16 results on '"Wallberg M"'
Search Results
2. Coagulation biomarkers and prediction of venous thromboembolism and survival in small cell lung cancer: A sub-study of RASTEN - A randomized trial with low molecular weight heparin
- Author
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Gezelius, E., primary, Flou Kristensen, A., additional, Bendahl, P. O., additional, Hisada, Y., additional, Risom Kristensen, S., additional, Ek, L., additional, Bergman, B., additional, Wallberg, M., additional, Falkmer, U., additional, Mackman, N., additional, Pedersen, S., additional, and Belting, M., additional
- Published
- 2018
- Full Text
- View/download PDF
3. Hyperglycemia does not affect antigen specific activation and cytolytic killing by CD8+ T cells in vivo
- Author
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Recino, A, Barkan, K, Ladds, G, Cooke, A, Wong, FS, Wallberg, M, Ladds, Graham [0000-0001-7320-9612], Cooke, Anne [0000-0003-3327-6081], and Apollo - University of Cambridge Repository
- Subjects
diabetes ,T-cells ,immunometabolism ,CD8-Positive T-Lymphocytes ,Lymphocyte Activation ,Diabetes Mellitus, Experimental ,Interferon-gamma ,Mice ,Oxygen Consumption ,Hyperglycemia ,Animals ,mouse models ,Antigens ,health care economics and organizations ,hyperglycaemia - Abstract
Metabolism is of central importance to T cell survival and differentiation. It is well known that T cells cannot function in the absence of glucose, but it is less clear how they respond to excessive levels of glucose. In this study we investigated how increasing levels of glucose affect T cell-mediated immune responses. We examined the effects of increased levels of glucose on CD8⁺ T cell behaviour in vitro by assessing activation and cytokine production, as well as oxygen consumption rate, extracellular acidification rate and intracellular signalling. In addition, we assessed in vivo proliferation, cytokine production and cytolytic activity of cells in chemically induced diabetic C57BL6 mice. Elevated levels of glucose in in vitro cultures had modest effects on proliferation and cytokine production, while in vivo hyperglycemia had no effect on CD8⁺ T cell proliferation, interferon gamma production or cytolytic killing., This work was supported by the NC3Rs [grant number NC/M001083/1]; the BBSRC [grant number BB/M00015X/2]; the Leverhulme Trust [grant number EM-2015-030]; the Medical Research Council [grant number G0901155]; the Diabetes U.K. [grant number BDA 09/0003840]; the BBSRC-funded Midlands Integrative Biosciences Training Partnership (MIBTP) (K.B.); and the Lollipop Foundation (M.W. as the main grant applicant and A.R. as a co-applicant).
- Published
- 2017
- Full Text
- View/download PDF
4. Activated Cdc42-associated kinase 1 (ACK1) binds the sterile α motif (SAM) domain of the adaptor SLP-76 and phosphorylates proximal tyrosines
- Author
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Thaker, YR, Recino, A, Raab, M, Jabeen, A, Wallberg, M, Fernandez, N, Rudd, CE, O'Neill, Luke A. J., and Apollo - University of Cambridge Repository
- Subjects
T-Lymphocytes ,Amino Acid Motifs ,cluster of differentiation 4 (CD4) ,non-receptor tyrosine kinase (nRTK) ,Mutation, Missense ,Receptors, Antigen, T-Cell ,T cell ,Protein-Tyrosine Kinases ,Lymphocyte Activation ,Phosphoproteins ,protein phosphorylation ,immunology ,Jurkat Cells ,Mice ,Amino Acid Substitution ,Protein Domains ,cell signaling ,Animals ,Humans ,Tyrosine ,ddc:610 ,sterile α motif (SAM) ,Phosphorylation ,Adaptor Proteins, Signal Transducing - Abstract
The adaptor protein Src homology 2 domain-containing leukocyte phosphoprotein of 76 kDa (SLP-76) plays a crucial role in T cell activation by linking antigen receptor (T cell receptor, TCR) signals to downstream pathways. At its N terminus, SLP-76 has three key tyrosines (Tyr-113, Tyr-128, and Tyr-145, "3Y") as well as a sterile α motif (SAM) domain whose function is unclear. We showed previously that the SAM domain has two binding regions that mediate dimer and oligomer formation. In this study, we have identified SAM domain-carrying non-receptor tyrosine kinase, activated Cdc42-associated tyrosine kinase 1 (ACK1; also known as Tnk2, tyrosine kinase non-receptor 2) as a novel binding partner of SLP-76. Co-precipitation, laser-scanning confocal microscopy, and in situ proximity analysis confirmed the binding of ACK1 to SLP-76. Further, the interaction was induced in response to the anti-TCR ligation and abrogated by the deletion of SLP-76 SAM domain (ΔSAM) or mutation of Tyr-113, Tyr-128, and Tyr-145 to phenylalanine (3Y3F). ACK1 induced phosphorylation of the SLP-76 N-terminal tyrosines (3Y) dependent on the SAM domain. Further, ACK1 promoted calcium flux and NFAT-AP1 promoter activity and decreased the motility of murine CD4(+) primary T cells on ICAM-1-coated plates, an event reversed by a small molecule inhibitor of ACK1 (AIM-100). These findings identify ACK1 as a novel SLP-76-associated protein-tyrosine kinase that modulates early activation events in T cells.
- Published
- 2017
5. P1.15-007 Randomized Phase III Trial of Enoxaparin in Addition to Standard Treatment in Small Cell Lung Cancer: The RASTEN Trial
- Author
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Gezelius, E., primary, Ek, L., additional, Bergman, B., additional, Bendahl, P., additional, Anderson, H., additional, Falkmer, U., additional, Verma, S., additional, Sundberg, J., additional, Wallberg, M., additional, and Belting, M., additional
- Published
- 2017
- Full Text
- View/download PDF
6. Hindering and enabling factors for young employees with common mental disorder to remain at or return to work affected by the Covid-19 pandemic - a qualitative interview study with young employees and managers.
- Author
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Wallberg M, Tinnerholm Ljungberg H, Björk Brämberg E, Nybergh L, Jensen I, and Olsson C
- Subjects
- Humans, Young Adult, Adult, Pandemics, Return to Work, Communicable Disease Control, Qualitative Research, Causality, COVID-19 epidemiology, Mental Disorders
- Abstract
Background: During the COVID-19 pandemic, changes in working life occurred, even in Sweden, where there was no general lockdown. The aim of this study was to examine how the COVID-19 pandemic was perceived as affecting the hindering and enabling factors among young employees with CMD to remain at or return to work, here as investigated from the perspective of young employees and managers., Material and Methods: A qualitative design was applied with semistructured interviews with 23 managers and 25 young employees (20-29 years old). The interviews were recorded and transcribed verbatim, and the parts of the interviews related to the aim of this article were analysed using conventional content analysis., Results: The hindering factors were changed working conditions, decreased well-being when spending more time at home, and uncertainty. The enabling factors were decreased demands, increased balance, and well-functioning work processes. For managers it is important to be aware of warning signals indicating blurred boundaries between work and private life, to create and maintain well-functioning communication, and leave room for recovery., Conclusion: The hindering and enabling factors can be described as two sides of the same coin. Changes in the working conditions during the pandemic led to difficulties for both young employees and managers when the margins of maneuver were insufficient., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Wallberg et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
- Published
- 2023
- Full Text
- View/download PDF
7. The Association Between Covariates, with Emphasis on Maternal Body Mass Index, and Duration of Exclusive and Total Breastfeeding.
- Author
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Claesson IM, Myrgård M, Wallberg M, and Blomberg M
- Subjects
- Adult, Breast Feeding statistics & numerical data, Female, Humans, Infant, Newborn, Longitudinal Studies, Mothers statistics & numerical data, Obesity epidemiology, Parity, Pregnancy, Socioeconomic Factors, Time Factors, Body Mass Index, Breast Feeding psychology, Maternal Behavior psychology, Mothers psychology, Obesity psychology
- Abstract
Objective: The aim of this study was to evaluate the association between possible covariates, with emphasis on maternal body mass index (BMI), on exclusive breastfeeding and on total breastfeeding during the first postnatal year. Design: A longitudinal study encompassing 723 women who were followed during the first postnatal year Methods: Data concerning pregnancy, delivery, neonatal period, and breastfeeding were extracted from respective medical records. Sociodemographic data on the participants were self-reported. The Cox Proportional Hazard Model was used for investigating the effects of different covariates. Results: Compared with women with BMI <25.0, obese women ran a higher risk of ceasing exclusive breastfeeding prematurely (Hazard ratio [HR] = 1.38, p = 0.009). Multiparous women had a lower risk of ceasing the exclusive breastfeeding prematurely, than primiparous women (HR = 0.78, p = 0.009). Concerning exclusive breastfeeding as well as total breastfeeding, the risk of prematurely ceasing the breastfeeding decreased with increasing age ( p = 0.028 and p ≤ 0.001, respectively). Median duration of exclusive breastfeeding was shorter among obese women compared with women with BMI <30.0 (3.0 months versus 6.0 months). Corresponding figures for total breastfeeding were 4.0 months versus 8.0 months. Concerning parity and exclusive breastfeeding, there was no difference in median duration between primiparous women and multiparous women (4.0 months), whereas multiparous women had a longer median duration of total breastfeeding, than primiparous women (8.0 months versus 7.0 months). Conclusion: The risk of ceasing exclusive breastfeeding prematurely is high among obese women. This result indicates the need for targeted supportive interventions, individualized according to BMI. With increasing age, the risk of ceasing breastfeeding prematurely decreases, and compared with primiparous women, multiparous women run a lower risk of ceasing exclusive breastfeeding prematurely.
- Published
- 2020
- Full Text
- View/download PDF
8. Pregnant women's intention to breastfeed; their estimated extent and duration of the forthcoming breastfeeding in relation to the actual breastfeeding in the first year postpartum-A Swedish cohort study.
- Author
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Claesson IM, Myrgård M, Wallberg M, and Blomberg M
- Subjects
- Adult, Analysis of Variance, Body Mass Index, Breast Feeding statistics & numerical data, Breast Feeding trends, Cohort Studies, Female, Humans, Pregnancy, Prospective Studies, Surveys and Questionnaires, Sweden, Breast Feeding psychology, Intention, Pregnant Women psychology, Time Factors
- Abstract
Objectives: To investigate the prenatal intention to breastfeed and the estimated extent and duration of the forthcoming breastfeeding among nulliparous and multiparous women in different Body Mass Index (BMI) classes. Furthermore, in a sub-group we study the actual breastfeeding in relation to the prenatal intended extent and duration., Design: A prospective cohort study., Methods: A total of 775 pregnant women answered a questionnaire concerning their intention to breastfeed and how they estimated the extent and duration of the forthcoming breastfeeding. In a sub-group of 174 women, data from the actual breastfeeding were obtained., Findings: There was no difference concerning intention to breastfeed among underweight and normal weight women, overweight or obese nulliparous or multiparous women. Fewer multiparous women with BMI <25 judged that the forthcoming breastfeeding would be partial, compared to multiparous women with overweight and obesity (p = 0.003). Furthermore, there was a significant difference within the group of nulliparous women concerning the prenatal intended extent and the actual breastfeeding at two weeks and five months postnatally (p = 0.000 and p = 0.041). There were more underweight and normal weight and overweight women who breastfed exclusively two weeks postnatally, compared with obese women. Additional, at five months postnatally there were more obese women who had ceased to breastfeed, than underweight and normal weight women., Conclusions: Among pregnant multiparous women there were more overweight and obese women who judged that the forthcoming breastfeeding would be partial, than pregnant underweight and normal weight women. The prenatal estimated extent of the forthcoming breastfeeding differed from the actually extent of breastfeeding among nulliparous women., Implications for Practice: The antenatal breastfeeding information and education should be tailored to prepare every woman/couple, irrespective of maternal body composition for the forthcoming task and furthermore, the continuum of care, from antenatal care to Child Health Service should offer a supportive atmosphere to protect and promote breastfeeding., (Copyright © 2019 Elsevier Ltd. All rights reserved.)
- Published
- 2019
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9. Immunosuppression overcomes insulin- and vector-specific immune responses that limit efficacy of AAV2/8-mediated insulin gene therapy in NOD mice.
- Author
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Recino A, Gan SU, Sia KC, Sawyer Y, Trendell J, Kay R, Gribble FM, Reimann F, Foale R, Notaridou M, Holmes N, Lever A, Lee KO, Nathwani A, Cooke A, Calne R, and Wallberg M
- Subjects
- Animals, CD4 Antigens immunology, Dependovirus genetics, Genetic Therapy methods, HEK293 Cells, Humans, Insulin genetics, Liver immunology, Male, Mice, Mice, Inbred C57BL, Mice, Inbred NOD, Mice, SCID, T-Lymphocytes immunology, Dependovirus immunology, Diabetes Mellitus, Experimental therapy, Genetic Therapy adverse effects, Immunosuppression Therapy methods, Insulin immunology
- Abstract
We report the restoration of euglycaemia in chemically induced diabetic C57BL/6 mice and spontaneously diabetic Non Obese Diabetic (NOD) mice by intravenous systemic administration of a single-stranded adeno-associated virus (ssAAV2/8) codon optimised (co) vector encoding furin cleavable human proinsulin under a liver-specific promoter. There were no immunological barriers to efficacy of insulin gene therapy in chemically induced C57BL/6 mice, which enjoyed long-lasting correction of hyperglycaemia after therapy, up to 250 days. Euglycaemia was also restored in spontaneously diabetic NOD mice, although these mice required a 7-10-fold higher dose of vector to achieve similar efficacy as the C57BL/6 mice and the immunodeficient NOD
scid mice. We detected CD8+ T cell reactivity to insulin and mild inflammatory infiltration in the livers of gene therapy recipient NOD mice, neither of which were observed in the treated C57BL/6 mice. Efficacy of the gene therapy in NOD mice was partially improved by targeting the immune system with anti-CD4 antibody treatment, while transfer of NOD mouse AAV2/8-reactive serum to recipients prevented successful restoration of euglycaemia in AAV2/8-HLP-hINSco-treated NODscid mice. Our data indicate that both immune cells and antibodies form a barrier to successful restoration of euglycaemia in autoimmune diabetic recipient mice with insulin gene therapy, but that this barrier can be overcome by increasing the dose of vector and by suppressing immune responses.- Published
- 2019
- Full Text
- View/download PDF
10. Anti-oncostatin M antibody inhibits the pro-malignant effects of oncostatin M receptor overexpression in squamous cell carcinoma.
- Author
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Kucia-Tran JA, Tulkki V, Scarpini CG, Smith S, Wallberg M, Paez-Ribes M, Araujo AM, Botthoff J, Feeney M, Hughes K, Caffarel MM, and Coleman N
- Subjects
- Animals, Autocrine Communication, Cell Line, Tumor, Female, Gene Expression Regulation, Neoplastic, Head and Neck Neoplasms immunology, Head and Neck Neoplasms metabolism, Head and Neck Neoplasms pathology, Humans, Janus Kinase 2 genetics, Janus Kinase 2 metabolism, Lung Neoplasms immunology, Lung Neoplasms metabolism, Lung Neoplasms pathology, Mice, Inbred NOD, Mice, SCID, Oncostatin M genetics, Oncostatin M metabolism, Oncostatin M Receptor beta Subunit genetics, Oncostatin M Receptor beta Subunit immunology, Oncostatin M Receptor beta Subunit metabolism, Phosphorylation, STAT3 Transcription Factor genetics, STAT3 Transcription Factor metabolism, Signal Transduction, Squamous Cell Carcinoma of Head and Neck immunology, Squamous Cell Carcinoma of Head and Neck metabolism, Squamous Cell Carcinoma of Head and Neck pathology, Suppressor of Cytokine Signaling 3 Protein genetics, Suppressor of Cytokine Signaling 3 Protein metabolism, Up-Regulation, Uterine Cervical Neoplasms immunology, Uterine Cervical Neoplasms metabolism, Uterine Cervical Neoplasms pathology, Xenograft Model Antitumor Assays, Antineoplastic Agents, Immunological pharmacology, Head and Neck Neoplasms drug therapy, Lung Neoplasms drug therapy, Oncostatin M Receptor beta Subunit antagonists & inhibitors, Squamous Cell Carcinoma of Head and Neck drug therapy, Uterine Cervical Neoplasms drug therapy
- Abstract
The oncostatin M (OSM) receptor (OSMR) shows frequent gene copy number gains and overexpression in cervical squamous cell carcinomas (SCCs), associated with adverse clinical outcomes. In SCC cells that overexpress OSMR, the major ligand OSM induces multiple pro-malignant effects, including invasion, secretion of angiogenic factors, and metastasis. Here, we demonstrate, for the first time, that OSMR overexpression in SCC cells activates cell-autonomous feed-forward signalling, via further expression of OSMR and OSM and sustained STAT3 activation, despite expression of the negative regulator suppressor of cytokine signalling 3 (SOCS3). The pro-malignant effects associated with OSMR overexpression are critically mediated by JAK-STAT3 activation, which is induced by exogenous OSM and also by autocrine OSM-OSMR interactions. Importantly, specific inhibition of OSM-OSMR interactions by neutralizing antibodies significantly inhibits STAT3 activation and feed-forward signalling, leading to reduced invasion, angiogenesis, and metastasis. Our findings are supported by data from 1254 clinical SCC samples, in which OSMR levels correlated with multiple cognate genes, including OSM, STAT3, and downstream targets. These data strongly support the development of OSM-OSMR-blocking antibodies as biologically targeted therapies against SCCs of the cervix and other anatomical sites. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd., (Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.)
- Published
- 2018
- Full Text
- View/download PDF
11. Hyperglycaemia does not affect antigen-specific activation and cytolytic killing by CD8 + T cells in vivo .
- Author
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Recino A, Barkan K, Wong FS, Ladds G, Cooke A, and Wallberg M
- Subjects
- Animals, CD8-Positive T-Lymphocytes pathology, Diabetes Mellitus, Experimental pathology, Hyperglycemia pathology, Interferon-gamma immunology, Mice, Oxygen Consumption immunology, Antigens immunology, CD8-Positive T-Lymphocytes immunology, Diabetes Mellitus, Experimental immunology, Hyperglycemia immunology, Lymphocyte Activation
- Abstract
Metabolism is of central importance for T cell survival and differentiation. It is well known that T cells cannot function in the absence of glucose, but it is less clear how they respond to excessive levels of glucose. In the present study, we investigated how increasing levels of glucose affect T-cell-mediated immune responses. We examined the effects of increased levels of glucose on CD8
+ T-cell behaviour in vitro by assessing activation and cytokine production, as well as oxygen consumption rate (OCR), extracellular acidification rate (ECAR) and intracellular signalling. In addition, we assessed in vivo proliferation, cytokine production and cytolytic activity of cells in chemically induced diabetic C57BL/6 mice. Elevated levels of glucose in in vitro cultures had modest effects on proliferation and cytokine production, while in vivo hyperglycaemia had no effect on CD8+ T-cell proliferation, interferon γ (IFNγ) production or cytolytic killing., (© 2017 The Author(s).)- Published
- 2017
- Full Text
- View/download PDF
12. Anti-CD3 treatment up-regulates programmed cell death protein-1 expression on activated effector T cells and severely impairs their inflammatory capacity.
- Author
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Wallberg M, Recino A, Phillips J, Howie D, Vienne M, Paluch C, Azuma M, Wong FS, Waldmann H, and Cooke A
- Subjects
- Animals, Female, Mice, Mice, Inbred NOD, Mice, SCID, Programmed Cell Death 1 Receptor biosynthesis, Programmed Cell Death 1 Receptor genetics, Programmed Cell Death 1 Receptor immunology, Antibodies immunology, CD3 Complex immunology, Inflammation immunology, Programmed Cell Death 1 Receptor metabolism, T-Lymphocytes immunology, T-Lymphocytes metabolism, Up-Regulation
- Abstract
T cells play a key role in the pathogenesis of type 1 diabetes, and targeting the CD3 component of the T-cell receptor complex provides one therapeutic approach. Anti-CD3 treatment can reverse overt disease in spontaneously diabetic non-obese diabetic mice, an effect proposed to, at least in part, be caused by a selective depletion of pathogenic cells. We have used a transfer model to further investigate the effects of anti-CD3 treatment on green fluorescent protein (GFP)
+ islet-specific effector T cells in vivo. The GFP expression allowed us to isolate the known effectors at different time-points during treatment to assess cell presence in various organs as well as gene expression and cytokine production. We find, in this model, that anti-CD3 treatment does not preferentially deplete the transferred effector cells, but instead inhibits their metabolic function and their production of interferon-γ. Programmed cell death protein 1 (PD-1) expression was up-regulated on the effector cells from anti-CD3-treated mice, and diabetes induced through anti-PD-L1 antibody could only be reversed with anti-CD3 antibody if the anti-CD3 treatment lasted beyond the point when the anti-PD-L1 antibody was washed out of the system. This suggests that PD-1/PD-L1 interaction plays an important role in the anti-CD3 antibody mediated protection. Our data demonstrate an additional mechanism by which anti-CD3 therapy can reverse diabetogenesis., (© 2017 The Authors. Immunology Published by John Wiley & Sons Ltd.)- Published
- 2017
- Full Text
- View/download PDF
13. Activated Cdc42-associated kinase 1 (ACK1) binds the sterile α motif (SAM) domain of the adaptor SLP-76 and phosphorylates proximal tyrosines.
- Author
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Thaker YR, Recino A, Raab M, Jabeen A, Wallberg M, Fernandez N, and Rudd CE
- Subjects
- Adaptor Proteins, Signal Transducing genetics, Adaptor Proteins, Signal Transducing immunology, Amino Acid Motifs, Amino Acid Substitution, Animals, Humans, Jurkat Cells, Mice, Mutation, Missense, Phosphoproteins genetics, Phosphoproteins immunology, Phosphorylation physiology, Protein Domains, Protein-Tyrosine Kinases genetics, Protein-Tyrosine Kinases immunology, Receptors, Antigen, T-Cell genetics, Receptors, Antigen, T-Cell immunology, T-Lymphocytes immunology, Tyrosine, Adaptor Proteins, Signal Transducing metabolism, Lymphocyte Activation physiology, Phosphoproteins metabolism, Protein-Tyrosine Kinases metabolism, Receptors, Antigen, T-Cell metabolism, T-Lymphocytes metabolism
- Abstract
The adaptor protein Src homology 2 domain-containing leukocyte phosphoprotein of 76 kDa (SLP-76) plays a crucial role in T cell activation by linking antigen receptor (T cell receptor, TCR) signals to downstream pathways. At its N terminus, SLP-76 has three key tyrosines (Tyr-113, Tyr-128, and Tyr-145, "3Y") as well as a sterile α motif (SAM) domain whose function is unclear. We showed previously that the SAM domain has two binding regions that mediate dimer and oligomer formation. In this study, we have identified SAM domain-carrying non-receptor tyrosine kinase, activated Cdc42-associated tyrosine kinase 1 (ACK1; also known as Tnk2, tyrosine kinase non-receptor 2) as a novel binding partner of SLP-76. Co-precipitation, laser-scanning confocal microscopy, and in situ proximity analysis confirmed the binding of ACK1 to SLP-76. Further, the interaction was induced in response to the anti-TCR ligation and abrogated by the deletion of SLP-76 SAM domain (ΔSAM) or mutation of Tyr-113, Tyr-128, and Tyr-145 to phenylalanine (3Y3F). ACK1 induced phosphorylation of the SLP-76 N-terminal tyrosines (3Y) dependent on the SAM domain. Further, ACK1 promoted calcium flux and NFAT-AP1 promoter activity and decreased the motility of murine CD4
+ primary T cells on ICAM-1-coated plates, an event reversed by a small molecule inhibitor of ACK1 (AIM-100). These findings identify ACK1 as a novel SLP-76-associated protein-tyrosine kinase that modulates early activation events in T cells., (© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.)- Published
- 2017
- Full Text
- View/download PDF
14. Promoter optimisation of lentiviral vectors for efficient insulin gene expression in canine mesenchymal stromal cells: potential surrogate beta cells.
- Author
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Gautam P, Recino A, Foale RD, Zhao J, Gan SU, Wallberg M, Calne R, and Lever AM
- Subjects
- Animals, Bone Marrow Cells metabolism, Cell Line, Tumor, Cells, Cultured, Dogs, Gene Transfer Techniques, Genetic Therapy methods, Genetic Vectors genetics, HEK293 Cells, Hepatocytes metabolism, Humans, Insulin metabolism, Insulin-Secreting Cells metabolism, Proinsulin genetics, Proinsulin metabolism, Gene Expression, Insulin genetics, Lentivirus genetics, Mesenchymal Stem Cells metabolism, Promoter Regions, Genetic genetics
- Abstract
Background: The lack of an ideal cell type that can be easily acquired, modified to produce insulin, and re-implanted has been a limitation for ex vivo insulin gene therapy. Canine diabetes is currently treated with human insulin and is a good model for human diabetes. Mesenchymal stromal cells (MSCs) are a promising candidate cell type for gene therapy. In the present study, we optimised insulin production using lentiviral transduced canine MSCs (cMSCs), aiming to evaluate their ability for use as surrogate beta cells., Methods: Canine MSCs were derived from bone marrow and validated by measuring the expression of MSC lineage specific markers. Lentivirus vectors encoding the proinsulin gene (with or without a Kozak sequence) under the control of spleen focus forming virus, cytomegalovirus, elongation factor 1α and simian virus 40 promotors were generated and used to transduce primary cMSCs and a hepatocyte cell line. The insulin-producing capacity of transduced primary cMSCs was assessed by measuring the concentration of C-peptide produced., Results: Primary cMSC could be readily expanded in culture and efficiently transduced using lentiviral vectors encoding proinsulin. Increasing the multiplicity of infection from 3 to 20 led to an increase in C-peptide secretion (from 1700 to 4000 pmol/l). The spleen focus forming virus promoter conferred the strongest transcriptional ability., Conclusions: The results of the present study suggest that optimised lentiviral transduction of the insulin gene into primary cMSCs renders these cells capable of secreting insulin over both the short- and long-term, in sufficient quantities in vitro to support their potential use in insulin gene therapy., (© 2016 The Authors. The Journal of Gene Medicine Published by John Wiley & Sons, Ltd.)
- Published
- 2016
- Full Text
- View/download PDF
15. Inhibition of Phosphoinositide 3-Kinase p110delta Does Not Affect T Cell Driven Development of Type 1 Diabetes Despite Significant Effects on Cytokine Production.
- Author
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Barbera Betancourt A, Emery JL, Recino A, Wong FS, Cooke A, Okkenhaug K, and Wallberg M
- Subjects
- Adenine pharmacology, Animals, Cell Differentiation immunology, Cells, Cultured, Class I Phosphatidylinositol 3-Kinases, Diabetes Mellitus, Experimental genetics, Diabetes Mellitus, Experimental immunology, Diabetes Mellitus, Experimental metabolism, Diabetes Mellitus, Type 1 genetics, Diabetes Mellitus, Type 1 metabolism, Female, Mice, Mice, Inbred C57BL, Mice, Inbred CBA, Mice, Inbred NOD, Mice, SCID, Mice, Transgenic, Phosphatidylinositol 3-Kinases genetics, Phosphatidylinositol 3-Kinases metabolism, T-Lymphocytes physiology, Adenine analogs & derivatives, Cell Differentiation drug effects, Cytokines biosynthesis, Diabetes Mellitus, Type 1 immunology, Phosphoinositide-3 Kinase Inhibitors, Quinazolines pharmacology, T-Lymphocytes drug effects
- Abstract
Type 1 diabetes is caused by the destruction of insulin producing beta cells by the immune system. The p110δ isoform of PI3K is expressed primarily in cells of haematopoietic origin and the catalytic activity of p110δ is important for the activation of these cells. Targeting of this pathway offers an opportunity to reduce immune cell activity without unwanted side effects. We have explored the effects of a specific p110δ isoform inhibitor, IC87114, on diabetogenic T cells both in vitro and in vivo, and find that although pharmacological inhibition of p110δ has a considerable impact on the production of pro-inflammatory cytokines, it does not delay the onset of diabetes after adoptive transfer of diabetogenic cells. Further, we demonstrate that combination treatment with CTLA4-Ig does not improve the efficacy of treatment, but instead attenuates the protective effects seen with CTLA4-Ig treatment alone. Our results suggest that decreased IL-10 production by Foxp3+ CD4+ T cells in the presence of IC87114 negates individual anti-inflammatory effects of IC8114 and CTLA4-Ig.
- Published
- 2016
- Full Text
- View/download PDF
16. User-driven development of a web-based tool for patient reporting of drug-related harm.
- Author
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Plöen M, Wallberg M, and Olsson S
- Subjects
- European Union, Humans, Internationality, Legislation, Drug, Pharmacovigilance, Pilot Projects, Self Report, Time Factors, Adverse Drug Reaction Reporting Systems organization & administration, Community Participation methods, Drug-Related Side Effects and Adverse Reactions epidemiology, Internet
- Abstract
Commissioned by the Monitoring Medicines project, the Uppsala Monitoring Centre (UMC) led the design and development of a web-based ADR (adverse drug reaction) reporting tool intended for use by patients. The software design was undertaken in close collaboration with representatives of national pharmacovigilance centres (NPCs) and with patient and consumer organizations. The web-based tool was developed by these participants through several telephone conferences, a workshop and site testing. The tool is directly compatible with the UMC's Individual Case Safety Report (ICSR) data management system VigiFlow(®) and is also compliant with the ICH-E2B(R2) format. The UMC team benefited by working closely with the end-users during the development process. A major challenge was to balance the need for detailed information required by the NPCs to be able to assess reports with the amount of detail patients are able and willing to provide. Needs, ideas and suggestions from the end users were valuable and were taken into account throughout the process of designing the tool.
- Published
- 2015
- Full Text
- View/download PDF
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