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Title: Granulocyte colony-stimulating factor induces in vitro lymphangiogenesis

Abstract

Highlights: •G-CSF induces tube formation, migration and proliferation of lymphatic cells. •G-CSF increases phosphorylation of MAPK and Akt in lymphatic endothelial cells. •MAPK and Akt pathways are linked to G-CSF-induced in vitro lymphangiogenesis. •G-CSF increases sprouting of a lymphatic ring. •G-CSF produces peritoneal lymphangiogenesis. -- Abstract: Granulocyte-colony stimulating factor (G-CSF) is reported to induce differentiation in cells of the monocyte lineage and angiogenesis in vascular endothelial cells, but its effects on lymphangiogenesis is uncertain. Here we examined the effects and the mechanisms of G-CSF-induced lymphangiogenesis using human lymphatic endothelial cells (hLECs). Our results showed that G-CSF induced capillary-like tube formation, migration and proliferation of hLECs in a dose- and time-dependent manner and enhanced sprouting of thoracic duct. G-CSF increased phosphorylation of Akt and ERK1/2 in hLECs. Supporting the observations, specific inhibitors of phosphatidylinositol 3′-kinase and MAPK suppressed the G-CSF-induced in vitro lymphangiogenesis and sprouting. Intraperitoneal administration of G-CSF to mice also stimulated peritoneal lymphangiogenesis. These findings suggest that G-CSF is a lymphangiogenic factor.

Authors:
; ; ; ;  [1]; ;  [1];  [2];  [1];  [2];  [1];  [2]
  1. Department of Internal Medicine, Chonbuk National University Medical School, Jeonju (Korea, Republic of)
  2. (Korea, Republic of)
Publication Date:
OSTI Identifier:
22239658
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 436; Journal Issue: 4; Other Information: Copyright (c) 2013 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ANGIOGENESIS; CAPILLARIES; CESIUM FLUORIDES; IN VITRO; LYMPH VESSELS; MICE; MONOCYTES; PHOSPHORYLATION; SPROUTING

Citation Formats

Lee, Ae Sin, Kim, Dal, Wagle, Susbin Raj, Lee, Jung Eun, Jung, Yu Jin, Kang, Kyung Pyo, Lee, Sik, Research Institute of Clinical Medicine of Chonbuk National University-Chonbuk National University Hospital, Chonbuk National University Medical School, Jeonju, Park, Sung Kwang, E-mail: parksk@jbnu.ac.kr, Research Institute of Clinical Medicine of Chonbuk National University-Chonbuk National University Hospital, Chonbuk National University Medical School, Jeonju, Kim, Won, E-mail: kwon@jbnu.ac.kr, and Research Institute of Clinical Medicine of Chonbuk National University-Chonbuk National University Hospital, Chonbuk National University Medical School, Jeonju. Granulocyte colony-stimulating factor induces in vitro lymphangiogenesis. United States: N. p., 2013. Web. doi:10.1016/J.BBRC.2013.05.062.
Lee, Ae Sin, Kim, Dal, Wagle, Susbin Raj, Lee, Jung Eun, Jung, Yu Jin, Kang, Kyung Pyo, Lee, Sik, Research Institute of Clinical Medicine of Chonbuk National University-Chonbuk National University Hospital, Chonbuk National University Medical School, Jeonju, Park, Sung Kwang, E-mail: parksk@jbnu.ac.kr, Research Institute of Clinical Medicine of Chonbuk National University-Chonbuk National University Hospital, Chonbuk National University Medical School, Jeonju, Kim, Won, E-mail: kwon@jbnu.ac.kr, & Research Institute of Clinical Medicine of Chonbuk National University-Chonbuk National University Hospital, Chonbuk National University Medical School, Jeonju. Granulocyte colony-stimulating factor induces in vitro lymphangiogenesis. United States. doi:10.1016/J.BBRC.2013.05.062.
Lee, Ae Sin, Kim, Dal, Wagle, Susbin Raj, Lee, Jung Eun, Jung, Yu Jin, Kang, Kyung Pyo, Lee, Sik, Research Institute of Clinical Medicine of Chonbuk National University-Chonbuk National University Hospital, Chonbuk National University Medical School, Jeonju, Park, Sung Kwang, E-mail: parksk@jbnu.ac.kr, Research Institute of Clinical Medicine of Chonbuk National University-Chonbuk National University Hospital, Chonbuk National University Medical School, Jeonju, Kim, Won, E-mail: kwon@jbnu.ac.kr, and Research Institute of Clinical Medicine of Chonbuk National University-Chonbuk National University Hospital, Chonbuk National University Medical School, Jeonju. 2013. "Granulocyte colony-stimulating factor induces in vitro lymphangiogenesis". United States. doi:10.1016/J.BBRC.2013.05.062.
@article{osti_22239658,
title = {Granulocyte colony-stimulating factor induces in vitro lymphangiogenesis},
author = {Lee, Ae Sin and Kim, Dal and Wagle, Susbin Raj and Lee, Jung Eun and Jung, Yu Jin and Kang, Kyung Pyo and Lee, Sik and Research Institute of Clinical Medicine of Chonbuk National University-Chonbuk National University Hospital, Chonbuk National University Medical School, Jeonju and Park, Sung Kwang, E-mail: parksk@jbnu.ac.kr and Research Institute of Clinical Medicine of Chonbuk National University-Chonbuk National University Hospital, Chonbuk National University Medical School, Jeonju and Kim, Won, E-mail: kwon@jbnu.ac.kr and Research Institute of Clinical Medicine of Chonbuk National University-Chonbuk National University Hospital, Chonbuk National University Medical School, Jeonju},
abstractNote = {Highlights: •G-CSF induces tube formation, migration and proliferation of lymphatic cells. •G-CSF increases phosphorylation of MAPK and Akt in lymphatic endothelial cells. •MAPK and Akt pathways are linked to G-CSF-induced in vitro lymphangiogenesis. •G-CSF increases sprouting of a lymphatic ring. •G-CSF produces peritoneal lymphangiogenesis. -- Abstract: Granulocyte-colony stimulating factor (G-CSF) is reported to induce differentiation in cells of the monocyte lineage and angiogenesis in vascular endothelial cells, but its effects on lymphangiogenesis is uncertain. Here we examined the effects and the mechanisms of G-CSF-induced lymphangiogenesis using human lymphatic endothelial cells (hLECs). Our results showed that G-CSF induced capillary-like tube formation, migration and proliferation of hLECs in a dose- and time-dependent manner and enhanced sprouting of thoracic duct. G-CSF increased phosphorylation of Akt and ERK1/2 in hLECs. Supporting the observations, specific inhibitors of phosphatidylinositol 3′-kinase and MAPK suppressed the G-CSF-induced in vitro lymphangiogenesis and sprouting. Intraperitoneal administration of G-CSF to mice also stimulated peritoneal lymphangiogenesis. These findings suggest that G-CSF is a lymphangiogenic factor.},
doi = {10.1016/J.BBRC.2013.05.062},
journal = {Biochemical and Biophysical Research Communications},
number = 4,
volume = 436,
place = {United States},
year = 2013,
month = 7
}
  • Interleukin 1 alpha (IL-1 alpha), tumor necrosis factor alpha (TNF alpha), granulocyte-colony-stimulating factor (G-CSF), and granulocyte-macrophage colony-stimulating factor (GM-CSF) are molecularly distinct cytokines acting on separate receptors. The release of these cytokines can be concomitantly induced by the same signal and from the same cellular source, suggesting that they may cooperate. Administered alone, human recombinant (hr)IL-1 alpha and hrTNF alpha protect lethally irradiated mice from death, whereas murine recombinant GM-CSF and hrG-CSF do not confer similar protection. On a dose basis, IL-1 alpha is a more efficient radioprotector than TNF alpha. At optimal doses, IL-1 alpha is a more radioprotectivemore » cytokine than TNF alpha in C57BL/6 and B6D2F1 mice and less effective than TNF alpha in C3H/HeN mice, suggesting that the relative effectiveness of TNF alpha and IL-1 alpha depends on the genetic makeup of the host. Administration of the two cytokines in combination results in additive radioprotection in all three strains. This suggests that the two cytokines act through different radioprotective pathways and argues against their apparent redundancy. Suboptimal, nonradioprotective doses of IL-1 alpha also synergize with GM-CSF or G-CSF to confer optimal radioprotection, suggesting that such an interaction may be necessary for radioprotection of hemopoietic progenitor cells.« less
  • We have attempted to evaluate in vivo effects of granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) on acute radiation hematopoietic injury in mice. BDF1 mice, irradiated with 7.5-Gy x-rays, were injected i.p. twice daily for 10 days with 10(5) U recombinant human G-CSF, 3.75 x 10(5) U recombinant murine GM-CSF, or a combination of both. G-CSF significantly enhanced the recovery of not only peripheral leukocytes but also platelets and hematocrit on days 14 and 21 after irradiation. GM-CSF significantly enhanced the recovery of platelets on day 14 and peripheral leukocytes on day 21. G-CSF markedly enhanced the recoverymore » of spleen colony-forming units (CFU-S), colony-forming units in culture (CFU-C), erythroid burst-forming units (BFU-E), and megakaryocyte colony-forming units (CFU-Meg) both in bone marrow and in the spleen. GM-CSF significantly enhanced the recovery of CFU-Meg in bone marrow on day 14. We found synergistic effects between G-CSF and GM-CSF on CFU-S, CFU-C, and CFU-Meg in the spleen on day 14, although we found antagonistic effects between G-CSF and GM-CSF on CFU-S and CFU-C in bone marrow on day 7, and on platelet counts on day 7. These results indicate that the administration of recombinant G-CSF and GM-CSF may be useful in accelerating hematopoietic recovery in patients with acute radiation hematopoietic injuries.« less
  • Myeloid cell kinetics in mice treated with pure hematopoietic growth factors have been investigated using tritiated thymidine labeling and autoradiography. Mice were injected subcutaneously with 125 micrograms/kg granulocyte colony-stimulating factor (G-CSF) (in some cases 5 micrograms/kg), or 10 micrograms/kg of granulocyte-macrophage CSF (GM-CSF), or interleukin-3 (IL-3) every 12 hours for 84 hours. {sup 3}HTdR labeling was performed in vivo after 3 days of treatment. G-CSF increased the peripheral neutrophil count 14-fold and increased the proportion and proliferation rate of neutrophilic cells in the marrow, suppressing erythropoiesis at the same time. Newly produced mature cells were released into the circulation withinmore » 24 hours of labeling, compared with a normal appearance time of about 96 hours. By contrast, GM-CSF and IL-3 had little effect on either marrow cell kinetics or on the rate of release of mature cells, although GM-CSF did stimulate a 50% increase in peripheral neutrophils. Monocyte production was also increased about eightfold by G-CSF and 1.5-fold by GM-CSF, but their peak release was only slightly accelerated. While the peripheral half-lives of the neutrophilic granulocytes were normal, those of the monocytes were dramatically reduced, perhaps due to sequestration in the tissues for functional purposes. The stimulated monocyte production in the case of G-CSF required an additional five cell cycles, a level that might have repercussions on the progenitor compartments.« less
  • Purified recombinant murine granulocyte/macrophage colony-stimulating factor (GM-CSF) was labeled with /sup 125/I and used to examine the GM-CSF receptor on unfractionated normal murine bone marrow cells, casein-induced peritoneal exudate cells, and highly purified murine granulocyte/macrophage progenitor cells (CFU-GM). CFU-GM were isolated from cyclophosphamide-treated mice by Ficoll-Hypaque density centrifugation followed by counterflow centrifugal elutriation. The resulting population had a cloning efficiency of 62-99% in cultures containing conditioned medium from pokeweed mitogen-stimulated spleen cells and 55-86% in the presence of a plateau concentration of purified recombinant murine GM-CSF. Equilibrium binding studies with /sup 125/I-labeled GM-CSF showed that normal bone marrow cells, casein-inducedmore » peritoneal exudate cells, and purified CFU-GM had a single class of high-affinity receptor. Affinity crosslinking studies demonstrated that /sup 125/I-labeled GM-CSF bound specifically to two species of M/sub r/ 180,000 and 70,000 on CFU-GM, normal bone marrow cells, and peritoneal exudate cells. The M/sub r/ 70,000 species is thought to be a proteolytic fragment of the intact M/sub r/ 180,000 receptor. The present studies indicate that the GM-CSF receptor expressed on CFU-GM and mature myeloid cells are structurally similar. In addition, the number of GM-CSF receptors on CFU-GM is twice the average number of receptors on casein-induced mature myeloid cells, suggesting that receptor number may decrease as CFU-GM mature.« less
  • Supernatants of COS-1 cells transfected with gibbon cDNA encoding interleukin 3 (IL-3) with homology to sequences for human IL-3 were tested for ability to promote growth of various human hemopoietic progenitors. The effect of these supernatants as a source of recombinant IL-3 was compared to that of recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) as well as to that of medium conditioned by phytohemagglutinin-stimulated leukocytes. The frequency of multilineage colonies, erythroid bursts, and megakaryocyte colonies in cultures containing the COS-1 cell supernatant was equivalent to the frequency observed in the controls and significantly higher than foundmore » in cultures plated with recombinant GM-CSF. G-CSF did not support the formation of multilineage colonies, erythroid bursts, and megakaryocyte colonies. In contrast, growth of granulocyte-macrophage colonies was best supported with GM-CSF, while recombinant IL-3 yielded colonies at lower or at best equivalent frequency. The simultaneous addition of higher concentrations of GM-CSF to cultures containing IL-3 in optimal amounts did not enhance the formation of multilineage colonies, erythroid bursts, and megakaryocyte colonies. However, the frequency of such colonies and bursts increased with GM-CSF when cultures were plated with suboptimal concentrations of IL-3. Growth of colonies within the granulocyte-macrophage lineage is optimally supported by GM-CSF and does not increase with further addition of IL-3.« less