Mechanical forces in lymphatic vascular development and disease

The lymphatic vasculature is essential for fluid homeostasis and transport of immune cells, inflammatory molecules, and dietary lipids. It is composed of a hierarchical network of blind-ended lymphatic capillaries and collecting lymphatic vessels, both lined by lymphatic endothelial cells (LECs). The low hydrostatic pressure in lymphatic capillaries, their loose intercellular junctions, and attachment to the surrounding extracellular matrix (ECM) permit passage of extravasated blood plasma from the interstitium into the lumen of the lymphatic capillaries. It is generally thought that interstitial fluid accumulation leads to a swelling of the ECM, to which the LECs of lymphatic capillaries adhere, for example via anchoring filaments. As a result, LECs are pulled away from the vascular lumen, the junctions open, and fluid enters the lymphatic vasculature. The collecting lymphatic vessels then gather the plasma fluid from the capillaries and carry it through the lymph nodes to the blood circulation. The collecting vessels contain intraluminal bicuspid valves that prevent fluid backflow, and are embraced by smooth muscle cells that contribute to fluid transport. Although the lymphatic vessels are regular subject to mechanical strain, our knowledge of its influence on lymphatic development and pathologies is scarce. Here, we discuss the mechanical forces and molecular mechanisms regulating lymphatic vascular growth and maturation in the developing mouse embryo. We also consider how the lymphatic vasculature might be affected by similar mechanomechanisms in two pathological processes, namely cancer cell dissemination and secondary lymphedema.     

Bovine pituitary intraglandular colloid fraction F5 localized in the rat endocrine pancreas

Bovine pituitary intraglandular colloid thought to be a waste product, is the holocrine secretion of intermediate lobe cells. It is housed in the intraglandular lumen (residual lumen) and is extruded into the venous circulation of the cavernous sinuses via clefts in the capsule of the gland aligned with the intraglandular lumen. Intraglandular colloid, fraction F5 (mol.wt 34,000), radiolabeled with (125I)Na and injected (0.15 ml) into the right internal jugular vein of male Wistar rats, accumulated in the endocrine pancreas. Autoradiographs showed that the material had specifically localized in the capillary network of the endocrine pancreas. Since the intermediate lobe is poorly vascularized, intraglandular colloid is considered to be the transport medium for intermediate lobe materials.     

Lysophosphatidic acid up-regulates vascular endothelial growth factor-C and lymphatic marker expressions in human endothelial cells

Lysophosphatidic acid (LPA) is a low-molecular-weight lipid growth factor, which binds to G-protein-coupled receptors. Previous studies have shown that LPA enhances vascular endothelial growth factor-A (VEGF-A) expression in cancer cells and promotes angiogenesis process. However, the roles of LPA in lymphatic vessel formation and lymphangiogenesis have not been investigated. Here, we demonstrated that LPA up-regulated VEGF-C mRNA and protein expressions in human umbilical vein endothelial cells (HUVECs). Furthermore, the expression levels of lymphatic markers, including Prox-1, LYVE-1 and podoplanin, were enhanced in LPA-stimulated tube forming endothelial cells in vitro and in vivo. Moreover, we showed that pretreatment with MAZ51, a VEGFR-3 kinase inhibitor, and introduction of VEGFR-3 siRNA suppressed LPA-induced HUVEC tube formation and lymphatic marker expressions. These results demonstrated that LPA enhances expression of lymphatic markers through activating VEGF-C receptors in endothelial cells. This study provides basic information that LPA might be a target for therapeutics against lymphangiogenesis and tumor metastasis.     

Bovine pituitary intraglandular colloid fraction F5 localized in the rat endocrine pancreas

Summary Bovine pituitary intraglandular colloid thought to be a waste product, is the holocrine secretion of intermediate lobe cells. It is housed in the intraglandular lumen (residual lumen) and is extruded into the venous circulation of the cavernous sinuses via clefts in the capsule of the gland aligned with the intraglandular lumen. Intraglandular colloid, fraction F₅ (mol.wt 34,000), radiolabeled with (¹²⁵I)Na and injected (0.15 ml) into the right internal jugular vein of male Wistar rats, accumulated in the endocrine pancreas. Autoradiographs showed that the material had specifically localized in the capillary network of the endocrine pancreas. Since the intermediate lobe is poorly vascularized, intraglandular colloid is considered to be the transport medium for intermediate lobe materials.This study was supported by a grant from the Natural Sciences and Engineering Research Council of Canada.I would like to thank S. Choles and J. Bennett for their technical expertise with the help of P. Galvin and W. Halina. I thank members of the Central Animal Facilities, McMaster University Medical Centre, Hamilton, Ontario, for labeling the intraglandular colloid and for counting the decay rate of the tissue. I would also like to thank Drs. A. deBold and J. Kraicer, Faculty of Medicine, Queen's University, Kingston, Ontario, for their interest in this study.     

Lymphatic metastasis of tumour; persistent transport of cells.

A model of lymphatic metastasis established by injecting Walker rat carcinoma cells into the rat footpad was used to study the output of tumour cells from the footpad. The lymphatic efferent from the footpad was cannulated in a group of rats with advanced neoplasm; it was shown that the output of tumour cells was continuous over periods up to 90 min and ranged from 10(2)-10(5) cells/min.     

Ultrastructural observation on the tips of growing vascular cords in the rat cerebral cortex

Summary The distal ends of the vascular cord in the cerebral cortex are investigated electron microscopically in rats at the 13th postnatal day. The tip of the vascular cord consists of central cuboidal cells (primitive endothelial cells) and surrounding flat cells (primitive pericytes), and has no lumen. The primitive endothelial cells (tip cells) possess several long tentacles which contain only fibrous structures and extend through the neuropile.This study was supported in part by a grant in aid for Scientific Research from the Ministry of Education of Japan (No. 448085).     

In situ observations of spontaneous contractions of the peripheral lymphatic vessels in the rat mesentery: Effects of temperature

Summary By means of an intracellular glass microelectrode, action potential changes were successfully recorded in situ from the endothelial cells of rat mesenteric lymphatics over the temperature range of 27–40°C. The frequency of action potential and the lymphatic contraction rate correlated well with temperature.     

Lymphatic metastasis of tumour; persistent transport of cells

Summary A model of lymphatic metastasis established by injecting Walker rat carcinoma cells into the rat footpad was used to study the output of tumour cells from the footpad. The lymphatic efferent from the footpad was cannulated in a group of rats with advanced neoplasm; it was shown that the output of tumou cells was continuou over periods up to 90 min and ranged from 10²–10⁵ cells/min.Acknowledgment. This work was supported by a grant from the National Cancer Institute of Canada to C.R.F. and I.C. We are grateful to Mrs W. Kao and Mr I. Etches for meticulous technical help and to the Photographic Section, Department of Pathology for illustrations. Dr V.S. Gupta of Veterinary Physiology, University of Saskatchewan supplied the tumour for transplantation.     

Formation of cardiovascular tubes in invertebrates and vertebrates

The cardiovascular system developed early in evolution and is pivotal for the transport of oxygen, nutrients, and waste products within the organism. It is composed of hollow tubular structures and has a high level of complexity in vertebrates. This complexity is, at least in part, due to the endothelial cell lining of vertebrate blood vessels. However, vascular lumen formation by endothelial cells is still controversially discussed. For example, it has been suggested that the lumen mainly forms via coalescence of large intracellular vacuoles generated by pinocytosis. Alternatively, it was proposed that the vascular lumen initiates extracellularly between adjacent apical endothelial cell surfaces. Here we discuss invertebrate and vertebrate cardiovascular lumen formation and highlight the possible modes of blood vessel formation. Finally, we point to the importance of a better understanding of vascular lumen formation for treating human pathologies, including cancer and coronary heart disease.     

Fine structural relation between pancreatic excretory ductules and intercellular spaces

Summary Horse-radish peroxidase injected into the femoral vein of intact rats, or infused at 30 cm H₂O pressure into the main pancreatic duct of intact dogs, entered easily the interstitial spaces surrounding acini and acinar cells. The latter are interconnected at their luminal segments by zonulae occludentes. These junctions form a barrier to tracer penetrating from the interstitium towards the lumen of terminal ductules. However, the intraductally infused peroxidase entered the interstitial spaces, probably through the pressure injured acinar cells, as did colloidal carbon particles when infused intraductally.     

Quantitative und qualitative Untersuchungen der Esteraseaktivität in normalen Granulozyten,normalen Lymphozyten und in leukämischen Zellen

Summary A quantitative and qualitative analysis of the carboxylic esterases of normal granulocytes, normal lymphocytes and leukemic cells was performed. In the myeloid leukemic cells and in the lymphatic leukemic cells, the esterase activity was significantly increased. By means of electrophoretic fractionation, four zones with esterase activity were detected in normal granulocytes and lymphocytes; the electrophoretic pattern of the leukemic cells was similar to that of the normal cells.

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Diese Forschung wurde vom Fonds des Lady Tata Memorial Trust unterstützt.     

Kristalloide Strukturen im Zytoplasma von lymphatischen Zellen verschiedener Affen

Summary In the endoplasmic reticulum of cells of spleen and lymphnodes of different monkeys, crystalloid inclusions were often found. These inclusions show a pattern of small balls with a diameter of 250 Å, which are connected by 100 Å long bridges. It is supposed that these crystalloid inclusions are produced by the lymphatic cells as a reaction to virus infection, which remains clinically non-apparent, because the same pattern was observed in cells, which were experimentally infected with yellow fever virus, West Nile virus, and rubella virus.     

TGFβ-induced protein mediates lymphatic endothelial cell adhesion to the extracellular matrix under low oxygen conditions

TGFbeta-induced protein (TGFBI) is an extracellular protein that mediates cell adhesion to collagen, laminin and fibronectin through its interaction with different beta integrins. We had previously reported that hypoxia-induced TGFBI mRNA expression in lymphatic endothelial cells (LEC). Here, we demonstrate that TGFBI can contribute to hypoxia-induced increases in LEC adhesion to the ECM. We show that while there are no changes in alpha1, alpha4, alphav, beta1, beta2, beta3, alpha5beta1, alphavbeta3, alphavbeta5 integrin expression on the LEC surface after hypoxia exposure, there exists an accumulation of TGFBI adaptor protein in LEC supernatants. We also demonstrate that hypoxia driven TGBFI expression is dependent on TGFbeta production by LEC. Furthermore, we show that TGFBI mediated LEC adhesion and migration through the ECM by its binding to the beta3 integrin. The identification of the specific mechanisms regulating LEC-ECM interactions may help us design new therapeutic applications for diseases in which lymphatic vessel function is compromised.     

Decrease of mast cells in regional lymph nodes in response to allogeneic antigens and syngeneic tumor antigens

Summary The absolute number of mast cells in regional lymph nodes decreases on the 5th day after stimulation by allogeneic lymphocytes and semisyngeneic leukaemic cells, despite the enlargement of stimulated lymph nodes. It is postulated that the reaction of lymphatic mast cells could be a sensitive test for tissue incompatibility, and probably also for the presence of tumor associated antigens.     

Morphogenesis of human colon cancer cells with fetal rat mesenchymes in organ culture

The morphogenesis and cytodifferentiation of human colon cancer cells (LS174T and HT29) were examined by combining cancer cells with fetal rat digestive-tract mesenchyme in organ culture. LS174T cells migrated into the mesenchyme to form glandular structures composed of single columnar cells with their nuclei oriented basally, while HT29 cells formed cell masses with little lumen formation. Immunohistochemical studies with antibodies against carcinoembryonic antigen and secretory components showed that the composition of cell surface glycoproteins was not necessarily reversed to the normal type, even when neoplastic cells exhibited normal glandular structures.     

In situ observations of spontaneous contractions of the peripheral lymphatic vessels in the rat mesentery: effects of temperature

By means of an intracellular glass microelectrode, action potential changes were successfully recorded in situ from the endothelial cells of rat mesenteric lymphatics over the temperature range of 27-40 degrees C. The frequency of action potential and the lymphatic contraction rate correlated well with temperature.     

The relation between capillaries and neurons in the caudal neurosecretory system of Pomatomus saltatrix.

The caudal neurosecretory system of Pomatomus saltatrix was examined. Particular interest was devoted to the perikarya of the neurosecretory cells. The majority of these cells were characterized by a close association with the capillary network. The cells appeared to ensheath the capillaries thus bringing the nucleus and perinuclear cytoplasm into close association with the capillaries.     

In vivo imaging of therapy-induced anti-cancer immune responses in humans

Immunotherapy aims to re-engage and revitalize the immune system in the fight against cancer. Research over the past decades has shown that the relationship between the immune system and human cancer is complex, highly dynamic, and variable between individuals. Considering the complexity, enormous effort and costs involved in optimizing immunotherapeutic approaches, clinically applicable tools to monitor therapy-induced immune responses in vivo are most warranted. However, the development of such tools is complicated by the fact that a developing immune response encompasses several body compartments, e.g., peripheral tissues, lymph nodes, lymphatic and vascular systems, as well as the tumor site itself. Moreover, the cells that comprise the immune system are not static but constantly circulate through the vascular and lymphatic system. Molecular imaging is considered the favorite candidate to fulfill this task. The progress in imaging technologies and modalities has provided a versatile toolbox to address these issues. This review focuses on the detection of therapy-induced anticancer immune responses in vivo and provides a comprehensive overview of clinically available imaging techniques as well as perspectives on future developments. In the discussion, we will focus on issues that specifically relate to imaging of the immune system and we will discuss the strengths and limitations of the current clinical imaging techniques. The last section provides future directions that we envision to be crucial for further development.     

The relation between capillaries and neurons in the caudal neurosecretory system ofPomatomus saltatrix

Summary The caudal neurosecretory system ofPomatomus saltatrix was examined. Particular interest was devoted to the perikarya of the neurosecretory cells. The majority of these cells were characterized by a close association with the capillary network. The cells appeared to ensheath the capillaries thus bringing the nucleus and perinuclear cytoplasm into close association with the capillaries.Supported byA. D. Williams No. 3558 (505), Medical College of Virginia. The authors are grateful to Dr.H. R. Seibel for reading this report.     

Zytophotometrische Untersuchungen an normalen Lymphknotenzellen des Schafes

Summary Cytophotometric investigations of lymphocytic populations of 4 sheeps were carried out to determine the diploid standard value for comparative investigations of lymph node cells from sheep with lymphatic leucosis. Inspite of exact and simultaneous preparation there were marked and significant differences in the histograms as well as in the diagrams of absorption level separation. Therefore, the control of several normal cell populations is essential before starting cytophotometric investigations of neoplastic cells.

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Diese Arbeit entstand im Rahmen des Sonderforschungsbereiches 47 (Virologie) an der Justus Liebig-Universität Giessen.