Ferritin: a novel mechanism for delivery of iron to the brain and other organs

2007 ◽  
Vol 293 (2) ◽  
pp. C641-C649 ◽  
Author(s):  
J. Fisher ◽  
K. Devraj ◽  
J. Ingram ◽  
B. Slagle-Webb ◽  
A. B. Madhankumar ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Protein A ◽  
Feedback Mechanism ◽  
Iron Storage ◽  
Multiple Organs ◽  
Cellular Iron ◽  
Ferritin Iron ◽  
A Cell ◽  
Iron Delivery ◽  
The Brain

Traditionally, transferrin has been considered the primary mechanism for cellular iron delivery, despite suggestive evidence for additional iron delivery mechanisms. In this study we examined ferritin, considered an iron storage protein, as a possible delivery protein. Ferritin consists of H- and L-subunits, and we demonstrated iron uptake by ferritin into multiple organs and that the uptake of iron is greater when the iron is delivered via H-ferritin compared with L-ferritin. The delivery of iron via H-ferritin but not L-ferritin was significantly decreased in mice with compromised iron storage compared with control, indicating that a feedback mechanism exists for H-ferritin iron delivery. To further evaluate the mechanism of ferritin iron delivery into the brain, we used a cell culture model of the blood-brain barrier to demonstrate that ferritin is transported across endothelial cells. There are receptors that prefer H-ferritin on the endothelial cells in culture and on rat brain microvasculature. These studies identify H-ferritin as an iron transport protein and suggest the presence of an H-ferritin receptor for mediating iron delivery. The relative amount of iron that could be delivered via H-ferritin could make this protein a predominant player in cellular iron delivery.

Presence of antibody in virus-infected brain cells of SSPE ferrets

1983 ◽  
Vol 41 ◽  
pp. 804-805
Author(s):  
Hannah R. Brown ◽  
Tammy L. Donato ◽  
Halldor Thormar
Keyword(s):  
Measles Virus ◽  
Plasma Cells ◽  
Subacute Sclerosing Panencephalitis ◽  
Protein A ◽  
Neutralizing Antibody ◽  
Brain Cells ◽  
Antibody Titers ◽  
A Cell ◽  
The Central Nervous System ◽  
The Brain

Measles virus specific immunoglobulin G (IgG) has been found in the brains of patients with subacute sclerosing panencephalitis (SSPE), a slowly progressing disease of the central nervous system (CNS) in children. IgG/albumin ratios indicate that the antibodies are synthesized within the CNS. Using the ferret as an animal model to study the disease, we have been attempting to localize the Ig's in the brains of animals inoculated with a cell associated strain of SSPE. In an earlier report, preliminary results using Protein A conjugated to horseradish peroxidase (PrAPx) (Dynatech Diagnostics Inc., South Windham, ME.) to detect antibodies revealed the presence of immunoglobulin mainly in antibody-producing plasma cells in inflammatory lesions and not in infected brain cells.In the present experiment we studied the brain of an SSPE ferret with neutralizing antibody titers of 1:1024 in serum and 1:512 in CSF at time of sacrifice 7 months after i.c. inoculation with SSPE measles virus-infected cells. The animal was perfused with saline and portions of the brain and spinal cord were immersed in periodate-lysine-paraformaldehyde (P-L-P) fixative. The ferret was not perfused with fixative because parts of the brain were used for virus isolation.

scholarly journals Iron storage in ferritin following intracellular hemoglobin denaturation in erythroleukemic cells

Blood ◽  
1983 ◽  
Vol 62 (4) ◽  
pp. 928-930 ◽  
Author(s):  
E Fibach ◽  
ER Bauminger ◽  
AM Konijn ◽  
S Ofer ◽  
EA Rachmilewitz
Keyword(s):  
Cell Lines ◽  
Storage Protein ◽  
Intracellular Iron ◽  
Iron Storage ◽  
Cellular Iron ◽  
Ferritin Iron ◽  
Induction Of Differentiation ◽  
K 562 Cells ◽  
Murine Erythroleukemia ◽  
Erythroleukemic Cells

Abstract Murine erythroleukemia (MEL) and human K-562 cell lines were cultured in the presence of 57Fe, and the quantities of cellular iron-containing compounds were determined with the aid of Mossbauer spectroscopy. Upon induction of differentiation, both ferritin-iron and hemoglobin (Hb) iron could be detected. Treatment of the cells with 0.01%-0.02% acetylphenylhydrazine (APH) resulted in gradual denaturation of Hb and incorporation of the released Hb-iron into ferritin. Following treatment with APH, the ratio of Hb-57Fe to ferritin-57Fe decreased from 2.6 to 0.2 in MEL cells and from 0.56 to 0.12 in K-562 cells. No change was observed in the total intracellular iron. Using fluorescence ELISA, an increased level of immunologically detectable ferritin was found in hemoglobinized K-562 cells treated with APH, as compared to the amount of ferritin found in untreated cells. Ferritin may thus function not only as an intermediate during Hb synthesis, but also as storage protein for iron released during Hb denaturation.

Abstract TMP113: Deletion of Alk1 Gene in Bone Marrow-Derived Endothelial Cells is Sufficient to Cause Brain Arteriovenous Malformation in Mice

Stroke ◽  
2017 ◽  
Vol 48 (suppl_1) ◽  
Author(s):  
Man Luo ◽  
Qiang Li ◽  
Li Ma ◽  
Rui Zhang ◽  
Lei Zhan ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Endothelial Cells ◽  
Viral Vector ◽  
Peripheral Blood Cell ◽  
Vascular Morphology ◽  
Multiple Organs ◽  
Adult Mice ◽  
Cell Counts ◽  
Brain Avm ◽  
The Brain

Introduction: In humans, Alk1 deficiency causes arteriovenous malformations (AVMs) in multiple organs. Endothelial deletion of Alk1 in adult mice leads to AVM formation in multiple organs and the brain angiogenic region, and endoglin-deficient bone marrow (BM) transmits abnormal brain vascular phenotype to wild-type (WT) mice. We hypothesize that Alk1 deletion in BM-derived endothelial cells (BMDECs) is sufficient to induce AVM in the brain angiogenic region in adult mice. Methods: Adult pdgf b-iCreER; Alk1 2f/2f mice that have Alk1 exons 4-6 flanked by loxP sites and a transgene expressing tamoxifen (TM)-inducible cre recombinase in the endothelial cells were used as BM donor. BM isolated from these mice was transplanted to lethally irradiated 8-week-old WT mice. Brain angiogenesis was induced through an intra-brain injection of an adeno-associated viral vector expressing VEGF 4 weeks after the BM transplantation. Two weeks later, Alk1 deletion was induced through an intra-peritoneal injection of TM (2.5 mg/20g body weight). Vascular morphology was analyzed using latex casting 6 weeks later. Results: Peripheral blood cell counts fully recovered in the recipients 4 weeks after BM transplantation. Mice transplanted with pdgfb -iCreER; Alk1 2f/2f BM developed AVM in the brain angiogenic region. BM-derived endothelial cells were detected in brain AVM vessels. Unlike pdgfb -iCreER; Alk1 2f/2f mice that died 2 weeks after TM treatment due to bleeding from intestinal AVM, mice with pdgfb -iCreER; Alk1 2f/2f BM stayed alive within a 6-week period after TM treatment, suggesting that their intestinal AVM was less severe than in pdgfb -iCreER; Alk1 2f/2f mice. Conclusion: BMDECs play a significant role in brain AVM development. Transfusion of endothelial progenitor cells or stem cell-derived endothelial cells could be a potential therapy for brain AVM patients.

scholarly journals Iron storage in ferritin following intracellular hemoglobin denaturation in erythroleukemic cells

Blood ◽  
1983 ◽  
Vol 62 (4) ◽  
pp. 928-930 ◽  
Author(s):  
E Fibach ◽  
ER Bauminger ◽  
AM Konijn ◽  
S Ofer ◽  
EA Rachmilewitz
Keyword(s):  
Cell Lines ◽  
Storage Protein ◽  
Intracellular Iron ◽  
Iron Storage ◽  
Cellular Iron ◽  
Ferritin Iron ◽  
Induction Of Differentiation ◽  
K 562 Cells ◽  
Murine Erythroleukemia ◽  
Erythroleukemic Cells

Murine erythroleukemia (MEL) and human K-562 cell lines were cultured in the presence of 57Fe, and the quantities of cellular iron-containing compounds were determined with the aid of Mossbauer spectroscopy. Upon induction of differentiation, both ferritin-iron and hemoglobin (Hb) iron could be detected. Treatment of the cells with 0.01%-0.02% acetylphenylhydrazine (APH) resulted in gradual denaturation of Hb and incorporation of the released Hb-iron into ferritin. Following treatment with APH, the ratio of Hb-57Fe to ferritin-57Fe decreased from 2.6 to 0.2 in MEL cells and from 0.56 to 0.12 in K-562 cells. No change was observed in the total intracellular iron. Using fluorescence ELISA, an increased level of immunologically detectable ferritin was found in hemoglobinized K-562 cells treated with APH, as compared to the amount of ferritin found in untreated cells. Ferritin may thus function not only as an intermediate during Hb synthesis, but also as storage protein for iron released during Hb denaturation.

scholarly journals Endothelial cells are critical regulators of iron transport in a model of the human blood–brain barrier

2018 ◽  
Vol 39 (11) ◽  
pp. 2117-2131 ◽  
Author(s):  
Brian Chiou ◽  
Emma H Neal ◽  
Aaron B Bowman ◽  
Ethan S Lippmann ◽  
Ian A Simpson ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Iron Deficiency ◽  
Blood Brain Barrier ◽  
Iron Transport ◽  
Brain Barrier ◽  
Brain Iron ◽  
Blood Brain ◽  
Iron Delivery ◽  
The Impact ◽  
The Brain

Iron delivery to the brain is essential for multiple neurological processes such as myelination, neurotransmitter synthesis, and energy production. Loss of brain iron homeostasis is a significant factor in multiple neurological disorders. Understanding the mechanism by which the transport of iron across the blood–brain barrier (BBB) is regulated is crucial to address the impact of iron deficiency on brain development and excessive accumulation of iron in neurodegenerative diseases. Using induced pluripotent stem cell (iPSC)-derived brain endothelial cells (huECs) as a human BBB model, we demonstrate the ability of transferrin, hepcidin, and DMT1 to impact iron transport and release. Our model reveals a new function for H-ferritin to transport iron across the BBB by binding to the T-cell immunoglobulin and mucin receptor 1. We show that huECs secrete both transferrin and H-ferritin, which can serve as iron sources for the brain. Based on our data, brain iron status can exert control of iron transport across the endothelial cells that constitute the BBB. These data address a number of pertinent questions such as how brain iron uptake is regulated at the regional level, the source of iron delivery to the brain, and the clinical strategies for attempting to treat brain iron deficiency.

Endotoxin Alteration of the Mucopolysaccharide Blood Vessel Coating in Rats

1974 ◽  
Vol 32 ◽  
pp. 148-149
Author(s):  
D. E. Philpott ◽  
A. Takahashi
Keyword(s):  
Skeletal Muscle ◽  
Endothelial Cells ◽  
Salivary Gland ◽  
Carotid Artery ◽  
Basement Membrane ◽  
Coronary Vessels ◽  
Ruthenium Red ◽  
E Coli ◽  
Old Rats ◽  
The Brain

Two month, eight month and two year old rats were treated with 10 or 20 mg/kg of E. Coli endotoxin I. P. The eight month old rats proved most resistant to the endotoxin. During fixation the aorta, carotid artery, basil arartery of the brain, coronary vessels of the heart, inner surfaces of the heart chambers, heart and skeletal muscle, lung, liver, kidney, spleen, brain, retina, trachae, intestine, salivary gland, adrenal gland and gingiva were treated with ruthenium red or alcian blue to preserve the mucopolysaccharide (MPS) coating. Five, 8 and 24 hrs of endotoxin treatment produced increasingly marked capillary damage, disappearance of the MPS coating, edema, destruction of endothelial cells and damage to the basement membrane in the liver, kidney and lung.

Use of Protein a Conjugated to Peroxidase to Localize Immunoglobulin G in SSPE Ferret Brain

1982 ◽  
Vol 40 ◽  
pp. 350-351
Author(s):  
Hannah R. Brown ◽  
Anthony F. Nostro ◽  
Halldor Thormar
Keyword(s):  
Immunoglobulin G ◽  
Human Disease ◽  
Measles Virus ◽  
Subacute Sclerosing Panencephalitis ◽  
Protein A ◽  
Inflammatory Lesions ◽  
Sspe Virus ◽  
Specific Immunoglobulin ◽  
Antibody Titers ◽  
The Brain

Subacute sclerosing panencephalitis (SSPE) is a slowly progressing disease of the CNS in children which is caused by measles virus. Ferrets immunized with measles virus prior to inoculation with the cell associated, syncytiogenic D.R. strain of SSPE virus exhibit characteristics very similar to the human disease. Measles virus nucleocapsids are present, high measles antibody titers are found in the sera and inflammatory lesions are prominent in the brains. Measles virus specific immunoglobulin G (IgG) is present in the brain,and IgG/ albumin ratios indicate that the antibodies are synthesized within the CNS.

scholarly journals Age influences susceptibility of brain capillary endothelial cells to La Crosse virus infection and cell death

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Rahul Basu ◽  
Vinod Nair ◽  
Clayton W. Winkler ◽  
Tyson A. Woods ◽  
Iain D. C. Fraser ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Death ◽  
Virus Infection ◽  
La Crosse Virus ◽  
Brain Capillary ◽  
Adult Mice ◽  
Age Related ◽  
Capillary Endothelial Cells ◽  
The Brain

Abstract Background A key factor in the development of viral encephalitis is a virus crossing the blood-brain barrier (BBB). We have previously shown that age-related susceptibility of mice to the La Crosse virus (LACV), the leading cause of pediatric arbovirus encephalitis in the USA, was associated with the ability of the virus to cross the BBB. LACV infection in weanling mice (aged around 3 weeks) results in vascular leakage in the olfactory bulb/tract (OB/OT) region of the brain, which is not observed in adult mice aged > 6–8 weeks. Thus, we studied age-specific differences in the response of brain capillary endothelial cells (BCECs) to LACV infection. Methods To examine mechanisms of LACV-induced BBB breakdown and infection of the CNS, we analyzed BCECs directly isolated from weanling and adult mice as well as established a model where these cells were infected in vitro and cultured for a short period to determine susceptibility to virus infection and cell death. Additionally, we utilized correlative light electron microscopy (CLEM) to examine whether changes in cell morphology and function were also observed in BCECs in vivo. Results BCECs from weanling, but not adult mice, had detectable infection after several days in culture when taken ex vivo from infected mice suggesting that these cells could be infected in vitro. Further analysis of BCECs from uninfected mice, infected in vitro, showed that weanling BCECs were more susceptible to virus infection than adult BCECs, with higher levels of infected cells, released virus as well as cytopathic effects (CPE) and cell death. Although direct LACV infection is not detected in the weanling BCECs, CLEM analysis of brain tissue from weanling mice indicated that LACV infection induced significant cerebrovascular damage which allowed virus-sized particles to enter the brain parenchyma. Conclusions These findings indicate that BCECs isolated from adult and weanling mice have differential viral load, infectivity, and susceptibility to LACV. These age-related differences in susceptibility may strongly influence LACV-induced BBB leakage and neurovascular damage allowing virus invasion of the CNS and the development of neurological disease.

scholarly journals Synthesis and pharmacokinetic characterisation of a fluorine-18 labelled brain shuttle peptide fusion dimeric affibody

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Takahiro Morito ◽  
Ryuichi Harada ◽  
Ren Iwata ◽  
Yiqing Du ◽  
Nobuyuki Okamura ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Post Injection ◽  
Affibody Molecule ◽  
Positron Emission ◽  
A Cell ◽  
Protein Binders ◽  
Rapid Clearance ◽  
Labelling Method ◽  
The Brain ◽  
Ex Vivo Study

AbstractBrain positron emission tomography (PET) imaging with radiolabelled proteins is an emerging concept that potentially enables visualization of unique molecular targets in the brain. However, the pharmacokinetics and protein radiolabelling methods remain challenging. Here, we report the performance of an engineered, blood–brain barrier (BBB)-permeable affibody molecule that exhibits rapid clearance from the brain, which was radiolabelled using a unique fluorine-18 labelling method, a cell-free protein radiosynthesis (CFPRS) system. AS69, a small (14 kDa) dimeric affibody molecule that binds to the monomeric and oligomeric states of α-synuclein, was newly designed for brain delivery with an apolipoprotein E (ApoE)-derived brain shuttle peptide as AS69-ApoE (22 kDa). The radiolabelled products 18F-AS69 and 18F-AS69-ApoE were successfully synthesised using the CFPRS system. Notably, 18F-AS69-ApoE showed higher BBB permeability than 18F-AS69 in an ex vivo study at 10 and 30 min post injection and was partially cleared from the brain at 120 min post injection. These results suggest that small, a brain shuttle peptide-fused fluorine-18 labelled protein binders can potentially be utilised for brain molecular imaging.

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