62 results on '"Zimmermann VS"'
Search Results
2. Requirement for dendritic cells in the establishment of anti-phospholipid antibodies
- Author
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BONDANZA, ATTILIO, ROVERE QUERINI , PATRIZIA, Zimmermann VS, Balestrieri G, Tincani A, Sabbadini MG, MANFREDI , ANGELO ANDREA M. A., Bondanza, Attilio, ROVERE QUERINI, Patrizia, Zimmermann, V, Balestrieri, G, Tincani, A, Sabbadini, Mg, and Manfredi, ANGELO ANDREA M. A.
- Published
- 2007
3. TNF-alpha coupled to membrane of apoptotic cells favors the cross-priming to melanoma antigens
- Author
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Zimmermann VS, BONDANZA , ATTILIO, Monno A, ROVERE QUERINI , PATRIZIA, CORTI , ANGELO, MANFREDI , ANGELO ANDREA M. A., Zimmermann, V, Bondanza, Attilio, Monno, A, ROVERE QUERINI, Patrizia, Corti, Angelo, and Manfredi, ANGELO ANDREA M. A.
- Abstract
The cross-presentation of Ags derived from apoptotic cell processing contributes to peripheral tolerance. Environmental signals possibly modify this default outcome, favoring cross-priming. In this study, we anchored via a biotin-avidin-biotin bridge soluble TNF-alpha to the membrane of apoptotic melanoma cells and studied in vivo and in vitro the interaction with Ag-presenting phagocytes. TNF-alpha-coated apoptotic melanoma cells injected s.c. had a faster and more efficient access to draining lymph nodes, with cross-priming of melanoma-specific CTLs and delayed outgrowth of melanomas in all treated animals. Twenty. percent of the animals, in the absence of further adjuvant, did not develop the tumor. Immature dendritic cells challenged with TNF-alpha-coated apoptotic melanoma cells secreted proinflammatory cytokines in an autocrine/paracrine fashion, efficiently matured, as assessed functionally and by flow cytometry and cross-presented with enhanced efficiency melanoma Ags to MHC class I- and II-restricted T cells. The results indicate that TNF-alpha targeted to apoptotic membranes, at concentrations that can be safely reached in growing tumors without undue systemic toxicity, influences the outcome of the disposal of dying cells and enhances tumor immunogenicity.
- Published
- 2004
4. Dissociation between autoimmune response and clinical disease after vaccination with dendritic cells
- Author
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BONDANZA , ATTILIO, Zimmermann VS, Dell'Antonio G, Dal Cin E, Capobianco A, Sabbadini MG, MANFREDI, ANGELO ANDREA M. A., ROVERE QUERINI , PATRIZIA, Bondanza, Attilio, Zimmermann, V, Dell'Antonio, G, Dal Cin, E, Capobianco, A, Sabbadini, Mg, Manfredi, ANGELO ANDREA M. A., and ROVERE QUERINI, Patrizia
- Published
- 2003
5. CUTTING EDGE: Dissociation between autoimmune response and clinical disease after vaccination with dendritic cells
- Author
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BONDANZA A, ZIMMERMANN VS, DELLANTONIO G, DAL CIN E, CAPOBIANCO A., SABBADINI MG, ROVERE QUERINI, PATRIZIA, MANFREDI , ANGELO ANDREA M. A., Bondanza, A, Zimmermann, V, Dellantonio, G, DAL CIN, E, Capobianco, A., Sabbadini, Mg, Manfredi, ANGELO ANDREA M. A., and ROVERE QUERINI, Patrizia
- Published
- 2003
6. Anti-beta 2 glycoprotein I antibodies cause inflammation and recruit dendritic cells in platelet clearance
- Author
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BONDANZA , ATTILIO, MANFREDI , ANGELO ANDREA M. A., Zimmermann VS, Iannacone M, Tincani A, Balestrieri G, Sabbadini MG, ROVERE QUERINI , PATRIZIA, RI Iannacone Matteo/B 3342 2008 Tincani Angela/E 7608 2010, Bondanza, Attilio, Manfredi, ANGELO ANDREA M. A., Zimmermann, V, Iannacone, M, Tincani, A, Balestrieri, G, Sabbadini, Mg, ROVERE QUERINI, Patrizia, and RI Iannacone Matteo/B 3342 2008 Tincani Angela/E 7608, 2010
- Subjects
Platelets ,business.industry ,Antiphospholipid antibodies ,Macrophages ,Antigen presentation ,Inflammation ,Hematology ,Dendritic cell ,Dendritic cells ,Proinflammatory cytokine ,Tumor necrosis factor-α ,Immunology ,medicine ,Macrophage ,Platelet ,Platelet activation ,medicine.symptom ,business ,Antigen-presenting cell - Abstract
SummaryScavenger phagocytes are mostly responsible for the in vivo clearance of activated or senescent platelets. In contrast to other particulate substrates, the phagocytosis of platelets does not incite pro-inflammatory responses in vivo. This study assessed the contribution of macrophages and dendritic cells (DCs) to the clearance of activated platelets. Furthermore, we verified whether antibodies against the β2 Glycoprotein I (β2GPI), which bind to activated platelets, influence the phenomenon. DCs did not per se internalise activated platelets. In contrast, macrophages efficiently phagocytosed platelets. In agreement with the uneventful nature of the clearance of platelets in vivo, phagocytosing macrophages did not release IL-1β, TNF-α or IL-10. β2GPI bound to activated platelets and was required for their recognition by anti-ββ2GPI antibodies. DCs internalised platelets opsonised by anti-ββ2GPI antibodies. The phagocytosis of opsonised platelets determined the release of TNF-α and IL-1β by DCs and macrophages. Phagocytosing macrophages, but not DCs, secreted the antiinflammatory cytokine IL-1β0. We conclude that anti-ββ2GPI antibodies cause inflammation during platelet clearance and shuttle platelet antigens to antigen presenting DCs.
- Published
- 2001
7. Anti-beta2 glycoprotein I antibodies cause inflammation and recruit dendritic cells in platelet clearance
- Author
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Bondanza, A, Manfredi, Aa, Zimmermann, Vs, Iannacone, M, Tincani, Angela, Balestrieri, G, Sabbadini, Mg, and Querini, P. R.
- Published
- 2001
8. Apoptotic cell-pulsed dendritic cells break tolerance and accelerate lupus in NZB/W F1 mice
- Author
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BONDANZA , ATTILIO, Sabbadini MG, Zimmermann VS, Dell'Antonio G, Tincani A, Balestrieri G, MANFREDI, ANGELO ANDREA M. A., ROVERE QUERINI , PATRIZIA, RI Tincani Angela/E 7608 2010, Bondanza, Attilio, Sabbadini, Mg, Zimmermann, V, Dell'Antonio, G, Tincani, A, Balestrieri, G, Manfredi, ANGELO ANDREA M. A., ROVERE QUERINI, Patrizia, and RI Tincani Angela/E 7608, 2010
- Published
- 2001
9. Remnants of suicidal cells fostering systemic autoaggression: apoptosis in the origin and maintenance of autoimmunity
- Author
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ROVERE QUERINI , PATRIZIA, Sabbadini MG, Fazzini F, BONDANZA , ATTILIO, Zimmermann VS, Rugarli C, MANFREDI , ANGELO ANDREA M. A., ROVERE QUERINI, Patrizia, Sabbadini, Mg, Fazzini, F, Bondanza, Attilio, Zimmermann, V, Rugarli, C, and Manfredi, ANGELO ANDREA M. A.
- Published
- 2000
10. Anti-beta2 glycoprotein I antibodies prevent the De-activation of platelets and sustain their phagocytic clearance
- Author
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Bondanza, A, Sabbadini, Mg, Pellegatta, F, Zimmermann, Vs, Tincani, Angela, Balestrieri, G, Manfredi, Aa, and Rovere, P.
- Published
- 2000
11. The MHC class II presentation pathway in invariant chain deficient mice: comparison between B lymphocytes and dendritic cells
- Author
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Zimmermann VS, Trucy J, Forquet F, Serre K, Machy P, Barois N, Leserman L, Davoust J., ROVERE QUERINI , PATRIZIA, Zimmermann, V, ROVERE QUERINI, Patrizia, Trucy, J, Forquet, F, Serre, K, Machy, P, Barois, N, Leserman, L, and Davoust, J.
- Published
- 1998
12. An apoptotic cell threshold for dendritic cell maturation and function
- Author
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ROVERE QUERINI , PATRIZIA, Vallinoto C, BONDANZA , ATTILIO, Zimmermann VS, Rescigno M, Riccardi Castagnoli P, Rugarli C, MANFREDI, ANGELO ANDREA M. A., ROVERE QUERINI, Patrizia, Vallinoto, C, Bondanza, Attilio, Zimmermann, V, Rescigno, M, Riccardi Castagnoli, P, Rugarli, C, and Manfredi, ANGELO ANDREA M. A.
- Published
- 1998
13. Anti-phospholipid antibodies opsonize activated platelets for uptake by dendritic cells: relevance to epitope spreading in platelet autoimmunity
- Author
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BONDANZA , ATTILIO, Pellegatta F, Zimmermann VS, Balestrieri G, Tincani A, Sabbadini MG, MANFREDI, ANGELO ANDREA M. A., ROVERE QUERINI , PATRIZIA, Bondanza, Attilio, Pellegatta, F, Zimmermann, V, Balestrieri, G, Tincani, A, Sabbadini, Mg, Manfredi, ANGELO ANDREA M. A., and ROVERE QUERINI, Patrizia
- Published
- 1998
14. Autoantibody opsonization of apoptotic cells facilitates antigen presentation by dendritic cells: implications for autoimmunity establishment in Systemic Lupus Erythematosus
- Author
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ROVERE QUERINI , PATRIZIA, Zimmermann VS, Vallinoto C, Crosti MC, Fascio U, Rescigno M, Ricciardi Castagnoli P, Sabbadini MG, MANFREDI, ANGELO ANDREA M. A., ROVERE QUERINI, Patrizia, Zimmermann, V, Vallinoto, C, Crosti, Mc, Fascio, U, Rescigno, M, Ricciardi Castagnoli, P, Sabbadini, Mg, and Manfredi, ANGELO ANDREA M. A.
- Published
- 1998
15. Tuft cell acetylcholine is released into the gut lumen to promote anti-helminth immunity.
- Author
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Ndjim M, Gasmi I, Herbert F, Joséphine C, Bas J, Lamrani A, Coutry N, Henry S, Zimmermann VS, Dardalhon V, Campillo Poveda M, Turtoi E, Thirard S, Forichon L, Giordano A, Ciancia C, Homayed Z, Pannequin J, Britton C, Devaney E, McNeilly TN, Berrard S, Turtoi A, Maizels RM, Gerbe F, and Jay P
- Subjects
- Animals, Mice, Choline O-Acetyltransferase metabolism, Interleukin-13 metabolism, Interleukin-13 immunology, Mice, Knockout, Mice, Inbred C57BL, Helminthiasis immunology, Helminthiasis parasitology, Epithelial Cells immunology, Epithelial Cells metabolism, Immunity, Innate, Nematospiroides dubius immunology, Tuft Cells, Acetylcholine metabolism, Intestinal Mucosa immunology, Intestinal Mucosa metabolism, Intestinal Mucosa parasitology
- Abstract
Upon parasitic helminth infection, activated intestinal tuft cells secrete interleukin-25 (IL-25), which initiates a type 2 immune response during which lamina propria type 2 innate lymphoid cells (ILC2s) produce IL-13. This causes epithelial remodeling, including tuft cell hyperplasia, the function of which is unknown. We identified a cholinergic effector function of tuft cells, which are the only epithelial cells that expressed choline acetyltransferase (ChAT). During parasite infection, mice with epithelial-specific deletion of ChAT had increased worm burden, fitness, and fecal egg counts, even though type 2 immune responses were comparable. Mechanistically, IL-13-amplified tuft cells release acetylcholine (ACh) into the gut lumen. Finally, we demonstrated a direct effect of ACh on worms, which reduced their fecundity via helminth-expressed muscarinic ACh receptors. Thus, tuft cells are sentinels in naive mice, and their amplification upon helminth infection provides an additional type 2 immune response effector function., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
- Full Text
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16. Combined immunodeficiency caused by pathogenic variants in the ZAP70 C-terminal SH2 domain.
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Mongellaz C, Vicente R, Noroski LM, Noraz N, Courgnaud V, Chinen J, Faria E, Zimmermann VS, and Taylor N
- Subjects
- Infant, Humans, src Homology Domains genetics, Protein-Tyrosine Kinases, Arginine, ZAP-70 Protein-Tyrosine Kinase genetics, Primary Immunodeficiency Diseases, Lymphopenia genetics
- Abstract
Introduction: ZAP-70, a protein tyrosine kinase recruited to the T cell receptor (TCR), initiates a TCR signaling cascade upon antigen stimulation. Mutations in the ZAP70 gene cause a combined immunodeficiency characterized by low or absent CD8+ T cells and nonfunctional CD4+ T cells. Most deleterious missense ZAP70 mutations in patients are located in the kinase domain but the impact of mutations in the SH2 domains, regulating ZAP-70 recruitment to the TCR, are not well understood., Methods: Genetic analyses were performed on four patients with CD8 lymphopenia and a high resolution melting screening for ZAP70 mutations was developed. The impact of SH2 domain mutations was evaluated by biochemical and functional analyses as well as by protein modeling., Results and Discussion: Genetic characterization of an infant who presented with pneumocystis pneumonia, mycobacterial infection, and an absence of CD8 T cells revealed a novel homozygous mutation in the C-terminal SH2 domain (SH2-C) of the ZAP70 gene (c.C343T, p.R170C). A distantly related second patient was found to be compound heterozygous for the R170C variant and a 13bp deletion in the ZAP70 kinase domain. While the R170C mutant was highly expressed, there was an absence of TCR-induced proliferation, associated with significantly attenuated TCR-induced ZAP-70 phosphorylation and a lack of binding of ZAP-70 to TCR-ζ. Moreover, a homozygous ZAP-70 R192W variant was identified in 2 siblings with combined immunodeficiency and CD8 lymphopenia, confirming the pathogenicity of this mutation. Structural modeling of this region revealed the critical nature of the arginines at positions 170 and 192, in concert with R190, forming a binding pocket for the phosphorylated TCR-ζ chain. Deleterious mutations in the SH2-C domain result in attenuated ZAP-70 function and clinical manifestations of immunodeficiency., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Mongellaz, Vicente, Noroski, Noraz, Courgnaud, Chinen, Faria, Zimmermann and Taylor.)
- Published
- 2023
- Full Text
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17. Arginine metabolism regulates human erythroid differentiation through hypusination of eIF5A.
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Gonzalez-Menendez P, Phadke I, Olive ME, Joly A, Papoin J, Yan H, Galtier J, Platon J, Kang SWS, McGraw KL, Daumur M, Pouzolles M, Kondo T, Boireau S, Paul F, Young DJ, Lamure S, Mirmira RG, Narla A, Cartron G, Dunbar CE, Boyer-Clavel M, Porat-Shliom N, Dardalhon V, Zimmermann VS, Sitbon M, Dever TE, Mohandas N, Da Costa L, Udeshi ND, Blanc L, Kinet S, and Taylor N
- Subjects
- Humans, Peptide Initiation Factors genetics, Cell Differentiation, Eukaryotic Translation Initiation Factor 5A, Spermidine metabolism, Proteomics
- Abstract
Metabolic programs contribute to hematopoietic stem and progenitor cell (HSPC) fate, but it is not known whether the metabolic regulation of protein synthesis controls HSPC differentiation. Here, we show that SLC7A1/cationic amino acid transporter 1-dependent arginine uptake and its catabolism to the polyamine spermidine control human erythroid specification of HSPCs via the activation of the eukaryotic translation initiation factor 5A (eIF5A). eIF5A activity is dependent on its hypusination, a posttranslational modification resulting from the conjugation of the aminobutyl moiety of spermidine to lysine. Notably, attenuation of hypusine synthesis in erythroid progenitors, by the inhibition of deoxyhypusine synthase, abrogates erythropoiesis but not myeloid cell differentiation. Proteomic profiling reveals mitochondrial translation to be a critical target of hypusinated eIF5A, and accordingly, progenitors with decreased hypusine activity exhibit diminished oxidative phosphorylation. This affected pathway is critical for eIF5A-regulated erythropoiesis, as interventions augmenting mitochondrial function partially rescue human erythropoiesis under conditions of attenuated hypusination. Levels of mitochondrial ribosomal proteins (RPs) were especially sensitive to the loss of hypusine, and we find that the ineffective erythropoiesis linked to haploinsufficiency of RPS14 in chromosome 5q deletions in myelodysplastic syndrome is associated with a diminished pool of hypusinated eIF5A. Moreover, patients with RPL11-haploinsufficient Diamond-Blackfan anemia as well as CD34+ progenitors with downregulated RPL11 exhibit a markedly decreased hypusination in erythroid progenitors, concomitant with a loss of mitochondrial metabolism. Thus, eIF5A-dependent protein synthesis regulates human erythropoiesis, and our data reveal a novel role for RPs in controlling eIF5A hypusination in HSPCs, synchronizing mitochondrial metabolism with erythroid differentiation.
- Published
- 2023
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18. Intrathymic AAV delivery results in therapeutic site-specific integration at TCR loci in mice.
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Calabria A, Cipriani C, Spinozzi G, Rudilosso L, Esposito S, Benedicenti F, Albertini A, Pouzolles M, Luoni M, Giannelli S, Broccoli V, Guilbaud M, Adjali O, Taylor N, Zimmermann VS, Montini E, and Cesana D
- Subjects
- Mice, Animals, Transgenes, Plasmids, Receptors, Antigen, T-Cell genetics, Dependovirus genetics, Virus Integration, Genetic Vectors genetics, Genetic Therapy methods
- Abstract
Adeno-associated virus (AAV) vectors have been successfully exploited in gene therapy applications for the treatment of several genetic disorders. AAV is considered an episomal vector, but it has been shown to integrate within the host cell genome after the generation of double-strand DNA breaks or nicks. Although AAV integration raises some safety concerns, it can also provide therapeutic benefit; the direct intrathymic injection of an AAV harboring a therapeutic transgene results in integration in T-cell progenitors and long-term T-cell immunity. To assess the mechanisms of AAV integration, we retrieved and analyzed hundreds of AAV integration sites from lymph node-derived mature T cells and compared these with liver and brain tissue from treated mice. Notably, we found that although AAV integrations in the liver and brain were distributed across the entire mouse genome, >90% of the integrations in T cells were clustered within the T-cell receptor α, β, and γ genes. More precisely, the insertion mapped to DNA breaks created by the enzymatic activity of recombination activating genes (RAGs) during variable, diversity, and joining recombination. Our data indicate that RAG activity during T-cell receptor maturation induces a site-specific integration of AAV genomes and opens new therapeutic avenues for achieving long-term AAV-mediated gene transfer in dividing cells., (© 2023 by The American Society of Hematology. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved.)
- Published
- 2023
- Full Text
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19. Vitamin C deficiency reveals developmental differences between neonatal and adult hematopoiesis.
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Phadke I, Pouzolles M, Machado A, Moraly J, Gonzalez-Menendez P, Zimmermann VS, Kinet S, Levine M, Violet PC, and Taylor N
- Subjects
- Animals, Ascorbic Acid pharmacology, Erythropoiesis, Female, Glucose, Humans, L-Gulonolactone Oxidase genetics, Mice, Pregnancy, Vitamins, Ascorbic Acid Deficiency genetics, Mustelidae, Scurvy
- Abstract
Hematopoiesis, a process that results in the differentiation of all blood lineages, is essential throughout life. The production of 1x10
12 blood cells per day, including 200x109 erythrocytes, is highly dependent on nutrient consumption. Notably though, the relative requirements for micronutrients during the perinatal period, a critical developmental window for immune cell and erythrocyte differentiation, have not been extensively studied. More specifically, the impact of the vitamin C/ascorbate micronutrient on perinatal as compared to adult hematopoiesis has been difficult to assess in animal models. Even though humans cannot synthesize ascorbate, due to a pseudogenization of the L-gulono-γ-lactone oxidase ( GULO ) gene, its generation from glucose is an ancestral mammalian trait. Taking advantage of a Gulo-/- mouse model, we show that ascorbic acid deficiency profoundly impacts perinatal hematopoiesis, resulting in a hypocellular bone marrow (BM) with a significant reduction in hematopoietic stem cells, multipotent progenitors, and hematopoietic progenitors. Furthermore, myeloid progenitors exhibited differential sensitivity to vitamin C levels; common myeloid progenitors and megakaryocyte-erythrocyte progenitors were markedly reduced in Gulo-/- pups following vitamin C depletion in the dams, whereas granulocyte-myeloid progenitors were spared, and their frequency was even augmented. Notably, hematopoietic cell subsets were rescued by vitamin C repletion. Consistent with these data, peripheral myeloid cells were maintained in ascorbate-deficient Gulo-/- pups while other lineage-committed hematopoietic cells were decreased. A reduction in B cell numbers was associated with a significantly reduced humoral immune response in ascorbate-depleted Gulo-/- pups but not adult mice. Erythropoiesis was particularly sensitive to vitamin C deprivation during both the perinatal and adult periods, with ascorbate-deficient Gulo-/- pups as well as adult mice exhibiting compensatory splenic differentiation. Furthermore, in the pathological context of hemolytic anemia, vitamin C-deficient adult Gulo-/- mice were not able to sufficiently increase their erythropoietic activity, resulting in a sustained anemia. Thus, vitamin C plays a pivotal role in the maintenance and differentiation of hematopoietic progenitors during the neonatal period and is required throughout life to sustain erythroid differentiation under stress conditions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Phadke, Pouzolles, Machado, Moraly, Gonzalez-Menendez, Zimmermann, Kinet, Levine, Violet and Taylor.)- Published
- 2022
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20. A boy with a novel homozygous ZAP70 mutation presenting with a dermatological phenotype and postnatal decrease in CD8 + T cells.
- Author
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Babayeva R, Mongellaz C, Karakus IS, Cansever M, Bilgic Eltan S, Catak MC, Bulutoglu A, Kendir Demirkol Y, Eser M, Karakoc-Aydiner E, Ozen A, Zimmermann VS, Taylor N, and Baris S
- Subjects
- Humans, Male, Mutation, Phenotype, CD8-Positive T-Lymphocytes cytology, ZAP-70 Protein-Tyrosine Kinase genetics
- Published
- 2022
- Full Text
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21. Regulatory T cell differentiation is controlled by αKG-induced alterations in mitochondrial metabolism and lipid homeostasis.
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Matias MI, Yong CS, Foroushani A, Goldsmith C, Mongellaz C, Sezgin E, Levental KR, Talebi A, Perrault J, Rivière A, Dehairs J, Delos O, Bertand-Michel J, Portais JC, Wong M, Marie JC, Kelekar A, Kinet S, Zimmermann VS, Levental I, Yvan-Charvet L, Swinnen JV, Muljo SA, Hernandez-Vargas H, Tardito S, Taylor N, and Dardalhon V
- Subjects
- Animals, Cells, Cultured, Cytokines genetics, Cytokines metabolism, Diacylglycerol O-Acyltransferase metabolism, Fibrosarcoma genetics, Fibrosarcoma immunology, Fibrosarcoma metabolism, Fibrosarcoma therapy, Forkhead Transcription Factors genetics, Forkhead Transcription Factors metabolism, Homeostasis, Humans, Immunotherapy, Adoptive, Mice, Inbred C57BL, Mice, Knockout, Mitochondria genetics, Mitochondria metabolism, Phenotype, Receptors, Chimeric Antigen genetics, Receptors, Chimeric Antigen metabolism, Signal Transduction, T-Lymphocytes, Regulatory immunology, T-Lymphocytes, Regulatory metabolism, T-Lymphocytes, Regulatory transplantation, Th1 Cells drug effects, Th1 Cells immunology, Th1 Cells metabolism, Mice, Cell Differentiation drug effects, Energy Metabolism drug effects, Ketoglutaric Acids pharmacology, Lipid Metabolism drug effects, Mitochondria drug effects, T-Lymphocytes, Regulatory drug effects
- Abstract
Suppressive regulatory T cell (Treg) differentiation is controlled by diverse immunometabolic signaling pathways and intracellular metabolites. Here we show that cell-permeable α-ketoglutarate (αKG) alters the DNA methylation profile of naive CD4 T cells activated under Treg polarizing conditions, markedly attenuating FoxP3+ Treg differentiation and increasing inflammatory cytokines. Adoptive transfer of these T cells into tumor-bearing mice results in enhanced tumor infiltration, decreased FoxP3 expression, and delayed tumor growth. Mechanistically, αKG leads to an energetic state that is reprogrammed toward a mitochondrial metabolism, with increased oxidative phosphorylation and expression of mitochondrial complex enzymes. Furthermore, carbons from ectopic αKG are directly utilized in the generation of fatty acids, associated with lipidome remodeling and increased triacylglyceride stores. Notably, inhibition of either mitochondrial complex II or DGAT2-mediated triacylglyceride synthesis restores Treg differentiation and decreases the αKG-induced inflammatory phenotype. Thus, we identify a crosstalk between αKG, mitochondrial metabolism and triacylglyceride synthesis that controls Treg fate., Competing Interests: Declaration of interests C.M., S.K., V.D., and N.T. are inventors on patents describing the use of ligands for detection of and modulation of metabolite transporters (N.T. gave up her rights), licensed to METAFORA-biosystems., (Copyright © 2021. Published by Elsevier Inc.)
- Published
- 2021
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22. Targeting of the Tec Kinase ITK Drives Resolution of T Cell-Mediated Colitis and Emerges as Potential Therapeutic Option in Ulcerative Colitis.
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Lechner K, Mott S, Al-Saifi R, Knipfer L, Wirtz S, Atreya R, Vieth M, Rath T, Fraass T, Winter Z, August A, Luban J, Zimmermann VS, Weigmann B, and Neurath MF
- Subjects
- Animals, Apoptosis drug effects, Cells, Cultured, Colitis, Ulcerative enzymology, Colitis, Ulcerative immunology, Colitis, Ulcerative pathology, Colon enzymology, Colon immunology, Colon pathology, Cyclosporine pharmacology, Cytokines metabolism, Disease Models, Animal, Humans, Intestinal Mucosa enzymology, Intestinal Mucosa immunology, Intestinal Mucosa pathology, Intraepithelial Lymphocytes enzymology, Intraepithelial Lymphocytes immunology, Intraepithelial Lymphocytes pathology, Mice, Knockout, Molecular Targeted Therapy, Phosphorylation, Protein-Tyrosine Kinases genetics, Protein-Tyrosine Kinases metabolism, Signal Transduction, Mice, Anti-Inflammatory Agents pharmacology, Colitis, Ulcerative prevention & control, Colon drug effects, Intestinal Mucosa drug effects, Intraepithelial Lymphocytes drug effects, Protein Kinase Inhibitors pharmacology, Protein-Tyrosine Kinases antagonists & inhibitors
- Abstract
Background & Aims: The molecular checkpoints driving T cell activation and cytokine responses in ulcerative colitis (UC) are incompletely understood. Here, we studied the Tec kinase ITK in UC., Methods: We analyzed patients with inflammatory bowel disease (n = 223) and evaluated ITK activity as well as the functional effects of cyclosporine-A (CsA). In addition, 3 independent murine colitis models were used to investigate the functional role of ITK. Finally, the activity of ITK was blocked via pharmacological inhibitors and genetically engineered mice. Readout parameters were mini-endoscopy, histopathology, mucosal T cell apoptosis, and cytokine production., Results: We found an expansion of pITK-expressing mucosal CD4
+ T cells in UC rather than Crohn's disease that correlated with disease severity. CsA suppressed activation of ITK in cultured CD4+ T cells and calcineurin-containing microclusters adjacent to the T cell receptor signaling complex. Functionally, the capacity of CsA to suppress activity of experimental colitis was critically dependent on ITK. Genetic inactivation of Itk via gene targeting or induction of allele-sensitive Itk mutants prevented experimental colitis in 3 colitis models, and treatment with pharmacological ITK blockers suppressed established colitis. In addition, ITK controlled apoptosis and activation of mucosal Th2 and Th17 lymphocytes via NFATc2 signaling pathways., Conclusions: ITK activation was detected in UC and could be down-regulated in cultured T cells by CsA administration. Selective targeting of ITK emerges as an attractive approach for treatment of chronic intestinal inflammation and potentially UC by driving resolution of mucosal inflammation., (Copyright © 2021 AGA Institute. Published by Elsevier Inc. All rights reserved.)- Published
- 2021
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23. An IDH1-vitamin C crosstalk drives human erythroid development by inhibiting pro-oxidant mitochondrial metabolism.
- Author
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Gonzalez-Menendez P, Romano M, Yan H, Deshmukh R, Papoin J, Oburoglu L, Daumur M, Dumé AS, Phadke I, Mongellaz C, Qu X, Bories PN, Fontenay M, An X, Dardalhon V, Sitbon M, Zimmermann VS, Gallagher PG, Tardito S, Blanc L, Mohandas N, Taylor N, and Kinet S
- Subjects
- Cell Differentiation, Humans, Ascorbic Acid metabolism, Erythropoiesis genetics, Isocitrate Dehydrogenase metabolism, Mitochondria metabolism
- Abstract
The metabolic changes controlling the stepwise differentiation of hematopoietic stem and progenitor cells (HSPCs) to mature erythrocytes are poorly understood. Here, we show that HSPC development to an erythroid-committed proerythroblast results in augmented glutaminolysis, generating alpha-ketoglutarate (αKG) and driving mitochondrial oxidative phosphorylation (OXPHOS). However, sequential late-stage erythropoiesis is dependent on decreasing αKG-driven OXPHOS, and we find that isocitrate dehydrogenase 1 (IDH1) plays a central role in this process. IDH1 downregulation augments mitochondrial oxidation of αKG and inhibits reticulocyte generation. Furthermore, IDH1 knockdown results in the generation of multinucleated erythroblasts, a morphological abnormality characteristic of myelodysplastic syndrome and congenital dyserythropoietic anemia. We identify vitamin C homeostasis as a critical regulator of ineffective erythropoiesis; oxidized ascorbate increases mitochondrial superoxide and significantly exacerbates the abnormal erythroblast phenotype of IDH1-downregulated progenitors, whereas vitamin C, scavenging reactive oxygen species (ROS) and reprogramming mitochondrial metabolism, rescues erythropoiesis. Thus, an IDH1-vitamin C crosstalk controls terminal steps of human erythroid differentiation., Competing Interests: Declaration of interests M.S. and S.K. are inventors on a patent describing the use of a ligand for evaluation of GLUT1 expression (N.T. gave up her rights). M.S. is a co-founder of METAFORA Biosystems, focusing on metabolite transporters under physiological and pathological conditions, and is head of the scientific board., (Published by Elsevier Inc.)
- Published
- 2021
- Full Text
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24. Perforin-deficient CAR T cells recapitulate late-onset inflammatory toxicities observed in patients.
- Author
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Ishii K, Pouzolles M, Chien CD, Erwin-Cohen RA, Kohler ME, Qin H, Lei H, Kuhn S, Ombrello AK, Dulau-Florea A, Eckhaus MA, Shalabi H, Yates B, Lichtenstein DA, Zimmermann VS, Kondo T, Shern JF, Young HA, Taylor N, Shah NN, and Fry TJ
- Subjects
- Animals, Cytokines biosynthesis, Disease Models, Animal, Humans, In Vitro Techniques, Inflammation Mediators metabolism, Lymphohistiocytosis, Hemophagocytic etiology, Lymphohistiocytosis, Hemophagocytic immunology, Lymphohistiocytosis, Hemophagocytic pathology, Macrophage Activation Syndrome etiology, Macrophage Activation Syndrome immunology, Macrophage Activation Syndrome pathology, Mice, Mice, Inbred C57BL, Mice, Knockout, Models, Immunological, Perforin genetics, T-Lymphocytes pathology, Immunotherapy, Adoptive adverse effects, Perforin deficiency, Receptors, Chimeric Antigen immunology, T-Lymphocytes immunology
- Abstract
Late-onset inflammatory toxicities resembling hemophagocytic lymphohistiocytosis (HLH) or macrophage activation syndrome (MAS) occur after chimeric antigen receptor T cell (CAR T cell) infusion and represent a therapeutic challenge. Given the established link between perforin deficiency and primary HLH, we investigated the role of perforin in anti-CD19 CAR T cell efficacy and HLH-like toxicities in a syngeneic murine model. Perforin contributed to both CD8+ and CD4+ CAR T cell cytotoxicity but was not required for in vitro or in vivo leukemia clearance. Upon CAR-mediated in vitro activation, perforin-deficient CAR T cells produced higher amounts of proinflammatory cytokines compared with WT CAR T cells. Following in vivo clearance of leukemia, perforin-deficient CAR T cells reexpanded, resulting in splenomegaly with disruption of normal splenic architecture and the presence of hemophagocytes, which are findings reminiscent of HLH. Notably, a substantial fraction of patients who received anti-CD22 CAR T cells also experienced biphasic inflammation, with the second phase occurring after the resolution of cytokine release syndrome, resembling clinical manifestations of HLH. Elevated inflammatory cytokines such as IL-1β and IL-18 and concurrent late CAR T cell expansion characterized the HLH-like syndromes occurring in the murine model and in humans. Thus, a murine model of perforin-deficient CAR T cells recapitulated late-onset inflammatory toxicities occurring in human CAR T cell recipients, providing therapeutically relevant mechanistic insights.
- Published
- 2020
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25. Phosphate Transporter Profiles in Murine and Human Thymi Identify Thymocytes at Distinct Stages of Differentiation.
- Author
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Machado A, Pouzolles M, Gailhac S, Fritz V, Craveiro M, López-Sánchez U, Kondo T, Pala F, Bosticardo M, Notarangelo LD, Petit V, Taylor N, and Zimmermann VS
- Subjects
- Animals, Biomarkers, Gene Expression Profiling, Glucose Transporter Type 1 genetics, Glucose Transporter Type 1 metabolism, Humans, Immunophenotyping, Lymphocyte Subsets immunology, Lymphocyte Subsets metabolism, Mice, Mice, Knockout, Phosphate Transport Proteins genetics, Sodium-Phosphate Cotransporter Proteins, Type III genetics, Sodium-Phosphate Cotransporter Proteins, Type III metabolism, Thymocytes cytology, Thymocytes immunology, Thymus Gland cytology, Thymus Gland immunology, Thymus Gland metabolism, Transcriptome, Cell Differentiation genetics, Phosphate Transport Proteins metabolism, Thymocytes metabolism
- Abstract
Thymocyte differentiation is dependent on the availability and transport of metabolites in the thymus niche. As expression of metabolite transporters is a rate-limiting step in nutrient utilization, cell surface transporter levels generally reflect the cell's metabolic state. The GLUT1 glucose transporter is upregulated on actively dividing thymocytes, identifying thymocytes with an increased metabolism. However, it is not clear whether transporters of essential elements such as phosphate are modulated during thymocyte differentiation. While PiT1 and PiT2 are both phosphate transporters in the SLC20 family, we show here that they exhibit distinct expression profiles on both murine and human thymocytes. PiT2 expression distinguishes thymocytes with high metabolic activity, identifying immature murine double negative (CD4
- CD8- ) DN3b and DN4 thymocyte blasts as well as immature single positive (ISP) CD8 thymocytes. Notably, the absence of PiT2 expression on RAG2-deficient thymocytes, blocked at the DN3a stage, strongly suggests that high PiT2 expression is restricted to thymocytes having undergone a productive TCRβ rearrangement at the DN3a/DN3b transition. Similarly, in the human thymus, PiT2 was upregulated on early post-β selection CD4+ ISP and TCRαβ- CD4hi DP thymocytes co-expressing the CD71 transferrin receptor, a marker of metabolic activity. In marked contrast, expression of the PiT1 phosphate importer was detected on mature CD3+ murine and human thymocytes. Notably, PiT1 expression on CD3+ DN thymocytes was identified as a biomarker of an aging thymus, increasing from 8.4 ± 1.5% to 42.4 ± 9.4% by 1 year of age ( p < 0.0001). We identified these cells as TCRγδ and, most significantly, NKT, representing 77 ± 9% of PiT1+ DN thymocytes by 1 year of age ( p < 0.001). Thus, metabolic activity and thymic aging are associated with distinct expression profiles of the PiT1 and PiT2 phosphate transporters., (Copyright © 2020 Machado, Pouzolles, Gailhac, Fritz, Craveiro, López-Sánchez, Kondo, Pala, Bosticardo, Notarangelo, Petit, Taylor and Zimmermann.)- Published
- 2020
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26. Intrathymic adeno-associated virus gene transfer rapidly restores thymic function and long-term persistence of gene-corrected T cells.
- Author
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Pouzolles M, Machado A, Guilbaud M, Irla M, Gailhac S, Barennes P, Cesana D, Calabria A, Benedicenti F, Sergé A, Raman I, Li QZ, Montini E, Klatzmann D, Adjali O, Taylor N, and Zimmermann VS
- Subjects
- Animals, Dependovirus, Genetic Vectors, Immunologic Deficiency Syndromes therapy, Mice, Mice, Knockout, Genetic Therapy methods, Thymocytes, Transduction, Genetic methods, ZAP-70 Protein-Tyrosine Kinase administration & dosage, ZAP-70 Protein-Tyrosine Kinase genetics
- Abstract
Background: Patients with T-cell immunodeficiencies are generally treated with allogeneic hematopoietic stem cell transplantation, but alternatives are needed for patients without matched donors. An innovative intrathymic gene therapy approach that directly targets the thymus might improve outcomes., Objective: We sought to determine the efficacy of intrathymic adeno-associated virus (AAV) serotypes to transduce thymocyte subsets and correct the T-cell immunodeficiency in a zeta-associated protein of 70 kDa (ZAP-70)-deficient murine model., Methods: AAV serotypes were injected intrathymically into wild-type mice, and gene transfer efficiency was monitored. ZAP-70
-/- mice were intrathymically injected with an AAV8 vector harboring the ZAP70 gene. Thymus structure, immunophenotyping, T-cell receptor clonotypes, T-cell function, immune responses to transgenes and autoantibodies, vector copy number, and integration were evaluated., Results: AAV8, AAV9, and AAV10 serotypes all transduced thymocyte subsets after in situ gene transfer, with transduction of up to 5% of cells. Intrathymic injection of an AAV8-ZAP-70 vector into ZAP-70-/- mice resulted in a rapid thymocyte differentiation associated with the development of a thymic medulla. Strikingly, medullary thymic epithelial cells expressing the autoimmune regulator were detected within 10 days of gene transfer, correlating with the presence of functional effector and regulatory T-cell subsets with diverse T-cell receptor clonotypes in the periphery. Although thymocyte reconstitution was transient, gene-corrected peripheral T cells harboring approximately 1 AAV genome per cell persisted for more than 40 weeks, and AAV vector integration was detected., Conclusions: Intrathymic AAV-transduced progenitors promote a rapid restoration of the thymic architecture, with a single wave of thymopoiesis generating long-term peripheral T-cell function., (Copyright © 2019 American Academy of Allergy, Asthma & Immunology. All rights reserved.)- Published
- 2020
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27. Single-cell mapping of the thymic stroma identifies IL-25-producing tuft epithelial cells.
- Author
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Bornstein C, Nevo S, Giladi A, Kadouri N, Pouzolles M, Gerbe F, David E, Machado A, Chuprin A, Tóth B, Goldberg O, Itzkovitz S, Taylor N, Jay P, Zimmermann VS, Abramson J, and Amit I
- Subjects
- Animals, Epigenesis, Genetic, Epithelial Cells immunology, Female, Humans, Interleukin-17 biosynthesis, Interleukins biosynthesis, Male, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Models, Molecular, Transcription Factors biosynthesis, Transcription Factors deficiency, Transcription Factors genetics, Transcription Factors metabolism, AIRE Protein, Epithelial Cells cytology, Epithelial Cells metabolism, Interleukin-17 metabolism, Interleukins metabolism, Single-Cell Analysis, Thymus Gland cytology, Thymus Gland immunology
- Abstract
T cell development and selection are coordinated in the thymus by a specialized niche of diverse stromal populations
1-3 . Although much progress has been made over the years in identifying the functions of the different cell types of the thymic stromal compartment, there is no comprehensive characterization of their diversity and heterogeneity. Here we combined massively parallel single-cell RNA-sequencing4,5 , spatial mapping, chromatin profiling and gene targeting to characterize de novo the entire stromal compartment of the mouse thymus. We identified dozens of cell states, with thymic epithelial cells (TECs) showing the highest degree of heterogeneity. Our analysis highlights four major medullary TEC (mTEC I-IV) populations, with distinct molecular functions, epigenetic landscapes and lineage regulators. Specifically, mTEC IV constitutes a new and highly divergent TEC lineage with molecular characteristics of the gut chemosensory epithelial tuft cells. Mice deficient in Pou2f3, a master regulator of tuft cells, have complete and specific depletion of mTEC IV cells, which results in increased levels of thymus-resident type-2 innate lymphoid cells. Overall, our study provides a comprehensive characterization of the thymic stroma and identifies a new tuft-like TEC population, which is critical for shaping the immune niche in the thymus.- Published
- 2018
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28. Resveratrol stimulates the metabolic reprogramming of human CD4 + T cells to enhance effector function.
- Author
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Craveiro M, Cretenet G, Mongellaz C, Matias MI, Caron O, de Lima MCP, Zimmermann VS, Solary E, Dardalhon V, Dulić V, and Taylor N
- Subjects
- Adult, Antioxidants pharmacology, Ataxia Telangiectasia Mutated Proteins genetics, Ataxia Telangiectasia Mutated Proteins metabolism, CD4-Positive T-Lymphocytes metabolism, Cells, Cultured, Cytokines metabolism, G1 Phase Cell Cycle Checkpoints genetics, Gene Expression Profiling methods, Glycolysis drug effects, Glycolysis genetics, Humans, Oxidative Phosphorylation drug effects, Phosphorylation drug effects, Resveratrol, Signal Transduction genetics, Tumor Suppressor Protein p53 genetics, Tumor Suppressor Protein p53 metabolism, CD4-Positive T-Lymphocytes drug effects, DNA Damage, G1 Phase Cell Cycle Checkpoints drug effects, Signal Transduction drug effects, Stilbenes pharmacology
- Abstract
The polyphenol resveratrol activates the deacetylase Sirt1, resulting in various antioxidant, chemoprotectant, neuroprotective, cardioprotective, and anti-inflammatory properties. We found that at high concentrations of resveratrol, human CD4
+ T cells showed defective antigen receptor signaling and arrest at the G1 stage of the cell cycle, whereas at low concentrations, cells were readily activated and exhibited enhanced Sirt1 deacetylase activity. Nevertheless, low-dose resveratrol rapidly stimulated genotoxic stress in the T cells, which resulted in engagement of a DNA damage response pathway that depended on the kinase ATR [ataxia telangiectasia-mutated (ATM) and Rad3-related], but not ATM, and subsequently in premitotic cell cycle arrest. The concomitant activation of p53 was coupled to the expression of gene products that regulate cell metabolism, leading to a metabolic reprogramming that was characterized by decreased glycolysis, increased glutamine consumption, and a shift to oxidative phosphorylation. These alterations in the bioenergetic homeostasis of CD4+ T cells resulted in enhanced effector function, with both naïve and memory CD4+ T cells secreting increased amounts of the inflammatory cytokine interferon-γ. Thus, our data highlight the wide range of metabolic adaptations that CD4+ T lymphocytes undergo in response to genomic stress., (Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)- Published
- 2017
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29. Hematopoietic stem cell lineage specification.
- Author
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Pouzolles M, Oburoglu L, Taylor N, and Zimmermann VS
- Subjects
- Animals, Cell Communication, Cell Differentiation genetics, Cell Self Renewal genetics, Gene Expression Regulation, Hematopoietic Stem Cells metabolism, Humans, Signal Transduction, Single-Cell Analysis, Stem Cell Niche, Transcription Factors metabolism, Cell Lineage genetics, Hematopoiesis, Hematopoietic Stem Cells cytology
- Abstract
Purpose of Review: Hematopoietic stem cells (HSCs) possess two fundamental characteristics, the capacity for self-renewal and the sustained production of all blood cell lineages. The fine balance between HSC expansion and lineage specification is dynamically regulated by the interplay between external and internal stimuli. This review introduces recent advances in the roles played by the stem cell niche, regulatory transcriptional networks, and metabolic pathways in governing HSC self-renewal, commitment, and lineage differentiation. We will further focus on discoveries made by studying hematopoiesis at single-cell resolution., Recent Findings: HSCs require the support of an interactive milieu with their physical position within the perivascular niche dynamically regulating HSC behavior. In these microenvironments, transcription factor networks and nutrient-mediated regulation of energy resources, signaling pathways, and epigenetic status govern HSC quiescence and differentiation. Once HSCs begin their lineage specification, single-cell analyses show that they do not become oligopotent but rather, differentiate directly into committed unipotent progenitors., Summary: The diversity of transcriptional networks and metabolic pathways in HSCs and their downstream progeny allows a high level of plasticity in blood differentiation. The intricate interactions between these pathways, within the perivascular niche, broaden the specification of HSCs in pathological and stressed conditions.
- Published
- 2016
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30. Intestinal epithelial tuft cells initiate type 2 mucosal immunity to helminth parasites.
- Author
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Gerbe F, Sidot E, Smyth DJ, Ohmoto M, Matsumoto I, Dardalhon V, Cesses P, Garnier L, Pouzolles M, Brulin B, Bruschi M, Harcus Y, Zimmermann VS, Taylor N, Maizels RM, and Jay P
- Subjects
- Animals, Cell Lineage, Cell Proliferation, Feedback, Physiological, Female, Goblet Cells cytology, Goblet Cells immunology, Interleukin-13 immunology, Interleukin-17 immunology, Interleukin-17 metabolism, Intestinal Mucosa metabolism, Male, Mice, Octamer Transcription Factors deficiency, Receptors, Interleukin-4 immunology, Signal Transduction immunology, Stem Cells cytology, Stem Cells immunology, Strongylida Infections immunology, Th2 Cells cytology, Th2 Cells immunology, Immunity, Mucosal immunology, Intestinal Mucosa cytology, Intestinal Mucosa immunology, Nippostrongylus immunology, Parasites immunology
- Abstract
Helminth parasitic infections are a major global health and social burden. The host defence against helminths such as Nippostrongylus brasiliensis is orchestrated by type 2 cell-mediated immunity. Induction of type 2 cytokines, including interleukins (IL) IL-4 and IL-13, induce goblet cell hyperplasia with mucus production, ultimately resulting in worm expulsion. However, the mechanisms underlying the initiation of type 2 responses remain incompletely understood. Here we show that tuft cells, a rare epithelial cell type in the steady-state intestinal epithelium, are responsible for initiating type 2 responses to parasites by a cytokine-mediated cellular relay. Tuft cells have a Th2-related gene expression signature and we demonstrate that they undergo a rapid and extensive IL-4Rα-dependent amplification following infection with helminth parasites, owing to direct differentiation of epithelial crypt progenitor cells. We find that the Pou2f3 gene is essential for tuft cell specification. Pou2f3(-/-) mice lack intestinal tuft cells and have defective mucosal type 2 responses to helminth infection; goblet cell hyperplasia is abrogated and worm expulsion is compromised. Notably, IL-4Rα signalling is sufficient to induce expansion of the tuft cell lineage, and ectopic stimulation of this signalling cascade obviates the need for tuft cells in the epithelial cell remodelling of the intestine. Moreover, tuft cells secrete IL-25, thereby regulating type 2 immune responses. Our data reveal a novel function of intestinal epithelial tuft cells and demonstrate a cellular relay required for initiating mucosal type 2 immunity to helminth infection.
- Published
- 2016
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31. Glucose and glutamine metabolism regulate human hematopoietic stem cell lineage specification.
- Author
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Oburoglu L, Tardito S, Fritz V, de Barros SC, Merida P, Craveiro M, Mamede J, Cretenet G, Mongellaz C, An X, Klysz D, Touhami J, Boyer-Clavel M, Battini JL, Dardalhon V, Zimmermann VS, Mohandas N, Gottlieb E, Sitbon M, Kinet S, and Taylor N
- Subjects
- ADP-ribosyl Cyclase 1 metabolism, Animals, Antigens, CD34 metabolism, Biological Transport, Cell Differentiation, Chromatography, Liquid, Erythrocytes cytology, Glycolysis, Green Fluorescent Proteins metabolism, Humans, Mass Spectrometry, Mice, Mice, Inbred C57BL, Minor Histocompatibility Antigens, RNA, Small Interfering metabolism, Amino Acid Transport System ASC metabolism, Cell Lineage, Gene Expression Regulation, Glucose metabolism, Glutamine metabolism, Hematopoietic Stem Cells cytology
- Abstract
The metabolic state of quiescent hematopoietic stem cells (HSCs) is an important regulator of self-renewal, but it is unclear whether or how metabolic parameters contribute to HSC lineage specification and commitment. Here, we show that the commitment of human and murine HSCs to the erythroid lineage is dependent upon glutamine metabolism. HSCs require the ASCT2 glutamine transporter and active glutamine metabolism for erythroid specification. Blocking this pathway diverts EPO-stimulated HSCs to differentiate into myelomonocytic fates, altering in vivo HSC responses and erythroid commitment under stress conditions such as hemolytic anemia. Mechanistically, erythroid specification of HSCs requires glutamine-dependent de novo nucleotide biosynthesis. Exogenous nucleosides rescue erythroid commitment of human HSCs under conditions of limited glutamine catabolism, and glucose-stimulated nucleotide biosynthesis further enhances erythroid specification. Thus, the availability of glutamine and glucose to provide fuel for nucleotide biosynthesis regulates HSC lineage commitment under conditions of metabolic stress., (Copyright © 2014 Elsevier Inc. All rights reserved.)
- Published
- 2014
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32. Ikaros' suspended flight in the thymus.
- Author
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Taylor N and Zimmermann VS
- Subjects
- Animals, Humans, Cell Differentiation immunology, Ikaros Transcription Factor immunology, Ikaros Transcription Factor metabolism, T-Lymphocytes cytology, Thymus Gland cytology
- Published
- 2013
- Full Text
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33. Concise review: hematopoietic stem cell transplantation: targeting the thymus.
- Author
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De Barros SC, Zimmermann VS, and Taylor N
- Subjects
- Animals, Cell Differentiation, Hematopoietic Stem Cells immunology, Humans, Mice, Mice, SCID, Thymus Gland immunology, Hematopoietic Stem Cell Transplantation methods, Hematopoietic Stem Cells cytology, Thymus Gland cytology
- Abstract
Allogeneic hematopoietic stem cell (HSC) transplantation can cure patients suffering from diverse genetic and acquired diseases as well as cancers. Nevertheless, under conditions where T-cell reconstitution is critical, the entry of donor progenitors into the thymus remains a major bottleneck. It is assumed that following the intravenous injection of HSC, they first home to the BM. More committed progenitors can then be exported to the thymus in response to a myriad of signals regulating thymus seeding. Notably although, the thymus is not continually receptive to the import of hematopoietic progenitors. Furthermore, as stem cells with self-renewing capacity do not take up residence in the thymus under physiological conditions, the periodic colonization of the thymus is essential for the sustained differentiation of T lymphocytes. As such, we and others have invested significant efforts into exploring avenues that might foster a long-term thymus-autonomous differentiation. Here, we review strategic approaches that have resulted in long-term T-cell differentiation in immunodeficient (SCID) mice, even across histocompatibility barriers. These include the forced thymic entry of BM precursors by their direct intrathymic injection as well as the transplantation of neonatal thymi. The capacity of the thymus to support hematopoietic progenitors with renewal potential will hopefully promote the development of new therapeutic strategies aimed at enhancing T-cell differentiation in patients undergoing HSC transplantation., (Copyright © 2013 AlphaMed Press.)
- Published
- 2013
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34. Intrathymic progenitor cell transplantation across histocompatibility barriers results in the persistence of early thymic progenitors and T-cell differentiation.
- Author
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de Barros SC, Vicente R, Chebli K, Jacquet C, Zimmermann VS, and Taylor N
- Subjects
- Animals, Cells, Cultured, Hematopoiesis immunology, Hematopoiesis physiology, Histocompatibility immunology, Histocompatibility Testing, Infusions, Intravenous, Lymphoid Progenitor Cells cytology, Mice, Mice, Inbred C57BL, Mice, Inbred CBA, Mice, Knockout, Severe Combined Immunodeficiency genetics, Severe Combined Immunodeficiency immunology, Transplantation Conditioning methods, ZAP-70 Protein-Tyrosine Kinase deficiency, ZAP-70 Protein-Tyrosine Kinase genetics, ZAP-70 Protein-Tyrosine Kinase immunology, Cell Differentiation immunology, Graft Survival immunology, Graft Survival physiology, Hematopoietic Stem Cell Transplantation methods, Histocompatibility physiology, Lymphoid Progenitor Cells physiology, T-Lymphocytes physiology, Thymus Gland cytology
- Abstract
Donor hematopoietic stem cells (HSCs) can correct T-cell deficiencies in patients with severe combined immunodeficiency by replacing resident thymus cells. However, as those progenitors that naturally migrate to the thymus are not capable of supporting long-term thymopoiesis, a successful transplant is thought to require the ongoing migration of donor progenitors. We previously showed that the forced intrathymic administration of histocompatible HSCs can sustain long-term thymopoiesis in ZAP-70-immunodeficient mice. However, it is not known whether T-cell reconstitution across histocompatibility barriers is modulated by intrathymic vs intravenous administration of HSCs. In the absence of conditioning, long-term thymopoiesis by semiallogeneic progenitors was detected in mice transplanted via the intrathymic, but not the intravenous, route. In intrathymic-transplanted mice, ongoing thymopoiesis was associated with a 10-fold higher level of early thymic progenitors (ETPs). The enhanced reconstitution capacity of these intrathymic-derived ETPs was corroborated by their significantly augmented myeloid lineage potential compared with endogenous ETPs. Notably, though, myeloablative conditioning resulted in a reduced expansion of intrathymic-administered donor ETPs. Thus, in the absence of conditioning, the forced thymic entry of HSCs results in a sustained T-cell development across histocompatibility barriers, highlighting the capacity of the thymus to support cells with long-term renewal potential.
- Published
- 2013
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35. Molecular and cellular basis of T cell lineage commitment.
- Author
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Vicente R, Swainson L, Marty-Grès S, De Barros SC, Kinet S, Zimmermann VS, and Taylor N
- Subjects
- Animals, Cell Differentiation, Humans, Interleukin-7 immunology, Interleukin-7 metabolism, Receptor, Notch1 immunology, Receptor, Notch1 metabolism, Cell Lineage, T-Lymphocytes cytology, T-Lymphocytes immunology
- Abstract
The thymus forms as an alymphoid thymic primordium with T cell differentiation requiring the seeding of this anlage. This review will focus on the characteristics of the hematopoietic progenitors which colonize the thymus and their subsequent commitment/differentiation, both in mice and men. Within the thymus, the interplay between Notch1 and IL-7 signals is crucial for the orchestration of T cell development, but the precise requirements for these factors in murine and human thympoeisis are not synonymous. Recent advances in our understanding of the mechanisms regulating precursor entry and their maintenance in the thymus will also be presented., (Copyright 2010 Elsevier Ltd. All rights reserved.)
- Published
- 2010
- Full Text
- View/download PDF
36. Intrathymic transplantation of bone marrow-derived progenitors provides long-term thymopoiesis.
- Author
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Vicente R, Adjali O, Jacquet C, Zimmermann VS, and Taylor N
- Subjects
- Animals, Cell Differentiation genetics, Cell Differentiation physiology, Cell Lineage genetics, Cells, Cultured, Infusions, Intravenous, Lymphocyte Count, Lymphoid Progenitor Cells physiology, Lymphopoiesis genetics, Mice, Mice, Inbred C57BL, Mice, Knockout, Stem Cell Niche cytology, T-Lymphocytes cytology, T-Lymphocytes physiology, Thymus Gland physiology, Time Factors, ZAP-70 Protein-Tyrosine Kinase genetics, Bone Marrow Transplantation methods, Bone Marrow Transplantation physiology, Lymphoid Progenitor Cells transplantation, Lymphopoiesis physiology, Thymus Gland cytology
- Abstract
The sustained differentiation of T cells in the thymus cannot be maintained by resident intrathymic (IT) precursors and requires that progenitors be replenished from the bone marrow (BM). In patients with severe combined immunodeficiency (SCID) treated by hematopoietic stem cell transplantation, late T-cell differentiation defects are thought to be due to an insufficient entry of donor BM progenitors into the thymus. Indeed, we find that the intravenous injection of BM progenitors into nonconditioned zeta-chain-associated protein kinase 70 (ZAP-70)-deficient mice with SCID supports short- but not long-term thymopoiesis. Remarkably, we now show that the IT administration of these progenitors produces a significant level of donor-derived thymopoiesis for more than 6 months after transplantation. In contrast to physiologic thymopoiesis, long-term donor thymopoiesis was not due to the continued recruitment of progenitors from the BM. Rather, IT transplantation resulted in the unique generation of a large population of early c-Kit(high) donor precursors within the thymus. These ZAP-70-deficient mice that received an IT transplant had a significantly increased prothymocyte niche compared with their untreated counterparts; this phenotype was associated with the generation of a medulla. Thus, IT administration of BM progenitors results in the filling of an expanded precursor niche and may represent a strategy for enhancing T-cell differentiation in patients with SCID.
- Published
- 2010
- Full Text
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37. Efficient intrathymic gene transfer following in situ administration of a rAAV serotype 8 vector in mice and nonhuman primates.
- Author
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Moreau A, Vicente R, Dubreil L, Adjali O, Podevin G, Jacquet C, Deschamps JY, Klatzmann D, Cherel Y, Taylor N, Moullier P, and Zimmermann VS
- Subjects
- Animals, Cell Differentiation, Cell Line, Cell Movement, Dependovirus classification, Genetic Vectors pharmacology, Genome, Viral genetics, Humans, Kinetics, Mice, Phenotype, Thymus Gland cytology, Dependovirus genetics, Genetic Vectors administration & dosage, Genetic Vectors genetics, Macaca fascicularis metabolism, Thymus Gland metabolism, Transgenes genetics
- Abstract
The thymus is the primary site of T-cell development and plays a key role in the induction of self-tolerance. We previously showed that the intrathymic (i.t.) injection of a transgene-expressing lentiviral vector (LV) in mice can result in the correction of a T cell-specific genetic defect. Nevertheless, the efficiency of thymocyte transduction did not exceed 0.1-0.3% and we were unable to detect any thymus transduction in macaques. As such, we initiated studies to assess the capacity of recombinant adeno-associated virus (rAAV) vectors to transduce murine and primate thymic cells. In vivo administration of AAV serotype 2-derived single-stranded AAV (ssAAV) and self-complementary AAV (scAAV) vectors pseudotyped with capsid proteins of serotypes 1, 2, 4, 5, and 8 demonstrated that murine thymus transduction was significantly enhanced by scAAV2/8. Transgene expression was detected in 5% of thymocytes and, notably, transduced cells represented 1% of peripheral T lymphocytes. Moreover, i.t. administration of scAAV2/8 particles in macaques, by endoscopic-mediated guidance, resulted in significant gene transfer. Thus, in healthy animals, where thymic gene transfer does not provide a selective advantage, scAAV2/8 is a unique tool promoting the in situ transduction of thymocytes with the subsequent export of gene-modified lymphocytes to the periphery.
- Published
- 2009
- Full Text
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38. Tumors hamper the immunogenic competence of CD4+ T cell-directed dendritic cell vaccination.
- Author
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Zimmermann VS, Casati A, Schiering C, Caserta S, Hess Michelini R, Basso V, and Mondino A
- Subjects
- Animals, CD11b Antigen analysis, CD4-Positive T-Lymphocytes immunology, Dendritic Cells transplantation, Immunologic Memory, Mice, Mice, Inbred BALB C, Myeloid Cells immunology, Neoplasms therapy, Peptides genetics, Peptides immunology, Receptors, Chemokine analysis, Antigens, Protozoan immunology, Dendritic Cells immunology, Histocompatibility Antigens Class II immunology, Neoplasms prevention & control, Protozoan Proteins immunology, Vaccination methods
- Abstract
Dendritic cells loaded with tumor-derived peptides induce protective CTL responses and are under evaluation in clinical trails. We report in this study that prophylactic administration of dendritic cells loaded with a MHC class II-restricted peptide derived from a model tumor Ag (Leishmania receptor for activated C kinase (LACK)) confers protection against LACK-expressing TS/A tumors, whereas therapeutic vaccination fails to cure tumor-bearing mice. Although CD4+ T cell-directed dendritic cell vaccination primed effector-like (CD44(high)CD62L(low), IL-2(+), IFN-gamma(+)) and central memory-like lymphocytes (CD44(high)CD62L(high), only IL-2(+)) in tumor-free mice, this was not the case in tumor-bearing animals in which both priming and persistence of CD4+ T cell memory were suppressed. Suppression was specific for the tumor-associated Ag LACK, and did not depend on CD25+ T cells. Because T cell help is needed for protective immunity, we speculate that the ability of tumors to limit vaccine-induced CD4+ T cell memory could provide a partial explanation for the limited efficacy of current strategies.
- Published
- 2007
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39. Requirement for dendritic cells in the establishment of anti-phospholipid antibodies.
- Author
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Bondanza A, Rovere-Querini P, Zimmermann VS, Balestrieri G, Tincani A, Sabbadini MG, and Manfredi AA
- Subjects
- Animals, Cattle, Dendritic Cells transplantation, Female, Immunization, Lupus Erythematosus, Systemic immunology, Mice, Transplantation, Heterologous, Transplantation, Isogeneic, Antibodies, Antiphospholipid immunology, Apoptosis immunology, Dendritic Cells immunology, Pinocytosis immunology, beta 2-Glycoprotein I immunology
- Abstract
Objective: The cross-presentation of cell-associated autoantigens contributes to systemic autoimmune diseases, including systemic lupus erythematosus (SLE). Little is known about the regulation of the immune response against soluble autoantigens targeted in these diseases., Methods: We immunized the offspring of New Zealand Black and New Zealand White mice (NZB x NZW F(1)) with syngeneic dendritic cells (DC) that had macropinocytosed beta2-glycoprotein 1 (beta(2)GPI) during propagation in normal mouse serum or that had phagocytosed apoptotic thymocytes with syngeneic (murine) or xenogeneic (bovine) beta(2)GPI, which was associated to plasma membrane of the cells. Mice were in parallel immunized with apoptotic thymocytes that had associated the beta(2)GPI to their membranes in the absence of DC. The development of anti-beta(2)GPI antibodies and clinical features were monitored., Results: Apoptotic cells alone, opsonized with beta(2)GPI, failed to induce anti-beta(2)GPI autoantibodies or clinical disease. In contrast, autoimmunity developed in the presence of DC. Furthermore, the syngeneic beta(2)GPI was a more effective antigen than the xenogeneic protein in re-boosted animals., Conclusions: DC effectively initiate in NZB x NZW F(1) mice self-sustaining autoimmunity against the beta(2)GPI, either associated to apoptotic cells or macropinocytosed from the serum.
- Published
- 2007
- Full Text
- View/download PDF
40. Intrathymic administration of hematopoietic progenitor cells enhances T cell reconstitution in ZAP-70 severe combined immunodeficiency.
- Author
-
Adjali O, Vicente RR, Ferrand C, Jacquet C, Mongellaz C, Tiberghien P, Chebli K, Zimmermann VS, and Taylor N
- Subjects
- Animals, Cell Differentiation immunology, Humans, Mice, Mice, Knockout, Severe Combined Immunodeficiency immunology, Thymus Gland immunology, Transplantation, Homologous, ZAP-70 Protein-Tyrosine Kinase, Hematopoietic Stem Cell Transplantation, Hematopoietic Stem Cells immunology, Protein-Tyrosine Kinases deficiency, Recovery of Function immunology, Severe Combined Immunodeficiency therapy, T-Lymphocytes immunology
- Abstract
Patients with severe combined immunodeficiency (SCID) present with opportunistic infections that are almost universally fatal in infancy. The mainstay treatment for these patients is allogeneic hematopoietic stem cell (HSC) transplantation, but sustained polyclonal T cell reconstitution is too often unsatisfactory. Although transplantation is conventionally performed by i.v. administration of HSC, we hypothesized that an intrathymic strategy would be superior. Indeed, several progenitor cell populations are incapable of homing to the thymus, the major site of T cell differentiation, and it appears that there are extensive time periods during which the thymus is refractory to progenitor cell import. To test this hypothesis, nonconditioned infant ZAP-70-deficient SCID mice were intrathymically injected with WT bone marrow progenitor cells, a procedure accomplished without surgical intervention. Upon intrathymic HSC injection, there was a more rapid T cell differentiation, with mature thymocytes detected by 4 weeks after transplantation. Intrathymic injection of HSC also resulted in significantly higher numbers of peripheral T cells, increased percentages of naïve T cells, and more diverse T cell receptor repertoires. Moreover, T cell reconstitution after intrathymic transplantation was obtained after injection of 10-fold fewer donor HSC. Thus, this intrathymic transplantation approach may improve the outcome of SCID patients by enhancing T cell reconstitution.
- Published
- 2005
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41. Phenotype and homing of CD4 tumor-specific T cells is modulated by tumor bulk.
- Author
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Benigni F, Zimmermann VS, Hugues S, Caserta S, Basso V, Rivino L, Ingulli E, Malherbe L, Glaichenhaus N, and Mondino A
- Subjects
- Adenocarcinoma genetics, Adenocarcinoma surgery, Amino Acid Sequence, Animals, Antigens, Protozoan biosynthesis, Antigens, Protozoan immunology, Breast Neoplasms genetics, Breast Neoplasms surgery, CD4-Positive T-Lymphocytes metabolism, CD4-Positive T-Lymphocytes pathology, Cell Differentiation immunology, Cell Line, Tumor, Cell Movement genetics, Dendritic Cells immunology, Dendritic Cells transplantation, Epitopes, T-Lymphocyte biosynthesis, Histocompatibility Antigens Class II biosynthesis, Immunologic Memory genetics, Interferon-gamma metabolism, Lymph Nodes immunology, Lymph Nodes metabolism, Lymph Nodes pathology, Mice, Mice, Inbred BALB C, Mice, Transgenic, Molecular Sequence Data, Neoplasm Transplantation, Organ Specificity genetics, Organ Specificity immunology, Protozoan Proteins biosynthesis, Protozoan Proteins immunology, Adenocarcinoma immunology, Adenocarcinoma pathology, Breast Neoplasms immunology, Breast Neoplasms pathology, CD4-Positive T-Lymphocytes immunology, Cell Movement immunology, Epitopes, T-Lymphocyte immunology, Immunophenotyping
- Abstract
Technical difficulties in tracking endogenous CD4 T lymphocytes have limited the characterization of tumor-specific CD4 T cell responses. Using fluorescent MHC class II/peptide multimers, we defined the fate of endogenous Leishmania receptor for activated C kinase (LACK)-specific CD4 T cells in mice bearing LACK-expressing TS/A tumors. LACK-specific CD44(high)CD62L(low) CD4 T cells accumulated in the draining lymph nodes and had characteristics of effector cells, secreting IL-2 and IFN-gamma upon Ag restimulation. Increased frequencies of CD44(high)CD62L(low) LACK-experienced cells were also detected in the spleen, lung, liver, and tumor itself, but not in nondraining lymph nodes, where the cells maintained a naive phenotype. The absence of systemic redistribution of LACK-specific memory T cells correlated with the presence of tumor. Indeed, LACK-specific CD4 T cells with central memory features (IL-2(+)IFN-gamma(-)CD44(high)CD62L(high) cells) accumulated in all peripheral lymph nodes of mice immunized with LACK-pulsed dendritic cells and after tumor resection. Together, our data demonstrate that although tumor-specific CD4 effector T cells producing IFN-gamma are continuously generated in the presence of tumor, central memory CD4 T cells accumulate only after tumor resection. Thus, the continuous stimulation of tumor-specific CD4 T cells in tumor-bearing mice appears to hinder the systemic accumulation of central memory CD4 T lymphocytes.
- Published
- 2005
- Full Text
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42. Immune surveillance and anti-tumor immune responses: an anatomical perspective.
- Author
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Zimmermann VS, Benigni F, and Mondino A
- Subjects
- Animals, Cancer Vaccines immunology, Humans, Immunotherapy, Active, Lymphocytes immunology, Lymphoid Tissue immunology, Neoplasms drug therapy, Antigens, Neoplasm immunology, Neoplasms immunology
- Abstract
The development of adaptive immune responses against infectious agents relies on the initiation of antigen specific immune responses in secondary lymphoid organs and on the migration of effector cells at the site of infection. Similarly, the development of anti-tumor immunity depends on the recognition of tumor-derived antigens by specific lymphocytes in the context of the lymphoid tissues and on the re-localisation of the cells to the site of cell transformation. Here, we will review the preclinical studies, which have defined the spatial and temporal organisation of anti-tumor immunity, and discuss the implications of these findings in active immunotherapy.
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- 2005
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43. The immunogenicity of dendritic cell-based vaccines is not hampered by doxorubicin and melphalan administration.
- Author
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Casati A, Zimmermann VS, Benigni F, Bertilaccio MT, Bellone M, and Mondino A
- Subjects
- Animals, Antibiotics, Antineoplastic administration & dosage, Antibiotics, Antineoplastic toxicity, Antineoplastic Agents, Alkylating administration & dosage, Antineoplastic Agents, Alkylating toxicity, CD4-Positive T-Lymphocytes cytology, CD4-Positive T-Lymphocytes drug effects, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes drug effects, CD8-Positive T-Lymphocytes immunology, Cell Differentiation drug effects, Cell Division drug effects, Doxorubicin toxicity, Humans, In Vitro Techniques, Lymphocyte Count, Melphalan toxicity, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Mice, Transgenic, Neoplasms, Experimental drug therapy, Neoplasms, Experimental immunology, Neoplasms, Experimental therapy, Receptors, Antigen, T-Cell, alpha-beta genetics, Cancer Vaccines administration & dosage, Cancer Vaccines immunology, Dendritic Cells immunology, Doxorubicin administration & dosage, Melphalan administration & dosage
- Abstract
Immunization of cancer patients is most effective in tumor-free conditions or in the presence of minimal residual disease. In the attempt to develop new strategies able to control tumor recurrence while allowing the development of protective immunity, we have investigated the immunogenic potential of two distinct vaccine formulations when provided alone or upon single and repeated treatment with chemotherapeutics drugs. Vaccine-induced T cell responses were first investigated by tracing Ag-specific T cell responses in mice bearing detectable frequencies of Ag-specific TCR transgenic CD4 and CD8 T cells. These studies indicated that immunization with peptide-pulsed dendritic cells and soluble Ag plus adjuvant elicited a comparable expansion and differentiation of CD4 and CD8 effector cells in the peripheral lymphoid tissues when provided alone or shortly after Doxorubicin or Melphalan administration. We also analyzed the potency of the combined vaccination in transgenic adenocarcinoma mouse prostate mice, which develop spontaneous prostate cancer. Dendritic cell-based vaccination elicited potent tumor-specific cytotoxic responses in mice bearing prostate intraepithelial neoplasia both in the absence and in the presence of Doxorubicin. Together our results indicate that Doxorubicin- or Melphalan-based chemotherapy and Ag-specific vaccination can be combined for adjuvant treatments of cancer patients.
- Published
- 2005
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- View/download PDF
44. Inhibition of phosphatidylserine recognition heightens the immunogenicity of irradiated lymphoma cells in vivo.
- Author
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Bondanza A, Zimmermann VS, Rovere-Querini P, Turnay J, Dumitriu IE, Stach CM, Voll RE, Gaipl US, Bertling W, Pöschl E, Kalden JR, Manfredi AA, and Herrmann M
- Subjects
- Animals, Annexin A5 metabolism, Dendritic Cells immunology, Enzyme-Linked Immunosorbent Assay, Female, Flow Cytometry, Macrophages immunology, Mice, Mice, Inbred C57BL, Phagocytosis immunology, Receptors, Cell Surface metabolism, Tumor Cells, Cultured, Annexin A5 immunology, Immunization, Lymphoma immunology, Lymphoma therapy, Receptors, Cell Surface immunology, Ultraviolet Rays
- Abstract
Strategies to enhance the immunogenicity of tumors are urgently needed. Although vaccination with irradiated dying lymphoma cells recruits a tumor-specific immune response, its efficiency as immunogen is poor. Annexin V (AxV) binds with high affinity to phosphatidylserine on the surface of apoptotic and necrotic cells and thereby impairs their uptake by macrophages. Here, we report that AxV preferentially targets irradiated lymphoma cells to CD8+ dendritic cells for in vivo clearance, elicits the release of proinflammatory cytokines and dramatically enhances the protection elicited against the tumor. The response was endowed with both memory, because protected animals rejected living lymphoma cells after 72 d, and specificity, because vaccinated animals failed to reject unrelated neoplasms. Finally, AxV-coupled irradiated cells induced the regression of growing tumors. These data indicate that endogenous adjuvants that bind to dying tumor cells can be exploited to target tumors for immune rejection.
- Published
- 2004
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45. Requirement of dying cells and environmental adjuvants for the induction of autoimmunity.
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Bondanza A, Zimmermann VS, Dell'Antonio G, Cin ED, Balestrieri G, Tincani A, Amoura Z, Piette JC, Sabbadini MG, Rovere-Querini P, and Manfredi AA
- Subjects
- Animals, Autoantibodies immunology, Dendritic Cells immunology, Dendritic Cells transplantation, Female, Freund's Adjuvant pharmacology, Glycoproteins immunology, Immunization, Mice, Mice, Inbred BALB C, Mice, Inbred NZB, Severity of Illness Index, Thymus Gland cytology, Thymus Gland immunology, beta 2-Glycoprotein I, Apoptosis immunology, Autoimmunity immunology, Lupus Erythematosus, Systemic immunology, Lupus Erythematosus, Systemic pathology
- Abstract
Objective: Cells commonly die without eliciting autoimmunity. However, dying cells are a potential initiating stimulus for systemic lupus erythematosus (SLE). Our goal was to verify whether immune adjuvants influence the autoimmunity induction that ensues following in vivo injection of dying cells., Methods: Mice were immunized with apoptotic thymocytes in the presence of artificial moieties, such as Freund's incomplete adjuvant (IFA), or natural adjuvants, such as dendritic cells (DCs). Renal involvement and the development of autoantibodies were monitored., Results: Apoptotic cells failed to induce clinical disease or to sustain production of autoantibodies in (NZB x NZW)F(1) mice. In contrast, autoimmunity developed in the presence of IFA or DCs. The characteristics of the adjuvant influenced the array of autoantibodies, the kinetics of their development, and the severity of the disease. DCs were required for induction of anti-beta(2)-glycoprotein I IgG. Adjuvants alone did not elicit disease., Conclusion: A "two-hit" signal composed of autoantigens and adjuvants initiates systemic autoimmunity. Moreover, environmental signals at the site of clearance of dead cells shape the features and the severity of the autoimmune disease. Strategies aimed at preventing the accumulation of dying cells and at modulating endogenous adjuvants may be beneficial for the treatment of SLE.
- Published
- 2004
- Full Text
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46. Characterisation of functional biotinylated TNF-alpha targeted to the membrane of apoptotic melanoma cells.
- Author
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Zimmermann VS, Bondanza A, Rovere-Querini P, Colombo B, Sacchi A, Fascio U, Corti A, and Manfredi AA
- Subjects
- Animals, Biotinylation, Cell Membrane chemistry, Cell Membrane drug effects, Cytokines biosynthesis, Drug Delivery Systems, Flow Cytometry, Macrophages immunology, Melanoma, Experimental immunology, Melanoma, Experimental pathology, Mice, Phagocytosis, Tumor Cells, Cultured, Tumor Necrosis Factor-alpha analysis, Tumor Necrosis Factor-alpha chemistry, Apoptosis, Melanoma, Experimental drug therapy, Tumor Necrosis Factor-alpha therapeutic use
- Abstract
The physiologic clearance of apoptotic cells prevents inflammation at the site of cell death and limits the immunogenicity of tumors. In this study we report the functional characterisation of biotinylated tumor necrosis factor-alpha (TNF-alpha) after anchorage to apoptotic melanoma cells via a biotin-avidin-biotin bridge. Flow cytometric and morphological analysis showed that biotinylated TNF-alpha efficiently bound to apoptotic membrane blebs of dying cells. Membrane-bound TNF-alpha (12 fg/cell) killed sensitive WEHI164 cells 250-fold more effectively than equivalent amounts of the soluble cytokine. Furthermore, macrophages engulfing apoptotic cells with membrane-bound TNF-alpha secreted significantly higher amounts of soluble TNF-alpha and lower amounts of interleukin-10 (IL-10). Therefore the bridging of TNF-alpha potentiates its biological function and influences the outcome of the phagocytic clearance of apoptotic tumor cells.
- Published
- 2003
- Full Text
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47. Anti-beta2 glycoprotein I antibodies cause inflammation and recruit dendritic cells in platelet clearance.
- Author
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Bondanza A, Manfredi AA, Zimmermann VS, Iannacone M, Tincani A, Balestrieri G, Sabbadini MG, and Querini PR
- Subjects
- Antibodies isolation & purification, Blood Platelets metabolism, Dendritic Cells drug effects, Dendritic Cells metabolism, Glycoproteins metabolism, Humans, Immunoglobulin G isolation & purification, Immunoglobulin G pharmacology, Interleukin-1 analysis, Interleukin-1 metabolism, Interleukin-10 analysis, Interleukin-10 metabolism, Macrophages immunology, Macrophages metabolism, Macrophages physiology, Phagocytosis immunology, Platelet Activation, Tumor Necrosis Factor-alpha analysis, Tumor Necrosis Factor-alpha metabolism, beta 2-Glycoprotein I, Antibodies pharmacology, Dendritic Cells immunology, Glycoproteins immunology, Inflammation chemically induced
- Abstract
Scavenger phagocytes are mostly responsible for the in vivo clearance of activated or senescent platelets. In contrast to other particulate substrates, the phagocytosis of platelets does not incite proinflammatory responses in vivo. This study assessed the contribution of macrophages and dendritic cells (DCs) to the clearance of activated platelets. Furthermore, we verified whether antibodies against the beta2 Glycoprotein I (beta2GPI), which bind to activated platelets, influence the phenomenon. DCs did not per se intemalise activated platelets. In contrast, macrophages efficiently phagocytosed platelets. In agreement with the uneventful nature of the clearance of platelets in vivo, phagocytosing macrophages did not release IL-1beta, TNF-alpha, or IL-10, beta2GPI bound to activated platelets and was required for their recognition by anti-beta2GPI antibodies. DCs internalised platelets opsonised by anti-beta2GPI antibodies. The phagocytosis of opsonised platelets determined the release of TNF-alpha and IL-1beta by DCs and macrophages. Phagocytosing macrophages, but not DCs, secreted the antiinflammatory cytokine IL-10. We conclude that anti-beta2GPI antibodies cause inflammation during platelet clearance and shuttle platelet antigens to antigen presenting DCs.
- Published
- 2001
48. Cytokine secretion associated with the clearance of apoptotic bodies in renal cell carcinoma patients.
- Author
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Bondanza A, Rovere P, Borri A, Caremoli ER, Guidetti A, Citterio G, Consogno G, Zimmermann VS, Rugarli C, and Manfredi AA
- Subjects
- Adult, Age Factors, Aged, Carcinoma, Renal Cell mortality, Carcinoma, Renal Cell therapy, Case-Control Studies, Disease-Free Survival, Female, Humans, Immunotherapy, Interleukin-10 biosynthesis, Jurkat Cells, Kidney Neoplasms mortality, Kidney Neoplasms therapy, Macrophages immunology, Macrophages metabolism, Male, Middle Aged, Neoplasm Metastasis, Phagocytosis, Treatment Outcome, Apoptosis, Carcinoma, Renal Cell metabolism, Cytokines biosynthesis, Kidney Neoplasms metabolism
- Abstract
The factors determining the outcome of immunotherapy in metastatic renal cell carcinoma (RCC) patients remain elusive. Macrophages from normal donors that phagocytose apoptotic cells secrete the immunosuppressive cytokine IL-10 in vitro. Conversely, IL-10 genetic deletion enhances the immunogenicity of apoptotic tumor cells in vivo. Elevated pre-treatment levels of IL-10 are associated with an unfavorable outcome of RCC. We examined whether the ability to release IL-10 by macrophages from RCC patients that phagocytosed apoptotic cells correlated with the outcome of immunotherapy. To this aim, we derived macrophages from 30 patients with metastatic RCC and from 21 healthy subjects (11 sex- and age-matched healthy controls and 10 younger donors). Patients either had a clinical response after immunotherapy, with a median survival after treatment of more than 18 months (n = 16), or were beginning immunotherapy after diagnosis of metastatic disease (n = 14). Macrophages from responding patients challenged with apoptotic cells released significantly less IL-10 than controls (p = 0.0075) and recently diagnosed patients (p = 0.0198), as ascertained by a 2-sided Student's t-test. This was not because macrophages from responding patients lost the ability to secrete IL-10, because antibody opsonization of apoptotic cells rescued IL-10 secretion. In contrast, macrophages from all groups of donors released similar amounts of TNF-alpha. The failure in IL-10 secretion by engulfing macrophages of responding subjects may exalt the immunogenicity of dying tumor cells, contributing to the success of immunotherapy., (Copyright 2001 Wiley-Liss, Inc.)
- Published
- 2001
- Full Text
- View/download PDF
49. The long pentraxin PTX3 binds to apoptotic cells and regulates their clearance by antigen-presenting dendritic cells.
- Author
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Rovere P, Peri G, Fazzini F, Bottazzi B, Doni A, Bondanza A, Zimmermann VS, Garlanda C, Fascio U, Sabbadini MG, Rugarli C, Mantovani A, and Manfredi AA
- Subjects
- Acute-Phase Reaction, Antigens, Nuclear, Cell Membrane metabolism, Dendritic Cells drug effects, Humans, Inflammation pathology, Jurkat Cells metabolism, Jurkat Cells radiation effects, Microscopy, Confocal, Necrosis, Neutrophils cytology, Neutrophils metabolism, Phagocytosis physiology, Protein Structure, Tertiary, Recombinant Fusion Proteins metabolism, T-Lymphocytes cytology, T-Lymphocytes metabolism, Time Factors, Tumor Necrosis Factor-alpha pharmacology, Ultraviolet Rays, fas Receptor physiology, Apoptosis physiology, C-Reactive Protein metabolism, Dendritic Cells physiology, Nuclear Proteins metabolism, Serum Amyloid P-Component metabolism
- Abstract
Pentraxins are acute-phase proteins produced in vivo during inflammatory reactions. Classical short pentraxins, C-reactive protein, and serum amyloid P component are generated in the liver in response to interleukin (IL)-6. The long pentraxin PTX3 is produced in tissues under the control of primary proinflammatory signals, such as lipopolysaccharide, IL-1 beta, and tumor necrosis factor-alpha, which also promote maturation of dendritic cells (DCs). Cell death commonly occurs during inflammatory reactions. In this study, it is shown that PTX3 specifically binds to dying cells. The binding was dose dependent and saturable. Recognition was restricted to extranuclear membrane domains and to a chronological window after UV irradiation or after CD95 cross-linking-induced or spontaneous cell death in vitro. PTX3 bound to necrotic cells to a lesser extent. Human DCs failed to internalize dying cells in the presence of PTX3, while they took up normally soluble or inert particulate substrates. These results suggest that PTX3 sequesters cell remnants from antigen-presenting cells, possibly contributing to preventing the onset of autoimmune reactions in inflamed tissues. (Blood. 2000;96:4300-4306)
- Published
- 2000
50. Anti-beta2 glycoprotein I antibodies prevent the De-activation of platelets and sustain their phagocytic clearance.
- Author
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Bondanza A, Sabbadini MG, Pellegatta F, Zimmermann VS, Tincani A, Balestrieri G, Manfredi AA, and Rovere P
- Subjects
- Humans, Immunoglobulin G immunology, Macrophages physiology, Phosphatidylserines pharmacology, Thrombin pharmacology, beta 2-Glycoprotein I, Antibodies immunology, Glycoproteins immunology, Phagocytosis, Platelet Activation
- Abstract
Exposure to phosphatidylserine (PS) tags dying and senescent cells for removal and identifies activated platelets. In this study we followed the fate of PS-exposing platelets in the presence of antibodies purified from Systemic Lupus Erythematosus (SLE) and primary Anti-phospholipid Syndrome (APS) patients' sera by beta2GPI affinity chromatography. Thrombin-activated platelets exposed PS and associated to beta2GPI. Both events were required for recognition by antibodies. Human monocyte-derived macrophages phagocytosed activated platelets only. Each macrophage internalized an average of 3.16+/-0.2 platelets after 60 min at 37 degrees C. Phagocytosis did not increase after longer incubations (4.65+/-0.26 platelets internalized by each macrophage after 300 min). Recognition of platelets by anti-beta2GPI antibodies significantly increased phagocytosis (P< 0.01). Upon withdrawal of thrombin, platelets downregulated PS (PS exposure t(1/2): 242 min) and the ability to be recognized by macrophages. Purified beta2GPI bound to PS-exposing platelets (association t(1/2): 250 min). Phosphatidyl serine exposure and beta2GPI association had virtually identical kinetics. Antibody binding prolonged the exposure of the beta2GPI/PS complex (t(1/2): >1200 min). The ability to phagocytose opsonized platelets was accordingly sustained (5.3+/-0.2 opsonized platelets were internalized by each macrophage after 60 min and 9.4+/-0.3 after 300 min). Anti-beta2GPI antibodies therefore poise activated platelets in a PS-exposing status, preventing the recycling of their function and favoring their phagocytic clearance., (Copyright 2000 Academic Press.)
- Published
- 2000
- Full Text
- View/download PDF
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