Immunohistology of lymph nodes draining local skin reactions (chancres) in sheep infected with Trypanosoma congolense.

Marked enlargement of lymph nodes draining local skin reactions (chancres) occurred in sheep following intradermal inoculation of cultured metacyclic forms of Trypanosoma congolense. Histologically, these lymph nodes were characterized by follicular hypertrophy and hyperplasia, compression and relative reduction of the paracortical areas and expansion of the medullary regions. Immunohistochemical staining with monoclonal antibodies to ovine lymphocyte subsets and Fc receptor (FcR) bearing macrophages, revealed increased expression of B cells (CD45R+), major histocompatibility complex (MHC) Class II, FcR+ macrophages, and CD1+ cells in the cortical and paracortical areas. The paracortical areas were found to be sparsely populated by CD5+, CD4+ and CD8+ cells, while the medullary areas contained numerous CD8+ cells and FcR+ macrophages. FcR+ macrophages were also present in cortical trabecular and subcapsular sinuses. As the chancre regressed, lymph node reactivity also subsided and fewer B cell follicles were observed and there was decreased expression of CD45R+ and MHC Class II+ cells.     

Rat thymus macrophages: an immunohistochemical study on fetal, neonatal and adult thymus

The pre- and postnatal development of the macrophage population of rat thymus is investigated applying enzyme-histochemical and immunohistochemical techniques, both on tissue sections and cell suspensions. A set of three monoclonal antibodies (ED1, ED2 and ED3), each of which recognizes cells of the monocyte-macrophage lineage in the rat, enabled us to distinguish between macrophages in the various compartments of the thymus. The medulla is characterized by ED1-positive dendritic cells, the corticomedullary region comprises numerous monocyte-like ED1-positive macrophages and the cortex contains a particular subpopulation of branched ED2-positive macrophages. Both the medullary dendritic cells and the cortical branched cells show Ia-membrane staining. ED3-positive cells are only occasionally present. During fetal life ED1-positive monocyte-like macrophages and dendritic cells are present. Just after birth ED2-positive cortical macrophages start to develop. Their number increases strongly during the first week after birth. The role of the various subpopulations of thymic macrophages is discussed.     

Immunohistochemical study of the inflammatory infiltrate associated with equine squamous cell carcinoma.

The distribution of T (CD3), B (CD79) lymphocytes, immunoglobulin (IgG, IgM and IgA)-producing plasma cells, macrophages (lysozyme, Mac387) and MHC Class II antigen was analysed in the inflammatory infiltrate associated with 19 equine squamous cell carcinomas (SCCs) and six cases of precancerous lesions (actinic keratosis). The SCCs came from the penis (11 cases), conjunctiva (four), skin (two), nasal cavity (one) and oral cavity (one). Seven cases were well-differentiated and 12 moderately differentiated. Nine cases showed no invasion of peritumoral deep tissues (locally invasive), whereas the remaining 10 cases were highly invasive. An abundant inflammatory infiltrate was associated with the majority of the SCCs and with lesions of actinic keratosis. This infiltrate was composed mainly of CD3(+)T lymphocytes, CD79(+)B cells and numerous IgG(+)plasma cells; IgM- and IgA-producing plasma cells were scarce and variable, respectively. Macrophages were usually numerous. Macrophages, lymphocytes, intra-epithelial dendritic cells and fibroblasts expressed MHC Class II antigen. No significant correlation was found between the nature of the inflammatory infiltrate and the SCC histological grade or degree of invasion, suggesting that the local anti-tumour immune response failed to prevent tumour invasion or metastasis. MHC Class II was expressed by a variable number of neoplastic epithelial cells in four SCCs, all of which were only locally invasive. In addition, in areas where SCC cells expressed Class II antigen, numerous CD3(+)T lymphocytes were present and some of them were associated with degenerate tumour cells. These findings suggest that the expression of MHC Class II by neoplastic cells induces an improved local anti-tumour immune response.     

小白鼠胸腺发育的组织化学观察

本文观察了BALB/C小鼠从胚胎至成年胸腺发育分化的组织化学变化特征。1.胚胎胸腺皮质外带大淋巴细胞含有糖原。生后1～21 d,皮质内带淋巴细胞内可见很多糖原颗粒;以后减少,成年为阴性。2.胚胎16 d开始,胸腺皮质及髓质内均有大量ANAE阳性的淋巴细胞,以后反应性渐增,出生时已接近成年水平。在皮质的阳性淋巴细胞占皮质淋巴细胞总数的23.90±0.66%;在髓质的为髓质淋巴细胞总数的61.20±1.99%;3.SDH在胚胎16 d的胸腺淋巴细胞内出现,以后活性逐渐增强;4.5′-Nsae和ATPase在胚胎胸腺淋巴细胞为阴性;生后7 d开始,髓质内一部分淋巴细胞变为弱阳性,以后活性渐增强;5.AcP活性以吞噬细胞中最强,部分胸腺淋巴细胞为弱阳性。     

Anti-vimentin monoclonal antibody recognizes a cell with dendritic appearance in the chicken's bursa of Fabricius.

The bursa of Fabricius was studied by immunohistochemical method using anti-vimentin monoclonal antibody (clone 3B4). This monoclonal antibody identified a vimentin positive cell in the medulla of the bursal follicle. During the first 2 weeks of life the vimentin positive cells located along the corticomedullary border and later became prominent in the medulla with the exception of a narrow zone adjacent to the corticomedullary border. After hatching the accumulation of vimentin-type intermediate filaments on one side of the nucleus endowed the vimentin positive cells with a polarized appearance. This "cap-like" vimentin positive area of the cytoplasm determined the position of the major cell process. Within the medulla the Ia positive secretory dendritic cells contained secretory granules in one of the cell processes. The distribution, shape, and polarized appearance of the vimentin positive cells were identical with that of the secretory dendritic cells. Therefore, the anti-vimentin monoclonal antibody proved to be useful for identification of the bursal secretory dendritic cells. During rapid bursal growth the number of secretory dendritic cells increased, possibly, by proliferation of vimentin negative secretory dendritic cell precursors located along the corticomedullary border.     

Anti-vimentin monocional antibody recognizes a cell with dendritic appearance in the chicken's bursa of fabricius

The bursa of Fabricius was studied by immunohistochemical method using anti-vimentin monoclonal antibody (clone 3B4). This monoclonal antibody indentified a vimentin positive cell in the medulla of the bursal follicle. During the first 2 weeks of life the vimentin positive cells located along the corticomedullary border and later became prominent in the medulla with the exception of a narrow zone adjacent to the corticomedullary border. After hatching the accumulation of vimentin-type intermediate filaments on one side of the nucleus endowed the vimentin positive cells with a polarized appearance. This “cap-like” vimentin positive area of the cytoplasm determined the position of the major cell process. Within the medulla the Ia positive secretory dendritic cells contained secretory granules in one of the cell processes. The distribution, shape, and polarized appearance of the vimentin positive cells were identical with that of the secretory dendritic cells. Therefore, the anti-vimentin monoclonal antibody proved to be useful for identification of the bursal secretory dendritic cells. During rapid bursal growth the number of secretory dendritic cells increased, possibly, by proliferation of vimentin negative secretory dendritic cell precursors located along the corticomedullary border.     

Immunohistochemical analysis of small plaque parapsoriasis: involvement of dendritic cells

Small plaque parapsoriasis (SPP) is one of the cutaneous T-cell lymphoproliferative disorders. The aim of the present study was to show the antigenic profile of a subset of dendritic cells and lymphocytes in SPP in comparison with normal cells to provide data on the role of these two cell types in the pathogenesis of SPP. Skin biopsy specimens of lesions were obtained from 8 patients with SPP. Biopsies of the healthy skin from 9 control individuals were also analyzed. Immunohistochemistry was performed on the frozen tissue sections to reveal binding of anti-HLA Class II, anti-CD1a, anti-CD4, anti-CD8, anti-CD44, anti-CD45, and anti-CD68 monoclonal antibodies. There was a statistically significant increase in the number of CD1a(+), Langerhans cells (LCs), HLA-DR-immunoreactive and, CD1a-positive dermal dendritic cells and CD68(+) macrophages in the SPP group (p=0.008, 0.008, 0.002 and <0.0009, respectively). The number of lymphocytes positive for CD4, CD8 and CD45 was significantly higher than normal in the SPP group (p=0.015, <0.0009 and <0.0009, respectively). Our study demonstrates that both peptide- and lipid-based antigens are involved in the persistent antigenic exposure in SPP. Dendritic cells play a pivotal role in SPP by presenting antigens by both LC and dermal dendritic cells via MHC Class II and CD1a molecules. The CD68(+) macrophages are thought to be involved in the immune response in this pathology as an antigen-presenting cell.     

Immunohistochemical analysis of synovial membranes from inflammatory and non-inflammatory arthritides: scarcity of CD5 positive B cells and IL2 receptor bearing T cells.

Rheumatoid Arthritis (RA) synovial membranes were examined by single and dual immunohistological techniques with a number of monoclonal antibodies against lymphocyte and macrophage related antigens. CD4 positive T lymphocytes frequently expressed MHC Class II antigens and were found in sublining collections in close association with activated macrophages as well as B lymphocytes. CD8 positive T cells surrounded these collections as well as being scattered throughout the membrane and also frequently expressed MHC Class II antigens. IL2 receptor (IL2r) expression on T cells and CD5 expression on B cells were rarely seen in these synovial membranes. Similar immunohistological architecture was found in synovial membranes from patients with psoriatic arthritis (PA) and Reiter's Syndrome (RS). Normal synovium contained few T cells, with few cells expressing MHC Class II antigens. Synovium from osteoarthritis (OA) patients also demonstrated similar immunohistological changes to those found in inflammatory arthritides, suggesting that there are only quantitative rather than qualitative differences between the synovial membrane immunohistological architecture from patients with inflammatory and noninflammatory arthritides.     

Antigen-presenting cells expressing glutamate decarboxylase 67 were identified as epithelial cells in glutamate decarboxylase 67-GFP knock-in mouse thymus

Glutamate decarboxylase (GAD), which has two isoforms, GAD65, and GAD67, is responsible for synthesis of the major inhibitory neurotransmitter, gamma-aminobutyric acid. GAD is expressed predominantly in the central nervous system; recent reports suggest that GAD is also expressed in non-neuronal organs including the pancreas. In the pancreatic islets, GAD serves as one of the autoantigens in type I diabetes mellitus. Recent flow cytometric analyses have shown that a variety of self-antigens, including GAD, are ectopically transcribed and expressed in particular cell populations of the thymus, although consensus concerning the cellular phenotype has not been obtained. The aim of this study was to clarify the localization and cellular phenotype of GAD67-expressing cells in the thymus at a cellular level with a novel approach using GAD67-green fluorescent protein (GFP) knock-in mice, in which GFP is expressed specifically in GAD67-positive cells. GFP-positive cells were detected in the thymic medulla and were identified as epithelial cells by immunohistochemistry. Almost all GFP-positive cells were positive for major histocompatibility complex (MHC) class II antigen staining and were positive for both cytokeratin and Ulex Europaeus Agglutinin I, markers of medullary thymic epithelial cells, but were negative for CD11c, Gr-1, and CD45, markers of dendritic cells, macrophages, and B-lymphocytes, respectively.     

人胎儿胸腺淋巴细胞的免疫表型分析

用免疫组织化学ＡＢＣ法观察了１４例人胎胸腺中六种Ｔ细胞和细胞表面抗原的表达。结果表明，ＣＤ４和性和ＣＤ８阳性的细胞主要见于皮质深化深层，髓质中较少。ＣＤ３阳性细胞多分布在髓质，皮质深层有少量阳性细胞。ＣＤｌａ阳性细胞见于皮质。ＣＤ２５阳性细胞多分布于髓质。该阳性细胞在胎儿胸腺中的分布反映了Ｔ细胞的分化。Ｂ细胞表面抗原阳性细胞主要分布在髓质，皮质中很少，在髓质及被膜下亦见少数较大并有突起的阳性细胞。     

Immunosuppression with cyclosporin A alters the thymic microenvironment

The effect of cyclosporin A (CyA) immunosuppression on the murine thymic microenvironment and T lymphocyte development has been analysed using monoclonal antibodies to epithelial and lymphocyte subpopulations, macrophages and major histocompatibility complex (MHC) class II antigens in immunohistochemistry and flow cytometry. The major microenvironmental target for CyA-induced damage was the thymic medulla, where a reduction in all epithelial cell subsets, dendritic cells and macrophages was observed. In contrast, the thymic cortex appeared essentially normal. CyA had no detectable effect on the intensity of microenvironmental expression of MHC class II molecules in either cortex or medulla, although the number of MHC class II positive medullary cells was reduced after CyA treatment. CyA also had a differential effect on the thymic lymphocyte populations where there was little change in the Thy-1 bright, CD5 dull, CD4+, CD8+ cortical thymocytes but a depletion of the Thy-1 dull, CD5 bright, CD4 or CD8 single-positive medullary cells. This lymphocyte loss may be due partly to increased migration from thymus to spleen and other peripheral lymphoid organs, and partly to a block in the differentiation stage from cortical to medullary lymphocyte. The thymic microenvironment and lymphocyte subpopulations recover rapidly after cessation of CyA treatment, although there may be longer term functional defects resulting from the CyA-induced injury.     

Leishmaniosis and generalized demodicosis in three dogs: a clinicopathological and immunohistochemical study

This paper describes the clinicopathological and immunohistochemical aspects of the skin lesions in three dogs with leishmaniosis and generalized demodicosis. Diffuse alopecia, crusts, folliculitis and furunculosis, as commonly seen in generalized demodicosis, were prominent in all the dogs. MicroIscopically, there was a diffuse and perifollicular superficial and deep granulomatous dermatitis and, in two dogs, both Copyright Demodex canis mites and Leishmania spp. amastigotes were observed in the same lesions. Numerous Mac387(+)macrophages were observed in the inflammatory infiltrates, but macrophages loaded with amastigotes were Mac387(-). In all cases, immunoreactive CD3 lymphocytes were sparse, both in the granulomatous and perifollicular infiltrates. There were numerous IgG+, IgG4(+)-secreting plasma cells in areas of folliculitis and furunculosis and fewer IgG2(+), IgG3(+), IgA+and IgM+-secreting plasma cells in the inflammatory infiltrate. In all cases, MHC Class II was expressed by the majority of dermal macrophages and dendritic cells, as well as by lymphocytes and fibroblasts. The paucity of CD3(+)lymphocytes, usually abundant in D. canis lesions, points to leishmania-induced cell-mediated immunosuppression as a predisposing factor for generalized demodicosis. 1999 W.B. Saunders and Company Ltd.     

人胸腺内Ghrelin的免疫组织化学定位

目的:观察Ghrelin在人胸腺的定位与分布,为深入探讨胸腺内Ghrelin的功能意义提供实验依据。方法:采用特异性抗Ghrelin血清,用免疫组织化学ABC法观察人胸腺内Ghrelin的定位与分布。结果:Ghrelin免疫反应阳性细胞在胸腺内分布广泛。胸腺皮质浅层的Ghrelin阳性细胞呈巨噬细胞形态特点;而皮髓质交界区有大量上皮性网状细胞呈Ghrelin免疫反应阳性;胸腺髓质内Ghrelin阳性信号主要定位于树突状细胞和上皮性网状细胞;胸腺小体呈Ghrelin免疫反应强阳性。结论:胸腺内Ghrelin分布广泛,定位于胸腺巨噬细胞、上皮性网状细胞、树突状细胞和胸腺小体。Ghrelin可能参与胸腺细胞分化和成熟的调节。     

DEC-205/CD205+ dendritic cells are abundant in the white pulp of the human spleen, including the border region between the red and white pulp

The distribution of dendritic cells (DCs) and macrophages in the human spleen has received less attention than that of lymphocytes. Here we have addressed this problem with the human DEC-205/CD205 marker ('DEC'), which is an endocytic receptor on DCs that mediates efficient presentation of antigens. DEC was abundant on dendritic profiles in the white pulp but absent from the red pulp, the latter defined with antibodies to two antigens, mannose receptor/CD206 on sinusoidal lining cells, and macrosialin/CD68 on macrophages. Double staining with anti-DEC and anti-CD3 showed the expected concentration of DEC+ cells in the relatively small T-cell areas of the human spleen. DEC+ cells were also found in other regions of the white pulp. In all regions, the DEC+ cells were positive for major histocompatibility complex (MHC) class II and the CD11c integrin but largely immature, with low expression of B7-2/CD86 costimulator and DC-lysosome-associated membrane protein (LAMP)/CD208. When we concentrated on the perifollicular region between the red pulp and the marginal zone, we found macrophages that stained with antibodies to sialoadhesin/CD169 and DC-specific ICAM-3 grabbing non-integrin (SIGN)/CD209, and just inside these cells were DEC+ profiles. The DEC+ DCs were intertwined with cells that stained for the vascular addressin mucosal addressin cell adhesion molecule (MAdCAM). Therefore, anti-DEC-205/CD205 antibodies are useful for identifying DCs in human splenic white pulp and its border region with the red pulp.     

Macrophages and interdigitating reticulum cells in normal thymus and in thymoma: an immunohistochemical study

The distribution and immunophenotype of macrophages and interdigitating reticulum cells were investigated on frozen sections of seven normal thymuses and 10 thymomas. In normal thymus, macrophages were mainly located in the cortex, were markedly PAM-1+/MAC+, weakly Leu-M3+ (CD14), T4+ (CD4), T9+ and OKM-1+ (CD11b). Interdigitating reticulum cells were mainly located in the medulla and were pan-Leu+ (CD45), T4+(CD4+), HLA-DR+; furthermore, they were also often TAC+ (CD25) and T9+. Thymomas were composed of cytokeratin-containing epithelial cells admixed with variable proportions of T6+ (CD1a) lymphocytes. As defined by the histological features two thymomas were lymphocyte-rich, five were mixed type and three were epithelial-rich; eight thymomas were mainly composed of cortical epithelial cells and two were composed of spindle epithelial cells suggesting a medullary origin. In all cases, thymoma-associated macrophages were markedly PAM-1+/MAC+; they were numerous, and regularly distributed throughout the tumour. The density of macrophages per unit area was similar to that of the normal thymus, and was not influenced by the histological type or by the lymphocyte content of the tumour. Interdigitating reticulum cells were few and were confined to the areas of medullary differentiation.     

Follicle-associated epithelium and medullary epithelial tissue of the bursa of Fabricius are two different compartments

The bursae of Fabricius from the chicken and turkey were studied by light and electron microscopy and immunohistochemical methods. The study focused on the relationship of follicle-associated epithelium to the medulla. The follicle-associated epithelium was supported by 3 to 5 layers of stratified epithelial cells which were a continuation of the corticomedullary epithelial cells. The follicle-associated epithelium consisted of M cells and scattered secretory dendritic cells. The network of the reticular epithelial cells of the medulla was filled with secretory dendritic cells, B cells, and a few T cells and macrophages. The cellular content of the follicle-associated epithelium and the medulla suggested that they were different cellular compartments. Communication between the follicle associated epithelium and medullary epithelial compartment occurred through the supporting cells of the follicle-associated epithelium. When the supporting layers of the follicle-associated epithelium infolded into the medulla, they formed lamellated epithelial bodies similar to the thymic Hassall bodies. The lamellated bodies enclosed secretory dendritic cells but not lymphocytes. The infolding of supporting cells varied from follicle to follicle. The asynchronization of infolding contributed to heterogeneity of follicle composition. Follicle heterogeneity was demonstrated by differences in reactivity with a battery of monoclonal antibodies. © 1992 Wiley-Liss, Inc.     

Ontogenic development of macrophage subpopulations and Ia-positive dendritic cells in fetal and neonatal rat spleen.

Development, differentiation, and distribution of macrophage subpopulations and Ia+ dendritic cells in the fetal and neonatal rat spleen were investigated by means of double immunohistochemical staining and immunoelectron microscopy. To characterize these cell populations, a panel of anti-rat macrophage monoclonal antibodies (RM-1, ED2, ED3, TRPM-3, Ki-M2R) and an anti-rat Ia antibody (OX6) were used. In the fetal rat spleen, macrophages were first detected by RM-1 at fetal day 15. ED2+ and/or Ki-M2R+ macrophages appeared at fetal day 16. TRPM-3+ and/or ED3+ macrophages appeared a day later. During the fetal and neonatal development, ED2+ and TRPM-3+ macrophages differentiated independently, maturing into red pulp macrophages and marginal metallophilic and marginal zone macrophages respectively. Intimate topographical relations were observed between ED2+ macrophages and hematopoietic cells and between TRPM-3+ macrophages and marginal zone lymphocytes. Ia+ cells were first observed around arterioles at fetal day 15. In the fetal and neonatal period, the number of Ia+ cells gradually increased, their shape became dendritic, and they matured into interdigitating cells in the inner periarteriolar lymphatic sheath. In ontogeny, Ia+ dendritic cells were not stained with ED2 or TRPM-3. These results suggest that ED2+ macrophages, TRPM-3+ macrophages, and Ia+ dendritic cells are distinct cell lines that pursue independent developmental process in spleen ontogeny.     

Follicle-associated epithelium and medullary epithelial tissue of the bursa of fabricius are two different compartments.

The bursae of Fabricius from the chicken and turkey were studied by light and electron microscopy and immunohistochemical methods. The study focused on the relationship of follicle-associated epithelium to the medulla. The follicle-associated epithelium was supported by 3 to 5 layers of stratified epithelial cells which were a continuation of the corticomedullary epithelial cells. The follicle-associated epithelium consisted of M cells and scattered secretory dendritic cells. The network of the reticular epithelial cells of the medulla was filled with secretory dendritic cells, B cells, and a few T cells and macrophages. The cellular content of the follicle-associated epithelium and the medulla suggested that they were different cellular compartments. Communication between the follicle associated epithelium and medullary epithelial compartment occurred through the supporting cells of the follicle-associated epithelium. When the supporting layers of the follicle-associated epithelium infolded into the medulla, they formed lamellated epithelial bodies similar to the thymic Hassall bodies. The lamellated bodies enclosed secretory dendritic cells but not lymphocytes. The infolding of supporting cells varied from follicle to follicle. The asynchronization of infolding contributed to heterogeneity of follicle composition. Follicle heterogeneity was demonstrated by differences in reactivity with a battery of monoclonal antibodies.