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PubMed2003

Development of the peritoneal lymphatic stomata and lymphatic vessels of the diaphragm in mice.

The generation and development of the peritoneal lymphatic stomata (PLS) and lymphatic vessels of the diaphragm were studied in mice at gestational ages from the embryonic to the postnatal period with TEM, SEM and enzyme histochemistry and the PLS data were quantitatively analyzed with computer-assisted image processing technology (Elescope image analysis software). The results showed that the diaphragmatic mesothelium was covered only by flattened mesothelial cells (FMC) at the 13th embryonic day (ED 13). At ED 15, some cuboidal mesothelial cells (CMC) and immature lymphatic stomata (NLS) were found scattered on the diaphragmatic mesothelium. The sub-peritoneal lymphatic capillaries did not appear until ED 18. However, no absorptive function was observed in NLS when trypan blue granules were injected into the peritoneal cavity. At postnatal day 1 (PND 1), the endothelial cytoplasm processes of the diaphragm lymphatic capillaries span the connective tissue fibers and the basal membrane of CMC to form the subperitoneal channels. These channels were connected with NLS and serve as the absorptive route between the peritoneal cavity and the sub-peritoneal lymphatic vessels. The trypan blue absorption test demonstrated that postnatal PLS possessed an absorptive function and had transformed to mature lymphatic stomata (MLS) by PND 1. Thus, NLS were renamed of MLS. At PND 5, the cuboidal mesothelial cell ridge (CMCR) appeared with increased CMC areas. At PND 10, CMCR were fused to form the band-like CMC area with much more MLS distributed in the muscular portion of the diaphragm. With distribution area and density of PLS increasing and growth of lymphatic vessels, an increased absorptive function from the peritoneal cavity was observed in the experiment.

Animals↗

PubMed1991

Lymphatic vessels in the developing diaphragm of the rat.

Diaphragms of fetal, neonatal and young albino rats have been observed both under light and electron microscopes to examine the presence and distribution of lymphatic vessels and their morphological features. In fetal diaphragms of between 18 and 22 days of gestation, no normal lymphatic vessels can be seen; only after birth, specifically in neonatal and 2-day-old rats, small lymphatic vessels appear; they are in close proximity to the blood vessels in the inner areas of the muscle. As the rats get older, lymphatic vessels are also observed in the subserosa where an abundant connective tissue is present. The fine structure of diaphragmatic lymphatic vessels is different at different ages. In neonatal rats of up to 2 days, the endothelial wall is very thin and often holed. The relationships between contiguous endothelial cells are characterized by simple end-to-end or overlapping structures. The basement membrane is virtually absent. Within the first week of life, the endothelial wall becomes more complex; along the wall, complex interdigitations between two contiguous endothelial cells often touch. A discontinuous basement membrane and collagen and elastic fibers surround the vessels. In the older rats (from 14 to 25 to 140 days), next to the complex interdigitations which characterize the junction between two contiguous endothelial cells, cellular flaps interdigitate forming a channel which opens out either to the exterior or the interior of the vessel. Dense bundles of elastic and collagen fibers are closely apposed to the endothelial wall.

Animals↗

PubMed2002

Lymphatic vessel activation in cancer.

Metastasis of most cancers occurs primarily through the lymphatic system, and the extent of lymph node involvement is the most important prognostic indicator. While the importance of the lymphatic system as a pathway for metastasis has been well recognized, there is very little information available about the mechanisms by which tumor cells interact with the lymphatics. Recently, production of the lymphangiogenic factor VEGF-C has been detected in tumors, and the significance of VEGF-C-mediated lymphangiogenesis for tumor metastasis has been demonstrated. Increased lymphatic vessel density has been found associated with certain tumors. The mechanisms by which tumor cells gain access to and enter lymphatic vessels are critical issues that need to be addressed in the future. In contrast to the prevailing view that has assigned to the lymphatic system a passive role in the metastatic process, our results indicate the importance of lymphatic vessel activation in tumor dissemination.

Angiogenesis Inducing Agents↗

PubMed2006

Distributional characteristics of lymphatic vessels in normal human nasal mucosa and sinus mucosa.

An immunohistochemical staining technique with the D2-40 antibody was undertaken to examine the functional and morphological features of lymphatic networks in tissue sections and whole-mount preparations of normal nasal mucosa and ethmoid sinus mucosa. In normal nasal mucosa, most lymphatic vessels were found in the superficial mucosa beneath the epithelial layer. Some of these vessels were dilated, whereas others were compressed and had a slit-like lumen. Whole-mount preparations revealed the extent of lymphatic vessels in normal ethmoid sinus mucosa. A network of lymphatic vessels was mainly found in the subepithelial layer, where lymphatic vessels represented rich networks, possessing antler-like branches and typical blind ends. However, these lymphatic networks were not arranged in the form of lymphangion chains, with each lymphangion consisting of a contractile compartment and valve. Thus, recognition of the distinctive features of the lymphatic network in normal nasal and sinus mucosa might aid investigations of lymphatic involvement in sinonasal diseases, such as rhinitis, sinusitis, and malignancy.

Adult↗

PubMed1982

Use of isosulfan blue for identification of lymphatic vessels: experimental and clinical evaluation.

Use of vital dyes for identification of lymphatic vessels before cannulation has heretofore not been approved by the Food and Drug Administration (FDA). The suitability of isosulfan blue, the 2,5 disulfonic acid isomer of Patent Blue, for this purpose was evaluated experimentally in the rat and clinically in 11 volunteers and 543 patients under an investigational new drug application. FDA approval for this drug has been obtained. Volunteers and patients received up to 15 mg of a 1% sterile, pyrogen-free solution per extremity (total dose of 0.4 mg/kg in the average patient). Excellent identification of lymphatic vessels was achieved in 100% of volunteers and in 97.4% of patients. In the other 2.6%, lymphatic vessels were not identified mainly due to congenital lymphatic vascular (Milroy) disease. Baseline blood chemistry in volunteers was not altered after administration of the dye. No adverse reactions were found in volunteers and minimal allergic reactions occurred in less than 1% of patients. Acute toxicity studies demonstrated an LD50 greater than 150 mg/kg in the rat. Isosulfan blue was excreted unchanged in the urine (7%) and feces. Comparable excretion was found in volunteers. Isosulfan blue is a safe and efficacious vital dye for lymphangiography.

Adult↗

PubMed2007

Lymphatic vessels assessment in feline mammary tumours.

BACKGROUND: The lymphatic vessels play a crucial role in a variety of human cancers since tumour cell lymphatic invasion significantly influences prognosis. It is not known if pre-existing lymphatics are enough for tumour dissemination or de novo development is necessary. VEGFR-3 is an angiogenetic mediator for both lymphatic and blood vessels during embryonic development, and only for lymphatics after birth. VEGF is a mediator of both vasculogenesis and angiogenesis, regulates the growth of lymphatics in various experimental models, and is produced in many solid tumours. CD44 mediates hyaluronic acid (HA)-dependent cell adhesion: besides promoting invasion, this interaction also supports neoangiogenesis that indirectly stimulates tumour cell proliferation. The expression of VEGF-C (Vascular Endothelial Growth Factor-C), its receptor VEGFR-3 and CD44, were studied on feline mammary samples to assess the importance of lymphangiogenesis and lymphangiotrophism in neoplasia. METHODS: Samples were taken from six normal mammary glands (NMG), ten benign (BT) and 32 malignant (MT) tumours. Immunohistochemical laminin/VEGFR-3 double stain, VEGF-C and CD44 stains were applied to 4 mum-thick sections, and their expression evaluated in intratumoral/extratumoral and intramammary/extramammary fields. RESULTS: All groups revealed a higher number of lymphatics in the extratumoral/extramammary areas. VEGF-C expression in the epithelium paralleled the number of positive vessels in the NMG, BT and MT, whereas VEGF-C higher expression was noted in the intratumoral fields only in infiltrating MT. CD44 score was lower in extratumoral than intratumoral fields in tumours and showed a significant increase in extramammary/extratumoral fields from NMG to MT. Pearson test showed a significant and inversely proportional correlation between CD44 expression and the number of lymphatic vessels with VEGFR-3 in malignant infiltrating tumours. CONCLUSION: The number of both VEGFR-3 positive and negative lymphatics in the extratumoral and extramammary stroma was significantly higher than intratumoral and intramammary fields respectively in the NMG, BT and MT. This suggests a scant biological importance of intratumoral lymphatics while their higher number is due to the concentration of existing vessels following compression of the extratumoral stroma in spite of a non demonstrable increase from NMG to MT. The tumour model employed provided no evidence of lymphangiogenesis, and metastasis in the regional lymph node develops following the spread through the pre-existing lymphatic network.

Animals↗

PubMed2004

Expression of cys-cys chemokine ligand 21 on human gingival lymphatic vessels.

The cys-cys (C-C) chemokine ligand 21 is a member of the C-C chemokines that constitute a group of heparin-binding cytokines with a pattern of four or six conserved cysteines. The CCL21 is known to be expressed in secondary lymphoid tissues, however it has rarely been reported for the expression on peripheral lymphatic vessels in somatic tissue. Here we investigated the expression of CCL21 on lymphatic vessels identified by anti-desmoplakin in uninflamed and inflamed human gingiva. In uninflamed tissue the expression of CCL21 was detected on lymphatic vessels in gingiva. In uninflamed gingiva the expression of CCL21 was detected on all lymphatic capillaries of the mucosal connective tissue papillae. There were two types of collecting lymphatic vessels in the lamina propria mucosae expressing CCL21 strongly or very weakly. In inflamed gingiva no expression of CCL21 was detected on lymphatic vessels. In all tissue sections no blood vessels expressing CCL21 were observed. These results may suggest that the expression of CCL21 is predominantly induced in the peripheral lymphatic endothelium of the uninflamed mucosal microcirculation, and that under inflamed conditions a reduction of CCL21 occurs in lymphatic endothelium.

Adolescent↗

PubMed2005

Immunohistochemical identification of lymphatic vessels in the periodontium of equine cheek teeth.

Immunohistochemical detection of lymphatic capillaries was performed in the periodontium of maxillary and mandibular cheek teeth from 6 horses (aged 3-23 years). Tissue sections of the periodontium were taken at 4 different horizontal levels along the long axis of the tooth. The specimens were processed for immunoreaction with anti-Prox1, in order to distinguish lymphatic endothelium from blood vascular endothelium. Lymphatic vessels were detected in all periodontal tissues except for the dental cementum. Lymphatic capillaries were most densely distributed in the gingiva compared to other tissues of the periodontium. Lymphatic capillaries were found most consistently in samples taken from the gingival and subgingival regions in all horses examined. Within these levels, the gingiva as well as the spongiosa of the maxillary and mandibular bone had the greatest incidence of lymphatic vessels. Considering the distinct distribution of the lymphatic capillaries in the periodontium of the maxillary and mandibular cheek teeth, two complementary lymphatic drainage pathways are proposed: (1) superficial lymph drainage via the gingiva, emptying into the mandibular lymph nodes; (2) deep lymph drainage via the mandibular and maxillary spongiosa, emptying into the mandibular and retropharyngeal lymph nodes, respectively.

Animals↗

PubMed2003

Lymphatic vessels in colorectal cancer and their relation with inflammatory infiltrate.

PURPOSE: The aim of this study was to determine why colorectal tumors confined to submucosa rarely metastasize. Under normal conditions, the submucosa contains many large lymphatic vessels with thin walls that would presumably favor the spread of cancer cells through the lymphatic system. METHODS: Specimens of colorectal cancer tissue, the border between tumor and normal tissue, and normal tissue were obtained from patients undergoing radical resection of colorectal cancer. The material was embedded in methacrylate resin for light microscopy and Epon for transmission electron microscopy examination. Light microscopy observations were routinely performed on serial sections. RESULTS: No lymphatic vessels were ever found in the tumor mass. The border area contained peritumoral inflammatory infiltrate of variable thickness. Where submucosal lymphatic vessels came into contact with peritumoral inflammatory infiltrate, they were profoundly altered: their endothelium was fragmented, and their walls were disrupted. These altered lymphatic vessels were almost always accompanied by mast cells, which were observed in the process of degranulating toward the lymphatic endothelium. No such alterations were detected in blood vessels. CONCLUSION: Our results suggest that mast cells, probably influenced by inflammatory infiltrate and/or colorectal cancer cells, destroy lymphatic vessels, which prevents cancer cells from spreading through the lymphatic system.

Aged↗

PubMed1985

Evidence for constriction of canine prenodal lymphatic vessels by vasoactive agents and carotid occlusion.

We measured pressure in a prenodal lymphatic in the canine forelimb during constant flow pump-perfusion of the brachial artery. We made bolus i.a. injections of 1.0 micrograms angiotensin II, norepinephrine, bombesin, or bradykinin, 20 micrograms 5-hydroxytryptamine (5HT), or occluded the carotid arteries. Norepinephrine, 5HT, or carotid occlusion produced regular rises in forelimb perfusion pressure and in lymphatic pressure. Angiotensin II increased forelimb arterial pressures but increased lymphatic pressure in only four experiments. Bombesin increased artery pressures but did not affect lymphatic pressure. Small vein pressure was increased by carotid occlusion, 5HT and norepinephrine. Increases in lymphatic pressure were coincident with increases in vein pressure but no related in magnitude. Bradykinin decreased forelimb arterial and venous pressures but did not affect lymphatic pressure. Either active constriction of lymphatic vessels or passive compression by movements of adjacent blood vessels could increase lymphatic pressure. These data do not preclude a passive component of pressure rise in the lymphatics nor do they support the concept. We conclude that active constriction of prenodal lymphatic vessels in the dog forelimb can occur in response to circulating vasoactive agents and bilateral carotid occlusion.

Angiotensin II↗

PubMedDate not supplied

Progression of lymphatic vessel dilatation in the presence of living adult Wuchereria bancrofti.

Bancroftian filariasis, a mosquito-transmitted disease commonly known as elephantiasis, is caused by infection with the parasite Wuchereria bancrofti. Infection with this parasite can induce a broad array of chronic debilitating and socially stigmatizing conditions, but the pathogenesis of this morbidity remains obscure. Recent evidence indicates that in filariasis-endemic areas the primary lesion is not lymphatic vessel obstruction but, rather, dilatation. To determine the extent to which lymphatic dilatation occurs in the presence of living adult W. bancrofti, we performed longitudinal ultrasonographic measurements in 80 men (mean age 24 years) in Brazil who had a total of 107 W. bancrofti nests detectable by ultrasound. Initial mean lymphatic vessel diameter at the site of the worms was 3.4 mm (range, 0.7-11.3), and was greater in men with 2 or more nests (3.9 mm) than in those with only one nest (3.0 mm, P = 0.003). During the study period (2-35 months, mean, 13.7), lymphatic vessel diameter increased at the site of 92 (86.0%) adult worm nests. Mean rate of increase of lymphatic vessel diameter was 1.2 mm per person-year (range, 0-0.93 mm per month). In a general linear model, no factors, including treatment with antifilarial drugs, were significantly associated with rate of vessel diameter increase. Thus, lymphatic vessel dilatation progress in the presence of living adult W. bancrofti; the rate of this progression is heterogeneous. These data suggest that lymphatic dilatation will continue to progress in most infected persons even after mass treatment with currently recommended antifilarial drugs. In addition to interrupting transmission, the global programme for elimination of lymphatic filariasis should address the potential for disease progression in persons who remain infected with adult W. bancrofti.

Adolescent↗

PubMed1996

An examination of the close relationship between lymphatic vessels and nerve fibers containing calcitonin gene-related peptide and substance P in rat skin.

The distribution of nerve fibers containing either calcitonin gene-related peptide (CGRP) and substance P (SP) was investigated in rat skin with special reference to their relationship to the lymphatic vessels. These nerve fibers exhibited a similar distribution pattern but the former were more numerous than the latter. In the dermis and subcutaneous layers, thin nerve fibers containing CGRP or SP were in abundance, and were observed running along the blood vessels as well as freely in the tissue. Nerve fibers with these peptides were often located close to lymphatic capillaries, and innervated lymphatic vessels in the subcutaneous layer, reaching smooth muscles of the tunica media. These findings suggest that some CGRP and SP may directly drain into lymphatic vessels when released under noxious stimulation from nerve fibers around the lymphatic vessels. When discharged from nerve fibers in the vicinity of blood vessels, both peptides may also drain into the lymphatic vessels after causing blood vascular dilation and an increase in permeability producing edema. These peptides may then be transported to the draining lymph nodes where they can modulate the functions of the immune system.

Animals↗

PubMed2004

Expression of VEGFR-3 and 5'-nase in regenerating lymphatic vessels of the cutaneous wound healing.

The vascular endothelial growth factor-C (VEGF-C), a specific lymphangiogenic growth factor, raises new questions and perspectives in studying lymphatic development and regeneration. Wound healing skins in mice were processed for 5'-nucleotidase (5'-Nase) and VEGFR-3 (the receptor of VEGF-C) histochemical staining to distinguish lymphatics from blood capillaries and to analyze lymphangiogenesis. In the wounds of 3-5 days after injury, anti-VEGFR-3 immunopositive signals unevenly appeared in 5'-Nase-positive lymphatic vessels in the subcutaneous tissue. A few small circular and irregular lymphatic-like structures with VEGFR-3 expression scattered in the dermal and subcutaneous tissues. Between days 7 and 15 of the wounds, numerous accumulated vasculatures were stained for 5'-Nase and PECAM-1, extending irregularly along the wound edge. Von Willebrand factor was expressed in the endothelial cells of blood vessels and lymphatics in the subcutaneous tissue. Ultrastructural changes of lymphatic vessels developed at different stages, from lymphatic-like structures to newly formed lymphatic vessels with an extremely thin and indented wall. Endothelial cells of the lymphatic vessel were eventually featured by typical intercellular junctions, which deposited with reaction products of VEGFR-3 and 5'-Nase-cerium but lacked VEGF-C expression. The present findings indicate that VEGF-C-induced lymphangiogenesis occurs from the subcutaneous to the dermis along the wound healing edge, especially in the dermal-subcutaneous transitional area, favorable to growth of regenerating lymphatic vessels.

5'-Nucleotidase↗

PubMed2005

Novel expression and characterization of lymphatic vessel endothelial hyaluronate receptor 1 (LYVE-1) by conjunctival cells.

PURPOSE: Lymphatic vessel endothelial hyaluronic acid receptor (LYVE-1) is a newly discovered lymphatic-specific marker. To date, there is no report of its expression on conjunctival cells. The purpose of this study was to investigate the expression of LYVE-1 in normal conjunctiva, to phenotype LYVE-1+ cells, and to study changes in their expression levels during corneal inflammation. METHODS: Flat-mounted conjunctivae or cross sections of eyeballs were harvested from BALB/c mice (6-8 weeks of age) for immunofluorescent confocal microscopic studies. RESULTS: The data demonstrate, for the first time, that in addition to its expression on lymphatic vessels, LYVE-1 was expressed on CD45+, CD11b+, and CD31- conjunctival cells, indicating a bone-marrow-derived monocytic lineage. Surprisingly, the number of cells that expressed LYVE-1 decreased during corneal inflammation, in conjunction with ingrowth of lymphatics into the cornea. CONCLUSIONS: A new population of monocytic cells has been found to express LYVE-1 in normal conjunctiva. These cells that normally express LYVE-1 may act as a reservoir for lymphangiogenesis and cell recruitment when the immune system is challenged.

Animals↗

PubMed1994

The correlation between tumor size and lymphatic vessel invasion in resected peripheral stage I non-small-cell lung cancer. A potential risk of limited resection.

We attempted to clarify whether a correlation exists between tumor size and the incidence of lymphatic vessel invasion in peripheral non-small-cell lung cancer without regional lymph node metastasis. The study included 212 resected non-small-cell lung cancers classified as pathologic stage I disease and located on the periphery of the lung. The incidence of lymphatic vessel invasion was relatively correlated with the maximum diameter of the tumor as follows: 25% (1/4) for tumor size 1.0 cm or less, 40% (19/48) for size 1.1 to 2.0 cm, 49% (28/58) for size 2.1 to 3.0 cm, and 57% (58/102) for tumor size 3.1 cm or more. The incidence of lymphatic vessel invasion of tumors measuring 3 cm or less in greatest dimension was 44% (48/110). The degree of lymphatic vessel invasion of 20 resected tumor samples measuring 3 cm or less in greatest diameter with hilar lymph node metastasis was also examined for comparison. This figure was as high as 85%. These observations suggest that even small peripheral tumors without any regional lymph node metastasis have a relatively high rate of lymphatic vessel invasion and thus pose a potential risk of local recurrence after a limited resection, especially in a wedge resection of the tumor.

Adenocarcinoma↗

PubMed1985

[Lymphatic vessels of the upper extremity and their relation to nodes in the axillary area in healthy subjects and lymph flow blockade].

Large and small lymphatic vessels have been studied roentgenologically on the medial, lateral, posterior and anterior surfaces of the upper extremity in 113 patients at the age of 19-63 years at blockade of the lymphatic stream. On the medial and lateral surfaces the lymphatic vessels are filled with the contrast substance via anatomical approaches from the palmar and dorsal sides of the forearm. With isolated contrasting of various large lymphatic vessels, zones in the skin and in the subcutaneous fatty layers drained by them are revealed, as well as distribution of small vessels in the forearm and shoulder in each region. Variants of large lymphatic vessels and their tributaries are defined; an essential variability of their inflow into the axillary lymph nodes from various anatomical areas of the upper extremity is found. Into every 1-4 groups of the lymph nodes of the axillary area, 1-3 large vessels inflow, through them the contrast substance switches from the same anatomical zone repeatedly.

Adult↗

PubMed2005

A quantitative analysis of lymphatic vessels in human breast cancer, based on LYVE-1 immunoreactivity.

This study was undertaken to determine the highly sensitive method for detecting tumour lymphatic vessels in all the fields of each slide (LV), lymphatic microvessel density (LMVD) and lymphatic vessel invasion (LVI) and to compare them with other prognostic parameters using immunohistochemical staining with polyclonal (PCAB) and monoclonal antibodies (MCAB) to the lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), and the pan-endothelial marker factor VIII in a series of 67 human breast cancers. In all LYVE-1-stained sections, LV (some of which contained red blood cells) were frequently found localised in extralobular stroma, dermis, connective tissue stroma and adjacent to artery and vein, but were rare within the intralobular stroma or the tumour body (3/67 cases) or areas of widespread invasion. In contrast small blood vessels were observed in intra- and extralobular stroma in the factor VIII-stained sections. Quantitation of vessel numbers revealed that LYVE-1/PCAB detected a significantly larger number of LV than either H&E or LYVE-1/MCAB (P<0.0001). LYVE-1/PCAB detected LVI in 25/67 cases (37.3%) and their presence was significantly associated with both lymph node metastasis (chi(2)=4.698, P=0.0248) and unfavourable overall survival (OS) (P=0.0453), while not relapse- free survival (RFS) (P=0.2948). LMVD had no influence for RFS and OS (P=0.4879, P=0.1463, respectively). Our study demonstrates that immunohistochemistry with LYVE-1/PCAB is a highly sensitive method for detecting tumour LV/LVI in breast cancer and LVI is a useful prognostic indicator for lymphatic tumour dissemination.

Adult↗

PubMed2005

Dendritic-cell trafficking to lymph nodes through lymphatic vessels.

Antigen-presenting dendritic cells often acquire foreign antigens in peripheral tissues such as the skin. Optimal encounter with naive T cells for the presentation of these antigens requires that the dendritic cells migrate to draining lymph nodes through lymphatic vessels. In this article, we review important aspects of what is known about dendritic-cell trafficking into and through lymphatic vessels to lymph nodes. We present these findings in the context of information about lymphatic-vessel biology. Gaining a better understanding of the crosstalk between dendritic cells and lymphatic vessels during the migration of dendritic cells to lymph nodes is essential for future advances in manipulating dendritic-cell migration as a means to fine-tune immune responses in clinical settings.

Animals↗