the cell surface. To drive the mobilization process further, cytochalasin B was introduced to increase the stimulatory effect of fMLP. No further increase in CR1 surface expression was obtained. However, we found a characteristic time course of surface appearance of CR1 and CR3 with a maximal surface expression within 1 minute, followed by a time-related down-regulation of CR1 but not CR3. In addition, the total pool of CR1 in cytochalasin B treated neutrophils was reduced after 15 minutes stimulation with fMLP measured by flow cytometry and immunoblotting, indicating degradation of CR1. The down-regulation of CR1 was concomitant with a translocation of azurophil granules, in terms of upregulation of CD63. Azurophil, but not specific nor secretory, granule fractions caused a down-regulation of CR1 on fMLP activated neutrophils. The presence of human sera and serine protease inhibitor protected CR1 from down-regulation. Together, these findings indicate that intracellular stored proteases, released in the late part of the sequential mobilization process, alters the expression of functional receptors mobilized in the early part of the mobilization process. The findings also focus on the importance of the microenvironment for the net outcome of neutrophil activation in terms of functional receptor expression.     

白细胞进入腹腔是引起蛋白质经腹腔丢失的关键

目的为了研究细菌性腹膜炎时经腹腔蛋白质丢失的机制,阐明白细胞移行与蛋白质丢失的关系。方法给兔腹腔内注射大肠杆菌(E.coil)4×106CFU+生理盐水(35ml/kg),做成急性腹膜炎动物模型。通过测定实验6或8h期间腹透液中白细胞总数(WBC)及中性粒细胞(PMNs)计数情况以及腹膜对蛋白质的通透性(蛋白质D/P比值)。设立了两个系列的实验系列一,应用氮介(mustine),1.2mg/kg,实验前3天静脉注射,以耗尽循环血中的白细胞;系列二,用单克隆抗体(mAb)60.32mg/kg,实验前5min静脉注射,以阻滞PMNs上的粘附分子CD18,从而抑制PMNs向腹腔移行。同时设立了阳性对照组(即腹膜炎组)及阴性对照组(无腹膜炎组)。结果系列一中mustine降低87%循环血中白细胞及93%循环血中PMNs,此时即使腹腔内注射细菌,白细胞向腹腔的移行及腹腔蛋白质的渗出均较未注射mustine的腹膜炎组明显降低,而与无腹膜炎的正常对照组结果相似。系列二,静脉注射mAb60.3同样也明显降低白细胞及PMNs向腹腔的移行及经腹腔蛋白质的丢失。结论急性细菌性腹膜炎时白细胞向腹腔移行导致了经腹腔蛋白质的丢失。     

Effect of acute exercise on some haematological parameters and neutrophil functions in active and inactive subjects

In this work we studied the possible effects of acute exercise on some haematological parameters and on some functions of neutrophils in seven active and six inactive subjects. Physical exercise (10 min on a cycle ergometer at a heart rate of 150 beats · min^–1) induced a significant increase in total leucocyte, lymphocyte and neutrophil concentrations in active subjects; serum iron and ferritin concentrations were lower in active compared to inactive subjects. Cellular adhesion, bactericidal activity and superoxide anion production did not change after exercise, while we also observed some differences between active and inactive subjects before exercise. In particular, the neutrophils from active subjects showed a significantly higher percentage of adhesion, higher bactericidal activity and lower superoxide anion production. In conclusion, the training induced changes in some neutrophil functions, while acute exercise influenced, overall, leucocyte concentrations.     

兔中性粒细胞防御素对HL—60细胞毒性及对病毒的直接灭活作用

本研究从新西兰家兔腹腔渗出中性粒细胞溶酶体内提取酸溶性组分，经制备性ＡＵ－ＰＡＧＥ获得纯化的防御素（ＮＰ１和ＮＰ２），ＳＤＳ－ＰＡＧＥ测定分子量为３ｋｄ左右。以ＨＬ－６０细胞作靶细胞，当ＮＰ１或ＮＰ２在１００μｇ／ｍｌ浓度时，可杀死６０％以上的ＨＬ－６０细胞。ＮＰ１（２６０μｇ／ｍｌ）还可直接灭活单纯疱疹病毒Ｉ型和乙型流感病毒，分别使病毒感染滴度降度９８．２％和９０％。本研究结果提示，中性粒细胞可     

当药水提物对大鼠中性白细胞活性氧生成和兔血小板聚集的影响

目的 研究当药水提物对大鼠中性白细胞性细胞内内Ca^2 浓度增加、活性氧生成及兔血小板聚集的影响。方法 用酵母多糖、FMLP和A23 187活化大鼠中性白细胞,用化学发光法测定活性氧,用荧光光度法测细胞内Ca^2 浓度,用比浊法测血小板聚集。结果 当药水提物浓度依赖性地抑制酵母多精、FMLP和A23 187诱导的大鼠中性白细胞内Ca^2 浓度增加及活性氧生成;也抑制花生四稀酸、胶原及ADP诱导的兔血小板聚集。和吲哚美辛比较,当药水提物抑制活性氧生成的作用较强而抑制血小板聚集的作用较弱,其作用远强于swertiamarin。结论 当药水提物是极强的中性白细胞活性氧生成抑制剂,其作用强于swertiamarin。     

Neutrophil extracellular traps in sterile inflammation: the story after dying?

Evidences accumulated that the death of neutrophils are not the end of their missions. The neutrophil extracellular traps (NETs), web-like structure, formed after neutrophils dying contribute greatly to immune defense, in both innate and adaptive immunity. Interestingly, previous studies revealed that the generation and activation of NETs do not only rely on bacteria induction, but also in patients with sterile inflammatory diseases, implying an undeniable correlation between NETs and these diseases. This review summarized the latest findings that the crucial roles of NETs in sterile inflammatory diseases, as well as novel targeted therapy based on these new discoveries.     

Impact of cancer cell-intrinsic features on neutrophil behavior

Neutrophils are multifaceted innate immune cells that play a significant role in the progression of cancer by exerting both pro- and anti-tumorigenic functions. The crosstalk between cancer cells and neutrophils is complex and emerging evidence is pointing at cancer cell-intrinsic programs regulating neutrophil abundance, phenotype and function. Cancer cell-derived soluble mediators are key players in modulating the interaction with neutrophils. Here, we review how intrinsic features of cancer cells, including cancer cell genetics, epigenetics, signaling, and metabolism, manipulate neutrophil behavior and how to target these processes to impact cancer progression. A molecular understanding of cancer cell-intrinsic properties that shape the crosstalk with neutrophils will provide novel therapeutic strategies for personalized immunomodulation in cancer patients.     

牙周炎和慢性阻塞性肺疾病间共同危险因素及相关机制研究进展

慢性阻塞性肺疾病和牙周病两者都是感染引起的慢性炎症反应性疾病，两者的临床表现均涉及结缔组织的破坏。该文就两者的流行病学情况、共同危险因素、相关的生物学机制的研究进展进行综述。文献复习结果表明牙周炎与慢性阻塞性肺疾病两者之间可相互影响，感染细菌同源性，炎症因子和中性粒细胞都参与了两种疾病的过程，两者的共同危险因素有年龄增长、社会经济状况差、吸烟等，但仍需进一步研究来明确牙周炎和慢性阻塞性肺疾病之间相互作用的具体机制。