胸腺实质内存在肥大细胞

胸腺是T淋巴细胞分化和发育的重要器官。胸腺被膜和小叶间隔的结缔组织内存在着肥大细胞已有许多报道,由长期培养的胸腺细胞中可分化出肥大细胞也有报道,但是在胸腺实质内是否存在肥大细胞,至今尚未见有肯定的报道。我们将胸腺组织经Carnoy液固定、石蜡切片或经骤冷制成冰冻切片,采用Alcian Blue-Safranin(ABS)染色、甲苯胺蓝染色、天青A染色、氯     

L783白血病小鼠体内的免疫活性细胞和免疫球蛋白的变化

本文报道沪白 1号(SW1)近交系小鼠腹腔接种L783白血病小鼠脾脏细胞悬液后,胸腺、脾脏和淋巴结中T和B淋巴细胞百分率以及血清中 IgG和 IgM含量的变化。结果表明,淋巴器官除胸腺外,脾脏和淋巴结中的T淋巴细胞百分率明显增高,而B淋巴细胞显著下降。血清中Ig 含量未见明显变化。上述免疫活性细胞有规律性的变化为进一步研究白血病发病机理提供线索,并有可能作为考核抗白血病药物疗效的指标之一。     

重症肌无力胸腺肥大细胞形态分布和INF—α的表达

目的：探讨重症肌无力（ＭＧ）时胸腺微环境的变化及其发病机制。方法：选择ＭＧ患者胸腺切除标本２８例，分别行常规病理制片、ＨＥ染色、光镜观察和甲苯胺蓝染色后对胸腺中肥大细胞（ＭＣ）作半定量分析。另用ＴＮＦ－α单克隆抗体进行免疫组化标记。结果：２８例中，胸腺滤泡性增生者１６例，非增生者１２例，组织学表现胸腺淋巴组织增生伴生发中心形成的淋巴滤泡，而非增生者表现为正常组织形态。在对ＭＣ定量分析后发现，胸腺滤     

胸腺神经内分泌功能与衰老

胸腺是人体的中枢淋巴器官,是培育T淋巴细胞的主要场所。T细胞是胸腺依赖性淋巴细胞(Thymus-dependent Lymphocyte)的简称,其来源于骨髓的多功能造血干细胞,T细胞是淋巴细胞的主要组分,它具有多种生物学功能,如直接杀伤靶细胞,     

小鼠淋巴结内肥大细胞生长抑素免疫组织化学观察

目的：观察小鼠淋巴结内肥大细胞的分布特点及神经肽性质。方法：采用甲苯胺蓝染色和免疫组织化学ABC方法。结果：淋巴结内肥大细胞多为圆形和椭圆形，主要分布于淋巴窦内；经与甲苯胺蓝邻片比较，肥大细胞呈生长抑素（SS）免疫反应性，可见免疫反应阳性颗粒充满于胞质内，细胞核为阴性反应。结论：小鼠淋巴结肥大细胞呈SS免疫反应性，表明肥大细胞内神经肽SS可能通过神经－内分泌－免疫网络系统调节免疫器官的功能活动。     

用抗溴脱氧尿苷单克隆抗体检测小鼠各种器官内S期细胞分布的研究

用溴脱氧尿苷（ＢｒｄＵ）腹腔注入正常小鼠（０．１５ｍｇ／ｇ体重），１ｈ后处死，取出多种器官的组织用Ｃａｒｎｏｙ液固定，石蜡包埋切片。用免疫组织化学技术，以抗－ＢｒｄＵ的单克隆抗体检测不同器官内被ＢｒｄＵ标记的Ｓ期细胞。结果证明，ＢｒｄＵ阳性细胞主要分布在细胞增殖活跃的组织，如舌与食管的基底层细胞、胃小凹处的颈粘液细胞，小、大肠的肠腺细胞和睾丸曲细精管内的精原细胞等。ＢｒｄＵ阳性细胞在淋巴器官内主要分布于胸腺皮质、脾脏的动脉周围淋巴鞘和脾小结的生发中心以及淋巴结的生发中心等。高度分化的细胞如舌与食管上皮的角化层或肝、肾等处的细胞呈阴性。     

大鼠胸腺的嗜金属性细胞

非淋巴细胞参与二级淋巴器官的免疫学反应。它们存在于脾脏白髓和淋巴结的不同部位,主要与T细胞和B细胞的微环境有关。根据形态学、酶和功能特征可把非淋巴细胞分为树突状网状细胞(DRC)、嵌合细胞(IDC)和金属性细胞。作者用鼠龄2.5月的Wistar大鼠8只研究了正常情况下大鼠胸腺中金属性细胞的分布和形态学特征。方法是:将胸腺用Bouin     

过氧化物酶体增殖剂PFOA对小鼠免疫系统的影响

目的： 研究过氧化物酶体增殖剂（PP）全氟辛酸（PFOA）对小鼠的免疫器官和免疫细胞、免疫功能的影响.方法： 喂养含PFOA的食物后, 观察C57B/6雄性小鼠胸腺和脾脏的重量及胸腺细胞和脾细胞数的变化.用碘化丙啶（PI）进行细胞DNA染色, 用小鼠单克隆抗体（mAb）进行细胞免疫荧光染色, 流式细胞术分析胸腺和脾细胞的细胞周期和细胞表型的变化.用蛋白A噬菌斑实验和ELISA法检测小鼠抗马红细胞的抗体生成B细胞和血清中IgM和IgG滴度的变化.用刀豆蛋白A（ConA）和脂多糖（LPS）分别刺激小鼠脾细胞, 用 3H-TdR掺入法检测淋巴细胞增殖活性的变化.结果： 强效的PP（如PFOA）能导致小鼠胸腺和脾脏严重萎缩;其脾脏中T和B细胞数减少50%, 胸腺细胞数减少〉90%, 以成熟的CD4＋ CD8＋细胞数的减少最显著, S和G2/M期细胞数的下降最明显.PFOA还可降低马红细胞诱导的IgM和IgG抗体生成B细胞的数量和血清中特异性IgM和IgG抗体的滴度, 以及T细胞活化剂ConA和B细胞活化剂LPS刺激的淋巴细胞增殖反应强度.停止喂养含PFOA的食物后, 上述指标迅速恢复正常.结论： 强效的PP能对小鼠免疫系统产生明显的抑制作用.     

银杏叶提取物增强大鼠脾脏和胸腺的免疫功能

目的研究银杏叶提取物(GBE)对大鼠脾脏和胸腺免疫功能的影响。方法给SD大鼠灌胃给予(40、120、360)mg/(kg·d)GBE,同时设置对照组。28 d后,水合氯醛麻醉处死大鼠,测量胸腺和脾脏的质量指数;MTT法检测伴刀豆球蛋白A(Con A)诱导的大鼠脾淋巴细胞增殖转化;中性红实验测定大鼠腹腔巨噬细胞吞噬功能;扫描电镜观察大鼠脾脏和胸腺的超微结构变化。结果不同剂量的GBE均可提高胸腺和脾脏的器官质量指数,不同剂量组之间无显著性差异;不同剂量的GBE均可增强大鼠腹腔内巨噬细胞的吞噬作用及各级淋巴细胞的增殖能力,且呈现一定的剂量依赖性。电镜观察发现不同剂量的GBE处理组,大鼠脾脏和胸腺中成熟淋巴细胞数均较对照组增多。结论银杏叶提取物可增强大鼠胸腺和脾脏的免疫功能。     

突触体素、S—100蛋白和神经特异性烯醇化酶免疫反应神经纤维在人免疫器官的分布

目的：探讨人免疫器官突触体素、S－100蛋白和神经特异性烯醇化酶（NSE）免疫反应神经纤维的支配和免疫反应细胞的分布，为神经内分泌和免疫系统相互作用提供形态学资料。方法：应用免疫组织化学ABC法观察正常免疫器官包括胸腺、脾脏、淋巴结各30例。10％ 福尔马林固定，石蜡包埋。结果：胸腺，突触体素、S－100蛋白和NSE免疫反应神经众胸腺被膜随小叶间隔和血管胸腺皮质，再延伸到髓质形成神经纤维网，在胸腺组织散在分布突触体素、S－100蛋白和NSE免疫反应细胞。淋巴结，免疫反应神经纤维沿被膜和门部结缔组织小梁及血管进入皮质后主要分布于副皮质区环绕淋巴滤泡，进一步分支到达髓质。在髓质髓窦内有NSE免疫反应细胞。脾脏，免疫反应神经纤维沿着血管的各级分支进入脾实质，主要沿着脾动脉的分支而分布在白髓、红髓和边缘区，穿插于淋巴细胞之间。结论：在人免疫器官可能在神经内分泌免疫相互作用和调控。     

Human intestinal mucosal mast cells: evaluation of fixation and staining techniques

The staining properties of tissue mast cells are influenced by the method of fixation. Differences in fixation and staining techniques may explain the contradictory results in the published reports on the number of human mucosal mast cells (MMC) in the gastrointestinal mucosa in health and disease. We have examined the influence of fixatives on the staining properties of human MMC in operative biopsy specimens of human jejunum. Specimens were divided into pieces, each of which was fixed in one of the following fixatives: Carnoy's, basic lead acetate (BLA), Baker's, Bouin's, isotonic formol-acetic-acid (IFAA), 10% neutral buffered formalin, formol sublimate, and formol saline. Thereafter, tissues were paraffin-embedded and 5 micron sections were cut and stained with either astra-blue/safranin pH 0.3, or toluidine blue pH 0.5. Counts of the number of MMC/mm2 were obtained for each fixation method. The results show a critical influence of the fixative on the number of mast cells identified after staining. For example with astra-blue/safranin the mean MMC/mm2 count was 40 in formol-saline-fixed specimens, and 268 in Carnoy's-fixed specimens. In biopsies fixed with formalin-based fixatives, mast cells were more readily stained with toluidine blue. It is recommended that Carnoy's or BLA be used as the fixative for any light microscopic study of human MMC.     

Mast cells in human keloid, small intestine, and lung by an immunoperoxidase technique using a murine monoclonal antibody against tryptase.

A murine monoclonal antibody (G5) against human lung mast cell tryptase was used for selective staining of human mast cells by an indirect immunoperoxidase method. Human tissues (keloid, small bowel, lung) were fixed in either Carnoy's fluid or neutral buffered formalin. In all three tissues the number and location of G5-stained mast cells corresponded closely with metachromatic toluidine blue-stained mast cells, although the immunospecific technique appeared to be more sensitive. In lung the average concentration of G5-positive mast cells after Carnoy's fixation was 15,695/cu mm of subepithelial tissue in bronchi and bronchioles and 26,580/cu mm of alveolar wall, in small bowel was 20,958/cu mm of mucosa and 8576/cu mm of submucosa, and in keloid was 3068/cu mm. Formalin fixation significantly reduced concentrations of G5-positive mast cells in all tissues except keloid.     

Immunohistochemical identification of mast cells in formaldehyde-fixed tissue using monoclonal antibodies specific for tryptase

An avidin-biotin enhanced immunoperoxidase procedure using monoclonal antibodies (AA1, AA3, and AA5) prepared against human mast cell tryptase resulted in intense staining of mast cells in paraffin-embedded tissue. The distribution of mast cells observed was similar to that seen when adjacent serial sections were stained using a standard procedure with toluidine blue, though the immunoperoxidase technique permitted the identification of significantly more mast cells. With monoclonal antibody AA1, immunostaining was entirely specific for mast cell granules, and there was negligible background staining in a range of tissues including lung, tonsil, colon, gastric mucosa, skin, and pituitary. There was no staining of antibody on basophils or on any other normal blood leukocyte. The technique was effective with tissue fixed in either Carnoy's or neutral buffered formalin, though the internal mast cell structure was better preserved with formaldehyde fixation. The immunoperoxidase staining procedure with monoclonal antibody AA1 is a highly specific and sensitive means for the detection of mast cells in routinely processed tissues.     

Histochemical heterogeneity of human mast cells: disease-related differences in mast cell subsets recovered by bronchoalveolar lavage.

Fixation and staining characteristics were studied for mast cells recovered by bronchoalveolar lavage from 67 patients being investigated for lung disease. The number of toluidine blue stained mast cells in formaldehyde-fixed cytocentrifuge preparations was consistently less than in specimens fixed in Carnoy's solution, though the counts were highly dependent on the period of fixation or staining. The cellular histamine content closely correlated with total mast cell numbers in bronchoalveolar lavage fluid, but was not related to the relative proportions of mast cells which were sensitive or resistant to formaldehyde fixation when using a standard protocol. Compared with normal subjects, the numbers of formaldehyde-sensitive mast cells were significantly elevated in patients with bronchial carcinoma, sarcoidosis, extrinsic allergic alveolitis, cryptogenic fibrosing alveolitis, and mycobacterial infection and were particularly high in the cases of interstitial lung disease. An even greater increase in numbers of formaldehyde-resistant mast cells was observed in the patients with sarcoidosis and extrinsic allergic alveolitis. The associations of these mast cell subsets with disease may reflect relationships between the expansion of the formaldehyde-sensitive population and lymphocyte infiltration and between proliferation of formaldehyde-resistant mast cells and tissue fibrosis.     

Identification of mast cells in bronchoalveolar lavage fluid

The present study was carried out to compare the effectiveness of different fixation and staining methods in the identification of mast cells obtained by bronchoalveolar lavage from patients with interstitial lung diseases. Cell preparations were fixed with formaldehyde or methanol. Mast cells were identified by metachromatic staining with May Grünwald Giemsa, Toluidine blue or Gallamine blue Giemsa. After formaldehyde fixation only a few mast cells were identified, regardless of the stain method used. In contrast, a significantly higher number of mast cells were observed after methanol fixation. Using this fixative, Toluidine blue stain showed a higher number of mast cells than May Grünwald Giemsa. However, there was no difference between Toluidine and Gallamine blue Giemsa in the number of cells observed. The easy identification of mast cells after staining with toluidine, combined with its easy application, suggest that Toluidine blue stain after methanol fixation is the most useful method for determining the presence of mast cells in lavage fluid.     

Differential staining of mast cells with toluidine blue

Abstract

Mast cells play a significant role in inflammatory diseases such as asthma, inflammatory bowel disease, and autoimmune diseases. Inhibition of c-kit receptor tyrosine kinase, a growth factor receptor, significantly reduces mast cell numbers. The purpose of this study was to determine the effect of Compound X (a c-kit inhibitor) on mast cell numbers in rats. Connective tissue mast cells (CTMCs) and mucosal mast cells (MMCs) have differing histochemical characteristics which presents a challenge when staining for quantification by semi-automated image analysis. CTMCs are present in tissues such as tongue and skin and will stain readily in tissues fixed routinely. In contrast, MMCs, such as those present in the intestinal mucosa, are sensitive to fixation. Brief fixation in Carnoy’s solution, although seldom used due to its composition (a mixture of ethanol, chloroform, and acetic acid), was employed to fix tissues for MMC staining, while tissues for CTMC demonstration were fixed in 10% neutral buffered formalin. An enzyme histochemistry method, napthol AS-D choloroacetate (specific esterase), was briefly considered for staining; however, granulocytes stained along with mast cells, requiring manual identification and exclusion, thereby rendering the method incompatible with automated means of quantification. Instead, staining was performed using two different toluidine blue methods which have proven conducive to semi-automated image analysis techniques. CTMCs were stained using Luna’s toluidine blue, while MMCs were stained with Matsson’s toluidine blue modification. In summary, the selected methods, based upon a conventional stain, were easy to do and successfully identified both populations of mast cells for quantification by image analysis.     

绵羊肥大细胞中类胰蛋白酶的证实

用甲苯胺蓝和阿尔辛蓝常规组织化学方法及特异性酶底物鉴定人肥大细胞类胰蛋白酶的酶组织化学技术，采用小鼠抗人肥大细胞类胰蛋白酶单克隆抗体，（ＡＡ１、ＡＡ３和ＡＡ５）通过间接免疫过的经物酶技术对绵羊肥大细胞的组织化学特性进行研究，酶组织化学技术及免疫组织化学技术均首次证实了绵羊肥大细胞颗粒中含有类胰蛋白酶，且该酶可作为绵羊肥大细胞的特异性标志，对于绵羊肥大细胞的常规组织化学染色，Ｃａｒｎｏｙ液及中性缓门     

Sequestration of eosinophil major basic protein in human mast cells.

Previous studies showed that lung and skin mast cells do not contain eosinophil granule major basic protein (MBP). However, MBP has been localized by immunofluorescence to mast cells from a recently established human mast cell line. Analysis of MBP in human mast cell-1 cell lysates by radioimmunoassay showed immunochemical similarity to eosinophil MBP as judged by comparison of dose-response regression lines. Based on these findings and other new information about mast cell heterogeneity, we tested whether mast cells contain MBP. Mast cells were preserved in Carnoy's fixative and were identified by staining with rhodamine-conjugated avidin or for chloroacetate esterase or aminocaproate esterase activity. MBP was localized by immunofluorescence to mast cells in 6 of 7 nasal polyps, 4 of 4 ileal tissue specimens, and 12 of 14 cutaneous mastocytosis specimens. Furthermore, by immunoelectron microscopy MBP was localized to mast cell granules in cutaneous mastocytosis lesions. In contrast, normal skin mast cells preserved in Carnoy's fixative did not contain MBP. After injection of MBP into normal skin and fixation in Carnoy's fluid, mast cells became MBP-positive within 3 minutes, suggesting that endocytosis of MBP by mast cells had occurred. These results suggest that human mast cells in several tissues may sequester toxic eosinophil proteins by endocytosis.     

Long-term cultured mouse mast cells: ultrastructure, histamine and leukotriene levels.

Interleukin 3 (IL3), dependent cells were obtained from bone marrow (9/10 experiments) and spleen cells (4/5 experiments), but not from the thymus. These cells were similar to mucosal mast cell toluidine blue staining and electron microscopy. They had heterogenous metachromatic granules, and some had large scroll-like structures. They also contained histamine (200-800 ng/10(6) cells) for the first 2-5 weeks, whose level diminished to less than 30 ng/10(6) cells by 10 weeks of culture. They also generated leukotriene (LT) C4/D4 (10-40 ng/10(6) cells) and LTB4 (2-5 ng/10(6) cells) for over 100 days of culture. In one experiment, bone marrow-derived mast cells after 150 days of culture began to produce an IL3-like substance and proliferated exponentially without exogenous IL3.