1) The study examined CD4+ T cells in the central nervous system (CNS) of mice during experimental allergic encephalomyelitis (EAE), both during active disease and remission.
2) Flow cytometry analysis revealed a 30-fold reduction in the number of CD4+ T cells in the CNS of mice in remission compared to those with active EAE.
3) However, the CD4+ T cells that remained in the CNS during remission maintained an activated memory/effector phenotype, suggesting remission is not due to downregulation of T cell function.
This document describes a study examining the effects of T cell vaccination on the immune response in mice protected against experimental allergic encephalomyelitis (EAE). The researchers found that vaccination with myelin basic protein (MBP)-reactive T cell lines protected SJL/J mice against EAE induced by rat spinal cord homogenate. Lymph node cells from vaccinated mice showed enhanced proliferative responses to MBP and an encephalitogenic peptide from proteolipid protein, compared to untreated mice. The augmentation of responses was not antigen-specific and also occurred for ovalbumin. This suggests vaccination led to non-specific enhancement of immune activation in peripheral lymphoid tissues.
This document discusses research on selectively activating the AT1 and AT2 receptors to induce proliferation and differentiation of human neural stem cells. The renin-angiotensin system regulates cardiovascular function and is involved in brain development and function. The AT1 receptor induces proliferation while the AT2 receptor induces differentiation. The study cultured human neural stem cells and treated them with selective agonists of each receptor. Results showed the AT1 agonist increased proliferation and the AT2 agonist increased differentiation. Further research could explore higher drug doses to better understand these effects and potential applications for brain injuries and diseases.
Creando el mapa de la susceptibilidad genética y un modelo de patogénesis en ...Fundación Ramón Areces
Ciclo de conferencias y debates en Ciencias.
Fundación Ramón Areces-Nature Publishing Group.
Jorge R. Oksenberg. Universidad de California, San Francisco, EE. UU.
Madrid, 2 de febrero de 2012
Tetrahydrohyperforin (IDN5706), a derivative of the active molecule hyperforin in St. John's Wort, was examined for its ability to prevent cognitive deficits and synaptic impairment in an Alzheimer's disease mouse model. Five-month old APPswe/PSEN1DE9 mice were treated with IDN5706 for 10 weeks. IDN5706 improved memory, prevented decreases in synaptic proteins and LTP, and reduced amyloid-beta plaque burden, tau hyperphosphorylation, and astrogliosis. In cell cultures, IDN5706 decreased amyloid precursor protein processing leading to amyloid-beta peptide generation. The results suggest IDN5706 may be a potential therapeutic for treating Alzheimer's
The document describes the establishment of immortalized human amniotic epithelial cell (iHAE) lines. HAE cells were extracted from placentas and infected with retroviruses containing HPV16 E6/E7 and hTERT genes to extend their lifespan. The iHAE lines showed extended proliferation ability and expression of stem cell markers. They maintained multipotent differentiation potential as demonstrated by their ability to differentiate into adipocytes, osteocytes, neurons, and cardiac cell types. The iHAE cells represent a promising new cell source for applications in regenerative medicine and cell therapy due to their immunosuppressive properties and differentiation potential.
In vivo effects of Interleukin 2 on Lymphocyte Subpopulations in a Patient wi...Elke Stein
This document describes a clinical trial using interleukin-2 (IL-2) to treat a 17-month-old boy with combined immunodeficiency. IL-2 was purified from leukocytes and administered subcutaneously over 50 days. Within 3 weeks, IL-2 treatment normalized the patient's lymphocytosis and improved his eosinophilia. Most striking was the normalization of the patient's abnormal OKT4/OKT8 ratio and increase in OKT3+ cells. Infectious episodes decreased during treatment. Some effects were transient, returning to pathological levels after treatment ended.
Temp-Sensitive Inhibition of Development in Dictyostelium - Dev Bio 251 18-26...James Silverman
1) The Dictyostelium mutant HSB1 is temperature-sensitive for development, aggregating and forming fruiting bodies below 18°C but not above.
2) HSB1 cells have a defective G protein-linked adenylyl cyclase that is not stimulated by GTPγS in vitro but can be rescued by adding wild-type cytosol.
3) Transfection with the wild-type piaA gene rescued the HSB1 mutant phenotype, and sequencing revealed a point mutation in the HSB1 piaA gene resulting in a single amino acid change.
This document describes a study examining the effects of T cell vaccination on the immune response in mice protected against experimental allergic encephalomyelitis (EAE). The researchers found that vaccination with myelin basic protein (MBP)-reactive T cell lines protected SJL/J mice against EAE induced by rat spinal cord homogenate. Lymph node cells from vaccinated mice showed enhanced proliferative responses to MBP and an encephalitogenic peptide from proteolipid protein, compared to untreated mice. The augmentation of responses was not antigen-specific and also occurred for ovalbumin. This suggests vaccination led to non-specific enhancement of immune activation in peripheral lymphoid tissues.
This document discusses research on selectively activating the AT1 and AT2 receptors to induce proliferation and differentiation of human neural stem cells. The renin-angiotensin system regulates cardiovascular function and is involved in brain development and function. The AT1 receptor induces proliferation while the AT2 receptor induces differentiation. The study cultured human neural stem cells and treated them with selective agonists of each receptor. Results showed the AT1 agonist increased proliferation and the AT2 agonist increased differentiation. Further research could explore higher drug doses to better understand these effects and potential applications for brain injuries and diseases.
Creando el mapa de la susceptibilidad genética y un modelo de patogénesis en ...Fundación Ramón Areces
Ciclo de conferencias y debates en Ciencias.
Fundación Ramón Areces-Nature Publishing Group.
Jorge R. Oksenberg. Universidad de California, San Francisco, EE. UU.
Madrid, 2 de febrero de 2012
Tetrahydrohyperforin (IDN5706), a derivative of the active molecule hyperforin in St. John's Wort, was examined for its ability to prevent cognitive deficits and synaptic impairment in an Alzheimer's disease mouse model. Five-month old APPswe/PSEN1DE9 mice were treated with IDN5706 for 10 weeks. IDN5706 improved memory, prevented decreases in synaptic proteins and LTP, and reduced amyloid-beta plaque burden, tau hyperphosphorylation, and astrogliosis. In cell cultures, IDN5706 decreased amyloid precursor protein processing leading to amyloid-beta peptide generation. The results suggest IDN5706 may be a potential therapeutic for treating Alzheimer's
The document describes the establishment of immortalized human amniotic epithelial cell (iHAE) lines. HAE cells were extracted from placentas and infected with retroviruses containing HPV16 E6/E7 and hTERT genes to extend their lifespan. The iHAE lines showed extended proliferation ability and expression of stem cell markers. They maintained multipotent differentiation potential as demonstrated by their ability to differentiate into adipocytes, osteocytes, neurons, and cardiac cell types. The iHAE cells represent a promising new cell source for applications in regenerative medicine and cell therapy due to their immunosuppressive properties and differentiation potential.
In vivo effects of Interleukin 2 on Lymphocyte Subpopulations in a Patient wi...Elke Stein
This document describes a clinical trial using interleukin-2 (IL-2) to treat a 17-month-old boy with combined immunodeficiency. IL-2 was purified from leukocytes and administered subcutaneously over 50 days. Within 3 weeks, IL-2 treatment normalized the patient's lymphocytosis and improved his eosinophilia. Most striking was the normalization of the patient's abnormal OKT4/OKT8 ratio and increase in OKT3+ cells. Infectious episodes decreased during treatment. Some effects were transient, returning to pathological levels after treatment ended.
Temp-Sensitive Inhibition of Development in Dictyostelium - Dev Bio 251 18-26...James Silverman
1) The Dictyostelium mutant HSB1 is temperature-sensitive for development, aggregating and forming fruiting bodies below 18°C but not above.
2) HSB1 cells have a defective G protein-linked adenylyl cyclase that is not stimulated by GTPγS in vitro but can be rescued by adding wild-type cytosol.
3) Transfection with the wild-type piaA gene rescued the HSB1 mutant phenotype, and sequencing revealed a point mutation in the HSB1 piaA gene resulting in a single amino acid change.
By: Edward M. Barbieri and James Shorter
TDP-43 forms cytoplasmic aggregates in degenerating neurons of frontotemporal dementia (FTD) patients.
Laferrière et al. now establish that TDP-43 assemblies from distinct FTD subtypes have different structures,
neurotoxicities, and seeding activities, which correlate with FTD severity. Thus, distinct pathological TDP-43
assemblies akin to prion strains might underpin distinct FTD subtypes.
Boronated Cetuximab CCR tumor targeting in BNCTkent.riley
This document describes a study evaluating boronated cetuximab (BD-C225) for boron neutron capture therapy (BNCT) of epidermal growth factor receptor (EGFR) positive gliomas. In vitro, BD-C225 showed preferential uptake in EGFR positive glioma cells compared to EGFR negative cells. In vivo, rats with EGFR positive glioma tumors received intracerebral BD-C225, achieving high boron levels in the tumors. BNCT with BD-C225 alone or combined with boronophenylalanine extended survival compared to controls. The results provide support for using molecularly targeted boron delivery agents like BD-C225 for BNCT of brain tumors.
This summarizes a study that developed a modified non-biotin polymerized horseradish peroxidase (HRP) immunohistochemical method for diagnosing canine distemper virus (CDV) infection from formalin-fixed tissue samples. The method confirmed CDV infection in seven of eight suspected cases. Labelled CDV antigen was observed in various tissues including brain, spinal cord, kidney, lungs, skin, and others. Compared to microwave pretreatment alone, autoclaving followed by microwave heating produced better labelling results. The non-biotin HRP detection system produced similar results to a conventional biotin-linked system.
This study characterized the function of ALL2, a homolog of ALL1, in the fungal pathogen Cryptococcus neoformans. The key findings were:
1) Unlike deletion of ALL1, deletion of ALL2 attenuated virulence in a pulmonary infection model.
2) The all2Δ mutant shed a less viscous exopolysaccharide and was more sensitive to hydrogen peroxide but more resistant to macrophage killing than the wild type.
3) Transcriptome analysis supported distinct functions for ALL1 and ALL2, with ALL2 uniquely involved in maintaining intracellular pH under low-pH conditions.
This study investigated the role of astrocytes and neuronal PrPC in prion-induced neurodegeneration. The researchers set up cocultures of prion-infected astrocytes and uninfected neurons from transgenic mice expressing different PrP variants. They found that interaction between neuronal PrPC and exogenous PrPSc was not sufficient to induce neuronal death, but efficient conversion of neuronal PrPC into PrPSc was required for prion-associated neurotoxicity. Prion-infected astrocytes accelerated neurodegeneration in homologous cocultures compared to infected single neuronal cultures, despite no detectable neurotoxin release. Accumulation of PrPSc in neurons led to neuritic damage and cell death potentiated by glut
This document summarizes research on nicotinic acetylcholine receptors (nAChRs) in mouse brain. The researchers developed assays to measure nAChR binding and function, including assays for dopamine, GABA, and ion flux. Comparing results across assays and between wildtype and nAChR subunit knockout mice revealed diversity in nAChR subtypes. At least six nAChR binding sites were identified with different pharmacological properties. Assays of function suggested different nAChR subtypes mediate dopamine vs. GABA release and ion flux. Deletion of the beta2 subunit eliminated most nAChR binding and function, confirming its role in major brain nAChR subtypes.
This study developed a sensitive sandwich enzyme-linked immunosorbent assay (ELISA) to detect tau proteins in cerebrospinal fluid (CSF) using the monoclonal antibody AT120. The assay had a detection limit of less than 5 pg/ml of CSF for tau. Using this assay, CSF tau levels were found to be significantly higher in Alzheimer's disease patients compared to controls. However, CSF tau levels overlapped between Alzheimer's patients and those with other neurological diseases. The study suggests CSF tau measurement may help diagnose Alzheimer's but cannot on its own distinguish it from other conditions.
1) Cultured human limbal epithelial cells were stored for one week at 31°C in organ culture medium, 23°C in organ culture medium, or 5°C in Optisol-GS. Cell death due to apoptosis was assessed using caspase-3 staining, TUNEL staining, and gene expression profiling of 84 apoptosis genes.
2) Minimal cell death was observed under all storage conditions, as indicated by low caspase-3 and TUNEL labeling. Gene expression analysis found pro-apoptotic genes were downregulated and anti-apoptotic genes were upregulated.
3) Organ culture storage at 23°C best preserved cell structure, while 31°C and 5°C storage were associated
This study identified an ethylene-responsive element (ERE) involved in regulating transcription of the carnation glutathione-S-transferase 1 (GST1) gene during petal senescence. Through deletion analysis and transient expression assays of GST1-GUS gene fusions delivered to carnation petals via particle bombardment, the researchers identified a 197 bp region between -667 and -470 bp upstream of the GST1 transcription start site that is necessary for ethylene-responsive expression. Within this region, a 126 bp sequence between -596 and -470 bp conferred ethylene-regulated expression to a minimal CaMV 35S promoter. Nuclear protein extracts from carnation petals were found to specifically bind
This study examined the immune response in patients with early cutaneous leishmaniasis (ECL), defined as illness duration of 60 days or less, compared to those with late cutaneous leishmaniasis (LCL) of over 12 months. The results showed that in ECL, 32% of patients had absent or low lymphoproliferative responses to Leishmania antigens and 41% had an absence of interferon (IFN)-γ production. IFN-γ levels were significantly lower in ECL compared to LCL, while interleukin (IL)-10 levels were significantly higher in ECL. The addition of anti-IL-10 or anti-IL-12 antibodies restored lymphoprolifer
This document summarizes research on the effects of temperature on longevity in C. elegans. Microarray analysis identified lysosomal protease genes that are differentially expressed at 15°C vs 25°C. Follow up experiments showed that lysosome structure and number are altered at elevated temperatures. RNAi of temperature-regulated proteases caused progressive paralysis specifically at 15°C, indicating a role in neuronal health. Further experiments traced this paralysis to dysfunction of motor neurons rather than muscles. The research suggests that proteostasis, particularly lysosomal function, is important for neuronal health under cool temperatures and influences longevity.
Detection of ehrlichia platys dna in brown dog ticksJosephine Huang
This document reports on a study that detected Ehrlichia platys DNA in brown dog ticks (Rhipicephalus sanguineus) collected from dogs in Okinawa Island, Japan. Key findings include:
- Partial 16S rRNA gene sequences of E. platys were detected in 3 out of 32 ticks collected from 2 dogs using PCR and sequencing.
- This represents the first detection of E. platys in Japan and the first report of detecting it in ticks.
- Sequence analysis showed the 3 positive ticks were highly similar (99.7-100% identical) to known E. platys sequences, confirming the detection.
- This suggests R. sanguineus ticks
This study examined apoptosis of lymphocytes in patients with systemic lupus erythematosus (SLE). The percentage of apoptotic lymphocytes from SLE patients was significantly higher than healthy donors after 36 hours of incubation, indicating SLE patients have more lymphocytes undergoing apoptosis. A positive correlation was also found between lymphocyte apoptosis in SLE and disease activity markers like anti-dsDNA antibodies and prednisolone dosage. The percentage of CD4+ T cells was significantly lower in SLE patients compared to healthy donors.
1) Acute rejection of a transplanted organ is characterized by intense inflammation within the graft. Prostanoids, which are classic mediators of inflammation such as prostaglandins and thromboxane, modulate inflammation and alloimmune responses during graft rejection but their role has been overlooked.
2) Local production of prostanoids increases within the allograft during an episode of acute rejection. Prostanoids can interfere with graft function by modulating vascular tone, capillary permeability, and platelet aggregation.
3) Experimental evidence also suggests prostanoids may participate in alloimmune responses by directly modulating T lymphocyte and antigen-presenting cell function. The paper provides an overview of alloimmune responses,
This study examined the effects of chronic exposure to nicotine on the function of human α4β2 nicotinic acetylcholine receptors (nAChRs), which play a major role in nicotine addiction. The key findings were:
1) Acetylcholine dose-response curves for human α4β2 nAChRs expressed in human cells were biphasic, consisting of both high- and low-affinity components.
2) Chronic exposure to nicotine or nicotinic antagonists increased the fraction of high-affinity receptors from 25% to around 70% and doubled the acetylcholine-evoked currents.
3) Upregulation of receptor function after chronic nicotine exposure occurred
This study examined the functional state of epidermal growth factor (EGF) receptors during internalization in A-431 cells. The key findings were:
1) EGF-receptor complexes retained their phosphorylated state and connection to EGF during internalization, as shown using photoaffinity labeling and antibodies to phosphotyrosine.
2) Internalized EGF receptors, like those in the plasma membrane, were able to phosphorylate synthetic peptide substrates introduced into the cell, indicating they maintained kinase activity.
3) Approximately 15% of internalized EGF-receptor complexes were phosphorylated on tyrosine residues, suggesting functional heterogeneity of EGF receptors in A-431 cells.
This document summarizes research on the Scr74 gene family in Phytophthora infestans and how different variants of this gene are recognized in potato genotypes. The key points are:
1. 27 variants of the Scr74 effector gene were identified from 12 P. infestans isolates. 4 novel variants were cloned.
2. 13 amino acid sites in Scr74 were found to be under positive diversifying selection, indicating these sites are important for pathogen recognition.
3. 17 Scr74 variants were screened using PVX assays on 12 potato genotypes. Some genotypes had strong cell death responses to specific Scr74 variants, suggesting recognition of these variants.
4. Scr74 variants Scr74-A
Presence of genetically modified organism genes in carica papaya, glycine max...valrivera
This document summarizes a study that aimed to detect the presence of genetically modified organism (GMO) genes in fruits from four plants: papaya, soybean, corn, and wheat. DNA was extracted from samples of each fruit and tested via polymerase chain reaction (PCR) and gel electrophoresis to detect two common GMO markers - the 35S promoter and NOS terminator. The results were inconclusive due to DNA degradation and possible human errors during experiments. As such, the study was unable to determine if the fruits contained GMO genes.
The researchers developed a novel non-viral single doxycycline inducible system that can efficiently regulate gene expression in stem cells both in vitro and in vivo. They generated stable clonal cell lines from mouse neuroblastoma cells (N2a) and human embryonic stem cells (hNSC-ESC) that responded to doxycycline by eliminating GFP expression within 3-8 days. GFP expression was restored 9-10 days after doxycycline removal. When transplanted hNSC-ESCs labeled with quantum dots were subjected to doxycycline in mice, GFP expression was reduced by 61-56% compared to controls. This suggests the system can control gene expression in stem cells without using viral vectors
Posted: https://ash.confex.com/ash/2011/webprogram/Paper42474.html
Abstract
Adult hematopoietic cells transition through a hemogenic endothelial (HE) intermediate during development, but the signaling pathways modulating this transition are incompletely characterized. Although the Hedgehog (Hh) pathway is hypothesized to play a role in blood and endothelial cell formation, early embryonic lethality of mice lacking Hedgehog signaling precludes such analysis. To determine a role for Hh signaling in HE patterning, we assessed the effect of altered Hh signaling in differentiating mouse embryonic stem cells (mESCs), cultured embryonic day 9.5 mouse embryos, and developing zebrafish embryos. In differentiating mESCs, addition of Indian Hh ligand (IHH) increased the number of CD41+c-Kit+ hematopoietic progenitors, whereas chemical inhibition of Hh signaling led to a decrease without affecting primitive-streak mesoderm gene expression. In the setting of Hh inhibition, Notch induction rescued hemogenic VE-cadherin+ cells, demonstrating that Notch expands HE. Scl/Tal1 (stem cell leukemia/T-cell associated leukemia 1) induction rescued VE-cadherin+CD41+ cells, demonstrating that Scl/Tal1 converts endothelial cells to hematopoietic tissue. Similar experiments using cultured mouse yolk sacs demonstrated that signaling pathways are conserved in vivo. Moreover, VE-cadherin+ cells isolated from the mouse yolk sac or paraaortic splanchnopleura, when virally transduced with Notch signaling or Scl, had increased hematopoietic colony-forming activity. Finally, ectopic Notch or Scl induction in zebrafish embryos rescued the expression of the prototypical hemogenic endothelium marker Runx1 in the absence of Hh signalling. Together, our results reveal that the Hh-Notch-Scl axis promotes embryonic hematopoiesis through endothelial-to-hematopoietic transition.
O documento apresenta as especialidades e serviços de consultoria em recursos humanos oferecidos por Rubens Caetano Luiz, como diagnóstico organizacional, planos de cargos e salários, pesquisa salarial, programas de remuneração variável, avaliação de desempenho e pesquisa de clima organizacional. O consultor tem mais de 20 anos de experiência em RH e busca assessorar empresas a melhorar resultados, produtividade e satisfação dos empregados.
The document analyzes the results of a questionnaire given to the document author's media class about their preferences for a short film. Most respondents were between 15-19 years old, with a target audience of 15-30. Responses were also analyzed based on gender, religion, political views, favorite and least favorite film genres, and preferences between short films and documentaries. The key question asked if the short film should focus on the experience of homosexuality or pride celebrations, with an even split in responses. As a result, the document author plans to incorporate both story elements into the short film.
By: Edward M. Barbieri and James Shorter
TDP-43 forms cytoplasmic aggregates in degenerating neurons of frontotemporal dementia (FTD) patients.
Laferrière et al. now establish that TDP-43 assemblies from distinct FTD subtypes have different structures,
neurotoxicities, and seeding activities, which correlate with FTD severity. Thus, distinct pathological TDP-43
assemblies akin to prion strains might underpin distinct FTD subtypes.
Boronated Cetuximab CCR tumor targeting in BNCTkent.riley
This document describes a study evaluating boronated cetuximab (BD-C225) for boron neutron capture therapy (BNCT) of epidermal growth factor receptor (EGFR) positive gliomas. In vitro, BD-C225 showed preferential uptake in EGFR positive glioma cells compared to EGFR negative cells. In vivo, rats with EGFR positive glioma tumors received intracerebral BD-C225, achieving high boron levels in the tumors. BNCT with BD-C225 alone or combined with boronophenylalanine extended survival compared to controls. The results provide support for using molecularly targeted boron delivery agents like BD-C225 for BNCT of brain tumors.
This summarizes a study that developed a modified non-biotin polymerized horseradish peroxidase (HRP) immunohistochemical method for diagnosing canine distemper virus (CDV) infection from formalin-fixed tissue samples. The method confirmed CDV infection in seven of eight suspected cases. Labelled CDV antigen was observed in various tissues including brain, spinal cord, kidney, lungs, skin, and others. Compared to microwave pretreatment alone, autoclaving followed by microwave heating produced better labelling results. The non-biotin HRP detection system produced similar results to a conventional biotin-linked system.
This study characterized the function of ALL2, a homolog of ALL1, in the fungal pathogen Cryptococcus neoformans. The key findings were:
1) Unlike deletion of ALL1, deletion of ALL2 attenuated virulence in a pulmonary infection model.
2) The all2Δ mutant shed a less viscous exopolysaccharide and was more sensitive to hydrogen peroxide but more resistant to macrophage killing than the wild type.
3) Transcriptome analysis supported distinct functions for ALL1 and ALL2, with ALL2 uniquely involved in maintaining intracellular pH under low-pH conditions.
This study investigated the role of astrocytes and neuronal PrPC in prion-induced neurodegeneration. The researchers set up cocultures of prion-infected astrocytes and uninfected neurons from transgenic mice expressing different PrP variants. They found that interaction between neuronal PrPC and exogenous PrPSc was not sufficient to induce neuronal death, but efficient conversion of neuronal PrPC into PrPSc was required for prion-associated neurotoxicity. Prion-infected astrocytes accelerated neurodegeneration in homologous cocultures compared to infected single neuronal cultures, despite no detectable neurotoxin release. Accumulation of PrPSc in neurons led to neuritic damage and cell death potentiated by glut
This document summarizes research on nicotinic acetylcholine receptors (nAChRs) in mouse brain. The researchers developed assays to measure nAChR binding and function, including assays for dopamine, GABA, and ion flux. Comparing results across assays and between wildtype and nAChR subunit knockout mice revealed diversity in nAChR subtypes. At least six nAChR binding sites were identified with different pharmacological properties. Assays of function suggested different nAChR subtypes mediate dopamine vs. GABA release and ion flux. Deletion of the beta2 subunit eliminated most nAChR binding and function, confirming its role in major brain nAChR subtypes.
This study developed a sensitive sandwich enzyme-linked immunosorbent assay (ELISA) to detect tau proteins in cerebrospinal fluid (CSF) using the monoclonal antibody AT120. The assay had a detection limit of less than 5 pg/ml of CSF for tau. Using this assay, CSF tau levels were found to be significantly higher in Alzheimer's disease patients compared to controls. However, CSF tau levels overlapped between Alzheimer's patients and those with other neurological diseases. The study suggests CSF tau measurement may help diagnose Alzheimer's but cannot on its own distinguish it from other conditions.
1) Cultured human limbal epithelial cells were stored for one week at 31°C in organ culture medium, 23°C in organ culture medium, or 5°C in Optisol-GS. Cell death due to apoptosis was assessed using caspase-3 staining, TUNEL staining, and gene expression profiling of 84 apoptosis genes.
2) Minimal cell death was observed under all storage conditions, as indicated by low caspase-3 and TUNEL labeling. Gene expression analysis found pro-apoptotic genes were downregulated and anti-apoptotic genes were upregulated.
3) Organ culture storage at 23°C best preserved cell structure, while 31°C and 5°C storage were associated
This study identified an ethylene-responsive element (ERE) involved in regulating transcription of the carnation glutathione-S-transferase 1 (GST1) gene during petal senescence. Through deletion analysis and transient expression assays of GST1-GUS gene fusions delivered to carnation petals via particle bombardment, the researchers identified a 197 bp region between -667 and -470 bp upstream of the GST1 transcription start site that is necessary for ethylene-responsive expression. Within this region, a 126 bp sequence between -596 and -470 bp conferred ethylene-regulated expression to a minimal CaMV 35S promoter. Nuclear protein extracts from carnation petals were found to specifically bind
This study examined the immune response in patients with early cutaneous leishmaniasis (ECL), defined as illness duration of 60 days or less, compared to those with late cutaneous leishmaniasis (LCL) of over 12 months. The results showed that in ECL, 32% of patients had absent or low lymphoproliferative responses to Leishmania antigens and 41% had an absence of interferon (IFN)-γ production. IFN-γ levels were significantly lower in ECL compared to LCL, while interleukin (IL)-10 levels were significantly higher in ECL. The addition of anti-IL-10 or anti-IL-12 antibodies restored lymphoprolifer
This document summarizes research on the effects of temperature on longevity in C. elegans. Microarray analysis identified lysosomal protease genes that are differentially expressed at 15°C vs 25°C. Follow up experiments showed that lysosome structure and number are altered at elevated temperatures. RNAi of temperature-regulated proteases caused progressive paralysis specifically at 15°C, indicating a role in neuronal health. Further experiments traced this paralysis to dysfunction of motor neurons rather than muscles. The research suggests that proteostasis, particularly lysosomal function, is important for neuronal health under cool temperatures and influences longevity.
Detection of ehrlichia platys dna in brown dog ticksJosephine Huang
This document reports on a study that detected Ehrlichia platys DNA in brown dog ticks (Rhipicephalus sanguineus) collected from dogs in Okinawa Island, Japan. Key findings include:
- Partial 16S rRNA gene sequences of E. platys were detected in 3 out of 32 ticks collected from 2 dogs using PCR and sequencing.
- This represents the first detection of E. platys in Japan and the first report of detecting it in ticks.
- Sequence analysis showed the 3 positive ticks were highly similar (99.7-100% identical) to known E. platys sequences, confirming the detection.
- This suggests R. sanguineus ticks
This study examined apoptosis of lymphocytes in patients with systemic lupus erythematosus (SLE). The percentage of apoptotic lymphocytes from SLE patients was significantly higher than healthy donors after 36 hours of incubation, indicating SLE patients have more lymphocytes undergoing apoptosis. A positive correlation was also found between lymphocyte apoptosis in SLE and disease activity markers like anti-dsDNA antibodies and prednisolone dosage. The percentage of CD4+ T cells was significantly lower in SLE patients compared to healthy donors.
1) Acute rejection of a transplanted organ is characterized by intense inflammation within the graft. Prostanoids, which are classic mediators of inflammation such as prostaglandins and thromboxane, modulate inflammation and alloimmune responses during graft rejection but their role has been overlooked.
2) Local production of prostanoids increases within the allograft during an episode of acute rejection. Prostanoids can interfere with graft function by modulating vascular tone, capillary permeability, and platelet aggregation.
3) Experimental evidence also suggests prostanoids may participate in alloimmune responses by directly modulating T lymphocyte and antigen-presenting cell function. The paper provides an overview of alloimmune responses,
This study examined the effects of chronic exposure to nicotine on the function of human α4β2 nicotinic acetylcholine receptors (nAChRs), which play a major role in nicotine addiction. The key findings were:
1) Acetylcholine dose-response curves for human α4β2 nAChRs expressed in human cells were biphasic, consisting of both high- and low-affinity components.
2) Chronic exposure to nicotine or nicotinic antagonists increased the fraction of high-affinity receptors from 25% to around 70% and doubled the acetylcholine-evoked currents.
3) Upregulation of receptor function after chronic nicotine exposure occurred
This study examined the functional state of epidermal growth factor (EGF) receptors during internalization in A-431 cells. The key findings were:
1) EGF-receptor complexes retained their phosphorylated state and connection to EGF during internalization, as shown using photoaffinity labeling and antibodies to phosphotyrosine.
2) Internalized EGF receptors, like those in the plasma membrane, were able to phosphorylate synthetic peptide substrates introduced into the cell, indicating they maintained kinase activity.
3) Approximately 15% of internalized EGF-receptor complexes were phosphorylated on tyrosine residues, suggesting functional heterogeneity of EGF receptors in A-431 cells.
This document summarizes research on the Scr74 gene family in Phytophthora infestans and how different variants of this gene are recognized in potato genotypes. The key points are:
1. 27 variants of the Scr74 effector gene were identified from 12 P. infestans isolates. 4 novel variants were cloned.
2. 13 amino acid sites in Scr74 were found to be under positive diversifying selection, indicating these sites are important for pathogen recognition.
3. 17 Scr74 variants were screened using PVX assays on 12 potato genotypes. Some genotypes had strong cell death responses to specific Scr74 variants, suggesting recognition of these variants.
4. Scr74 variants Scr74-A
Presence of genetically modified organism genes in carica papaya, glycine max...valrivera
This document summarizes a study that aimed to detect the presence of genetically modified organism (GMO) genes in fruits from four plants: papaya, soybean, corn, and wheat. DNA was extracted from samples of each fruit and tested via polymerase chain reaction (PCR) and gel electrophoresis to detect two common GMO markers - the 35S promoter and NOS terminator. The results were inconclusive due to DNA degradation and possible human errors during experiments. As such, the study was unable to determine if the fruits contained GMO genes.
The researchers developed a novel non-viral single doxycycline inducible system that can efficiently regulate gene expression in stem cells both in vitro and in vivo. They generated stable clonal cell lines from mouse neuroblastoma cells (N2a) and human embryonic stem cells (hNSC-ESC) that responded to doxycycline by eliminating GFP expression within 3-8 days. GFP expression was restored 9-10 days after doxycycline removal. When transplanted hNSC-ESCs labeled with quantum dots were subjected to doxycycline in mice, GFP expression was reduced by 61-56% compared to controls. This suggests the system can control gene expression in stem cells without using viral vectors
Posted: https://ash.confex.com/ash/2011/webprogram/Paper42474.html
Abstract
Adult hematopoietic cells transition through a hemogenic endothelial (HE) intermediate during development, but the signaling pathways modulating this transition are incompletely characterized. Although the Hedgehog (Hh) pathway is hypothesized to play a role in blood and endothelial cell formation, early embryonic lethality of mice lacking Hedgehog signaling precludes such analysis. To determine a role for Hh signaling in HE patterning, we assessed the effect of altered Hh signaling in differentiating mouse embryonic stem cells (mESCs), cultured embryonic day 9.5 mouse embryos, and developing zebrafish embryos. In differentiating mESCs, addition of Indian Hh ligand (IHH) increased the number of CD41+c-Kit+ hematopoietic progenitors, whereas chemical inhibition of Hh signaling led to a decrease without affecting primitive-streak mesoderm gene expression. In the setting of Hh inhibition, Notch induction rescued hemogenic VE-cadherin+ cells, demonstrating that Notch expands HE. Scl/Tal1 (stem cell leukemia/T-cell associated leukemia 1) induction rescued VE-cadherin+CD41+ cells, demonstrating that Scl/Tal1 converts endothelial cells to hematopoietic tissue. Similar experiments using cultured mouse yolk sacs demonstrated that signaling pathways are conserved in vivo. Moreover, VE-cadherin+ cells isolated from the mouse yolk sac or paraaortic splanchnopleura, when virally transduced with Notch signaling or Scl, had increased hematopoietic colony-forming activity. Finally, ectopic Notch or Scl induction in zebrafish embryos rescued the expression of the prototypical hemogenic endothelium marker Runx1 in the absence of Hh signalling. Together, our results reveal that the Hh-Notch-Scl axis promotes embryonic hematopoiesis through endothelial-to-hematopoietic transition.
O documento apresenta as especialidades e serviços de consultoria em recursos humanos oferecidos por Rubens Caetano Luiz, como diagnóstico organizacional, planos de cargos e salários, pesquisa salarial, programas de remuneração variável, avaliação de desempenho e pesquisa de clima organizacional. O consultor tem mais de 20 anos de experiência em RH e busca assessorar empresas a melhorar resultados, produtividade e satisfação dos empregados.
The document analyzes the results of a questionnaire given to the document author's media class about their preferences for a short film. Most respondents were between 15-19 years old, with a target audience of 15-30. Responses were also analyzed based on gender, religion, political views, favorite and least favorite film genres, and preferences between short films and documentaries. The key question asked if the short film should focus on the experience of homosexuality or pride celebrations, with an even split in responses. As a result, the document author plans to incorporate both story elements into the short film.
El documento habla sobre la historia y tipos de software. Explica que durante los primeros años de las computadoras, el software se veía como un agregado y su desarrollo era sin planificación. Luego, los costos aumentaron y se adoptaron métodos más sistemáticos. Describe los tipos principales de software como software de sistema, de programación y de aplicación. También explica conceptos como ingeniería de software, prototipos y requerimientos.
Network Softwarization on KREONET: KREONET-SDongkyun Kim
KREONET is deploying an SD-WAN called KREONET-S* based on ONOS to softwarize its network. It aims to provide a virtualized, dynamic and flexible environment compared to its previous hardware-based fixed network. KREONET-S* is being rolled out in phases starting with two regional centers in 2015 and expanding from there. It uses ONOS controllers and OpenFlow/OpenStack technologies. Initial applications include Virtual Dedicated Networks and user visibility tools. The goal is to provide advanced services, user-defined networks, and new user experiences through the programmable SDN infrastructure.
This document provides instructions for preparing a balance sheet in 8 steps: write the heading, prepare the assets, liabilities, and owner's equity sections, add and compare the totals, rule single lines below the sections, write the totals, and rule double lines below the totals.
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El documento trata sobre la violencia de género. Explica que la violencia de género incluye cualquier acto que discrimine, ignore, someta o subordine a las mujeres. Luego describe los tres tipos principales de violencia de género: física, psicológica y sexual. También cuestiona si el género solo se refiere a lo femenino y masculino o también incluye otras identidades. Finalmente, honra a varias mujeres importantes que marcaron la historia.
O documento descreve a história e operações da empresa Omnilife, que atua na venda direta de suplementos nutricionais. A empresa cresceu de 6 empresários e 1 fábrica em 1991 para operar em 19 países em 2016 com 7 milhões de empresários. O documento também explica os benefícios dos produtos, formas de ganhar dinheiro como empresário e estratégias de sucesso para construir o negócio.
La Unión Europea ha acordado un paquete de sanciones contra Rusia por su invasión de Ucrania. Las sanciones incluyen restricciones a las transacciones con bancos rusos clave y la prohibición de la venta de aviones y equipos a Rusia. Los líderes de la UE también acordaron excluir a varios bancos rusos del sistema SWIFT de mensajería financiera.
En 20071 el mercado de la moda depende colombiatex 2017 - enero de 2017Camilo Herrera
El documento presenta información sobre el gasto de los hogares colombianos en 2016. Señala que el gasto creció en pesos pero cayó en unidades, y que hubo una desaceleración del gasto debido a factores como la alta inflación, la baja en la creación de empleo y la incertidumbre política. El gasto en alimentos y moda aumentó, mientras que en otros rubros como vivienda y transporte disminuyó.
Chromatin remodeling refers to dynamic changes in chromatin structure that occur in cells. There are several classes of chromatin remodeling complexes that use ATP hydrolysis to alter nucleosome positioning in different ways: 1) Nucleosome sliding, 2) Ejection of histone dimers or octamers, and 3) Replacement of core histones with histone variants. Disorders can result if chromatin remodeling is disrupted. Techniques like chromatin immunoprecipitation sequencing are used to study chromatin structure.
This study directly analyzed the infiltration of the central nervous system (CNS) by PKH2-labeled, myelin basic protein (MBP)-reactive CD4+ T cells in mice with experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. The researchers found that about 45% of CD4+ T cells in the CNS at disease onset were PKH2-labeled cells that had infiltrated from the periphery. Nearly all of these cells in the CNS exhibited a memory/effector phenotype of being CD44high and CD45RBlow and responded to MBP in vitro, indicating antigen recognition promoted their retention in the CNS. In contrast, few PK
1. T-cell infiltration is increased in the brains of transgenic mouse models of Alzheimer's disease-like cerebral amyloidosis compared to non-transgenic littermates. However, the infiltrating T-cells do not co-localize with amyloid plaques and produce less of the effector cytokine interferon-gamma.
2. Antigen-presenting cells in the brains of transgenic mice show an immature phenotype with accumulation of MHC-II in intracellular compartments, suggesting impaired antigen presentation.
3. The findings indicate that cerebral amyloidosis promotes T-cell infiltration but interferes with local T-cell activation and antigen presentation, which may contribute to amyloid accumulation in Alzheimer's disease progression.
Kshivets O. Lung Cancer: Early Detection and Diagnosis Oleg Kshivets
This document discusses a study examining the role of the immune system in the early detection and diagnosis of lung cancer. The study analyzed immune cell and humoral factors in 533 lung cancer patients and 282 patients with non-malignant lung conditions. The results showed that early detection of stage I-II lung cancer depended significantly on immune cell levels, monocyte/macrophage levels, and humoral immunity levels compared to non-malignant conditions. Diagnosis of lung cancer at all stages also depended significantly on immune cell subpopulations, macrophage levels, humoral factors, and neutrophil levels compared to non-malignant conditions. The immune system may thus provide valuable data to help with early detection and differential diagnosis of
1) WASp-deficient dendritic cells (DCs) show impaired activation of naive CD8+ T cells, especially at low antigen doses.
2) This is partly due to altered trafficking of antigen-bearing DCs from the periphery to lymph nodes. However, correcting DC migration does not fully rescue T cell activation.
3) In vitro and in vivo imaging revealed that cytoskeletal alterations in WASp-null DCs reduce their ability to form and maintain conjugates with naive CD8+ T cells in lymph nodes, contributing to defective T cell priming.
TP receptors augment cellular immune responses and inflammatory tissue injury. Mice lacking the TP receptor, which binds thromboxane A2, show reduced T cell proliferation in response to mitogens and alloantigens. They also show attenuated tissue injury in a cardiac transplant model of inflammation. Thus, thromboxane signaling through the TP receptor enhances immune responses and inflammation, suggesting that specific inhibition of this receptor may reduce inflammation without the side effects of broad COX inhibition by NSAIDs.
This document describes the development of a method to detect human CD4+ T cells specific for influenza A virus using soluble human MHC class II tetramers. Key points:
1) Soluble MHC class II tetramers containing an immunodominant peptide from influenza hemagglutinin were constructed and used to identify antigen-specific T cells from influenza-immune individuals.
2) After 7 days of proliferation in response to stimulation by influenza peptide or vaccine, tetramer-positive cells had undergone extensive division and expressed markers of activated CD4+ T cells.
3) The influenza peptide-specific T cells represented a major subset of proliferating CD4+ T cells responding to whole influenza vaccine.
4)
Sipuleucel-T (Provenge) is an autologous cellular immunotherapy for asymptomatic metastatic prostate cancer. It consists of autologous dendritic cells activated ex vivo with a recombinant fusion protein and infused back to the patient to stimulate an immune response against prostate cancer cells. Clinical trials demonstrated a significant survival benefit for patients treated with Sipuleucel-T compared to placebo. Dendreon is preparing for the commercial launch of Provenge in the US and EU, which will require a major manufacturing and logistics effort to process and deliver the personalized immunotherapy to thousands of patients.
The document discusses a study examining the effect of Mycobacterium avium infection on T cell differentiation in the thymus. The study found that (1) the thymi of infected mice retained the ability to generate new T cells, but (2) T cells differentiated in infected thymi had an impaired ability to protect against M. avium in peripheral organs compared to those differentiated in non-infected thymi. Specifically, T cells from infected thymi showed a reduced ability to produce IFN-gamma in response to M. avium antigens. The results suggest that infection induces central tolerance specifically to the infecting pathogen.
1. Sipuleucel-T (Provenge) is an autologous cellular immunotherapy for asymptomatic metastatic prostate cancer that works by activating antigen-presenting cells and T-cells against prostatic acid phosphatase.
2. Clinical trials showed Provenge improved overall survival in metastatic castration-resistant prostate cancer patients.
3. Manufacturing and delivering Provenge presents logistical challenges due to its personalized nature that Dendreon aims to address through an advanced planning system and partnerships.
This study examines the interactions between dendritic cells (DCs) and naive T cells, and how the maturation state of DCs impacts those interactions. The key findings are:
1) Mature DCs form stable conjugates with naive T cells and induce organized immune synapses, while immature DCs form few stable conjugates without organized synapses.
2) Mature DCs can sustain long-lasting interactions with naive T cells even without antigen, while immature DCs only establish short intermittent contacts.
3) The premature termination of interactions by immature DCs prevents the formation of organized immune synapses and full T cell activation.
Endothelial Cell Mediated Delay of Blood Brain Barrier Recovery Following Tra...Arthur Stem
TBI is the leading cause of death among young adults and children in the developed world, accounting for over 50,000 deaths per year. [12] TBI results in a sleuth of poor health outcomes, including hemorrhaging, seizures, neural edema, neural inflammation, and cognitive and emotional disabilities. All of these outcomes are a direct result of fundamental degradation of the BBB over a time course post TBI. [1] [12] The BBB is an integral structure that forms around the microvascular of the cerebral cavity. Endothelial cells form the basal membrane through which strictly controlled movement of molecules is observed between the extravascular and intravascular space across this basal membrane. This basal membrane is maintained by endothelial cells, having tight junctions between them to make up the pores through which transport of molecules can occur between the brain and microvasculature. These tight junctions are maintained through cross-talk between the endothelial cells and supporting neurons such as astrocytes and pericytes. [2] A multitude of proteins make up the tight junctions between the endothelial cells, including six main scaffolding structures Claudins 1, 3, and 5, ZO-1, Occludins, and Cadherins. [3] VEGF release following trauma induces endothelial cells to release matrix metalloproteinases (MMPs), in particular MMP9, which can catalyze the N-terminal amino acids that compose the tight junction protein ZO-1. [10] [11] MMP9 when in circulation is also known to activate tumor necrosis factor alpha (TNFɑ) which in turn upregulates transcription of MMP9, creating a positive feedback loop. [11] The management of MMP production is three fold, transcription, proenzyme activation, and substrate inhibition. [11] In our study, it is proenzyme activation via TP that is the focus and how that affects the overall transcription levels of the tight junction proteins within the endothelial cells and astrocytes.
The document discusses the role of invariant natural killer T (iNKT) cells in allergic asthma. It finds that iNKT cells produce cytokines like IL-4 and IL-13 that promote Th2 responses and airway hyperreactivity (AHR), and mice lacking iNKT cells do not develop AHR. Studies in humans also find increased iNKT cells in the lungs of asthmatic patients compared to healthy controls. However, the number of iNKT cells may not relate directly to asthma severity. While iNKT cells contribute to asthma pathogenesis, their therapeutic potential is still controversial.
Cytokines play an important role in regulating lymphocyte development and differentiation. The authors identified a cytokine-inducible protein called Cybr that binds to and regulates the activity of cytohesin-1. Cybr expression is increased by IL-2 and IL-12 and is most abundant in hematopoietic cells and tissues. Cybr physically interacts with cytohesin-1 through their coiled-coil domains and enhances cytohesin-1's ability to catalyze guanine nucleotide exchange on ARF GTPases. As Cybr modifies the activity of cytohesin-1, the authors designate it as a cytohesin binder and regulator.
This document describes a case study of a 5-year-old female Argentine patient with a mutation in the CD25 gene resulting in CD25 deficiency. Key findings include chronic inflammatory lung disease (follicular bronchiolitis), eczema, infections, extremely low levels of regulatory T cells, and a homozygous missense mutation (c.122a>c; p.Y41S) in the CD25αR gene. The mutation is predicted to compromise protein function and cause immune dysregulation similar to IPEX syndrome. CD25 deficiency should be considered in patients presenting with these clinical and laboratory features to facilitate early diagnosis and treatment.
This study aimed to differentiate between invasive squamous cell carcinoma (SCC) and carcinoma-in-situ (CIS) of the uterine cervix using immunohistochemical markers. Tissue samples from 37 patients were stained for CD3, CD4, CD8, CD20, and CD138. Scoring showed decreased expression of CD8, CD20, and CD138 in invasive SCC compared to CIS, indicating differences in immune cell infiltration between invasive and non-invasive lesions. The results provide potential markers for differentiating invasive SCC from CIS of the cervix.
1. The document discusses mechanisms of central and peripheral tolerance that prevent autoimmune diseases. Central tolerance involves positive and negative selection in the thymus to eliminate self-reactive T cells, aided by the AIRE gene.
2. Peripheral tolerance mechanisms help control self-reactive T cells that escape thymic selection. These include clonal deletion, anergy, and regulation by Treg cells. Anergy involves a lack of response upon re-exposure to self-antigen.
3. Tolerance is important physiologically for organ transplants, maternal-fetal tolerance, and tolerance of commensal gut microbes. A breakdown in tolerance can lead to autoimmune diseases.
Fifth Annual Mitchell Memorial Lecture, October 6, 2014, at UC San Diego, featuring Dr. Jonathan Karn of Case Western Reserve University speaking on "Lessons Learned from models for HIV latency helping to formulate virus eradication strategies."
In 3 sentences or less:
The document analyzes how anthrax and plague biowarfare vaccines interact with human dendritic cells. It finds that while the plague vaccine strongly induces dendritic cell maturation and stimulates immune responses, the anthrax vaccine is a poor stimulator of dendritic cell maturation and cytokine production. However, the anthrax vaccine can stimulate T cell responses when combined with an adjuvant like pertussis, supporting the use of adjuvants to enhance the immune response to suboptimal vaccines.
This document summarizes a study examining the use of biodegradable nanoparticles loaded with TGF-β and IL-2 cytokines and targeted to CD4+ cells for inducing and maintaining regulatory T cells (Tregs). The nanoparticles were able to induce CD4+ Tregs in vitro and expand their numbers in vivo. Nanoparticle-induced Tregs demonstrated enhanced stability and retained their suppressive phenotype even in inflammatory conditions, highlighting the potential of this nanoparticle approach for stabilizing Tregs to treat autoimmune disease and inflammation.
Similar to Zeine & Owens, J. Neuroimmunology 1993 (20)
This document discusses challenges faced by women in medicine and academia. It provides statistics showing fewer women reach higher ranks like full professor compared to men. A study of over 4,500 faculty found women reported lower sense of belonging, self-efficacy for career advancement, and perception of gender equity and family-friendly culture. Another study identified personal factors like marital status and work environment factors like incentives and research partnerships as affecting women's research productivity in academia.
Analysis of external adaptability (agility) as a measure of organizational Effectiveness in Higher Education and recommendation of diagnostic testing based on the performance triangle model
Zeine et al. 2011 Organizational Culture in Higher Education, in Kazeroony, H...Rana ZEINE, MD, PhD, MBA
The organizational culture of higher education institutions was analyzed using a survey. The results showed that behavioral norms associated with passive/defensive and aggressive/defensive cultures were overrepresented, while constructive norms were underrepresented compared to an ideal profile. This indicates a focus on tasks over people and lower-order needs over higher-order needs. Gaps between current and ideal profiles were identified to help target areas for cultural change in higher education institutions.
This study investigated the role of estrogen receptor-beta (ESR2) in regulating cyclooxygenase-2 (COX-2) expression and prostanoid levels in human placental villous endothelial cells. The researchers found that knocking down ESR2 led to decreased COX-2 mRNA and protein levels as well as diminished prostacyclin and thromboxane concentrations in the cells, both in the presence and absence of estradiol. This suggests that ESR2 mediates COX-2 expression and prostanoid levels in a ligand-independent manner, playing a role in fetoplacental vascular function. Further investigation of ESR2 regulation of prostanoid biosynthesis and its effects on the fetop
Zeine Seminar 2010, Cancer Associated Fibroblasts and Microvascular Prolifera...Rana ZEINE, MD, PhD, MBA
World Cancer Congress 2010
Presence of Cancer-Associated Fibroblasts correlates with Microvascular Proliferation which is a Poor Prognostic Factor in Neuroblastoma Tumors
This study analyzed cancer-associated fibroblasts in 60 primary neuroblastoma tumors and a neuroblastoma xenograft model. Cancer-associated fibroblasts were characterized by expression of α-smooth muscle actin but not high-molecular weight caldesmon. High numbers of cancer-associated fibroblasts were associated with Schwannian stroma-poor histology and microvascular proliferation in human tumors. In xenografts with infiltrating Schwann cells, cancer-associated fibroblasts were approximately sevenfold less than controls without Schwann cells, suggesting Schwann cells may prevent fibroblast activation.
This study examined microvascular proliferation (MVP) in neuroblastoma tumors to determine its clinical significance. MVP, including glomeruloid MVP, was significantly associated with poor prognosis in two independent cohorts of neuroblastoma patients. MVP was also significantly associated with the clinically aggressive Schwannian stroma-poor histology of neuroblastomas. These findings provide further evidence that angiogenesis plays an important role in neuroblastoma pathogenesis and behavior, and suggest angiogenesis is regulated differently depending on the amount of Schwannian stroma in the tumor.
This research article examines the combination of ABT-510, a peptide derivative of the natural angiogenic inhibitor thrombospondin-1, and valproic acid (VPA), a histone deacetylase inhibitor, as a potential antiangiogenic treatment strategy for high-risk neuroblastoma in children. In vitro, only VPA was able to inhibit neuroblastoma cell proliferation and induce apoptosis, but both ABT-510 and VPA significantly suppressed the growth of neuroblastoma xenografts in mice. Combination therapy more effectively inhibited tumor growth than single agents and achieved total cessation of tumor growth in some mice with large xenografts. The microvascular density and number of abnormal vessels in
This study examined the effects of perforin, a pore-forming protein, on oligodendrocytes cultured from SJL mice. Oligodendrocytes were exposed to perforin for up to 2.5 hours and examined using microscopy. The results showed that the majority of oligodendrocytes were killed within 60-90 minutes via pore expansion and membrane disruption. Structural features included cell body swelling, fenestration and fragmentation of membranes and processes, cytoplasmic vacuolation, and breakdown of the nuclear envelope. These patterns of damage resembled those seen in multiple sclerosis lesions. The findings suggest that perforin may play an important role in demyelination in multiple sclerosis.
1. The study examines the mechanism by which gd T cells induce cytotoxicity of human oligodendrocytes, which are relevant to multiple sclerosis.
2. The results show that gd T cells from MS patients utilize both the Fas-mediated and perforin-based pathways to exert cytotoxic effects on oligodendrocytes.
3. Blocking perforin release completely inhibited killing of targets expressing high levels of heat shock proteins, but additional blocking of Fas ligand was required to fully inhibit killing of Fas-expressing targets and fresh oligodendrocytes.
This document summarizes a study examining the presence of cancer-associated fibroblasts (CAFs) in different subtypes of neuroblastoma tumors. The study found that CAFs, identified by alpha-smooth muscle actin expression, were abundant in schwannian stroma-poor neuroblastoma regions and associated with microvascular proliferation, but were less prevalent in schwannian stroma-rich regions. This suggests CAFs may promote angiogenesis in aggressive, schwannian stroma-poor neuroblastomas.
Zeine et al. Poster 2009 Tumor Stromal Interactions in Neuroblastoma CancersRana ZEINE, MD, PhD, MBA
1) Cancer-associated fibroblasts (CAFs) were found to colocalize with microvascular proliferation (MVP), a marker of angiogenesis, in human neuroblastoma (NB) tumors. CAF levels inversely correlated with Schwannian stroma (SS), which indicates less tumor infiltration by supportive nerve cells.
2) In a mouse model of NB, xenografts engrafted inside the sciatic nerve, where Schwann cells were present, had significantly lower levels of CAFs compared to controls without Schwann cells.
3) The results suggest that Schwann cells may inhibit CAF accumulation in NB tumors by reducing angiogenesis as reflected by decreased MVP and CAF levels.
Cell Therapy Expansion and Challenges in Autoimmune DiseaseHealth Advances
There is increasing confidence that cell therapies will soon play a role in the treatment of autoimmune disorders, but the extent of this impact remains to be seen. Early readouts on autologous CAR-Ts in lupus are encouraging, but manufacturing and cost limitations are likely to restrict access to highly refractory patients. Allogeneic CAR-Ts have the potential to broaden access to earlier lines of treatment due to their inherent cost benefits, however they will need to demonstrate comparable or improved efficacy to established modalities.
In addition to infrastructure and capacity constraints, CAR-Ts face a very different risk-benefit dynamic in autoimmune compared to oncology, highlighting the need for tolerable therapies with low adverse event risk. CAR-NK and Treg-based therapies are also being developed in certain autoimmune disorders and may demonstrate favorable safety profiles. Several novel non-cell therapies such as bispecific antibodies, nanobodies, and RNAi drugs, may also offer future alternative competitive solutions with variable value propositions.
Widespread adoption of cell therapies will not only require strong efficacy and safety data, but also adapted pricing and access strategies. At oncology-based price points, CAR-Ts are unlikely to achieve broad market access in autoimmune disorders, with eligible patient populations that are potentially orders of magnitude greater than the number of currently addressable cancer patients. Developers have made strides towards reducing cell therapy COGS while improving manufacturing efficiency, but payors will inevitably restrict access until more sustainable pricing is achieved.
Despite these headwinds, industry leaders and investors remain confident that cell therapies are poised to address significant unmet need in patients suffering from autoimmune disorders. However, the extent of this impact on the treatment landscape remains to be seen, as the industry rapidly approaches an inflection point.
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Local Advanced Lung Cancer: Artificial Intelligence, Synergetics, Complex Sys...Oleg Kshivets
Overall life span (LS) was 1671.7±1721.6 days and cumulative 5YS reached 62.4%, 10 years – 50.4%, 20 years – 44.6%. 94 LCP lived more than 5 years without cancer (LS=2958.6±1723.6 days), 22 – more than 10 years (LS=5571±1841.8 days). 67 LCP died because of LC (LS=471.9±344 days). AT significantly improved 5YS (68% vs. 53.7%) (P=0.028 by log-rank test). Cox modeling displayed that 5YS of LCP significantly depended on: N0-N12, T3-4, blood cell circuit, cell ratio factors (ratio between cancer cells-CC and blood cells subpopulations), LC cell dynamics, recalcification time, heparin tolerance, prothrombin index, protein, AT, procedure type (P=0.000-0.031). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and N0-12 (rank=1), thrombocytes/CC (rank=2), segmented neutrophils/CC (3), eosinophils/CC (4), erythrocytes/CC (5), healthy cells/CC (6), lymphocytes/CC (7), stick neutrophils/CC (8), leucocytes/CC (9), monocytes/CC (10). Correct prediction of 5YS was 100% by neural networks computing (error=0.000; area under ROC curve=1.0).
Muktapishti is a traditional Ayurvedic preparation made from Shoditha Mukta (Purified Pearl), is believed to help regulate thyroid function and reduce symptoms of hyperthyroidism due to its cooling and balancing properties. Clinical evidence on its efficacy remains limited, necessitating further research to validate its therapeutic benefits.
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Here is the updated list of Top Best Ayurvedic medicine for Gas and Indigestion and those are Gas-O-Go Syp for Dyspepsia | Lavizyme Syrup for Acidity | Yumzyme Hepatoprotective Capsules etc
Basavarajeeyam is a Sreshta Sangraha grantha (Compiled book ), written by Neelkanta kotturu Basavaraja Virachita. It contains 25 Prakaranas, First 24 Chapters related to Rogas& 25th to Rasadravyas.
2. 194
CNS of remitted animals might be inhibited or altered. signs of EAE around day 16. In experiments designed
This would predict downregulation of cytokine produc- to study CNS T cells from remitted animals, symp-
tion by CNS T cells during remission as has been tomatic mice were selected and allowed to recover
suggested by one report (Merrill et al., 1992). T cells normal function.
retained within the CNS in active disease have been
shown to be of the CD44 high, CD45RB l°w memory/ef- Isolation of LNC and mononuclear cells from CNS
lector phenotype (Jensen et al., 1992; Zeine and Owens, CNS infiltrates were collected by discontinuous den-
1992). CD44, also known as Pgp-1, is a polymorphic sity gradient centrifugation (Zeine and Owens, 1992).
integral membrane glycoprotein (Trowbridge et al., Mice were anaesthetized with chloral hydrate (3.5 g
1982) which has a role in matrix adhesion, lymphocyte kg -~) and perfused through the heart with 100 ml of
activation and lymph node homing, and has been shown PBS. The brains, spinal cords, and LN were then
to be the principal cell surface receptor for hyaluronate collected (brains that were poorly perfused were dis-
(Aruffo et al., 1990). Elevated expression of CD44 is carded), and dissociated by passing through a nylon or
characteristic of memory T cells (Butterfield et al., stainless steel mesh, respectively. The nervous tissue
1989). CD45 is a family of leukocyte specific membrane was centrifuged at 200 × g for 10 min and then resus-
proteins with protein-tyrosine phosphatase activity. pended in 4 ml of 70% isotonic Percoll (Pharmacia,
CD45 isoforms of various molecular masses are pro- Montreal, Quebec) in RPMI 1640 medium. This was
duced by alternative splicing and usage of three exons then overlaid by equal volumes of 37% and 30% iso-
that encode the N-terminal portion of the external tonic Percoll, and the gradient was centrifuged at 500
domain (Barclay et al., 1987; Johnson et al., 1989). × g for 15 min. Mononuclear cells were collected from
Prolonged activation of CD4 + T cells in vitro leads to a the 37% : 70% interface, washed in medium containing
reduction in the level of high M r CD45R expression 10% FCS (ICN Biomedicals) and counted.
(Birkeland et al., 1989). The memory/effector pheno-
type is associated with active cytokine production by T Flow cytometry
cells (Bottomly, 1988). Whether this phenotype is Surface staining for CD4, CD8, CD3,, CD45,
maintained following remission is therefore a question CD45RB, CD44, and TcRa/3 was performed as previ-
that is relevant to the mechanism of remission. ously described (Zeine and Owens; 1992). CD2 expres-
In this study we have isolated CNS mononuclear sion was detected using 12.15A (Altevogt et al., 1989),
cells from mice at various intervals following disease and anti-TCRy/~ mAb was obtained from PharMin-
onset, and used flow cytometry to describe the kinetics gen (San Diego, CA). Where indicated mAbs were
of CD4 + T cell changes in the CNS following remis- purified by Protein G-sepharose affinity chromatogra-
sion, and to determine their surface phenotype. We phy (Pharmacia) and either coupled with biotin by
show loss of CD4 + T cells from the CNS during incubation with biotinamidocaproate N-hydroxysuc-
clinical remission. However, those CD4 + T cells that cinimide ester (Sigma) or fluorescinated by incubation
remained within the neural tissue, maintained an acti- with FITC-Celite (Sigma). Cells (5 × 105-106) were
vated, memory/effector surface phenotype up to 12 incubated with antibody at 4°C for 20 min and then
weeks after the initial attack. washed. Primary rat mAbs that were used as hy-
bridoma supernatants were visualized by using FITC-
goat anti-rat Ig (Southern Biotechnology, Birmingham,
Materials and Methods AL). Biotinylated primary Abs were visualized with
either FITC-coupled streptavidin (Bio-Can Scientific)
Mice or phycoerythrin-coupled streptavidin. Non-specific
Female SJL/J mice (5-8 weeks) were obtained from binding to goat anti-rat Ig was blocked by pre-incuba-
Harlan-Sprague Dawley (Indianapolis, IN). tion with rat Ig (100 /xg ml -~) (Bio-Can Scientific,
Toronto, Ontario), before incubation with PE-CD4
EAE induction, assessment and remission a n d / o r PE-CD8. Surface staining was analysed using a
EAE was induced by s.c. injections on day 0 and day FACScan (Becton Dickinson). Dead cells were ex-
7, of either 0.5 mg rat spinal cord homogenate (RSCH) cluded by propidium iodide staining.
or 400 /xg bovine myelin basic protein (MBP) (Sigma,
St. Louis, MO) in CFA (50 ~g Mycobacterium tubercu-
losis H37RA (Difco, Detroit, MI) per mouse). Mice Results
were monitored daily for symptoms and assigned clini-
cal scores as follows: 0 (no signs), 1 (flaccid tail, clumsi- Correlation between clinical remission and the number of
ness), 2 (moderate paresis), 3 (severe paresis or unilat- CNS CD4 + cells
eral hind limb paralysis), 4 (bilateral hindlimb paraly- Mononuclear cells were isolated by discontinuous
sis). Between 50 and 70% of animals developed clinical density gradient centrifugation (Zeine and Owens,
3. 195
1992) from the pooled brains and spinal cords of SJL//J
female mice. During the active phase of E A E (day 17
post-immunization), the proportion of the CNS 10 3
mononuclear cells that were CD3 ÷ and expressed high CD8
levels of CD4 was 4-5-fold greater than in remitted or 10 2
naive mice (Fig. 1). The small proportion of CD3 ÷
C D 4 - cells seen in Fig. 1, represents CD8 ÷ T ceils,
101
which did not increase in remitted mice (Fig. 1). Maxi-
mal CD8 to CD4 ratios within the CNS were obtained
during the active phase of E A E and did not exceed 101 10 2 10 3
0.37 (Fig. 2). This and previous work show that CD4 ÷
and CD8 ÷ ceils isolated from CNS are all CD3 ÷ T C D 3
cells. Macrophages/microglia could be excluded from Fig. 2. Isolation of CD8 + T cells from the CNS of mice with active
EAE. CNS mononuclear cells were isolated from ten mice in the
analysis by their 10-fold lower expression of CD4 active phase of E A E (day 17 post-immunization), and stained with
(Sedgwick et al., 1991). biotinylated-anti-CD8 and FITC-CD3. Anti-CD8 was visualized with
phycoerythrin-coupled streptavidin. The figure shows CD8 plotted
against CD3 expression on CNS mononuclear cells.
8.9%
In order to assess the state of CNS infiltration
10 3
CD4 I "'?-" "~" "
EAE during clinical remission, groups of mice with E A E
were allowed to recover normal motor function and the
10 2 number and phenotype of CNS-derived mononuclear
cells were analyzed through remission. The number of
101 cells that could be obtained from individual mice was
less than required for flow cytometric analysis, so
pooled samples from groups of ten mice were analysed.
A strong correlation between number of CD4 ÷ T cells
i 1.7%
I
and disease progression was observed. The mean num-
I ber of mononuclear cells obtained per mouse CNS was
10 3 I
CD4
I NAIVE only slightly reduced in remitted mice (8.6 x 104 -t- 0.2,
' • I ,." :,,:~'.]._ "
n = 4) as compared to symptomatic mice (13.7 x 10 4 -I-
10 2 .__ _ _ _ ~.=S - : ~ . . . . . . . 0.2, n = 3) The absolute number of CD4 ÷ T cells was
.....,+.: :+++24-,+ " .
dramatically reduced from 77.1 x 103 on day 17 to
101 2.6 x 103 on day 28 (Fig. 3). The proportion of blasts
- :';~N ~.~ • •
~'~ ;i" ~ -': • amongst the CNS CD4 ÷ cells also decreased from 30%
on day 17 to 3% at day 55. The kinetics of CNS
infiltration correlated with the mean clinical scores
i z s%
from groups of ten mice at each time point and in the
experiment done 105 days after immunization there
10 3 i
I • .
CD4
REMITTED was a slight rise in both the mean score and the
number of CNS CD4 ÷ cells (Fig. 3). In a previously
1 0 ~. _ :_ _ :., -.a_.a~::-'... "
published study of passively transferred E A E we
: .'~!~ ~ ' " " Z~'~ . " . .
showed that the percent of CD4 + T cells increased
10
:!:~:: ~;. from 8% 2 days after onset of EAE to 25.8% 3 days
after onset and decreased to 3.5% 6 days after onset
(see Table 1 in Zeine and Owens, 1992). Given that the
101 10 2 10 3 day of onset in active E A E is about 14 days, it can be
appreciated that the results from both active and pas-
CD3
Fig. 1. Isolation of CD4 + CD3 ÷ cells from the CNS of naive mice, sive EAE are consistent. It was further possible to
mice with active EAE, and mice in remission. E A E was induced by select cells that expressed a high level of CD4 (T cells)
immunization with R S C H in CFA, and mononuclear cells were for analysis by gating and double staining.
isolated from the CNS of ten mice either at the onset of clinical signs
(day 17 post-immunization) (top) or following remission (bottom).
CNS mononuclear cells were also isolated from naive mice (center).
T cell phenotype of CNS CD4 + cells isolated from
Cells were stained with PE-anti-CD4 and FITC-anti-CD3. The figure clinically remitted mice
shows CD4 plotted against CD3 expression on CNS mononuclear The CD4 ÷ cells isolated from the CNS of mice with
cells. E A E were all CD2 ÷, CD45 +, and CD3 ÷ TCRa/[3 +.
4. 196
84
(1.~ et al., 1989). In one study immunocytochemical staining
of CNS frozen sections showed decreased numbers of
Number 70
(x 10" ~) infiltrating CD4 ÷ T cells in remitted mice (Cannella et
CD4 T 56 al., 1990). The kinetics of T cell loss which we have
cells per now described (Fig. 3) suggest that the majority of
mouse 42
CNS
CD4 ÷ T cells that accumulate within the CNS at the
) onset of disease are lost from the CNS within 48 h. A
28 (o.s)
number of groups have shown that T cells, which are
capable of recognizing a CNS antigen, are either re-
14 l Io,,
Io, (oZ Io)
0 17 18 1; 20 2; 2; 55 1(;5
Days after first immunization I A
Fig. 3. Kinetics of CNS infiltration by CD4 ÷ T cells during EAE. I
I
E A E was induced in groups of mice by s.c. injections of either R S C H 10 3
or MBP in C F A on days 0 and 7. The mice exhibited signs of clinical
I. CNS
CD44
. . . . i -: " "2 ....
E A E between days 16 and 19. After day 20, all the mice had
remitted. In each experiment the brains and spinal cords from ten 10 2
mice were pooled. CNS mononuclear cells were isolated at various - - - ~_~.-,~--=~'-~-7. - - - .
•
time points and stained with PE-anti-CD4 for analysis by FACS. The
graph represents the n u m b e r of CD4 + T cells obtained per mouse 101
CNS in each experiment. N u m b e r s in parentheses are the mean
clinical scores on the day each experiment was done.
:1 itCg 3%
TCR3~/8 T cells were not detected in the CNS of mice
in the active phase of the disease nor on day 28
post-immunization (not shown).
CD44
10 3
10 2
j '
• .
'
~?.
:1-.
f...:[--...
"" •
"" .
•
Lymph
Node
"::~:i ri" :" :'i .:"
Memory / effector phenotype of CNS CD4 ÷ T cells 101
More than 70% of CNS CD4 ÷ cells, both in active
E A E and in remission, expressed high levels of Pgp-
t ..... 'i~r~g ~, , . . L ...........
1/CD44, whereas less than 20% of LN CD4 ÷ T ceils 200 400 600 800
were CD44 high (Fig. 4A). The level of expression of
Forward Scatter
CD44 on blasts (defined by forward scatter) was also
high, and was similar between CNS and LN (Fig. 4A). B
The majority of CD4 ÷ cells from LN and blood
expressed high levels of CD45RB. By contrast, more
than 60% of CD4 ÷ T cells in CNS were CD45RB ]°w. J~
The high proportion of CD45RB l°w at disease peak E
-j
was also seen in passively transferred E A E (Zeine and
,c
Owens, 1992) and in other studies (Jensen et al., 1992). =.-
O
Similar proportions of CD45RB ~°w CD4 ÷ T cells were ¢D
found in naive mice (not shown), mice in the active .>
phase of E A E and in remitted mice (Fig. 4B). Despite 4}
~r
some variability between times of analysis, the propor- |
i ' ' " " 1 ' ' ''""1 '
i
''""'|
tion of CNS CD4 ÷ T cells that were CD45RB ~°w re- 10 1 10 2 10 3
mained greater than 60%, and there was no trend in
the variation that could be correlated with disease CD45RB
Fig. 4. P g p - 1 / C D 4 4 and CD45RB expression on CD4 + T cells
progression (Fig. 5).
isolated from CNS and LN of mice in clinical remission. Cells were
isolated from CNS and L N after remission (Day 28 post immuniza-
tion) and stained with either biotinylated anti-CD44 or biotinylated
Discussion anti-CD45RB, the binding of which was visualized using FITC-
streptavidin. PE-anti-CD4 was used to gate on CD4 + cells. (A)
Panels show CD44 plotted against forward scatter for CD4 ÷ cells
Correlation of the onset of clinical signs of E A E from CNS (top) and LN (bottom); (B) Profiles show the distribution
with CNS infiltration by autoreactive helper T cells has of CD45RB expression on CD4 + T cells from CNS (solid line) and
been well documented (Mokhtarian et el., 1984; Lyman LN (stippled line).
5. 197
I oo tiple sclerosis (Brennan et aI., 1989; Kjeldsen-Kragh et
al., 1990; Viney et al., 1990; Wucherpfennig et al.,
~ 80 1991). y-~ T cells are capable of recognizing antigens
m
expressed by oligodendrocytes and have been shown to
m 60
cause lysis of oligodendrocytes in culture (Freedman et
to
40
al., 1991; Selmaj et al., 1992). However, our flow-cyto-
metric analysis of T cells from mice with EAE revealed
Q
no significant proportions of TCRTt~-bearing T cells
a.
within the CNS. 3,-t~ T cells may play a role in chronic
o inflammation, but such conditions are distinct from the
17 23 28 55 105
early stages of EAE that we have studied.
B a y s after immunization
The CD45RB l°w phenotype defines T cells that have
Fig. 5. Proportion of CD45RB l°w CD4 + T cells isolated from the
been activated through antigen recognition (Bottomly,
CNS of EAE and remitted mice. Cells were isolated from CNS of
groups of mice at various times following immunization for EAE
1988). Reversion from CD45RB l°w to CD45RB high has
induction. The cells were double stained with PE-CD4 and anti- been shown to occur (Bell and Sparshot, 1990). One
CD45RB. 23G2 was visualized with either FITC-goat-anti-rat Ig or might predict that T cells in remitted animals would
FITC-streptavidin. Levels of CD45RB expression were defined by not express the CD45RB l°w phenotype, as a conse-
correspondence to the two populations in Fig. 3B. Each histogram
quence either of downregulation by regulatory cells,
shows a separate experiment.
a n d / o r of decreased T C R / C D 3 signalling. Remission
also might be induced by or coincide with the entry to
the CNS of regulatory cells with a naive CD45RB high
tained in the tissue or cyclically re-enter to initiate phenotype. Our results, however, demonstrate that the
a n d / o r perpetuate inflammation (Hickey et al., 1991; majority of CNS CD4 + T cells from remitted mice
Zeine and Owens., 1992). Cells of irrelevant specificity were CD45RB l°w. Indeed, four out of the five groups
were not found within 1-2 days of their entry into the represented in Fig. 4 were in remission and all con-
CNS (Hickey et al., 1991). This argues for antigen tained as high or higher proportions of CD45RB =°w
recognition as a stimulus for T cell retention in the CD4 ÷ T cells as seen at peak EAE. This does not
CNS, but does not explain loss of CD4 ÷ T cells given exclude phenotypic interconversion or immigration of
that myelin antigen concentration does not diminish. It naive cells, but since CD45RB high cells never consti-
has been proposed that suppressor or immunoregula- tuted more than 40% of the CNS T cell populations,
tory CD8 ÷ T cells, a n d / o r the secretion of inhibitory the dynamic equilibrium always favours the activated
cytokines such as TGF-/3 play a role in remission phenotype. The most important difference, however,
(Miller et al., 1991; Jiang et al., 1992; Koh et al., 1992). between mice in the active phase of EAE and remitted
Our results, however, present evidence against any mice was in the number, not in the activation state, of
increase in the number of CNS CD8 ÷ T cells during CNS T cells.
remission phases of EAE (Fig. 1). In summary, we have isolated CD2 ÷ CD45 ÷ CD3 ÷
A role for CD4 ÷ suppressor T cells in the regulation TCRa/3 + CD4 ÷ cells from the CNS of S J L / J mice
of EAE was first demonstrated by the use of suppres- during the remission phase of EAE. There were no
sor cell lines generated in vitro from recovered rats TCRT~ + cells in the CNS, and there was no increase
(Ellerman et al., 1988). Protection against EAE can be in the proportion of CD8 + T cells during remission.
passively transferred by a combination of MBP-primed We have shown a reduction in the numbers of CD4 ÷ T
B cells and a nylon wool adherent subpopulation of cells during remission, but no change in their CD44 high,
CD4 ÷ T cells isolated from recovered rats (Karpus and CD45RB =°w, memory/effector phenotype. These find-
Swanborg, 1991). CD4 + T suppressor cells isolated ings argue against downregulation of T cell function as
from recovered rats had been shown to selectively a mechanism for remission, and instead suggest overt T
inhibit the in vitro production of IFN3, by effector cells cell loss to be the cause.
from rats with EAE (Karpus and Swanborg, 1988). The
CD4 + T cells which we have shown within the CNS of
recovered mice (Fig. 3) could have included suppres- Acknowledgements
sors. However, even if this were the case, they could
not be distinguished from presumed effectors by their We thank Dr. Jia-You Lin for technical assistance
CD45R phenotype. and Dr. Philippe Poussier, at the McGill Nutrition
Elevated proportions of CD3 ÷ TcR78 have been Center in Montreal, for Provision of PE-CD8. This
observed in a number of studies at sites of inflamma- work was funded by The Multiple Sclerosis Society of
tion in some autoimmune diseases such as rheumatoid Canada. T.O. is an MRC Canada Scholar. R.Z. was
arthritis, Sjogren's syndrome, coeliac disease and mul- supported by The Multiple Sclerosis Society of Canada.
6. 198
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