Dermal Lymphatic Vessels Research Articles

Human Langerhans cell in a dermal lymphatic vessel.

Human Langerhans cell in a dermal lymphatic vessel.

Visualization and Characterization of Migratory Langerhans Cells in Murine Skin and Lymph Nodes by Antibodies Against Langerin/CD207

Dendritic cells are professional antigen-presenting cells that initiate primary immunity. Migration from sites of antigen uptake to lymphoid organs is crucial for the generation of immune responses. We investigated the migratory pathways specifically of epidermal Langerhans cells by tracing them from the epidermis to the draining lymph nodes. This was possible with a new monoclonal antibody, directed against murine Langerin/CD207, a type II lectin specific for Langerhans cells. In situ, resident, and activated Langerhans cells express Langerin in the epidermis and on their way through dermal lymphatic vessels. Both emigrated and trypsinization-derived Langerhans cells expressed high levels of Langerin intracellularly but reduced it upon prolonged culture periods. Sizeable numbers of Langerin+ cells were found in skin draining lymph nodes but not in mesenteric nodes. Langerin+ cells localized to the T cells areas and rarely to B cell zones. Numbers of Langerin-expressing cells increased after application of a contact sensitizer. In the steady state, Langerhans cells in the skin-draining nodes expressed maturation markers, such as 2A1 and costimulatory molecules CD86 and CD40. These molecules, CD86 and CD40, were further upregulated upon inflammatory stimuli such as contact sensitization. Thus, the novel anti-Langerin monoclonal antibody permits the unequivocal visualization of migratory Langerhans cells in the lymph nodes for the first time and thereby allows to dissect the relative immunogenic or tolerogenic contributions of Langerhans cells and other types of dendritic cells.

Migration of dendritic cells into the lymphatics: the Langerhans cell example

Dendritic cells, including Langerhans cells of the epidermis and the mucous membranes, are key leukocytes for the initiation of adaptive immune responses as well as for the maintenance of peripheral tolerance. In the former regard they may well play an important, but as yet unrecognized role in the pathogenesis of Behcet's disease. Epidermal Langerhans cells may serve as a paradigm for their counterparts in the mucosae. These cells efficiently take up (microbial) antigens, they process them into immunogenic MHC–peptide complexes, and they transport this form of antigen to the lymph nodes via lymphatic vessels. Depending on the milieu where Langerhans cells have encountered antigen (inflammatory versus non-inflammatory/steady-state), they make T cells proliferate and acquire effector functions (immunity) or render them unresponsive or even delete them (tolerance), respectively. In addition, plasmacytoid dendritic cells, a recently characterized type of dendritic cells, may also directly trigger innate responses (e.g. by secretion of type I interferons in response to virus). Studying the pathways and the regulation of dendritic cell migration might help to unravel a possible involvement of dendritic cells in Behcet's disease. We present observations on the physical obstacles that dendritic cells migrating in the skin have to overcome until they reach dermal lymphatic vessels. Furthermore, we show that migration is critically dependent on the function of matrix metalloproteinases, in particular MMP-2 and MMP-9. It becomes evident that Langerhans cells indeed carry antigens (including self antigens such as melanosomes or apoptotoic bodies or tumor antigens such as particular cytokeratins) through the lymphatics. Given these observations it may be worth studying Langerhans cells and dermal dendritic cells in Behcet's disease.

高度リンパ管浸潤を伴う陰嚢原発の有きょく細胞癌の１例

A-75-year-old man developed a rapidly growing tumor followed by swelling of regional lymph nodes, on the left side of his scrotum. Histopathological examination of the tumor revealed a deep invasion of atypical squamous cells associated with embolism of dermal lymphatic vessels by tumor cells, as far as 4cm apart from the tumor. Wide local excision of the tumor including total scrotal skin and bilateral ilioinguinal lymph node dissection was performed. Subsequently five courses of chemotherapy with cisplatin and fluorouracil and radiotherapy were added. In spite of these treatments, he had a recurrence located on the posterior border of the grafted skin of scrotum one year after the operation. [Skin Cancer (Japan) 2001; 16: 47-50]

Migration of Langerhans cells and dermal dendritic cells in skin organ cultures: augmentation by TNF-α and IL-1 β

Migration from sites of antigen encounter to lymphoid organs is essential to the strong immunogenic function of dendritic cells (DC). In the skin, migration proceeds through dermal lymphatic vessels and is regulated in an incompletely understood way by inflammatory mediators. We studied the effects of tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL-1beta) in mouse skin organ cultures by direct enumeration of migrating DC and by immunohistochemistry. (1) Neutralizing antibodies to TNF-alpha and IL-1beta inhibited migration of DC, also in human skin explants (TNF-alpha). (2) TNF-alpha at low concentrations (50 U/mL) and IL-1beta (50-3000 U/mL) augmented migration to about 150% of spontaneous migration. (3) High concentrations of TNF-alpha (5000 U/mL) inhibited migration by approximately 50%. (4) DC migration from skin explants of TNF-alpha/lymphotoxin-alpha double-deficient mice and TNF-receptor type 1 and 2 double knockout mice was not impaired. (5) TNF-alpha effects were neutralized by anti-IL-1beta, and vice versa. We conclude that in normal animals both TNF-alpha and IL-1beta are required for DC migration to occur. In the complete absence of one cytokine (TNF-alpha), however, backup mechanisms step in.

A Case Control Study on Risk Factors Involved in Inflammatory Breast Recurrence afterBreast-Conserving Surgery

Recurrence that poses the biggest problem after breast-conserving surgery is local recurrence. Particularly, in the case of inflammatory breast recurrence which is rare but has a specific pathologic nature, it is important to elucidate the pathology and risk factors and to consider appropriate countermeasures. In the present study, we classified 133 cases of recurrence following breast-conserving surgery, collected from 18 key hospitals/institutes in Japan. Recurrence types were divided into three groups, namely, inflammatory breast recurrence, noninflammatory breast recurrence and distant metastasis only, and the risk factors involved in recurrence were investigated by the case control study allotting 2 controls to each case. The study population consisted of 9 cases of the inflammatory type, 64 cases of the noninflammatory type and 60 cases of distant metastasis. The significant risk factor for inflammatory breast recurrence was positive lymph node metastasis, which was significantly more frequent in lymphatic invasion-positive cases unlike in the distant metastasis group. The positive surgical margin and nonradiation therapy which have been shown to be significant risk factors for noninflammatory breast recurrence were entirely unrelated with inflammatory breast recurrence. In addition, the inflammatory-type recurrence time was as short as about 12 months irrespective of whether radiation therapy was performed or not. The inflammatory type was accompanied with local wide extension (cancerous embolus of the dermal lymphatic vessels), and distant metastasis (lymphangitis carcinomatosa) at the time of recurrence, and further surgery was impossible in most cases, with a significantly poorer prognosis than the other recurrence types. These findings suggest that this recurrence corresponds to the so-called ‘occult’ case of primary inflammatory breast carcinoma. We think it important to predict this recurrence by close pathological examination, particularly in patients with lymph node metastasis, and to consider appropriate measures.

Expression of the Vascular Endothelial Growth Factor C Receptor VEGFR-3 in Lymphatic Endothelium of the Skin and in Vascular Tumors

It is difficult to identify lymph vessels in tissue sections by histochemical staining, and thus a specific marker for lymphatic endothelial cells would be more practical in histopathological diagnostics. Here we have applied a specific antigenic marker for lymphatic endothelial cells in the human skin, the vascular endothelial growth factor receptor-3 (VEGFR-3), and show that it identifies a distinct vessel population both in fetal and adult skin, which has properties of lymphatic vessels. The expression of VEGFR-3 was studied in normal human skin by in situ hybridization, iodinated ligand binding, and immunohistochemistry. A subset of developing vessels expressed the VEGFR-3 mRNA in fetal skin as shown by in situ hybridization and radioiodinated vascular endothelial growth factor (VEGF)-C bound selectively to a subset of vessels in adult skin that had morphological characteristics of lymphatic vessels. Monoclonal antibodies against the extracellular domain of VEGFR-3 stained specifically endothelial cells of dermal lymph vessels, in contrast to PAL-E antibodies, which stained only blood vessel endothelia. In addition, staining for VEGFR-3 was strongly positive in the endothelium of cutaneous lymphangiomatosis, but staining of endothelial cells in cutaneous hemangiomas was weaker. These results establish the utility of anti-VEGFR-3 antibodies in the identification of lymphovascular channels in the skin and in the differential diagnosis of skin lesions involving lymphatic or blood vascular endothelium.

A physiologic function for p-glycoprotein (MDR-1) during the migration of dendritic cells from skin via afferent lymphatic vessels.

P-glycoprotein (MDR-1) is a well-known transporter that mediates efflux of chemotherapeutic agents from the intracellular milieu and thereby contributes to drug resistance. MDR-1 also is expressed by nonmalignant cells, including leukocytes, but physiologic functions for MDR-1 are poorly defined. Using an initial screening assay that included >100 mAbs, we observed that neutralizing mAbs MRK16, UIC2, and 4E3 against MDR-1 specifically and potently blocked basal-to-apical transendothelial migration of mononuclear phagocytes, a process that may mimic their migration into lymphatic vessels. Antagonists of MDR-1 then were used in a model of authentic lymphatic clearance. In this model, antigen-presenting dendritic cells (DC) migrate out of explants of cultured human skin and into the culture medium via dermal lymphatic vessels. DC and T cells derived from skin expressed MDR-1 on their surfaces. Addition of anti-MDR-1 mAbs MRK16, UIC2, or the MDR-1 antagonist verapamil to skin explants at the onset of culture inhibited the appearance of DC, and accompanying T cells, in the culture medium by approximately 70%. Isotype-matched control mAbs against other DC molecules including CD18, CD31, and major histocompatibility complex I did not block. In the presence of MDR-1 antagonists, epidermal DC were retained in the epidermis, in contrast to control conditions. In summary, this work identifies a physiologic function for MDR-1 during the mobilization of DC and begins to elucidate how these critical antigen-presenting cells migrate from the periphery to lymph nodes to initiate T lymphocyte-mediated immunity.

Ｅｔｏｐｏｓｉｄｅが奏効した外陰部Ｐａｇｅｔ癌の１例

Chemotherapy for extramammary Paget's carcinoma has not been established yet. We report a effective case with oral low dose etoposide therapy.A 69 year-old male visited our hospital with a scaling erythema and a subsequent nodule on his pubic lesion. Biopsy specimen revealed extramammary Paget's carcinoma. Tumor resection and left inguinal lymphadenectomy were performed. 8 months after operation, purpuric erythema developed on his left thigh and tumor cells were histolopathogically proven in the dermal lymph vessels. Oral tegafur uracil and tamoxifen were tried, but not effective.Chronic administration of low dose oral etoposide was tried to control the spreading of cutaneous tumors and lymphnode metastasis. The tumors regressed gradually and serum CEA level decreased to almost normal level. Only slight myelosuppression was found, and no severe adverse reaction was seen. After 5 months from administration of oral etoposide, nodules on the left thigh enlarged again. The patient died about 2 years after operation, but we conclude oral low dose etoposide therapy is worth trying.

Demonstration of Initial Lymphatics in Excised Human Skin Using an Extension Technique and Dye Injection

Using a newly developed technique (extension technique) we succeeded in presenting dermal lymph vessel networks by subepidermal injection of an aqueous Berlin blue solution in excised human skin. Of 94 skin specimens taken from the safety margin of excised melanomas it was possible to demonstrate 32 networks and 14 single dermal lymph vessels. The networks can be documented by macroscopic photographs. Acetic fixation enables us to produce transparent preparations suitable for investigations with a stereo-microscope. The technique described here for macroscopically demonstrating dermal lymph vessels makes it possible to investigate systemically structural variations of lymph vessel networks in human skin.

Indirect lymphography with Isovist-300 in various forms of lymphedema

Twenty patients with various forms of lymphedema were studied by indirect lymphography. Isovist, a new non-ionic, water-soluble, dimeric contrast media, was used for intradermal infusion. Simultaneous indirect lymphography showed characteristic patterns of lymphatics. The initial lymphatics were visualized by intradermal infusion of the newly developed isovist. However, they were not opacified with direct lymphography unless in the presence of valvular insufficiency. No systemic side-effects were observed except for a feeling of discomfort at the injection site. The results showed that indirect lymphography with intradermal infusion of Isovist is useful for demonstrating dermal lymph vessels and for the diagnosis of lymphedema.

Commentary on the Langerhans Cell in Contact Hypersensitivity

The origin and function of Langerhans cells remained elusive for many years after their discovery by Paul Langerhans in 1868. It was not until 1961 that an ultrastructural marker of Langerhans cells in the form of a unique organelle was described [1]. Subsequently, while conducting an electronmicroscope study of contact allergic reactions in humans [2], Dr. Inga Silberberg-Sinakin and her colleagues noted apposition of lymphocytes to Langerhans cells, suggesting a role for Langerhans cells in immunologic reactions. This paper summarizes an interview with Drs. Inga SilberbergSinakin and Rudolf Baer early in 1988, discussing their reminiscences concerning this seminal observation and its implications for Langerhans cell research. During her early studies of the topical application of mercuric chloride in humans, Dr. Silberberg-Sinakin observed that there was apposition of lymphocyte-like cells to Langerhans cells in the epidermis of individuals with contact allergic reactions but not of those individuals with irritant reactions or of normal subjects [2]. She remembers that it was a troublesome finding to evaluate because findings observed on electron microscopy are difficult to statistically quantitate. Therefore, she proceeded to study a variety of contact allergens in humans [3] and to seek and exploit an animal model [4]. In studies of guinea pigs sensitized to dinitrochlorobenzene (DNCB), subsequent passive transfer experiments with a time-course analysis demonstrated Langerhans cells in dermal lymphatics [5,6], a finding that strengthened the interpretation of an immunologic role for the Langerhans cell. At that time, very little was known about the biologic functions of the Langerhans cell. As background, Dr. Baer recalled that as early as 1875, Ranvier speculated that Langerhans cells were derived from lymphocytes. However, subsequent informed opinion was that Langerhans cells were effete melanocytes. Then it was recognized that Langerhans cells could be identified by electron microscopy based on the key feature of a distinctive cytoplasmic organelle [1], that the cell membrane of Langerhans cells contained ATPase [7]; and that Langerhans cell granules were endocytotic organelles [8]. Dr. Silberberg-Sinakin went on to say that the initial recognition of the morphologic observations led her and her collaborators at New York University School of Medicine (Dr. Jeanette Thorbecke and Dr. Baer) to suggest that the Langerhans cell was the potential antigen-presenting cell in the skin that people had been seeking. Dr. Silberberg-Sinakin added her perspective on the controversies involved in the acceptance of this work. She stated that there was support within the Department of Dermatology at New York University School of Medicine and among immunologists and cell biologists. Within the general dermatologic community, however, there was reluctance to accept this new immunologic function of the Langerhans cell. Both Dr. Silberberg-Sinakin and Dr. Baer went on to state that the studies which involved the uptake of ferratin by Langerhans cells and their migration through dermal lymph vessels to the lymph node were not only of great importance in elucidating the fate of antigens taken up in the epidermis and how they migrate to the regional lymph nodes [9,10], but also was the first work that showed a applicability to contact allergy [11]. Although contact allergy has been a model for studying the fate of many different kinds of antigens, it also suggested that in delayed-type hypersensitivity, reactions in general Langerhans cells could serve the same functions. Presumably, larger molecules, such as tumor antigens and microbial antigens such as viruses and fungal antigens, are taken up by Langerhans cells [12]. This function also applies to transplantation antigens, and it has been shown that Langerhans cells play a role in graft rejection [13] in the skin. Moreover, Langerhans cells could be target cells [14], and the damaging effects could be caused by lymphocytes or immune complexes plus complement [14]. In addition, certain physical agents and substances are capable of depressing the functional capacity of Langerhans cells [I5–19]; notably ultraviolet A and ultraviolet B and the topical or systemic administration of glucocorticosteroids [20]. There are also different numbers of Langerhans cells in various skin sites. Areas with diminished numbers of Langerhans cells are not able to mount contact reactions [21]. After the observation that Langerhans cells were antigenbearing cells, Dr. Silberberg-Sinakin said that she and her colleagues reviewed the literature on antigen-bearing or antigen-presenting cells (such as indeterminant dendritic cells, interdigitating reticular cells, follicular dendritic reticular cells described by Nossal, and dendritic cells described by Steinman and Cohn) in other organs such as lymph node,

Inflammatory Metastatic Carcinoma of the Parotid

Cutaneous metastases may assume a variety of distinctive morphologic patterns.¹We describe a man with parotid adenocarcinoma and an erythematous patch on the anterior side of the chest. The latter condition resulted from lymphatic cutaneous spread of his adenocarcinoma, producing so-called inflammatory metastatic carcinoma or carcinoma erysipelatoides. The pattern is almost always associated with breast cancer. To our knowledge, this is the first report of inflammatory metastatic carcinoma associated with parotid adenocarcinoma. <h3>Report of a Case</h3> A man, aged 60 years, was seen in August 1979 with a four-month history of inflammation over the left parotid gland. An examination disclosed that the gland was swollen and indurated. A left parotidectomy and radical neck dissection were done, followed by a pectoralis flap reconstruction. The parotid was found to be diffusely infiltrated with a tumor that also involved the dermal lymphatic vessels of the overlying skin, the masseter muscle, and 13

In situ identification of T6-positive cells in normal human dermis by immunoelectron microscopy.

Monoclonal anti-T6 antibody, which reacts with the majority of cortical thymocytes but not peripheral T cells, also reacts with human epidermal Langerhans cells, as shown by a four-step immunoperoxidase method and immunoelectron microscopy. To define whether T6-positive cells are also present in normal human dermis, we used these techniques to demonstrate two immunologically distinct populations of histiocyte-like cells in normal human dermis. The first population contains cells devoid of phagolysosomes or Birbeck granules. These cells react with anti-T6 antibody, but not with monoclonal anti-T3 antibody which defines peripheral T cells, and are found predominantly in and around dermal lymphatic vessels. The second is composed of phagolysosome-containing cells which do not react with anti-T6 antibody or anti-T3 antibody. Because to date, Langerhans cells are the only cells in normal human epidermis that react with anti-T6 antibody, these data provide immunological evidence for a specific link between Langerhans cells and a T6-positive dermal mononuclear cell, possibly the so-called indeterminate cell. In addition, application of these techniques should, for the first time, permit the immunological distinction of these T6-positive mononuclear cells from other cells bearing Ia antigens, such as dermal histiocytes and certain lymphocytes, in normal and diseased skin.

Prognostic factors in cutaneous malignant melanoma: A comparative study of long term and short term survivors

The clinical presentation and histologic characteristics of primary melanomas of a consecutive series of 100 short term (fatal within five years) and 100 long term survivors (disease-free for ten years or longer) were studied. The clinical parameters analyzed included the history of pre-existent mole, duration of changes before diagnosis, site, elevation, ulceration, extent of pigmentation, and satellitosis around the primary lesion. Study of histologic features included analysis of exact measurements of the primary melanoma, depth of dermal invasion, tumor cell morphology, mitotic rate, intracellular pigmentation, presence or absence of giant tumor cells, blood vessel or lymphatic invasion, lateral junctional activity, and presence and extent of lymphoid response to the primary lesion. Statistically significant differences were found between the long term and short term survivors with respect to several variables. It is one of the conclusions of this study that an elective node dissection would seem to be indicated in all patients with stage I melanoma who present with poor prognostic signs, that is, patients who belong to any combination of the following categories: male, primary lesions>1 cm., which has ulcerated, with a deeper level of dermal penetration, with dermal lymphatic or blood vessel invasion, and a lack of lymphoid reaction around the primary lesion.