柱状噬纤维菌免疫后兴国红鲤免疫细胞数量变化的研究

用柱状噬纤维菌灭活菌苗免疫兴国红鲤,于免疫后2周,4周分析对其外周血液和胸腺,脾脏,头肾和体肾中的免疫细胞数量变化进行了测定,结果显示：外周血液中,淋巴细胞有显著的增加,粒细胞和单核细胞有明显的下降,胸腺中淋巴细胞数目略有增加,脾脏中淋巴细胞有显著的增加,而粒细胞和单核细胞正相反,头肾中淋巴细胞有较多的增加,粒细胞数目增加不明显,单核细胞有大幅下降,体肾中,粒细胞和淋巴细胞大幅上升,单核细胞下降。     

斜带石斑鱼淋巴器官个体发育的组织学

本文应用连续组织切片技术和组织学观察,对出膜后1～60天的斜带石斑鱼(Epinephelus coioides)各期仔鱼、稚鱼和幼鱼的淋巴器官组织进行了研究,描述了淋巴器官的个体发育过程和组织学结构特征。研究表明：实验水温为22．0～27．8℃时,孵化后第10天出现头肾原基。头肾原基由未分化的造血干细胞组成。随着鱼体的生长,头肾原基的造血干细胞很快分化成不同类型的细胞;头肾主要由网状内皮系统支持下的淋巴造血组织构成。第11天出现脾脏原基。脾脏原基由造血细胞组成,淋巴化速度相对较慢。脾脏在整个发育过程中,红细胞和类红细胞占优势,没有红髓和白髓之分。第13天出现胸腺原基。胸腺发育速度较快,是明显的淋巴器官。胸腺主要由胸腺细胞(淋巴细胞)和上皮细胞组成,外区和内区没有明显的界限,但很容易区分。胸腺外被单层的上皮细胞层与咽腔相隔,保持浅表的位置,并且在整个发育过程中,胸腺与头肾是独立分开的。免疫器官原基出现顺序是头肾、脾脏和胸腺;而免疫器官淋巴化的顺序是胸腺,头肾和脾脏。和其它硬骨鱼类一样,斜带石斑鱼在早期发育阶段,淋巴器官的发育较迟,出现相对滞后的现象[动物学报49(6)：819～828,2003]。     

卵形鲳鲹血细胞发生的观察

对卵形鲳鲹(Trachinotus ovatus)肝脏、脾脏、头肾和体肾4种组织器官内各种血细胞的发生和外周血液的血细胞分类组成进行了观察。结果表明:卵形鲳鲹的红细胞主要在脾脏、体肾和头肾中发生;淋巴细胞主要在体肾、头肾和脾脏中发生;粒细胞主要在头肾和体肾中发生;单核细胞在肝脏、头肾和脾脏中均有发生;而血栓细胞的发生在这4种组织中均未观察到。卵形鲳鲹在外周血液中的白细胞包括淋巴细胞、血栓细胞、嗜中性粒细胞和单核细胞。在这几种白细胞中,单核细胞数量最少,很难观察到,数量少于1%;嗜中性粒细胞稍多,有(9.25±5.64)%;血栓细胞占(38.23±21.86)%;淋巴细胞最多,达(52.52±21.65)%。     

瓦氏黄颡鱼和岩原鲤脾脏的组织学观察

运用组织学方法对瓦氏黄颡鱼和岩原鲤脾脏的显微结构进行了研究.瓦氏黄颡鱼和岩原鲤脾脏被膜薄,仅由一层单层扁平上皮细胞和结缔组织纤维构成,脾小梁明显,红髓和白髓混合,无明显的分界,鞘毛细血管发达;瓦氏黄颡鱼脾小梁细而少,白髓中淋巴细胞密集,在靠近被膜的脾实质边缘区域密集的淋巴细胞形成外形类似淋巴小结的结构,巨噬细胞聚集形成明显的巨噬细胞中心;岩原鲤脾小梁发达,伸入脾实质将脾脏分隔成一个个的小叶,白髓中密集的淋巴细胞较少,未见类似淋巴小结的结构,巨噬细胞分散存在,无明显的巨噬细胞中心.     

汞、铬离子对黄鳝头肾组织结构的影响

采用毒性实验方法,用不同浓度的汞离子(Hg2 )、铬离子(cr6 )分别处理黄鳝(Monopterus albus),经1、2、4、8 d后光镜观察黄鳝头肾组织结构及免疫细胞数量的变化.结果表明,对照组黄鳝头肾实质内未见肾单位结构,主要由淋巴组织、造血组织和血窦构成.淋巴组织主要由淋巴细胞密集排列成淋巴索或聚集成团状;造血组织主要由红血细胞密集排列成细胞索或聚集成团状.淋巴组织和造血组织在头肾实质中各占据一定的区域或相间分布,且分别被血窦所分隔.黑色素巨噬细胞分散于淋巴细胞之间,数量较少,排列较松散,尚未集结成明显的球状体.经两种重金属离子分别染毒后的黄鳝头肾与对照组相比,组织结构表现出相似的变化,即随着重金属离子浓度的增加和染毒时间的延长,头肾组织中的黑色素巨噬细胞聚集形成黑色素巨噬细胞中心,并逐渐增大、增多,最后减少;淋巴组织逐渐松散,排列稀疏混乱,淋巴细胞界限逐渐不清晰,呈退化趋势,数量减少.红血细胞大量破坏,血窦扩张.与Cr6 相比,Hg2 对头肾组织结构的毒性影响更大,病变现象更为明显.     

鳜鱼头肾的组织发生及成鱼头肾B淋巴细胞的分布

通过整体连续切片,研究了鳜鱼不同发育时期的头肾结构,并利用原位PCR方法检测了B淋巴细胞在鳜鱼头肾中的分布。在孵化后第1d观察到了肾组织,主要由肾小管组成。尔后头肾的发育经历了三个结构和功能的转变。第一个阶段为孵化后第1d到第7d,头肾作为滤过性器官存在,由肾小管及少量淋巴细胞组成。第二个阶段从第8d到第36d,是一个功能混合型阶段,头肾中既有肾小管,又有造血组织;随时间推移,肾小管数量减少,淋巴细胞数量剧增。紧接着进入第三个阶段：肾小管完全消失,头肾中开始出现大量的嗜铬细胞,头肾作为淋巴-肾上腺组织而存在。肾上腺首先出现在头肾前端,随发育成熟,集中分布于头肾门静脉周围。IgM在鳜头肾中大量表达,IgM分泌细胞分布于整个头肾组织,在血管周围有集中趋势[动物学报51(3)：440—446,20051。     

鱼类染色体的白细胞-PHA活体内处理制片及其组型观察

目前,鱼类染色体标本制备的方法主要是由Yamamoto等建立的肾细胞-PHA体外短期培养法和Ojima等提出的外周血淋巴细胞离体培养法,以及活体注射PHA直接用肾组织的细胞制片等。用短期细胞培养研究鱼类染色体有较突出的优点,它能保持染色体的自然状态,染色体的形态特征较易于显示出来,中     

南方鲇头肾的组织学和超微结构

采用解剖学、组织学、组织化学方法,通过电镜观察,研究了南方鲇(Silurus meridionulis Chen)头肾的形态和超微结构。南方鲇左右头肾不相连,与肾明显分离。头肾组织包括血管系统、淋巴细胞聚集区、粒细胞聚集区、内分泌组织区。血管系统由头肾动脉、头肾静脉、后主静脉及其分枝所组成。淋巴区细胞染色深,紧密聚集成网状结构,主要包括大、小淋巴细胞等细胞类型。粒细胞区域狭窄,染色浅,主要为粒细胞聚集。内分泌组织区域细胞大型,常规染色极浅,包括肾间组织细胞、肾上组织细胞。肾间组织细胞有丰富的线粒体、内质网及分泌颗粒。肾上组织细胞主要分布于后主静脉及其分枝的周围、静脉壁中,细胞中有密集的分泌颗粒。肾上组织细胞和肾间组织细胞集中于头肾,内分泌功能增强,是与其运动能力强、快速捕食相适应的特征。     

中华鳖造血和免疫器官的个体发育

采用常规孵化的中华鳖胚胎为材料,对不同发育时期造血和免疫器官进行了组织学研究,描述了卵黄囊、胸腺、肝、脾、肾以及骨髓的形态结构变化。发现胚胎期首先出现的造血器官是卵黄囊。此后,卵黄囊的造血干细胞出现在胚体的血循环中,造血功能相继在胚胎胸腺、肝、脾、骨髓(可能还包括肾)中产生。胸腺是中华鳖免疫系统发育的第一个淋巴器官,来自卵黄囊的干细胞在此先分化成小淋巴细胞,然后再迁移至脾脏。脾脏发育中首先出现各发育阶段的红细胞、嗜酸性的细胞和少量粒细胞,淋巴细胞出现较晚,未发现淋巴小结。在胚胎期肝脏发育过程中可见不同发育时期的红细胞和嗜酸性的细胞。在肾的发育过程中,尚可观察到嗜酸性的细胞和类似头肾组织的细胞团。直至出壳前,骨髓内方可见各发育阶段的各系细胞[动物学报49(2)：238—247,2003]。     

大豆异黄酮对电离辐射小鼠淋巴细胞的防护作用

目的研究大豆异黄酮(SI)对小鼠淋巴细胞的辐射防护作用。方法24只雄性昆明小鼠,随机分为正常对照组、辐射对照组和辐射补充0.5%SI组,喂养2周后,4.0Gy照射。照射后24 h处死小鼠,取血、胸腺和脾脏分离淋巴细胞,进行血淋巴细胞计数、观察DNA损伤情况;培养胸腺和脾脏淋巴细胞,检测3H-dT掺入量,观察淋巴细胞的增殖能力,计算淋巴细胞的增殖指数。结果辐射使小鼠胸腺和脾脏淋巴细胞数明显减少、胸腺淋巴细胞增殖能力和脾脏淋巴细胞转化指数降低、血淋巴细胞DNA损伤增加,这些变化均具有统计学意义;补充SI可降低胸腺和脾脏淋巴细胞数的减少幅度,降低胸腺淋巴细胞增殖能力和脾脏淋巴细胞转化指数下降幅度,减少辐射对血淋巴细胞DNA损伤程度,其中SI对胸腺淋巴细胞增殖能力和对血淋巴细胞DNA损伤程度的防护作用与辐射对照组相比有统计学意义。结论大豆异黄酮可对小鼠的血、胸腺和脾脏淋巴细胞有一定的辐射防护作用。     

Proteome Profiles of Head Kidney and Spleen of Rainbow Trout (Oncorhynchus Mykiss)

The head kidney and spleen are major lymphoid organs of the teleost fish. The authors identify proteome profiles of head kidney and spleen of rainbow trout (Oncorhynchus mykiss) using a shotgun proteomic approach. Gene ontology annotation of proteins is predicted using bioinformatic tools. This study represents detailed proteome profiles of head kidney and spleen of rainbow trout, with a total of 3241 and 2542 proteins identified, respectively. It is found that lymphoid organs are equipped with a variety of functional proteins related to defense, receptor, signal transduction, antioxidant, cytoskeleton, transport, binding, and metabolic processes. The identified proteome profiles will serve as a template for understanding lymphoid organ functions in salmonids and will increase the amount of spectra information of rainbow trout proteins in the public data repository PRIDE. This data can be accessed via ProteomeXchange with identifiers PXD008473 and PXD008478.     

Detection of mitogen induced stimulation of leukocytes from the rainbow trout (Oncorhynchus mykiss) by flow cytometric analysis of intracellular calcium

Flow cytometric analysis of the forward/side light scatter (FSC/SSC) of density gradient-separated head kidney cells of the rainbow trout revealed three distinctly separated populations, which we defined as population 1, 2 and 3. In spleen cells, populations 1 and 2 were also found, whereas population 3 was not detected. Further characterization regarding the surface Ig (sIg) revealed that population 2 of the head kidney and spleen contained 37.4 and 34.4% sIg⁺-cells, respectively. Incubation of the head kidney and spleen cells with different concentrations of concanavalin A (ConA), phytohemagglutinin (PHA) and [PWM] induced a pronounced intracellular calcium increase only in cells of population 2. This reaction was concentration dependent and caused by a release of intracellular Ca²⁺-stores. FMLP, a chemotactic peptide, had no effect on intracellular calcium response in all three populations. Similarly, the stimulation with PMA had no effect. This indicates that population 2 of the head kidney as well as the spleen is characterized by a high forward and low side light scatter and contains both subpopulation of lymphocytes, B- and T-cells. We demonstrated that the analysis of intracellular calcium increase due to mitogens is a suitable approach to identify lymphocytes in fish and enables further functional studies in these cells.     

The development of the lymphoid organs of flounder, Paralichthys olivaceus, from hatching to 13 months

The growth of the lymphoid organs, such as head kidney, spleen and thymus were studied in flounder, Paralichthys olivaceus Temminck & Schlegel, from hatching to 13 months of age. Except for the thymus, all organs grew as the fish grew. By 2 months of age the lymphoid organs attained their maximum relative weight. The organ weight showed a closer correlation to body weight than they did to age. The total number of leucocytes in the lymphoid organs increased with age, but the number per milligram of lymphoid organ remained constant. A micro and ultrastructural study of the lymphoid organs showed that the full development of the lymphoid organs was not achieved until the juvenile stage. The spleen and head kidney had mixed populations of "red" and "white" cells. The head kidney was more lymphoid than the spleen. The thymus involuted quickly during the first 6 months. The blood components had no obvious relationship with age or season during the period studied.     

鲶鱼小窝器官的后期发育及形态的研究

电感官包括结节器官(tuberous organ)和壶形器官(ampullary organ)二大类。前者仅存在于电鱼上,后者则在电鱼及某些非电鱼都有发现。对于淡水鲶鱼而言,其壶形器官又称为小窝器官(small pit organ)。行为实验表明某些鱼类的小窝器官有极高的电敏感性,能检测0.5—0.8 nA/cm~2的电流刺激(Peters,1974;Kalmijn,1976),同时有不少报告描述了它的显微和亚显微结构(Sato,1949,1956,1969;Wachtel,1969;Roth,1969)。但是对这种器官的发育过程的研究报告还不多。     

Distribution of intravenously injected A-layer protein and lipopolysaccharide (LPS) from Aeromonas salmonicida in Atlantic salmon, Salmo salar L.

The distribution of intravenously injected A-layer protein and lipopolysaccharide (LPS) purified from the outer surface of the fish pathogen Aeromonas salmonicida, was studied in Atlantic salmon. Radiolabelling was achieved by conjugating the antigens to tyramine cellobiose (TC) or fluorescein isothiocyanate (FITC) which were radioiodinated either before or after conjugation. Since both TC and FITC are trapped intralysosomally at the cellular site of uptake, the ligands are advantageous in studies on tissue distribution of antigens. Injection of TC-A-layer protein and TC-LPS resulted in high specific radioactivity (cpm g⁻¹tissue) in both head kidney and trunk kidney. In contrast, only low specific radioactivity was recovered in spleen, heart and liver. Surprisingly, use of FITC-LPS as the antigen changed the uptake to be high in both spleen and head kidney. Radiolabelled (¹²⁵I-TC-) LPS and A-protein, administered by a dorsal aorta catheterisation technique, were cleared from the blood within 24 h. In immunised fish, the antibody activity against the A-layer protein was diminished even within 10 min after administration, in contrast to the level of anti-LPS antibodies which remained high. These results suggest that immune-complex formation took place at least with the A-layer protein, but the uptake of A-layer protein in the various tissues did not differ significantly in vaccinated (A. salmonicida bacterin) and non-vaccinated fish.     

Non-specific cellular responses of rainbow trout to Vibrio anguillarum and its extracellular products (ECPs)

We have studied the early inflammatory response induced by Vibrio anguillarum and by its extracellular products (ECPs) in rainbow trout after intraperitoneal injection. The results showed a very similar inflammatory response which included leucopenia, mainly due to lymphopenia, neutrophilia and an increase in the number of circulating monocytes. Melanomacrophages as well as immature leucocytes were frequently observed circulating in the blood of injected rainbow trout. Monocytes often contain phagocytosed bacteria and other, altered cells including erythrocytes and leucocytes. However, neutrophils only occasionally phagocytosed bacteria. Many circulating leucocytes showed important structural alterations. Neutrophils of trout injected with bacteria and ECPs also showed stronger PAS-staining than those of control trout as well as Döhle bodies and swollen granules. A marked vasodilatation was observed in the kidney and spleen which was coincidental with a mobilization of eosinophilic granular cells and an hypertrophy of sinusoidal endothelial cells showing an increase in the number of cytoplasmic granules. An increase in the number of macrophages and melanomacrophages in the kidney and spleen as well as oedema and leucocyte infiltration in the liver and gills were also noted.     

Comparison and definition of spleen and lymph node: a phylogenetic analysis

The hitherto largely unsolved problem with a biological definition of spleen versus lymph node seems possible to solve from a phylogenetic point of view. Thus, it is suggested that the spleen be defined as a hemopoietic organ which is able to filter blood with sinusoids. In contradistinction, a lymph node is defined as a hemopoietic organ which is able to filter lymph with sinusoids. Comparative anatomical studies show that the spleen appears as a condensation of the lymphomyeloid complex in the spiral fold of the gut in cyclostomes. The spiral fold spleen vanishes with the bony fishes, while in cartilaginous fishes a similar spleen appears in the dorsal mesentery. The dorsal spleen remains in a retroperitoneal position in higher vertebrates and is regarded as a specialized blood vessel compartment closely connected with the blood stream. In "higher" vertebrates the spleen is a stagnated organ because splenic functions are gradually transferred to other sites. The bone marrow takes over the erythro-, thrombo- and granulocytopoiesis while the lymph nodes take over the lymphocytopoiesis. This transfer of the splenic functions is first seen in anurans and seems to be a marvelous adaptation to life on land where the need for local defence against a large number of antigens is necessary before spread of the antigens to central parts of the body. In higher vertebrates, the great number of lymph nodes at peripheral positions, derived from the lymphatic vessels, are able to do so. It is demonstrated that the definitions of spleen and lymph nodes as hemopoietic organs which by their sinusoids are able to filter blood and lymph, respectively, are not only of semantic interest but also useful in regard the immunohematological system as an entity.     

A model for acute iron overload in sea bass (Dicentrarchus labrax L.)

Evaluation of several parameters involved in iron metabolism was carried out after intraperitoneal (i.p.) injection with iron dextran (IDx) in sea bass (Dicentrarchus labrax L.). After treatment, a rapid mobilization of IDx from the peritoneal cavity to other organs was observed. This was followed by a modification of normal peripheral blood iron parameters. Total iron (TI) and transferrin saturation (TS) rose rapidly, to 4.14 microg/ml and 83.7%, respectively, on day 3. In contrast, unsaturated iron binding capacity (UIBC) dropped from 3.19 microg/ml (at day 0) to 0.90 microg/ml on day 3. Tissue iron content was determined by atomic absorption spectometry (AAS). Three days post-IDx injection, values of iron concentration in liver, spleen and head kidney were significantly higher than control values (15, 6 and 9-fold increase, respectively). Samples of liver, spleen and head kidney were processed for routine histology, and the Perl's method was used for iron staining. Histological sections of the IDx-treated animals showed iron deposition in all tissues studied. In the liver, the iron was evenly distributed over the whole organ, being present in the hepatocytes. In the head kidney and spleen, the iron deposition was mainly observed in the melanomacrophage centres (MMCs). The present study characterizes several parameters involved in iron metabolism, and develops a fish model, of iron overload, which can be used in further studies of iron toxicity and iron-induced susceptibility to bacterial infections.