欧洲鳗鲡外周血细胞的显微和超微结构

利用光镜和透射电镜技术研究了欧洲鳗鲡（Anguilla anguilla）外周血细胞的显微和亚显微结构。结果表明：在外周血细胞中可以区分出红细胞、单核细胞、大淋巴细胞、小淋巴细胞、嗜中性粒细胞和血栓细胞;嗜中性粒细胞内的特殊颗粒包括A、B、C三型;嗜中性粒细胞分为Ⅰ型粒细胞（内含A、B、C三种特殊颗粒）、Ⅱ型粒细胞（内含A型和C型两种特殊颗粒）和Ⅲ型粒细胞（内含C型特殊颗粒）。还见到幼稚的和正在分裂的红细胞和单核细胞,提示示幼稚的红细胞能直接进入外周血流中,嗜酸性粒细胞和嗜碱性粒细胞无论在血涂片上或超薄切片上均未见到。描述了上述各种血细胞在光镜和电镜观察下的形态和细微结构。血栓细胞体积最小,嗜中性粒细胞体积最大;淋巴细胞数目最多,嗜中性粒细胞数目最少。     

临床型乳房炎奶牛与健康奶牛嗜中性粒细胞差异蛋白质组的表达分析

嗜中性粒细胞是防止病原入侵的第一道防线,且已有研究表明嗜中性粒细胞在奶牛乳腺免疫中发挥着关键作用。文章运用双向凝胶电泳方法对临床型乳房炎奶牛与健康奶牛嗜中性粒细胞差异表达蛋白质组进行分析,成功获得分辨率较高、重复性较好的奶牛嗜中性粒细胞双向电泳凝胶图谱,并通过MALDI-TOF MS鉴定获得差异表达的蛋白质7种,主要涉及细胞代谢、氧化应激、炎症反应等相关蛋白通路。实验获得的临床型乳房炎奶牛与健康奶牛嗜中性粒细胞差异表达蛋白有望为今后奶牛乳房炎的抗病育种研究提供理论依据。     

扬子鳄外周血细胞的超微结构

对扬子鳄血细胞亚显微结构进行透射电镜观察。结果表明：在外周血细胞中可区分出红细胞、嗜中性粒细胞、嗜酸性粒细胞、嗜碱性粒细胞、单核细胞、淋巴细胞以及血栓细胞。电镜图像显示：与其它爬行动物相比,各类血细胞体积较小并具有不同程度的变形运动及吞噬能力。红细胞的特征是长梭形,其长短径之比值超过3.0。在外周血液中发现了多种未成熟型血细胞;进入外周血液中的红细胞普遍已分化到一定程度,但有的仍须在外周血液中完成其最后的发育;单核细胞在外周血液中经历了从幼单核细胞到成熟单核细胞的过程,并描述了从未成熟到成熟阶段细胞的超微结构的变化;嗜中性粒细胞在外周血液中经历了从早幼粒细胞、中幼粒细胞,晚幼粒细胞到成熟粒细胞的过程,描述了嗜中性粒细胞各个发育阶段细胞的超微结构及特征,并讨论了嗜天青粒细胞和嗜中性粒细胞的关系。     

黄石爬鮡外周血液指标特征的观察

黄石爬鮡（Euchiloglanis kishinouyei）属于较为原始的高原鱼类。通过生化分析仪测定红细胞数及血红蛋白含量,Wright’s染色及细胞化学方法,即过碘酸雪夫（PAS）、酚氧化酶（PO）、苏丹黑B（SBB）和过氧化物（POX）染色,观察其外周血细胞的显微结构及细胞化学特征。黄石爬鮡的红细胞数为（0.55 ± 0.06）×1012 个/L,血红蛋白含量为（73.00 ± 5.57） g/L;血细胞中红细胞占98.03%,且其细胞体积较大,白细胞中血栓细胞占比例最多,为37.06%,异嗜性粒细胞最少,为9.64%,异嗜性粒细胞分为Ⅰ型、Ⅱ型和Ⅲ型。细胞化学染色显示,红细胞均呈阴性,白细胞存在染色特性差异,其中白细胞过碘酸雪夫（PAS）染色均为阳性,过氧化物（POX）染色除I型和III型异嗜性粒细胞外均为阴性。     

黄石爬(鱼兆)外周血液指标特征的观察

黄石爬(鱼兆)(Euchiloglanis kishinouyei)属于较为原始的高原鱼类。通过生化分析仪测定红细胞数及血红蛋白含量,Wright′s染色及细胞化学方法,即过碘酸雪夫(PAS)、酚氧化酶(PO)、苏丹黑B(SBB)和过氧化物(POX)染色,观察其外周血细胞的显微结构及细胞化学特征。黄石爬(鱼兆)的红细胞数为(0.55±0.06)×1012个/L,血红蛋白含量为(73.00±5.57)g/L;血细胞中红细胞占98.03%,且其细胞体积较大,白细胞中血栓细胞占比例最多,为37.06%,异嗜性粒细胞最少,为9.64%,异嗜性粒细胞分为Ⅰ型、Ⅱ型和Ⅲ型。细胞化学染色显示,红细胞均呈阴性,白细胞存在染色特性差异,其中白细胞过碘酸雪夫(PAS)染色均为阳性,过氧化物(POX)染色除Ⅰ型和Ⅲ型异嗜性粒细胞外均为阴性。     

趋化因子CCL-18 在慢性鼻- 鼻窦炎中的表达及意义

目的:分析趋化因子CCL-18在不同组织病理特征慢性鼻-鼻窦炎和正常鼻黏膜的表达差异,探讨CCL-18在慢性鼻-鼻窦炎中的表达及意义。方法:采用苏木精-伊红染色(HE),Masson染色及过碘酸-雪夫(PAS)染色对慢性鼻-鼻窦炎组织进行病理分析。采用Western blot检测CCL-18蛋白水平在不同组织病理特征慢性鼻-鼻窦炎和正常鼻黏膜组织中的表达差异。结果:CCL-18蛋白水平在伴鼻息肉和不伴有鼻息肉慢性鼻窦炎均较正常鼻黏膜组织中显著上调(P<0.05)。CCL-18蛋白水平在嗜酸性粒细胞慢性鼻窦炎的表达水平明显高于非嗜酸性粒细胞慢性鼻窦炎(P<0.05)。腺体型,纤维炎症型及水肿型慢性鼻-鼻窦炎中CCL-18表达水平均高于正常鼻黏膜,以水肿型表达最为显著(P<0.05)。结论:CCL-18在嗜酸性粒细胞和水肿型慢性鼻窦炎中高度表达,提示CCL-18可能参与慢性鼻-鼻窦炎中嗜酸性粒细胞的浸润这一基本病理过程。     

丹顶鹤粒细胞性白血病三例

禽类白血病是一种造血系统的疾病,易感动物有鸡、火鸡和雉,亦发现于野鸭,国内外已有许多报道。但对鹤形目丹顶鹤粒细胞性白血病报道却甚少。今将所见丹顶鹤粒细胞性白血病三例报告如下。 (一) 病例介绍三只丹顶鹤均为幼年鹤,其中1号鹤患病期间嗜泥土,三只均生长迟缓,机体消瘦。临床血液学检查,生前采取翼下静脉剖检后采心血制成血涂片,姬姆萨或端氏染色。镜下可见原始粒细胞占81—88％(其中幼粒和晚幼粒占多数)。细胞特征:型大,型状不规则,     

大口黑鲈外周血细胞显微结构、细胞化学特征及吞噬作用的研究

通过对大口黑鲈外周血细胞的显微结构及细胞化学染色特征和吞噬作用的研究,为大口黑鲈免疫学研究积累资料。Wright's染色表明:大口黑鲈外周血细胞分为红细胞、粒细胞、单核细胞、淋巴细胞和血栓细胞,其中粒细胞又分为Ⅰ型、Ⅱ型和Ⅲ型。外周血中成熟红细胞最多,占血细胞总数的98.12%,其次为淋巴细胞和血栓细胞,占白细胞中的比例分别为60.92%和22.99%;单核细胞最大,大小为(9.89±0.70)μm×(8.72±0.68)μm;小淋巴细胞最小,大小为(3.88±0.46)μm×(3.48±0.40)μm。大口黑鲈外周血细胞的免疫相关酶ACP、AKP、PO、POX及细胞成分PAS反应、SBB染色的结果表明,所有红细胞的细胞化学染色均呈阴性,不同白细胞的细胞化学染色特征存在差异。其中所有白细胞的AKP、POX染色均呈阴性,PAS反应均呈阳性,除Ⅱ型粒细胞外所有白细胞的PO、ACP染色均呈阳性,除单核细胞外所有白细胞的SBB染色均呈阳性。大口黑鲈外周血细胞吞噬酵母菌的实验表明,红细胞能够吞噬酵母菌,其吞噬率为(15.70±1.07)%,也观察到部分白细胞吞噬酵母的现象。PO和ACP及脂类和糖类可能是大口黑鲈外周血细胞吞噬作用的重要酶类和能量来源。     

中性粒细胞mGluRI的激活促进其与内皮细胞相互黏附

本文旨在探讨I组代谢型谷氨酸受体(group I metabotropic glutamate receptor,mGluRI)在中性粒细胞上的表达,以及该受体的激活对中性粒细胞与内皮细胞相互黏附的影响。取健康人新鲜静脉血,Ficoll-Hypaque密度梯度离心法分离中性粒细胞,免疫细胞化学法和real-time PCR法检测中性粒细胞mGluRI(包括mGluR1和mGluR5)的表达,分别应用不同浓度的mGluRI特异性激动剂S-3,5-二羟基苯甘氨酸(S-3,5-dihydroxy-phenylglycine,S-DHPG)处理中性粒细胞不同时间,通过比色法检测中性粒细胞和人正常脐静脉内皮细胞(human normal umbilical vein endothelial cells,HUVE-12)黏附率,采用流式细胞术测定中性粒细胞黏附分子CD11a表达的变化。结果证实中性粒细胞表达mGluRI(mGluR1/5);S-DHPG在1×10-8~1×10-6mol/L浓度范围内呈剂量依赖性地提高中性粒细胞与内皮细胞之间的黏附率(P0.05或P0.01);1×10-6mol/L S-DHPG单独作用于中性粒细胞0.5h,即可促进中性粒细胞黏附于内皮细胞(P0.01),但没有时间依赖性;1×10-6mol/L S-DHPG单独作用于中性粒细胞可促进CD11a表达(P0.01);mGluRI拮抗剂(RS)-α-甲基-4-羧基苯甘氨酸[(RS)-α-methyl-4-carboxyphenylglycine,(±)-MCPG](0.5mmol/L)可以显著阻断激动剂S-DHPG(1×10-6mol/L,1h)的促黏附效应(P0.01)。上述结果证实了中性粒细胞膜上mGluRI的激活可增强中性粒细胞表面黏附分子CD11a的表达,促进中性粒细胞与内皮细胞的黏附。     

淡水石斑外周血细胞显微结构观察

淡水石斑(Cichlasoma managuense)外周血细胞可区分出红血细胞、嗜中性粒细胞、嗜酸性粒细胞、单核细胞、淋巴细胞和血栓细胞,未发现嗜碱性粒细胞。外周血液中还存在少量未成熟的和正在分裂的红血细胞。白细胞中,血栓细胞体积最小,嗜中性粒细胞体积最大;数量上,血栓细胞最多,而嗜酸性粒细胞则最少。     

REGULATION OF MATURATION RATE OF MOUSE GRANULOCYTES

Regenerating mouse bone marrow cells were cultured i.p. in diffusion chambers (DC) to study factors affecting the maturation rate of granulocyte precursors. One day after exposing 3-day-old DC cultures to ³H-thymidine the cultures were harvested, and labelled proliferative and non-proliferative granulocytes were counted in radioautographs. The relative maturation rate—defined as the fraction of proliferative precursors maturing into the non-proliferative compartment per unit time—could be increased by different experimental procedures that inhibit cell production. Inhibition was obtained (a) by increasing culture cellularity; (b) by implanting DC into normal rats or rats with huge s.c. chloroma tumours rather than into mice; and (c) by treating the cells with leucocyte extracts (granulocyte chalone) during the last day of culture. Furthermore, a sudden inhibition of rapidly proliferating granulocytes by leucocyte extracts resulted in an increase (apparently transient) in the absolute number of labelled non-proliferative granulocytes. Such an increase was not detected in experiments involving a stronger or sustained inhibition of granulopoiesis, evidently because the size of the precursor population had been markedly reduced.     

The cytology and cytochemistry of the hemocytes of Mytilus edulis and their responses to experimentally injected carbon particles

Hemocytes of Mytilus edulis were examined cytologically and cytochemically. On the basis of structure, staining reactions, and phagocytic behavior, they were divided into two main groups: basophilic hemocytes and eosinophilic granular hemocytes (granulocytes). The basophilic cells were further divided into small lymphocytes and larger phagocytic macrophages reactive for lysosomal hydrolases. Mitosis was observed in granulocytes and in small lymphoid cells, believed to be the stem cells for the basophilic cell line. A few cells appeared to be intermediate between lymphocytes and small granulocytes. Macrophages were the main cell type involved in the clearance of injected carbon particles. However, granulocytes did show some phagocytic activity. Brown cells displaying apparent amoebocytic behavior were found to contain Fe³⁺ and Pb²⁺ in cytoplasmic inclusions, some of which were also reactive for β-glucuronidase and glucosaminidase. These cells appear to have a separate origin from the hemocytes.     

Cytochemical and functional characterization of blood and inflammatory cells from the lizard Ameiva ameiva

The fine structure and differential cell count of blood and coelomic exudate leukocytes were studied with the aim to identify granulocytes from Ameiva ameiva, a lizard distributed in the tropical regions of the Americas. Blood leukocytes were separated with a Percoll cushion and coelomic exudate cells were obtained 24 h after intracoelomic thioglycollate injection. In the blood, erythrocytes, monocytes, thrombocytes, lymphocytes, plasma cells and four types of granulocytes were identified based on their morphology and cytochemistry. Types I and III granulocytes had round intracytoplasmic granules with the same basic morphology; however, type III granulocyte had a bilobued nucleus and higher amounts of heterochromatin suggesting an advance stage of maturation. Type II granulocytes had fusiformic granules and more mitochondria. Type IV granulocytes were classified as the basophil mammalian counterpart based on their morphology and relative number. Macrophages and granulocytes type III were found in the normal coelomic cavity. However, after the thioglycollate injection the number of type III granulocyte increased. Granulocytes found in the coelomic cavity were related to type III blood granulocyte based on the morphology and cytochemical localization of alkaline phosphatase and basic proteins in their intracytoplasmic granules. Differential blood leukocyte counts showed a predominance of type III granulocyte followed by lymphocyte, type I granulocyte, type II granulocyte, monocyte and type IV granulocyte. Taken together, these results indicate that types I and III granulocytes correspond to the mammalian neutrophils/heterophils and type II to the eosinophil granulocytes.     

Highly biased type 1 immune responses in mice deficient in LFA-1 in Listeria monocytogenes infection are caused by elevated IL-12 production by granulocytes

LFA-1 (CD11a/CD18) plays a key role in various inflammatory responses. Here we show that the acquired immune response to Listeria monocytogenes is highly biased toward type 1 in the absence of LFA-1. At the early stage of listeriosis, numbers of IFN-gamma producers in the liver and spleen of LFA-1(-/-) mice were markedly increased compared with heterozygous littermates and Valpha14(+)NKT cell-deficient mice, and NK cells were major IFN-gamma producers. Numbers of IL-12 producers were also markedly elevated in LFA-1(-/-) mice compared with heterozygous littermates, and endogenous IL-12 neutralization impaired IFN-gamma production by NK cells. Granulocyte depletion diminished numbers of IL-12 producers and IFN-gamma-secreting NK cells in the liver of LFA-1(-/-) mice. Granulocytes from the liver of L. monocytogenes-infected LFA-1(-/-) mice were potent IL-12 producers. Thus, in the absence of LFA-1, granulocytes are a major source of IL-12 at the early stage of listeriosis. We assume that highly biased type 1 immune responses in LFA-1(-/-) mice are caused by increased levels of IL-12 from granulocytes and that granulocytes play a major role in IFN-gamma secretion by NK cells. In conclusion, LFA-1 regulates type 1 immune responses by controlling prompt infiltration of IL-12-producing granulocytes into sites of inflammation.     

Leukocyte kinetics in the microcirculation

The transport of leukocytes in the microcirculation is specific for the type, size, and the rheological and adhesive properties, the microanatomy of the host organ, and the hemodynamics. The adhesion to the endothelium is determined largely by the degree of activation via chemotactic factors. Leukocyte motion differs from that of red cells or platelets in several respects. When granulocytes enter into capillaries, they are deformed just like red cells. Under normal flow conditions, the time to deform at the entry to capillaries is typically 1,000 times larger than for the red cell, leading to temporary obstruction of the capillaries. After entry, granulocytes move with lower velocity than red cells which causes a cell train formation inside the capillary. At the venular side, the granulocyte is displaced from the center stream toward the endothelium by faster moving red cells. This process leads to systematic attachment of the granulocytes to the endothelium. At a reduced perfusion pressure or in the presence of locally elevated levels of chemotactic factors, the granulocytes may not be able to pass through the capillary network, which leads to microvascular obstruction. Organs with a narrow capillary network may thereby become filters for circulating granulocytes. This event is accompanied in many situations with damage to the host organ.     

The haematopoietic supraneural organ of adult, sexually immature river lampreys (Lampetra fluviatilis [L.] Gray) with particular reference to azurophil leucocytes.

The haematopoietic tissue in the supraneural organ of the freshwater river lamprey (Lampetra fluviatilis L. Gray) was studied in sexually immature animals. Besides erythro- and granulopoietic elements, macrophages, reticular cells, fibroblasts and glycogen-rich fat cells were seen. Developing granulocytes of the lamprey contain one type of azurophil granules originating from small cytoplasmic (Golgi) vesicles. The lamprey's azurophil granulocytes seem to be homologous with those of fishes. However, the granulocytes of fishes, studied thus far, show granules with only one type of inclusion, whereas in lamprey the granulocyte inclusions are variable in size and shape. Thus, lamprey granulocytes are, in this respect, reminiscent of similar cells of higher vertebrates. The PAS and alkaline phosphatase reactions, common markers of vertebrate neutrophil leucocytes, are very weak in the haematopoietic tissue granulocytes of the lamprey, and intense in the blood cells of the same animal. Lamprey granulocytes, similarly to the granulocytes of Chondrostei and Elasmobranchiata, do not stain with peroxidase, naphthol-AS-D-chloroacetate esterase and sudan black B. The haematopoietic tissue contains a relatively high number of degenerated granulocytes.     

REGULATION OF BONE MARROW CELL GROWTH IN DIFFUSION CHAMBERS: THE EFFECT OF GRANULOCYTE EXTRACTS

Hypotonic lysis of mature human blood granulocytes yielded an extract which reduced granulopoiesis and enhanced macrophage formation of mouse bone marrow cells cultured for 7 days in diffusion chambers (DC). The low molecular weight fraction (MW < 15,000–25,000 Daltons) obtained by Amicon filtration of the extract, reduced granulopoiesis without affecting macrophage formation. The high molecular weight fraction (MW > 15,000–25,000 Daltons) reduced the number of granulocytes and increased the number macrophages. Erythrocyte extract increased the macrophage formation in DC but did not alter the number of granulocytes. The spleen colony assay showed that the granulocyte extract increased the number of CFU-S in DC. It is suggested that the granulocyte extract contain an inhibitor of stem cell differentiation to myeloid cells thereby reducing the number of proliferative granulocytes in DC 7 days later. The inhibitor of differentiation may lead to an increased self renewal of the stem cell in the DC system.     

THE IN VITRO DIFFERENTIATION OF DENSITY SUB-POPULATIONS OF COLONY-FORMING CELLS UNDER THE INFLUENCE OF DIFFERENT TYPES OF COLONY-STIMULATING FACTOR

The in vitro proliferation and differentiation of myeloid progenitor cells (CFU-c) in agar culture from CBA/Ca mouse bone marrow cells was studied. Density sub-populations of marrow cells were obtained by equilibrium centrifugation in continuous albumin density gradients. The formation of colonies of granulocytes and/or macrophages was studied under the influence of three types of colony-stimulating factor (CSF) from mouse lung conditioned medium CSF_MLCM), post-endotoxin mouse serum (CSF_ES) and from human urine (CSF_Hu). The effect of the sulphydryl reagent mercaptoethanol on colony development was also examined. The density distribution of CFU-c was dependent on the type of CSF. Functional heterogeneity was found among CFU-c with partial discrimination between progenitor cells forming pure granulocytic colonies and those forming pure macro-phage colonies. Mercaptoethanol increased colony incidence but had no apparent effect on colony morphology or the density distribution of CFU-c.     

Sydney rock oyster (Saccostrea glomerata) hemocytes: morphology and function

In this study, three major hemocyte types were identified in the Sydney rock oyster. They were characterized primarily by light and electron microscopy based on the presence or absence of granules and nucleus to cytoplasm ratios. Hemoblast-like cells were the smallest cell type 4.0+/-0.4microm and comprised 15+/-3% of the hemocyte population. They had large nuclei and scanty basic cytoplasm. This cell type also had some endoplasmic reticuli and mitochondria. The second major type were hyalinocytes. Hyalinocytes represented 46+/-6% of all hemocytes. They were large cells (7.1+/-1.0microm) that had low nucleus:cytoplasm ratios and agranular basic or acidic cytoplasm. Hyalinocytes had the ability to phagocytose yeast cells and formed the core of hemocyte aggregates associated with agglutination. Four discrete sub-populations of hyalinocytes were identified. The third major cell type were the granulocytes, comprising 38+/-1% of the hemocyte population. These cells were large (9.3+/-0.3microm) and were characterized by cytoplasm containing many acidic or basic granules. Granulocytes were more phagocytic than hyalinocytes and they formed the inner layer of hemocytes during the encapsulation of fungal hyphae. Five discrete sub-populations of granulocytes were identified based on the types of granules in their cytoplasm. Flow cytometry showed that the hemocytes of rock oysters could be divided into between two and four major cell types based on their light scattering properties. The most common of the cell types identified by flow cytometry corresponded to hyalinocytes and granulocytes. Cytochemical assays showed that most enzymes associated with immunological activity were localized in granulocytes. Their granules contained acid phosphatase, peroxidase, phenoloxidase, superoxide and melanin. Hyalinocytes were positive only for acid phosphatase. All of these observations suggest that Sydney rock oysters have a broad variety of functionally specialized hemocytes, many of which are involved in host defense.