المؤلفون: Kathrin Balz, Magali Grange, Uta Pegel, Zain A. Karamya, Marielle Mello, Xiaoying Zhou, Thilo Berger, Konstantin Bloch, Diane Dunham, Sharon Chinthrajah, Kari Nadeau, Hervé Luche, Chrysanthi Skevaki

المصدر: Frontiers in Cellular and Infection Microbiology, Vol 13 (2024)

مصطلحات موضوعية: T-cell activation, CyTOF, sample barcoding, T-cell stimulation assay, method, Microbiology, QR1-502

الوصف: Understanding antigen-specific T-cell responses, for example, following virus infections or allergen exposure, is of high relevance for the development of vaccines and therapeutics. We aimed on optimizing immunophenotyping of T cells after antigen stimulation by improving staining procedures for flow and mass cytometry. Our method can be used for primary cells of both mouse and human origin for the detection of low-frequency T-cell response using a dual-barcoding system for individual samples and conditions. First, live-cell barcoding was performed using anti-CD45 antibodies prior to an in vitro T-cell stimulation assay. Second, to discriminate between stimulation conditions and prevent cell loss, sample barcoding was combined with a commercial barcoding solution. This dual-barcoding approach is cell sparing and, therefore, particularly relevant for samples with low cell numbers. To further reduce cell loss and to increase debarcoding efficiency of multiplexed samples, we combined our dual-barcoding approach with a new centrifugation-free washing system by laminar flow (Curiox™). Finally, to demonstrate the benefits of our established protocol, we assayed virus-specific T-cell response in SARS-CoV-2–vaccinated and SARS-CoV-2–infected patients and compared with healthy non-exposed individuals by a high-parameter CyTOF analysis. We could reveal a heterogeneity of phenotypes among responding CD4, CD8, and gd-T cells following antigen-specific stimulations. Our protocol allows to assay antigen-specific responses of minute populations of T cells to virus-derived peptides, allergens, or other antigens from the same donor sample, in order to investigate qualitative and quantitative differences.

وصف الملف: electronic resource

العلاقة: https://www.frontiersin.org/articles/10.3389/fcimb.2023.1336489/fullTest; https://doaj.org/toc/2235-2988Test

الوصول الحر: https://doaj.org/article/cd666241a2b54fa2b38d3d9aa950af30Test

المؤلفون: Konstantin Bloch, Shay Henry Hornfeld, Shira Dar, Alexey Vanichkin, Irit Gil-Ad, Pnina Vardi, Abraham Weizman

المصدر: PLoS ONE, Vol 15, Iss 1, p e0227879 (2020)

مصطلحات موضوعية: Medicine, Science

الوصف: Accumulating evidence suggests that Alzheimer's disease is associated with brain insulin resistance, as are some other types of dementia. Intranasal insulin administration has been suggested as a potential approach to overcoming brain insulin resistance and improving cognitive functions. Islet transplantation into the cranial subarachnoid cavity was used as an alternative route for insulin delivery into the brain. Recently, the authors showed the short-term beneficial cognitive effect of a small number of intracranially grafted islets in rats with cognitive dysfunction induced by intracerebroventricular administration of streptozotocin (icv-STZ). This was associated with continuous and safe insulin delivery to the rat brain. The current study investigated the long-term effect of intracranial grafting of islets on cognitive functioning in icv-STZ rats. Severe dementia, associated with obesity and cerebral amyloid-β angiopathy, was induced in Lewis inbred rats by icv-STZ. Two months after icv-STZ, one hundred syngeneic islets were transplanted into the cranial subarachnoid space. Two and six months later, cognitive alterations were assessed by Morris water-maze tests. Islet graft survival was evaluated by immunohistochemical and biochemical assays. Improvement was found in spatial learning and memory of grafted rats as opposed to the sham-operated icv-STZ rats. The grafted islets showed intact morphology, intensive expression of insulin, glucagon and glucose transporter 2. Normoglycemic obesity and cerebral amyloid-β angiopathy were found in both grafted and sham-operated icv-STZ rats. In conclusion, islet grafting into cranial subarachnoid space provides an efficient and safe approach for insulin delivery to the brain, leading to a long-term attenuation of icv-STZ-induced cognitive dysfunction.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/1932-6203Test

الوصول الحر: https://doaj.org/article/6b21641591c6463ebcc010eae6f955dbTest

المؤلفون: Uriel Barkai Ph.D., Gordon C. Weir, Clark K. Colton, Barbara Ludwig, Stefan R. Bornstein, Mathias D. Brendel, Tova Neufeld, Chezi Bremer, Assaf Leon, Yoav Evron, Karina Yavriyants, Dimitri Azarov, Baruch Zimermann, Shiri Maimon, Noa Shabtay, Maria Balyura, Tania Rozenshtein, Pnina Vardi, Konstantin Bloch, Paul De Vos, Avi Rotem

المصدر: Cell Transplantation, Vol 22 (2013)

مصطلحات موضوعية: Medicine

الوصف: The current epidemic of diabetes with its overwhelming burden on our healthcare system requires better therapeutic strategies. Here we present a promising novel approach for a curative strategy that may be accessible for all insulin-dependent diabetes patients. We designed a subcutaneous implantable bioartificial pancreas (BAP)—the “β-Air”—that is able to overcome critical challenges in current clinical islet transplantation protocols: adequate oxygen supply to the graft and protection of donor islets against the host immune system. The system consists of islets of Langerhans immobilized in an alginate hydrogel, a gas chamber, a gas permeable membrane, an external membrane, and a mechanical support. The minimally invasive implantable device, refueled with oxygen via subdermally implanted access ports, completely normalized diabetic indicators of glycemic control (blood glucose intravenous glucose tolerance test and HbA1c) in streptozotocin-induced diabetic rats for periods up to 6 months. The functionality of the device was dependent on oxygen supply to the device as the grafts failed when oxygen supply was ceased. In addition, we showed that the device is immunoprotective as it allowed for survival of not only isografts but also of allografts. Histological examination of the explanted devices demonstrated morphologically and functionally intact islets; the surrounding tissue was without signs of inflammation and showed visual evidence of vasculature at the site of implantation. Further increase in islets loading density will justify the translation of the system to clinical trials, opening up the potential for a novel approach in diabetes therapy.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/0963-6897Test; https://doaj.org/toc/1555-3892Test

الوصول الحر: https://doaj.org/article/16b3b5a5d3cc465aa8cfdf841bc10acfTest

المؤلفون: Kathrin Balz, Magali Grange, Uta Pegel, Zain A. Karamya, Marielle Mello, Xiaoying Zhou, Thilo Berger, Konstantin Bloch, Diane Dunham, Sharon Chinthrajah, Kari Nadeau, Hervé Luche, Chrysanthi Skevaki

مصطلحات موضوعية: Clinical Microbiology, Medical Bacteriology, Medical Infection Agents (incl. Prions), Medical Virology, T-cell activation, CyTOF, sample barcoding, T-cell stimulation assay, method

الوصف: Understanding antigen-specific T-cell responses, for example, following virus infections or allergen exposure, is of high relevance for the development of vaccines and therapeutics. We aimed on optimizing immunophenotyping of T cells after antigen stimulation by improving staining procedures for flow and mass cytometry. Our method can be used for primary cells of both mouse and human origin for the detection of low-frequency T-cell response using a dual-barcoding system for individual samples and conditions. First, live-cell barcoding was performed using anti-CD45 antibodies prior to an in vitro T-cell stimulation assay. Second, to discriminate between stimulation conditions and prevent cell loss, sample barcoding was combined with a commercial barcoding solution. This dual-barcoding approach is cell sparing and, therefore, particularly relevant for samples with low cell numbers. To further reduce cell loss and to increase debarcoding efficiency of multiplexed samples, we combined our dual-barcoding approach with a new centrifugation-free washing system by laminar flow (Curiox™). Finally, to demonstrate the benefits of our established protocol, we assayed virus-specific T-cell response in SARS-CoV-2–vaccinated and SARS-CoV-2–infected patients and compared with healthy non-exposed individuals by a high-parameter CyTOF analysis. We could reveal a heterogeneity of phenotypes among responding CD4, CD8, and gd-T cells following antigen-specific stimulations. Our protocol allows to assay antigen-specific responses of minute populations of T cells to virus-derived peptides, allergens, or other antigens from the same donor sample, in order to investigate qualitative and quantitative differences.

العلاقة: https://figshare.com/articles/figure/Image_2_A_novel_mass_cytometry_protocol_optimized_for_immunophenotyping_of_low-frequency_antigen-specific_T_cells_jpeg/24997655Test

الإتاحة: https://doi.org/10.3389/fcimb.2023.1336489.s002Test
https://figshare.com/articles/figure/Image_2_A_novel_mass_cytometry_protocol_optimized_for_immunophenotyping_of_low-frequency_antigen-specific_T_cells_jpeg/24997655Test

المؤلفون: Shay Henry Hornfeld, Alexey Vanichkin, Irit Gil-Ad, Abraham Weizman, Konstantin Bloch, Pnina Vardi, Shira Dar

المصدر: PLoS ONE, Vol 15, Iss 1, p e0227879 (2020)
PLoS ONE

مصطلحات موضوعية: 0301 basic medicine, endocrine system diseases, Physiology, medicine.medical_treatment, Islets of Langerhans Transplantation, Alzheimer's Disease, Hippocampus, Biochemistry, Nervous System, 0302 clinical medicine, Cerebrospinal fluid, Cognition, Endocrinology, Medicine and Health Sciences, Insulin, Enzyme-Linked Immunoassays, Cerebrospinal Fluid, Cognitive Impairment, Multidisciplinary, geography.geographical_feature_category, Cognitive Neurology, Brain, Neurodegenerative Diseases, Islet, Body Fluids, Neurology, Physiological Parameters, Medicine, Anatomy, medicine.drug, Research Article, medicine.medical_specialty, Cognitive Neuroscience, Science, Research and Analysis Methods, Angiopathy, 03 medical and health sciences, Islets of Langerhans, Insulin resistance, Alzheimer Disease, Memory, Internal medicine, Mental Health and Psychiatry, medicine, Dementia, Animals, Humans, Obesity, Maze Learning, Immunoassays, Diabetic Endocrinology, geography, business.industry, Body Weight, Biology and Life Sciences, medicine.disease, Streptozotocin, Animal Cognition, Hormones, Rats, Transplantation, Disease Models, Animal, 030104 developmental biology, Immunologic Techniques, Cognitive Science, business, Cognition Disorders, Zoology, 030217 neurology & neurosurgery, Neuroscience

الوصف: Accumulating evidence suggests that Alzheimer's disease is associated with brain insulin resistance, as are some other types of dementia. Intranasal insulin administration has been suggested as a potential approach to overcoming brain insulin resistance and improving cognitive functions. Islet transplantation into the cranial subarachnoid cavity was used as an alternative route for insulin delivery into the brain. Recently, the authors showed the short-term beneficial cognitive effect of a small number of intracranially grafted islets in rats with cognitive dysfunction induced by intracerebroventricular administration of streptozotocin (icv-STZ). This was associated with continuous and safe insulin delivery to the rat brain. The current study investigated the long-term effect of intracranial grafting of islets on cognitive functioning in icv-STZ rats. Severe dementia, associated with obesity and cerebral amyloid-β angiopathy, was induced in Lewis inbred rats by icv-STZ. Two months after icv-STZ, one hundred syngeneic islets were transplanted into the cranial subarachnoid space. Two and six months later, cognitive alterations were assessed by Morris water-maze tests. Islet graft survival was evaluated by immunohistochemical and biochemical assays. Improvement was found in spatial learning and memory of grafted rats as opposed to the sham-operated icv-STZ rats. The grafted islets showed intact morphology, intensive expression of insulin, glucagon and glucose transporter 2. Normoglycemic obesity and cerebral amyloid-β angiopathy were found in both grafted and sham-operated icv-STZ rats. In conclusion, islet grafting into cranial subarachnoid space provides an efficient and safe approach for insulin delivery to the brain, leading to a long-term attenuation of icv-STZ-induced cognitive dysfunction.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::750f98b7637bce3abbe0dd9e0eb7cbd9Test
https://doaj.org/article/6b21641591c6463ebcc010eae6f955dbTest

المؤلفون: Abraham Weizman, Alexey Vanichkin, Pnina Vardi, Michal Taler, Shay Henry Hornfeld, Nickolay Koroukhov, Irit Gil-Ad, Konstantin Bloch

المصدر: Journal of Alzheimer's Disease. 60:121-136

مصطلحات موضوعية: Blood Glucose, Male, 0301 basic medicine, endocrine system diseases, Morris water navigation task, 0302 clinical medicine, Glucose homeostasis, Cognitive decline, Antibiotics, Antineoplastic, General Neuroscience, Leptin, digestive, oral, and skin physiology, Fatty liver, Brain, General Medicine, Psychiatry and Mental health, Clinical Psychology, Liver, Encephalitis, Locomotion, hormones, hormone substitutes, and hormone antagonists, medicine.drug, medicine.medical_specialty, Streptozocin, 03 medical and health sciences, Internal medicine, medicine, Animals, Dementia, Obesity, Maze Learning, Pancreas, Amyloid beta-Peptides, Adiponectin, business.industry, Body Weight, medicine.disease, Streptozotocin, Hormones, Rats, Disease Models, Animal, 030104 developmental biology, Endocrinology, nervous system, Rats, Inbred Lew, Exploratory Behavior, Geriatrics and Gerontology, business, 030217 neurology & neurosurgery

الوصف: Background Animal models of dementia associated with metabolic abnormalities play an important role in understanding the bidirectional relationships between these pathologies. Rodent strains develop cognitive dysfunctions without alteration of peripheral metabolism following intracerebroventricular administration of streptozotocin (icv-STZ). Objective We aimed to estimate the effect of icv-STZ on cognitive functions and peripheral metabolism in Lewis rats, which are rarely used for the induction of cognitive abnormalities. Methods Inbred adult Lewis rats were treated with single icv-STZ (3 mg/kg). Cognitive functions were assessed using Morris water maze (MWM) test and locomotion by the Open Field test. Metabolic alterations were studied using histological and biochemical analysis of brain and peripheral tissues. Results The icv-STZ induced rapid weight decline during the first two weeks. Thereafter, the rats showed an accelerated weight gain. Three months after the icv-STZ treatment, the rats were severely obese and revealed fatty liver, pancreatic islet hypertrophy, significantly elevated levels of blood insulin, leptin, and adiponectin, but intact peripheral glucose homeostasis. The icv-STZ rats expressed amyloid-β deposits in blood vessels of leptomeningeal area, microgliosis, astrogliosis, and spongiosis in fimbria-fornix area of hippocampus. Locomotor activities of icv-STZ treated and sham-operated rats were similar. In the MWM test, the icv-STZ treated rats demonstrated severely impaired spatial learning during both acquisition and reversal phases. Conclusions Icv-STZ treated Lewis rats develop severe dementia associated with obesity and peripheral metabolic abnormalities. This animal model may be useful for exploring the pathophysiological relationship between obesity and dementia and provides a new tool for development of effective therapy.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::0e79fd4efde4b8a0514bb81c7b4a69c9Test
https://doi.org/10.3233/jad-161289Test

المؤلفون: Abraham Weizman, Alexey Vanichkin, Irit Gil-Ad, Konstantin Bloch, Pnina Vardi

المصدر: Journal of Tissue Engineering and Regenerative Medicine. 11:3263-3272

مصطلحات موضوعية: 0301 basic medicine, medicine.medical_specialty, medicine.medical_treatment, Biomedical Engineering, Medicine (miscellaneous), Biomaterials, 03 medical and health sciences, 0302 clinical medicine, Insulin resistance, Internal medicine, medicine, Glucose homeostasis, geography, geography.geographical_feature_category, business.industry, Insulin, Pancreatic islets, medicine.disease, Islet, Transplantation, 030104 developmental biology, Endocrinology, medicine.anatomical_structure, Nasal administration, business, 030217 neurology & neurosurgery, Hormone

الوصف: There is increasing evidence supporting a link between cognitive dysfunctions and impaired brain insulin signalling. Insulin therapy has previously been tested as an approach to ameliorate brain insulin resistance and deficiency in patients with various brain disorders. However, current strategies for insulin delivery to the brain may induce severe hypoglycaemia when injected peripherally or show poor uptake when delivered intranasally. Recently, we have shown that intracranial transplantation of naked pancreatic islets increased insulin content in the brain and attenuated cognitive dysfunctions without altering peripheral glucose homeostasis in rats with schizophrenia-like syndrome. In this study, we show that intracranial implantation of 50 pancreatic islets encapsulated in disc-shaped alginate is sufficient to elevate insulin content in the rat brain. Three weeks after implantation, the islets displayed intact morphology, intensive hormone staining and glucose-sensitive insulin release. The ultrapure alginate with high guluronic acid content used for islet encapsulation demonstrated good biocompatibility and stability after intracranial implantation. All implanted animals were normoglycaemic and normoinsulinaemic. In conclusion, the intracranial implantation of a small amount of alginate-encapsulated islets is an efficient tool for metabolically regulated insulin delivery to the brain. Copyright © 2017 John Wiley & Sons, Ltd.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::606258becda71f090579681cdffd625dTest
https://doi.org/10.1002/term.2235Test

المؤلفون: Alexey Vanichkin, Shira Dar, Irit Gil-Ad, Konstantin Bloch, Pnina Vardi, Abraham Weizman

المصدر: Transplantation. 103(6)

مصطلحات موضوعية: Transplantation, Pathology, medicine.medical_specialty, Time Factors, Behavior, Animal, business.industry, Pancreatic islets, Islets of Langerhans Transplantation, Brain lateral ventricles, Cognition, Tropism, Streptozocin, Disease Models, Animal, Islets of Langerhans, medicine.anatomical_structure, Alzheimer Disease, Lateral Ventricles, medicine, Animals, business

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::df2147ab7ef4d5ef4fa8d9e2fd018896Test
https://pubmed.ncbi.nlm.nih.gov/30801525Test

المؤلفون: Konstantin Bloch, Irit Gil-Ad, Alexey Vanichkin, Dmitry Azarov, Michal Taler, Pnina Vardi, Abraham Weizman, Shay Henry Hornfeld, Shira Dar

المصدر: Journal of Alzheimer's disease : JAD. 65(4)

مصطلحات موضوعية: 0301 basic medicine, Blood Glucose, Male, endocrine system diseases, medicine.medical_treatment, Islets of Langerhans Transplantation, Eating, 0302 clinical medicine, Medicine, Glucose homeostasis, Cognitive decline, General Neuroscience, digestive, oral, and skin physiology, Brain, General Medicine, Psychiatry and Mental health, Clinical Psychology, medicine.anatomical_structure, hormones, hormone substitutes, and hormone antagonists, medicine.drug, medicine.medical_specialty, Drinking, Streptozocin, 03 medical and health sciences, Insulin resistance, Metabolic Diseases, Alzheimer Disease, Internal medicine, Animals, Rats, Wistar, Pancreas, business.industry, Insulin, Pancreatic islets, Body Weight, Streptozotocin, medicine.disease, Rats, Transplantation, Disease Models, Animal, 030104 developmental biology, Endocrinology, Metabolism, Rats, Inbred Lew, Pancreatic islet transplantation, Geriatrics and Gerontology, business, Cognition Disorders, 030217 neurology & neurosurgery, Follow-Up Studies

الوصف: Background Alzheimer's disease (AD) is often associated with brain insulin resistance and peripheral metabolic dysfunctions. Recently, we developed a model of sporadic AD associated with obesity-related peripheral metabolic abnormalities in Lewis rats using intracerebroventricular administration of streptozotocin (icv-STZ). Objective We aimed to assess the effect of intracranially grafted pancreatic islets on cognitive and peripheral metabolic dysfunctions in the icv-STZ Lewis rats. Methods AD-like dementia associated with obesity was induced in inbred Lewis rats using a single icv-STZ. Two months after icv-STZ, syngeneic islets (100 islets per recipient) were implanted in the cranial subarachnoid cavity of icv-STZ rats. Morris water maze and marble burying tests were used for studying cognitive and behavioral functions. Central and peripheral metabolic alterations were assessed by histological and biochemical assays. Results The icv-STZ induced increases in food intake, body weight, and blood levels of insulin and leptin without alteration of glucose homeostasis. Grafted islets reduced body weight gain, food consumption, peripheral insulin resistance, and hyperleptinemia. Biochemical and histological analysis of the brain revealed viable grafted islets expressing insulin and glucagon. The grafted islets did not affect expression of brain insulin receptors and peripheral glucose homeostasis. Two months after islet transplantation, cognitive and behavioral functioning in transplanted rats were significantly better than the sham-operated icv-STZ rats. No significant differences in the locomotor activity between transplanted and non-transplanted icv-STZ rats were found. Conclusions Intracranial islet transplantation attenuates cognitive decline and peripheral metabolic dysfunctions providing a novel therapeutic approach for sporadic AD associated with peripheral metabolic dysfunctions.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::bd8961dd5b2742328bae5b574d4585d0Test
https://pubmed.ncbi.nlm.nih.gov/30175977Test

المؤلفون: Konstantin Bloch, Pnina Vardi, Daniel Lazard

المصدر: Diabetes/Metabolism Research and Reviews. 32:11-20

مصطلحات موضوعية: 0301 basic medicine, endocrine system, medicine.medical_specialty, endocrine system diseases, Endocrinology, Diabetes and Metabolism, medicine.medical_treatment, Central nervous system, Neuroprotection, 03 medical and health sciences, 0302 clinical medicine, Endocrinology, Diabetes mellitus, Internal medicine, Internal Medicine, medicine, geography, geography.geographical_feature_category, business.industry, Insulin, Pancreatic islets, Islet, medicine.disease, Transplantation, 030104 developmental biology, medicine.anatomical_structure, Pancreatic islet transplantation, business, 030217 neurology & neurosurgery

الوصف: During the last decades, the central nervous system (CNS) was intensively tested as a site for islet transplantation in different animal models of diabetes. Immunoprivilege properties of intracranial and intrathecal sites were found to delay and reduce rejection of transplanted allo-islets and xeno-islets, especially in the form of dispersed single cells. Insulin released from islets grafted in CNS was shown to cross the blood-brain barrier and to act as a regulator of peripheral glucose metabolism. In diabetic animals, sufficient nutrition and oxygen supply to islets grafted in the CNS provide adequate insulin response to increase glucose level resulting in rapid normoglycemia. In addition to insulin, pancreatic islets produce and secrete several other hormones, as well as neurotrophic and angiogenic factors with potential neuroprotective properties. Recent experimental studies and clinical trials provide a strong support for delivery of islet-derived macromolecules to CNS as a promising strategy to treat various brain disorders. This review article focuses mainly on analysis of current status of intracranial and intrathecal islet transplantations for treatment of experimental diabetes and discusses the possible neuroprotective properties of grafted islets into CNS as a novel therapeutic approach to brain disorders with cognitive dysfunctions characterized by impaired brain insulin signalling. Copyright © 2015 John Wiley & Sons, Ltd.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::bf20c1788739682dae486d8282b86dacTest
https://doi.org/10.1002/dmrr.2644Test