Elevated endothelial microparticle—monocyte complexes induced by multiple sclerosis plasma and the inhibitory effects of interferon-β1b on release of endothelial microparticles, formation and transendothelial migration of monocyte-endothelial microparticle complexes

2005 ◽  
Vol 11 (3) ◽  
pp. 310-315 ◽  
Author(s):  
Joaquin J Jimenez ◽  
Wenche Jy ◽  
Lucia M Mauro ◽  
Lawrence L Horstman ◽  
Eugene R Ahn ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Transendothelial Migration ◽  
Endothelial Microparticles ◽  
Functional Roles ◽  
Monocyte Migration ◽  
Therapeutic Mechanism ◽  
Demyelinating Lesions ◽  
Vitro Model ◽  
And Control

Monocyte migration through the disrupted cerebral endothelial cell (EC) junctions plays an essential role in formation of multiple sclerosis (MS) demyelinating lesions. During pathogenesis of MS, activated ECs release endothelial microparticles (EMP), which possibly facilitate transendothelial migration (TEMIG) of monocytes. To assess functional roles of EMP in MS, specifically, their (i) interaction with monocytes, (ii) effect on monocyte TEMIG in an in vitro model of the brain microvascular endothelial cells (BMVEC), (iii) phenotypic profiles of EMP elicited by MS plasma and (iv) the effects of IFN-b1b on release of EMP and on TEMIG of monocytes (mono) and monocytes:EMP complexes (mono:EMP) through the BMVEC. The effect of IFN-b1b on the release of EMP and the TEMIG of mono and mono:EMP was assessed by preincubating BMVEC cultures of IFN-b1b prior to addition of plasma. Three EMP phenotypes, CD54, CD62E and CD31 were assayed. Plasma specimens from 20 patients with relapsing—remitting MS (11 in exacerbation, MS-E, and 9 in remission, ME-R) and 10 healthy controls were studied. Incubation of BMVEC with MS-E plasma yielded elevated levels of EMPCD54, EMP62E and EMPCD31 relative to MS-R and control plasmas. MS-E but not MS-R or control plasma also augmented TEMIG of monocytes, respectively. Mono:EMP complexes further augmented TEMIG relative to mono alone, but only in the presence of MS-E plasma; there was no significant effect with MS-R or control plasmas. The presence of IFN-b1b inhibited TEMIG of mono and mono:EMP by 20% and 30%, respectively. MS-E but not MS-R plasma elicited release of activation-derived EMP and enhanced TEMIG of mono and mono:EMP. IFN-b1b inhibited TEMIG and release of EMP, suggesting a role of EMP and a novel therapeutic mechanism for IFN-β1b in MS.

Remyelination of Demyelinated CNS Axons by Transplanted Human Schwann Cells: The Deleterious Effect of Contaminating Fibroblasts

2001 ◽  
Vol 10 (3) ◽  
pp. 305-315 ◽  
Author(s):  
C. M. H. Brierley ◽  
A. J. Crang ◽  
Y. Iwashita ◽  
J. M. Gilson ◽  
N. J. Scolding ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Schwann Cell ◽  
Schwann Cells ◽  
Axonal Degeneration ◽  
Autologous Transplantation ◽  
Growth Factor Receptor ◽  
Adult Rats ◽  
Demyelinating Lesions ◽  
Cell Implantation

Areas of demyelination can be remyelinated by transplanting myelin-forming cells. Schwann cells are the naturally remyelinating cells of the peripheral nervous system and have a number of features that may make them attractive for cell implantation therapies in multiple sclerosis, in which spontaneous but limited Schwann cell remyelination has been well documented. Schwann cells can be expanded in vitro, potentially affording the opportunity of autologous transplantation; and they might also be spared the demyelinating process in multiple sclerosis. Although rat, cat, and monkey Schwann cells have been transplanted into rodent demyelinating lesions, the behavior of transplanted human Schwann cells has not been evaluated. In this study we examined the consequences of injecting human Schwann cells into areas of acute demyelination in the spinal cords of adult rats. We found that transplants containing significant fibroblast contamination resulted in deposition of large amounts of collagen and extensive axonal degeneration. However, Schwann cell preparations that had been purified by positive immunoselection using antibodies to human low-affinity nerve growth factor receptor containing less than 10% fibroblasts were associated with remyelination. This result indicates that fibroblast contamination of human Schwann cells represents a greater problem than would have been appreciated from previous studies.

scholarly journals THE CHANGES IN DISTRIBUTION OF NMDA- AND AMPA-RECEPTORS IN RETINOCOLLICULAR SYNAPSES

2021 ◽  
Vol 67 (3) ◽  
pp. 10-16
Author(s):  
H.V. Dumanska ◽  
◽  
O.V. Rikchalsky ◽  
N.S. Veselovsky ◽  
◽  
...  
Keyword(s):  
Ampa Receptors ◽  
In Vitro Model ◽  
Ganglion Cells ◽  
Signal Transmission ◽  
Main Role ◽  
Retinal Ganglion ◽  
Dynamic Changes ◽  
Functional Roles ◽  
Vitro Model

We investigated the changes in distribution of the NMDA- and AMPA-receptors in the synapses in the in vitro model of the retinocollicular pathway. The model was a primary coculture of the retinal cells and superficial superior colliculus (SSC) neurons. Evoked postsynaptic currents (ePSCs) were recorded in SSC neurons in response to local electrical extracellular stimulation of the afferent retinal ganglion cells’ (RGC) axons. We analyzed the changes in the kinetic characteristics of ePSCs at different holding potentials during cocultivation. The results obtained reflect that NMDA receptors play an essential role in the formation of the retinocollicular synapses. After the formation of retinocollicular connections, the main role in the sensory signal transmission belongs to AMPA receptors. Thus, the data obtained indicate the specific dynamic changes of the functional roles of NMDA- and AMPA-receptors in the forma- tion and development of the retinocollicular synaptic contacts.

The zebrafish brain in research and teaching: a simple in vivo and in vitro model for the study of spontaneous neural activity

2011 ◽  
Vol 35 (2) ◽  
pp. 188-196 ◽  
Author(s):  
R. Vargas ◽  
I. þ. Jóhannesdóttir ◽  
B. Sigurgeirsson ◽  
H. þorsteinsson ◽  
K. Æ. Karlsson
Keyword(s):  
Neural Development ◽  
Neural Circuits ◽  
Brain Slices ◽  
Functional Roles ◽  
Zebrafish Brain ◽  
Vitro Model ◽  
Immunohistochemical Techniques ◽  
Neuronal Groups

Recently, the zebrafish ( Danio rerio ) has been established as a key animal model in neuroscience. Behavioral, genetic, and immunohistochemical techniques have been used to describe the connectivity of diverse neural circuits. However, few studies have used zebrafish to understand the function of cerebral structures or to study neural circuits. Information about the techniques used to obtain a workable preparation is not readily available. Here, we describe a complete protocol for obtaining in vitro and in vivo zebrafish brain preparations. In addition, we performed extracellular recordings in the whole brain, brain slices, and immobilized nonanesthetized larval zebrafish to evaluate the viability of the tissue. Each type of preparation can be used to detect spontaneous activity, to determine patterns of activity in specific brain areas with unknown functions, or to assess the functional roles of different neuronal groups during brain development in zebrafish. The technique described offers a guide that will provide innovative and broad opportunities to beginner students and researchers who are interested in the functional analysis of neuronal activity, plasticity, and neural development in the zebrafish brain.

scholarly journals CELLULAR IMMUNITY IN VITRO

1971 ◽  
Vol 133 (6) ◽  
pp. 1377-1389 ◽  
Author(s):  
Harvey B. Simon ◽  
John N. Sheagren
Keyword(s):  
Cellular Immunity ◽  
In Vitro Model ◽  
Specific Antigen ◽  
Cell Types ◽  
Intracellular Bacteria ◽  
Migration Inhibition ◽  
Vitro Model ◽  
And Control ◽  
Macrophage Migration Inhibition

An in vitro model of cellular immunity in the guinea pig was established. Animals were immunized with tubercle bacilli, bovine gamma globulin, or picrylated human serum albumin in complete Freund's adjuvant. Oil-induced peritoneal exudates from immune and control animals were cultured overnight with and without specific antigen. The cultures were washed and the macrophage monolayers were infected with Listeria monocytogenes. At intervals the monolayers were lysed and the numbers of viable intracellular bacteria were quantitated by pour plate cultures. Random monolayers were also evaluated in sequence by visually counting the intracellular bacteria on Gram-stained plates. Both methods demonstrated that the macrophages from immune animals had markedly enhanced listericidal activity when the peritoneal exudates were cultured with antigen before infection. Macrophage migration inhibition was also demonstrated under these conditions. The experiments reported here describe an in vitro model of cellular immunity which will allow separation and recombination of cell types and direct assay of cell products in efforts to elucidate further the mechanisms of the immunologically mediated enhancement of macrophage bactericidal capacity.

scholarly journals RNA-binding protein altered expression and mislocalization in multiple sclerosis

2019 ◽  
Author(s):  
Katsuhisa Masaki ◽  
Yoshifumi Sonobe ◽  
Ghanashyam Ghadge ◽  
Peter Pytel ◽  
Paula Lépine ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Binding Protein ◽  
Metabolic Stress ◽  
Nucleocytoplasmic Transport ◽  
Altered Expression ◽  
Polypyrimidine Tract Binding Protein ◽  
Demyelinating Lesions

AbstractObjectiveNuclear depletion and mislocalization of RNA-binding proteins (RBPs) trans-activation response DNA-binding protein of 43 kDa (TDP-43) and fused in sarcoma (FUS) are thought to contribute to the pathogenesis of a number of disorders, including amyotrophic lateral sclerosis (ALS). We recently found that TDP-43 as well as polypyrimidine tract binding protein (PTB) have decreased expression and mislocalization in oligodendrocytes in demyelinated lesions in an experimental mouse model of multiple sclerosis (MS) caused by Theiler’s murine encephalomyelitis virus infection.MethodsThe latter finding prompted us to investigate TDP-43, FUS, and PTB in the demyelinated lesions of MS and in in vitro cultured human brain-derived oligodendrocytes.ResultsWe found: i) mislocalized TDP-43 in oligodendrocytes in active lesions in some MS patients; ii) decreased PTB1 expression in oligodendrocytes in mixed active/inactive demyelinating lesions; iii) decreased nuclear expression of PTB2 in neurons in cortical demyelinating lesions; iv) nuclear depletion of TDP-43 in oligodendrocytes under metabolic stress induced by low glucose/low nutrient conditions compared to optimal culture conditions.ConclusionTDP-43 has been found to have a key role in oligodendrocyte function and viability, while PTB is important in neuronal differentiation, suggesting that altered expression and mislocalization of these RBPs in MS lesions may contribute to the pathogenesis of demyelination and neurodegeneration. Our findings also identify nucleocytoplasmic transport as a target for treatment.

scholarly journals Infection of Human Endothelial Cells with Chlamydia pneumoniae Stimulates Transendothelial Migration of Neutrophils and Monocytes

1999 ◽  
Vol 67 (3) ◽  
pp. 1323-1330 ◽  
Author(s):  
Robert E. Molestina ◽  
Richard D. Miller ◽  
Julio A. Ramirez ◽  
James T. Summersgill
Keyword(s):  
Endothelial Cells ◽  
Chlamydia Pneumoniae ◽  
Umbilical Vein ◽  
Neutrophil Migration ◽  
Transendothelial Migration ◽  
Serial Passage ◽  
Human Endothelial Cells ◽  
Monocyte Migration ◽  
Low Passage

ABSTRACT We have previously shown that different isolates of Chlamydia pneumoniae display heterogeneity in the in vitro stimulation of chemokines and adhesion molecules from infected human endothelial cells. In the present study, we examined the ability of different isolates of C. pneumoniae to promote transendothelial migration of neutrophils and monocytes. Human umbilical vein endothelial cells (HUVEC) were infected with low (<15)-passageC. pneumoniae isolates A-03, PS-32, and BR-393 and high (>40)-passage isolates BAL-16, TW-183, and T-2634, and levels of neutrophil and monocyte transendothelial migration were determined following 24 h of infection. Compared to mock-infected controls, significant increases in neutrophil migration were observed in response to most C. pneumoniae isolates examined (P < 0.001). Levels of monocyte migration were significantly increased in response to TW-183 and T-2634 (P < 0.001). Serial passage (>40 times) of the three low-passage isolates in HEp-2 cell cultures prior to infection of HUVEC generally resulted in the promotion of higher levels of neutrophil and monocyte transendothelial migration. These findings were compatible with differences observed in the extent of interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1) stimulation between low- and high-passage A-03, PS-32, and BR-393. As opposed toC. pneumoniae, infection with C. trachomatis L2 caused only a slight increase in neutrophil transendothelial migration, which correlated with the lack of measurable IL-8 levels by this species. However, significant levels of monocyte migration were induced in response to C. trachomatis L2 despite a lack of measurable MCP-1 stimulation.C. trachomatis serovars A and E also failed to induce IL-8 and MCP-1 production in HUVEC. Results from this study indicate that the passage history of C. pneumoniae may play a role in the divergence of stimulatory activities observed among isolates in human endothelial cells. In addition, the differences observed between this organism and C. trachomatissuggest that the upregulation of IL-8 and MCP-1 in endothelial cells may be unique to C. pneumoniae.

PDX validation of a 3D microtumor platform.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. 3029-3029
Author(s):  
Ellen Sampson ◽  
Katya Nikolov ◽  
Paul T. Henderson ◽  
Christian Apfel ◽  
Chong-xian Pan ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Tumor Growth ◽  
Saline Control ◽  
Patient Derived Xenograft ◽  
Pdx Model ◽  
End Of Treatment ◽  
Observational Cohort ◽  
Vitro Model ◽  
And Control

3029 Title: Patient-derived xenograft validation of a 3D microtumor platform Background: Patient-derived xenograft (PDX) mouse models are thought to most closely reflect the biology of a patient’s cancer. Unfortunately, growing sufficient tumor in a PDX model takes several months and more often than not, the tumor fails to grows at all. The SAGE Direct Platform, an in-vitro model, can create hundreds of live microtumors from virtually every patient’s viable biopsy and test a panel of clinically relevant drugs within no more than 1 week. Thus, concordance of results from a PDX model with results of the SAGE Direct Platform would support a rational for the platform to be potentially useful to predict tumor response in cancer patients. Methods: A bladder cancer from a 77 year old female was used to establish a PDX model. Mice were divided into three groups receiving either saline (control), cisplatin, or gemcitabine intraperitoneal on the days 1, 8, and 13, and tumor growth was observed. One tumor sample was used to create 3D microtumors and those were tested using the same drugs. Results: Tumor growth (exceeding 1,000 mm3) was similar after cisplatin compared to control (4.8 vs. 3.7 weeks). After gemcitabine tumors initially shrank and only started growing a couple of weeks after the end of treatment so that 1,000 mm3 was only reached after 10.2 weeks (p<0.001 compared to cisplatin and control). In the SAGE Direct Platform the EC50 of cisplatin was 97.3 µM and thus two orders of magnitudes higher than the EC50 of gemcitabine, which was 0.7 µM. Conclusions: Both the PDX model and the SAGE Direct Platform have shown this bladder cancer to be virtually resistant to cisplatin while very sensitive to gemcitabine. The next steps of these preliminary data could be to repeat this experimental design with other tumors and/or to start an observational cohort study in patients correlating the SAGE Direct Platform results to patient outcomes.

scholarly journals Effects of Axotomy on Cultured Sensory Neurons of Aplysia: Long-Term Injury-Induced Changes in Excitability and Morphology Are Mediated by Different Signaling Pathways

2008 ◽  
Vol 100 (6) ◽  
pp. 3209-3224 ◽  
Author(s):  
Supinder S. Bedi ◽  
Diancai Cai ◽  
David L. Glanzman
Keyword(s):  
Signaling Pathways ◽  
Sensory Neurons ◽  
Specific Inhibitor ◽  
Distinct Pattern ◽  
Extracellular Signal ◽  
Vitro Model ◽  
Induced Changes ◽  
And Control

To facilitate an understanding of injury-induced changes within the nervous system, we used a single-cell, in vitro model of axonal injury. Sensory neurons were individually dissociated from the CNS of Aplysia and placed into cell culture. The major neurite of some neurons was then transected (axotomized neurons). Axotomy in hemolymph-containing culture medium produced long-term hyperexcitability (LTH-E) and enhanced neuritic sprouting (long-term hypermorphogenesis [LTH-M]). Axotomy in the absence of hemolymph induced LTH-E, but not LTH-M. Hemolymph-derived growth factors may activate tyrosine receptor kinase (Trk) receptors in sensory neurons. To examine this possibility, we treated uninjured (control) and axotomized sensory neurons with K252a, an inhibitor of Trk receptor activity. K252a depressed the excitability of both axotomized and control neurons. K252a also produced a distinct pattern of arborizing outgrowth of neurites in both axotomized and control neurons. Protein kinase C (PKC) is an intracellular signal downstream of Trk; accordingly, we tested the effects of bisindolylmaleimide I (Bis-I), a specific inhibitor of PKC, on the axotomy-induced cellular changes. Bis-I blocked LTH-E, but did not disrupt LTH-M. Finally, because Trk activates the extracellular signal regulated kinase pathway in Aplysia sensory neurons, we examined whether this pathway mediates the injury-induced changes. Sensory neurons were axotomized in the presence of U0126, an inhibitor of mitogen-activated/extracellular receptor-regulated kinase. U0126 blocked the LTH-M due to axotomy, but did not impair LTH-E. Therefore distinct cellular signaling pathways mediate the induction of LTH-E and LTH-M in the sensory neurons.

Synergy between paclitaxel plus an exogenous methyl donor in the suppression of murine demyelinating diseases

2007 ◽  
Vol 13 (5) ◽  
pp. 596-609 ◽  
Author(s):  
FG Mastronardi ◽  
H. Tsui ◽  
S. Winer ◽  
DD Wood ◽  
T. Selvanantham ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Combination Therapy ◽  
Clinical Signs ◽  
Demyelinating Diseases ◽  
Model Systems ◽  
Methyl Donor ◽  
Negative Balance ◽  
Vitro Model

Progressive demyelination in multiple sclerosis (MS) reflects the negative balance between myelin damage and repair due to physical and molecular barriers, such as astrocytic glial scars, between oligodendrocytes and target neurons. In this paper, we show that combination therapy with paclitaxel (Taxol®) plus the universal methyl-donor, vitamin B12CN (B12CN), dramatically limits progressive demyelination, and enhances remyelination in several independent, immune and nonimmune, in vivo and in vitro model systems. Combination therapy significantly reduced clinical signs of EAE in SJL mice, as well as the spontaneously demyelinating ND4 transgenic mouse. Astrocytosis was normalised in parallel to ultrastructural and biochemical evidence of remyelination. The combination therapy suppressed T cell expansion, reduced IFN-gamma, while enhancing IFN-beta and STAT-1 expression, STAT-1 phosphorylation and methylation of STAT-1 and MBP in the brain. Paclitaxel/B12CN has nearly identical effects to the previously described combination of IFN-beta/ B12CN, whose clinical usefulness is transient because of IFN-neutralising antibodies, not observed (or expected) with the present drug combination. This report provides a mechanistic foundation for the development of a new therapeutic strategy in humans with MS. Multiple Sclerosis 2007; 13: 596-609. http://msj.sagepub.com

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