75 results on '"Ramstedt U"'
Search Results
2. Methods of treating orthomyxoviral infections
- Author
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Ramstedt, U, Klose, B, Zitzmann, N, Dwek, R, and Butters, T
- Abstract
Provided are novel iminosugars and methods of treating and/or preventing a disease or condition caused by or associated with a virus belonging to the Orthomyxoviridae family using iminosugars, such as DNJ derivatives.
- Published
- 2012
3. Iminosugars and methods of treating viral diseases
- Author
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Ramstedt, U, Klose, B, Zitzmann, N, Dwek, R, and Butters, T
- Subjects
viruses - Abstract
Provided are methods of treating or preventing viral infections caused by or associated with a Dengue virus using iminosugars.
- Published
- 2011
4. Methods of treating Poxviral Infections
- Author
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Ramstedt, U, Klose, B, Zitzmann, N, Dwek, R, and Butters, T
- Subjects
viruses - Abstract
The present application relates to iminosugars and methods of treating viral infections with iminosugars and, in particular, to the use of iminosugars for treatment and/or prevention of viral infections caused by or associated with a virus belonging to the Poxviridae family.
- Published
- 2011
5. Iminosugars and methods of treating Filoviral diseases
- Author
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Ramstedt, U, Klose, B, Zitzmann, N, Dwek, R, and Butters, T
- Subjects
viruses - Abstract
The present application relates to iminosugars and methods of treating viral infections with iminosugars and, in particular, to the use of iminosugars for treatment and/or prevention of viral infections caused by or associated with a virus belonging to the Filoviridae family.
- Published
- 2011
6. Iminosugars and methods of treating Bunyaviral and Togaviral diseases
- Author
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Ramstedt, U, Klose, B, Zitzmann, N, Dwek, R, and Butters, T
- Subjects
viruses - Abstract
The present application relates to iminosugars and methods of treating viral infections with iminosugars and, in particular, to the use of iminosugars for treatment and/or prevention of viral infections caused by or associated with a virus belonging to the Bunyaviral or Togaviral family.
- Published
- 2010
7. Iminosugars and methods of treating Arenaviral infections
- Author
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Ramstedt, U, Klose, B, Zitzmann, N, Dwek, R, and Butters, T
- Subjects
viruses - Abstract
The present application relates to iminosugars and methods of treating viral infections with iminosugars and, in particular, to the use of iminosugars for treatment and/or prevention of viral infections caused by or associated with a virus belonging to the Arenaviridae family.
- Published
- 2010
8. Role of human T-cell leukemia virus type 1 X region proteins in immortalization of primary human lymphocytes in culture
- Author
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Grassmann, R, primary, Berchtold, S, additional, Radant, I, additional, Alt, M, additional, Fleckenstein, B, additional, Sodroski, J G, additional, Haseltine, W A, additional, and Ramstedt, U, additional
- Published
- 1992
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- View/download PDF
9. Impact of 90Sr on Mouse Natural Killer Cells and their Regulation by Alpha Interferon and Interleukin 2
- Author
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GIDLUND, M., primary, BIERKE, P., additional, ÖRN, A., additional, AXBERG, I., additional, RAMSTEDT, U., additional, and WIGZELL, H., additional
- Published
- 1990
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10. Impact of 90Sr on Mouse Natural Killer Cells and their Regulation by Alpha Interferon and Interleukin 2.
- Author
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Gidlund, M., Bierke, P., Örn, A., Axberg, I., Ramstedt, U., and Wigzell, H.
- Subjects
KILLER cells ,IMMUNOCOMPETENT cells ,INTERLEUKIN-2 ,ALLOCATION of organs, tissues, etc. ,INTERFERONS ,IONIZING radiation - Abstract
Male CBA/SU mice were exposed to ionizing radiation by intraperitoneal injection of the bone-seeking β-emitter
90 Sr. NK-cell lytic activities in spleen, peripheral blood, and lymph nodes were severely depressed or completely abolished. In contrast, production of the NK regulatory proteins alpha interferon (IFN-α) and interleukin 2 (IL-2) was normal 5-8 weeks after90 Sr injection. IFN-α, produced in vivo or in vitro by cells from injected mice, was able to enhance strongly NK lytic activities. These data indicate that90 Sr acts on the bone marrow, where it interferes with the maturation and seeding of NK precursor cells. The mechanisms regulating NK activities in peripheral organs remained relatively unchanged. Finally, we did not detect any major organ redistribution of NK cells as a result of90 Sr irradiation. [ABSTRACT FROM AUTHOR]- Published
- 1990
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11. Inhibition of Natural Killer Cell Cytotoxicity by a Monoclonal Antibody Directed against Adhesion-Mediating Protein gp 90 (CD 18).
- Author
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Axberg, I., Ramstedt, U., Patarroyo, M., Beatty, P., and Wigzell, H.
- Subjects
KILLER cells ,IMMUNOCOMPETENT cells ,LYMPHOCYTES ,MOLECULES ,IMMUNOGLOBULINS ,MONOCLONAL antibodies - Abstract
Contact is required between the effector natural killer (NK) or cytotoxic killer T cell and its corresponding target in order for efficient lysis to occur. Several surface molecules of different types are involved in this process. Here we could show that Fab fragments from a murine monoclonal antibody reacting with gp 90. The human leucocyte common antigen CD18 are extremely efficient in blocking human NK of killer T cells, regardless of whether the target has or does not have the antigen. In contrast, no impact of the antibody was observed when the effector cells were of murine origin, again regardless of whether the target cell did or did not display the leucocyte common antigen. The inhibition could be shown to occur at the level of blockage of target-conjugate formation. This means that the functional display of effector/ target gp 90 on the effector but not the target cell is necessary for efficient lysis to occur both in NK and killer T cell systems, [ABSTRACT FROM AUTHOR]
- Published
- 1987
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12. Human Peripheral Blood Null Lymphocytes Stimulated by Staphylococcus aureus Cowan I Produce Atypical Acid-Labile Interferon in Vitro.
- Author
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Funa, K., Ramstedt, U., and Rönnblom, L.
- Subjects
LYMPHOCYTES ,INTERFERONS ,STAPHYLOCOCCUS aureus ,BLOOD cells ,STAPHYLOCOCCUS ,CELLS ,LYMPHOKINES ,ANTINEOPLASTIC agents - Abstract
Peripheral blood mononuclear leucocytes (PBLs) stimulated in vitro by heat-killed formaldehyde-fixed Staphylococcus aureus Cowan I (SACol) produced acid-labile alpha interferon (IFN-α) and, to various extents, also IFN-γ. The IFN producers resided in nylon wool-nonadherent cells, and monocytes suppressed SACol-induced IFN responses. Further separation of nonadherent PBLs in accordance with expression of surface antigenic markers was performed with a ‘panning’ technique. The SACol induced production of IFN in cells that carried neither surface immunoglobulins nor OKT3-defined antigens. These cells were also characterized as OKM1- and OKT10-negative. In contrast, cells with natural killer (NK) activity against K562 erythroleukaemia cells were located in both OKM1- and OKT10-positive and -negative cells. At centrifugation on Percoll density gradients, cells with NK activity and IFN response against SACol were recovered from light gradient fractions that contained mainly large granular lymphocytes (LGL). Furthermore, the IFN producers were enriched by removal of sheep erythrocyte-rosetting T cells from the Percoll fractions. These SACol-induced IFN-producing PBLs are LGL but lack certain antigens that are frequently found on NK cells. [ABSTRACT FROM AUTHOR]
- Published
- 1985
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13. Free Oxygen Radicals Are Not Detectable by Chemiluminescence during Human Natural Killer Cell Cytotoxicity.
- Author
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Ramstedt, U., Rossi, P., Kullman, C., Warren, E., Palmblad, J., and Jondal, M.
- Subjects
KILLER cells ,SUPEROXIDE dismutase ,CELL culture ,FREE radicals ,REACTIVE oxygen species - Abstract
Mononuclear cells isolated from peripheral blood of normal donors produce free oxygen radicals (FR), detectable by chemiluminsecence (CL), when interacting with target cells during natural killer (NK) cell lysis. FR-producing cells were found to have monocyte characteristics and gave a positive CL reaction when mixed at low concentration (0.5%) with purified NK cells. No correlation was found between susceptibility to NK cell lysis and capacity to induce CL with different target cell lines. Using high and low molecular FR scavengers, no NK cell inhibition was seen with superoxide dismutase, cytochrome c, and catalase, whereas some inhibition was seen with 4,5-dihydroxy-m-benzenedisulphonic acid (tiron*) and 2,3- dihydroxybenzoate. These compounds, however, required higher concentrations than used for inhibition of CL, suggesting an alternative action of these compounds. Normal levels of NK cell activity were found in two patients with chronic gramulomatous disease, who were genetically incapable of producing detectable amounts of FR. As a result, it is concluded that human NK cells do not produce large amounts of FR during killing and that FR are unlikely to be the lytic end product. Nevertheless, neither a low degree of FR formation in NK cells nor a more subtle signal-transmitting role of FR during NK cell triggering can be excluded. [ABSTRACT FROM AUTHOR]
- Published
- 1984
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14. Pigment Mutations in the Mouse Which Also Affect Lysosomal Functions Lead to Suppressed Natural Killer Cell Activity.
- Author
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Örn, A., Håkansson, E. M., Gidlund, M., Ramstedt, U., Axberg, I., Wigzell, H., and Lundin, L. -G.
- Subjects
GENETIC mutation ,LYSOSOMES ,ENZYMES ,KILLER cells ,KIDNEYS ,CHROMOSOMES - Abstract
The impact of five pigment mutations in the mouse on natural killer (NK) activity was examined in inbred strains congenic for the respective mutation. Whereas the nature of pigmentation disorder was similar in the five mutant strains (beige, pallid, reduced pigmentation, pale ear, and sepia), all mutations except sepia also led to a significant change in lysosomal enzyme activities in the kidney. A significant reduction in NK activity was observed in the four strains with lysosomal impact, whereas homozygous sepia mice displayed normal NK activity. The pigment mutations analysed are located on different chromosomes and fail to cross-interact negatively with each other in the heterozygous mice. This would indicate that pigment mutations with a parallel impact on lysosomal enzyme activities probably always result in a reduction in natural killer cell activity. [ABSTRACT FROM AUTHOR]
- Published
- 1982
- Full Text
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15. Inhibition of human natural killer cell activity by (14R,15S)-14,15-dihydroxy-5Z,8Z,10E,12E- icosatetraenoic acid.
- Author
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Ramstedt, U, Serhan, C N, Lundberg, U, Wigzell, H, and Samuelsson, B
- Abstract
The interactions between products of the 15-lipoxygenase cascade and human natural killer (NK) cell activity have been studied. Addition of human leukocyte-derived (14R,15S)-14,15-dihydroxy-5Z,8Z,10E,12E-ic osatetraenoic acid (14,15-DiHETE) to the NK cytotoxicity assay against K562 target cells resulted in inhibition of NK cell activity, whereas addition of other 15-lipoxygenase-associated metabolites [i.e., (15S)-15-hydroperoxy-5Z,8Z,11Z,13E-icosatetra eno ic acid, (15S)-15-hydroxy-5Z,8Z,11Z,13E-icosatetraenoic acid, and (8R,15S)- and (8S,15S)-8,15-dihydroxy-5Z,9E,-11E,13E-icosat etr aenoic acid isomers] resulted in little or no inhibition of NK function. Dose-response studies indicate that leukocyte-derived 14,15-DiHETE and 14,15-DiHETE methyl ester, at micromolar concentrations, inhibit NK function even in the presence of 2.5% fetal calf serum. Synthetic 14,15-DiHETE prepared by total organic synthesis displayed similar biological activities over identical dose ranges. These icosanoids do not inhibit NK target cell binding and they exert only a variable effect in either antibody-dependent cytotoxicity or cytotoxic T-lymphocyte assays. These results demonstrate that the 14,15-DiHETE inhibits NK cell function in vitro. Moreover, they suggest that activation of the 15-lipoxygenase cascade and formation of 14,15-DiHETE in vivo may provide a mode of immune regulation.
- Published
- 1984
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16. Selective inhibition of human T cell cytotoxicity at levels of target recognition or initiation of lysis by monoclonal OKT3 and Leu-2a antibodies.
- Author
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Landegren, U, Ramstedt, U, Axberg, I, Ullberg, M, Jondal, M, and Wigzell, H
- Abstract
Out of a panel of seven monoclonal antibodies with affinity for human lymphoid cells, three were shown to prevent cytotoxic T cell activity, whereas none affected natural killer cell activity when applied without complement. Anti-OKT3 and anti-Leu-2a, with affinity for all T cells and the cytotoxic/suppressive subset, respectively were both shown to inhibit T killing by their interaction with the effector cell. For anti-OKT3, the inhibition remained after free antibody was washed away. Anti-Leu-2a, in contrast, induced a rapidly reversible inhibition. Using a single cell assay, anti-OKT3 was shown to reduce the lytic ability without affecting target cell binding, whereas anti-Leu-2a prevented the effectors from binding target cells.
- Published
- 1982
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17. Properties of human natural interferon-producing cells stimulated by tumorcell lines.
- Author
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Ronnblom, L, Ramstedt, U, Alm, GV, Ronnblom, L, Ramstedt, U, and Alm, GV
- Published
- 1983
18. Human peripheral blood null lymphocytes stimulated by Staphylococcusaureus Cowan I produce atypical acid-labile interferon in vitro.
- Author
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Funa, K, Ramstedt, U, Ronnblom, L, Alm, GV, Funa, K, Ramstedt, U, Ronnblom, L, and Alm, GV
- Published
- 1985
19. Mutant monoclonal antibodies with select alteration in complement activation ability. Impact on immune complex functions in vivo
- Author
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Nose, M., Okuda, T., Magnus Gidlund, Ramstedt, U., Okada, N., Okada, H., Heyman, B., Kyogoku, M., and Wigzell, H.
- Subjects
Mice, Inbred BALB C ,Granuloma ,Complement Pathway, Alternative ,Kidney Glomerulus ,Immunology ,Antibody-Dependent Cell Cytotoxicity ,Antibodies, Monoclonal ,Antigen-Antibody Complex ,Receptors, Fc ,Mice ,Immunoglobulin G ,Mice, Inbred CBA ,Animals ,Immunology and Allergy ,Binding Sites, Antibody ,Staphylococcal Protein A ,Complement Activation ,Mutagens - Abstract
Mutagenesis of mAb is a useful means for studying the biologic and pathologic functions of immune complexes. Treatment of the Hy-1.2 hybridoma-producing IgG2a-anti-TNP antibodies with ethylmethanesulfonate provided us with a mutant clone, producing antibodies with reduced capacity for C activation. The antibodies retained normal Ag-binding capacity, staphylococcal protein A reactivity, and association to FcR for IgG on murine macrophages. No significant polypeptide deletion or class-switch was observed, but a significant change in clonotype was revealed by IEF. Intravenous injection of the mutant antibodies in immune complex form induced different tissue distributions of Ag in mice; i.e., more in kidneys and less in spleen, and developed more mesangial deposits in renal glomeruli compared with those of the wild type. Moreover, the production of granulomatous lesions in vivo caused by immune complexes of TNP-Sepharose was augmented by using mutant antibodies. These lesions demonstrated an enhanced accumulation of macrophages with multinucleated giant cells. Availability of this kind of mutant mAb is thus helpful in the elucidation of the biologic functions and consequences of immune complexes.
20. Action of novel eicosanoids lipoxin A and B on human natural killer cell cytotoxicity: effects on intracellular cAMP and target cell binding.
- Author
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Ramstedt, U, primary, Ng, J, additional, Wigzell, H, additional, Serhan, C N, additional, and Samuelsson, B, additional
- Published
- 1985
- Full Text
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21. Lipoxin A-induced inhibition of human natural killer cell cytotoxicity: studies on stereospecificity of inhibition and mode of action.
- Author
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Ramstedt, U, primary, Serhan, C N, additional, Nicolaou, K C, additional, Webber, S E, additional, Wigzell, H, additional, and Samuelsson, B, additional
- Published
- 1987
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22. Mutant monoclonal antibodies with select alteration in complement activation ability. Impact on immune complex functions in vivo.
- Author
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Nose, M, primary, Okuda, T, additional, Gidlund, M, additional, Ramstedt, U, additional, Okada, N, additional, Okada, H, additional, Heyman, B, additional, Kyogoku, M, additional, and Wigzell, H, additional
- Published
- 1988
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23. A specific assay measuring binding of 125I-Gp 120 from HIV to T4+/CD4+ cells
- Author
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Lundin, K., primary, Nygren, A., additional, Arthur, L.O., additional, Robey, W.G., additional, Morein, B., additional, Ramstedt, U., additional, Gidlund, M., additional, and Wigzell, H., additional
- Published
- 1987
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24. NK cells stimulating properties of a non capsulated klebsiella pneumoniae biotype A membranal proteoglycan
- Author
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Pinel, A.M., Normier, G., Dussourd D'Hinterland, L., Ramstedt, U., and Wigzell, H.
- Published
- 1985
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25. Reproductive and developmental toxicology studies of iminosugar UV-4.
- Author
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Shearer J, Wolfe G, Khaliq M, Kaufman B, Sampath A, Warfield KL, Ramstedt U, and Treston A
- Subjects
- Pregnancy, Female, Humans, Male, Rats, Rabbits, Animals, Rats, Sprague-Dawley, Dose-Response Relationship, Drug, Fertility, Body Weight, Mammals, Sperm Motility, Reproduction
- Abstract
UV-4 (N-(9-methoxynonyl)-1-deoxynojirimycin) is a host-targeted antiviral agent, which targets mammalian proteins (endoplasmic reticulum glucosidases) rather than virally encoded proteins. This mechanism confers both broad-spectrum activity and low potential for generation of viral drug resistance mutations. Reproductive and developmental studies of UV-4 evaluated effects on fertility and early embryonic development in rats, embryo-fetal development in rats and rabbits, and pre- and postnatal development including maternal function in rats. All reproductive and developmental studies conducted achieved dose levels where parental toxicity (generally decreased body weight, decreased food consumption and adverse clinical signs) were observed. Toxicokinetic evaluations confirmed UV-4 crossed the placenta exposing fetal rats and rabbits in utero. Adverse findings in reproductive and developmental studies included decreases in sperm motility with histopathology correlates, visceral and skeletal malformations, changes in eye opening, air drop reflex, vaginal opening and preputial separation. The combined results of the fertility and early embryonic developmental study and pre- and postnatal study suggested that there may be an increased risk for male fertility. These effects are similar to those reported in pre-clinical studies of the structurally related drug Miglustat (N-butyl-1-deoxynojirimycin), therefore UV-4 may have risk of developmental or reproductive adverse outcomes in humans similar to existing approved agents in this drug class., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Jeffry Shearer: Data compilation from study reports, writing and review of the manuscript; Gary Wolfe: overall study design and protocol development for all studies, study execution, result analysis and reporting, manuscript review; Mansoora Khaliq: study manager for reproductive and developmental toxicology nonclinical program under NIAID contract, protocol development, study execution, result analysis and reporting, manuscript review; Brian Kaufman: protocol development of all studies, analytical validation, result analysis and reporting and Quality oversight for GLP studies, manuscript review; Aruna Sampath: study manager for nonclinical program under NIAID contract, protocol development, study execution, result analysis and reporting, manuscript review; Kelly Warfield: study design and protocol development, dose decisions, study execution, manuscript writing and review; Urban Ramstedt: protocol development, dose decisions, result analysis and reporting and Principal Investigator of NIAID contract during nonclinical development, manuscript review; Anthony Treston: bioanalytical method development and validation, pharmacokinetics, all protocol development, dose decision, result analysis and reporting, manuscript writing and review. Conflicts of Interest Kelly L Warfield is an employee of Emergent BioSolutions Inc. Mansoora Khaliq and Anthony Treston carried out the work for this program on behalf of Emergent BioSolutions Inc. and the work is unrelated to their current employers., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2022
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26. Randomized single oral dose phase 1 study of safety, tolerability, and pharmacokinetics of Iminosugar UV-4 Hydrochloride (UV-4B) in healthy subjects.
- Author
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Callahan M, Treston AM, Lin G, Smith M, Kaufman B, Khaliq M, Evans DeWald L, Spurgers K, Warfield KL, Lowe P, Duchars M, Sampath A, and Ramstedt U
- Subjects
- 1-Deoxynojirimycin adverse effects, Antiviral Agents pharmacology, Area Under Curve, Dose-Response Relationship, Drug, Double-Blind Method, Healthy Volunteers, Humans, Dengue drug therapy, alpha-Glucosidases metabolism, alpha-Glucosidases therapeutic use
- Abstract
Background: UV-4 (N-(9'-methoxynonyl)-1-deoxynojirimycin, also called MON-DNJ) is an iminosugar small-molecule oral drug candidate with in vitro antiviral activity against diverse viruses including dengue, influenza, and filoviruses and demonstrated in vivo efficacy against both dengue and influenza viruses. The antiviral mechanism of action of UV-4 is through inhibition of the host endoplasmic reticulum-resident α-glucosidase 1 and α-glucosidase 2 enzymes. This inhibition prevents proper glycan processing and folding of virus glycoproteins, thereby impacting virus assembly, secretion, and the fitness of nascent virions., Methodology/principal Findings: Here we report a first-in-human, single ascending dose Phase 1a study to evaluate the safety, tolerability, and pharmacokinetics of UV-4 hydrochloride (UV-4B) in healthy subjects (ClinicalTrials.gov Identifier NCT02061358). Sixty-four subjects received single oral doses of UV-4 as the hydrochloride salt equivalent to 3, 10, 30, 90, 180, 360, 720, or 1000 mg of UV-4 (6 subjects per cohort), or placebo (2 subjects per cohort). Single doses of UV-4 hydrochloride were well tolerated with no serious adverse events or dose-dependent increases in adverse events observed. Clinical laboratory results, vital signs, and physical examination data did not reveal any safety signals. Dose-limiting toxicity was not observed; the maximum tolerated dose of UV-4 hydrochloride in humans has not yet been determined (>1000 mg). UV-4 was rapidly absorbed and distributed after dosing with the oral solution formulation used in this study. Median time to reach maximum plasma concentration ranged from 0.5-1 hour and appeared to be independent of dose. Exposure increased approximately in proportion with dose over the 333-fold dose range. UV-4 was quantifiable in pooled urine over the entire collection interval for all doses., Conclusions/significance: UV-4 is a host-targeted broad-spectrum antiviral drug candidate. At doses in humans up to 1000 mg there were no serious adverse events reported and no subjects were withdrawn from the study due to treatment-emergent adverse events. These data suggest that therapeutically relevant drug levels of UV-4 can be safely administered to humans and support further clinical development of UV-4 hydrochloride or other candidate antivirals in the iminosugar class., Trial Registration: ClinicalTrials.gov NCT02061358 https://clinicaltrials.gov/ct2/show/NCT02061358., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: L.DW, K.S, K.L.W, and P.L are employees of Emergent BioSolutions Inc. A.T and M.K carried out the work for this program on behalf of Emergent BioSolutions Inc. and the work is unrelated to current employers.
- Published
- 2022
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27. Investigational New Drug Enabling Nonclinical Safety Pharmacology Assessment of the Iminosugar UV-4, a Broad-Spectrum Host-Targeted Antiviral Agent.
- Author
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Shearer J, Wolfe G, Sampath A, Warfield KL, Kaufman B, Ramstedt U, and Treston A
- Subjects
- Animals, Antiviral Agents toxicity, Dogs, Drug Evaluation, Preclinical, Drugs, Investigational, Mice, Telemetry, Cardiovascular System, Drug-Related Side Effects and Adverse Reactions
- Abstract
UV-4 (N-(9-methoxynonyl)-1-deoxynojirimycin) is a broad-spectrum antiviral drug candidate with demonstrated activity in vitro and in vivo against multiple, diverse viruses. Nonclinical safety pharmacology studies were conducted to support the filing of an Investigational New Drug (IND) application. Preliminary in vitro pharmacology testing evaluating potential for binding to "off-target" receptors and enzymes indicated no significant liability for advanced development of UV-4. The safety pharmacology of UV-4 was evaluated in the in vitro human ether-à-go-go-related gene (hERG) assay, in a central nervous system (CNS) study in the mouse (modified Irwin test), in a respiratory safety study in conscious mice using whole body plethysmography, and in a cardiovascular safety study in conscious, radiotelemetry-instrumented beagle dogs. There were no observed adverse treatment-related effects following administration of UV-4 as the hydrochloride salt in the hERG potassium channel assay, on respiratory function, in the CNS study, or in the cardiovascular assessment. Treatment-related cardiovascular effect of decreased arterial pulse pressure after 50 or 200 mg of UV-4/kg was the only change outside the normal range, and all hemodynamic parameters returned to control levels by the end of the telemetry recording period. These nonclinical safety pharmacology assessments support the evaluation of this host-targeted broad-spectrum antiviral drug candidate in clinical studies.
- Published
- 2022
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28. Investigational New Drug Enabling Nonclinical Safety Assessment of the Iminosugar UV-4, a Broad-Spectrum Host-Targeted Antiviral Agent.
- Author
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Shearer J, Wolfe G, Sampath A, Warfield KL, Kaufman B, Ramstedt U, and Treston A
- Subjects
- Administration, Oral, Animals, Dogs, Maximum Tolerated Dose, Mice, Rats, Antiviral Agents therapeutic use, Antiviral Agents toxicity, Drugs, Investigational toxicity
- Abstract
The iminosugar UV-4 is a broad-spectrum antiviral drug candidate with activity in vitro and in vivo against multiple, diverse viruses. The toxicological profile of UV-4, dosed as the hydrochloride salt, was evaluated in single-dose and repeat-dose oral toxicity studies in mice, rats, dogs, and non-human primates (NHP). No moribundity or deaths were associated with the drug up to the maximum tolerated dose. No treatment-related adverse effects were observed following single oral doses in dogs, rats, and mice up to 250, 400, 1000 mg/kg, respectively, and in NHP up to 180 mg/kg administered three times daily for 10 days. UV-4-related findings were generally seen at higher doses after 7- or 14-day exposure. The most common clinical pathology findings (increase in aspartate aminotransferase and decreased platelet count) were consistently found across species and each appeared dose related. The kidney, mesenteric lymph nodes, stomach including gastrointestinal tract, and thymus were identified as target organs in mice, rats, and dogs. In 14-day repeat-dose toxicology studies in mice and dogs conducted in compliance with Good Laboratory Practice regulations, the dog was considered to be the most sensitive species to UV-4 exposure based on the treatment-related adverse effects noted in the identified target organs. The results of these studies demonstrate the safety profile of UV-4 hydrochloride and supported the selection of starting and maximal doses for a single ascending dose first-in-human clinical study.
- Published
- 2022
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29. Lack of selective resistance of influenza A virus in presence of host-targeted antiviral, UV-4B.
- Author
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Warfield KL, Schaaf KR, DeWald LE, Spurgers KB, Wang W, Stavale E, Mendenhall M, Shilts MH, Stockwell TB, Barnard DL, Ramstedt U, and Das SR
- Subjects
- Animals, Female, Genome, Viral, Influenza A virus drug effects, Mice, Mice, Inbred BALB C, Mutation, Recombination, Genetic, Selection, Genetic, Antiviral Agents pharmacology, Drug Resistance, Viral genetics, Glycoside Hydrolase Inhibitors pharmacology, Influenza A virus genetics, Orthomyxoviridae Infections virology
- Abstract
Development of antiviral drug resistance is a continuous concern for viruses with high mutation rates such as influenza. The use of antiviral drugs targeting host proteins required for viral replication is less likely to result in the selection of resistant viruses than treating with direct-acting antivirals. The iminosugar UV-4B is a host-targeted glucomimetic that inhibits endoplasmic reticulum α-glucosidase I and II enzymes resulting in improper glycosylation and misfolding of viral glycoproteins. UV-4B has broad-spectrum antiviral activity against diverse viruses including dengue and influenza. To examine the ability of influenza virus to develop resistance against UV-4B, mouse-adapted influenza virus was passaged in mice in the presence or absence of UV-4B and virus isolated from lungs was used to infect the next cohort of mice, for five successive passages. Deep sequencing was performed to identify changes in the viral genome during passaging in the presence or absence of UV-4B. Relatively few minor variants were identified within each virus and the ratio of nonsynonymous to synonymous (dN/dS) substitutions of minor variants confirmed no apparent positive selection following sustained exposure to UV-4B. Three substitutions (one synonymous in PB2, one nonsynonymous in M and PA each) were specifically enriched (>3%) in UV-4B-treated groups at passage five. Recombinant viruses containing each individual or combinations of these nonsynonymous mutations remained sensitive to UV-4B treatment in mice. Overall, these data provide evidence that there is a high genetic barrier to the generation and selection of escape mutants following exposure to host-targeted iminosugar antivirals.
- Published
- 2019
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30. Assessment of the potential for host-targeted iminosugars UV-4 and UV-5 activity against filovirus infections in vitro and in vivo.
- Author
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Warfield KL, Warren TK, Qiu X, Wells J, Mire CE, Geisbert JB, Stuthman KS, Garza NL, Van Tongeren SA, Shurtleff AC, Agans KN, Wong G, Callahan MV, Geisbert TW, Klose B, Ramstedt U, and Treston AM
- Subjects
- 1-Deoxynojirimycin administration & dosage, 1-Deoxynojirimycin agonists, 1-Deoxynojirimycin analogs & derivatives, 1-Deoxynojirimycin pharmacology, 1-Deoxynojirimycin therapeutic use, Animals, Antiviral Agents administration & dosage, Antiviral Agents chemistry, Chlorocebus aethiops, Disease Models, Animal, Dose-Response Relationship, Drug, Imino Sugars administration & dosage, Imino Sugars chemistry, Macaca, Mice, Models, Animal, Vero Cells, Antiviral Agents pharmacology, Antiviral Agents therapeutic use, Ebolavirus drug effects, Imino Sugars pharmacology, Imino Sugars therapeutic use, Marburgvirus drug effects
- Abstract
Iminosugars are host-directed antivirals with broad-spectrum activity. The iminosugar, N-butyl-deoxynojirimycin (NB-DNJ or Miglustat
® ), is used in humans for treatment of Gaucher's disease and has mild antiviral properties. More potent analogs of NB-DNJ have been generated and have demonstrated activity against a variety of viruses including flaviviruses, influenza, herpesviruses and filoviruses. In the current study, a panel of analogs based on NB-DNJ was analyzed for activity against Ebola (EBOV) and Marburg viruses (MARV). The antiviral activity of NB-DNJ (UV-1), UV-2, UV-3, UV-4 and UV-5 against both EBOV and MARV was demonstrated in Vero cells. Subsequent studies to examine the activity of UV-4 and UV-5 using rodent models of EBOV and MARV were performed. In vivo efficacy studies provided inconsistent data following treatment with iminosugars using filovirus mouse models. A tolerability study in nonhuman primates demonstrated that UV-4 could be administered at much higher dose levels than rodents. Since UV-4 was active in vitro, had been demonstrated to be active against influenza and dengue in vivo, and was being tested in a Phase 1 clinical trial, a small proof-of-concept nonhuman primate trial was performed to determine whether this antiviral candidate could provide clinical benefit to EBOV-infected individuals. Administration of UV-4B did not provide a clinical or survival benefit to macaques infected with EBOV-Makona; however, dosing of animals was not optimal in this study. Efficacy may be improved by thrice daily dosing (e.g. by nasogastric tube feeding) to match the efficacious dosing regimens demonstrated against dengue and influenza viruses., (Copyright © 2016 Elsevier B.V. All rights reserved.)- Published
- 2017
- Full Text
- View/download PDF
31. Inhibition of endoplasmic reticulum glucosidases is required for in vitro and in vivo dengue antiviral activity by the iminosugar UV-4.
- Author
-
Warfield KL, Plummer EM, Sayce AC, Alonzi DS, Tang W, Tyrrell BE, Hill ML, Caputo AT, Killingbeck SS, Beatty PR, Harris E, Iwaki R, Kinami K, Ide D, Kiappes JL, Kato A, Buck MD, King K, Eddy W, Khaliq M, Sampath A, Treston AM, Dwek RA, Enterlein SG, Miller JL, Zitzmann N, Ramstedt U, and Shresta S
- Subjects
- 1-Deoxynojirimycin administration & dosage, 1-Deoxynojirimycin pharmacology, 1-Deoxynojirimycin therapeutic use, Animals, Antibodies, Viral blood, Antibody-Dependent Enhancement drug effects, Antiviral Agents administration & dosage, Antiviral Agents therapeutic use, Cells, Cultured, Chlorocebus aethiops, Clinical Trials as Topic, Disease Models, Animal, Drugs, Investigational, Endoplasmic Reticulum drug effects, Endoplasmic Reticulum enzymology, Glycoside Hydrolase Inhibitors administration & dosage, Glycoside Hydrolase Inhibitors chemistry, Glycoside Hydrolase Inhibitors therapeutic use, Humans, Inhibitory Concentration 50, Mice, Monocytes virology, Receptors, Interferon deficiency, Serogroup, Severe Dengue virology, Vero Cells, 1-Deoxynojirimycin analogs & derivatives, Antiviral Agents pharmacology, Dengue Virus drug effects, Glycoside Hydrolase Inhibitors pharmacology, Severe Dengue drug therapy, alpha-Glucosidases metabolism
- Abstract
The antiviral activity of UV-4 was previously demonstrated against dengue virus serotype 2 (DENV2) in multiple mouse models. Herein, step-wise minimal effective dose and therapeutic window of efficacy studies of UV-4B (UV-4 hydrochloride salt) were conducted in an antibody-dependent enhancement (ADE) mouse model of severe DENV2 infection in AG129 mice lacking types I and II interferon receptors. Significant survival benefit was demonstrated with 10-20 mg/kg of UV-4B administered thrice daily (TID) for seven days with initiation of treatment up to 48 h after infection. UV-4B also reduced infectious virus production in in vitro antiviral activity assays against all four DENV serotypes, including clinical isolates. A set of purified enzyme, in vitro, and in vivo studies demonstrated that inhibition of endoplasmic reticulum (ER) α-glucosidases and not the glycosphingolipid pathway appears to be responsible for the antiviral activity of UV-4B against DENV. Along with a comprehensive safety package, these and previously published data provided support for an Investigational New Drug (IND) filing and Phases 1 and 2 clinical trials for UV-4B with an indication of acute dengue disease., (Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
32. The Iminosugar UV-4 is a Broad Inhibitor of Influenza A and B Viruses ex Vivo and in Mice.
- Author
-
Warfield KL, Barnard DL, Enterlein SG, Smee DF, Khaliq M, Sampath A, Callahan MV, Ramstedt U, and Day CW
- Subjects
- 1-Deoxynojirimycin administration & dosage, 1-Deoxynojirimycin pharmacology, Animals, Antiviral Agents pharmacology, Body Weight, Cells, Cultured, Disease Models, Animal, Epithelial Cells virology, Humans, Mice, Orthomyxoviridae Infections pathology, Orthomyxoviridae Infections virology, Survival Analysis, Treatment Outcome, 1-Deoxynojirimycin analogs & derivatives, Antiviral Agents administration & dosage, Influenza A virus drug effects, Influenza B virus drug effects, Orthomyxoviridae Infections drug therapy
- Abstract
Iminosugars that are competitive inhibitors of endoplasmic reticulum (ER) α-glucosidases have been demonstrated to have antiviral activity against a diverse set of viruses. A novel iminosugar, UV-4B, has recently been shown to provide protection against lethal infections with dengue and influenza A (H1N1) viruses in mice. In the current study, the breadth of activity of UV-4B against influenza was examined ex vivo and in vivo. Efficacy of UV-4B against influenza A and B viruses was shown in primary human bronchial epithelial cells, a principal target tissue for influenza. Efficacy of UV-4B against influenza A (H1N1 and H3N2 subtypes) and influenza B was demonstrated using multiple lethal mouse models with readouts including mortality and weight loss. Clinical trials are ongoing to demonstrate safety of UV-4B and future studies to evaluate antiviral activity against influenza in humans are planned.
- Published
- 2016
- Full Text
- View/download PDF
33. A Novel Iminosugar UV-12 with Activity against the Diverse Viruses Influenza and Dengue (Novel Iminosugar Antiviral for Influenza and Dengue).
- Author
-
Warfield KL, Plummer E, Alonzi DS, Wolfe GW, Sampath A, Nguyen T, Butters TD, Enterlein SG, Stavale EJ, Shresta S, and Ramstedt U
- Subjects
- Animals, Antiviral Agents pharmacology, Dengue Virus drug effects, Disease Models, Animal, Female, Guinea Pigs, Imino Sugars pharmacology, Male, Mice, Microbial Sensitivity Tests, Orthomyxoviridae drug effects, Treatment Outcome, Antiviral Agents therapeutic use, Dengue drug therapy, Imino Sugars therapeutic use, Orthomyxoviridae Infections drug therapy
- Abstract
Iminosugars are capable of targeting the life cycles of multiple viruses by blocking host endoplasmic reticulum α-glucosidase enzymes that are required for competent replication of a variety of enveloped, glycosylated viruses. Iminosugars as a class are approved for use in humans with diseases such as diabetes and Gaucher's disease, providing evidence for safety of this class of compounds. The in vitro antiviral activity of iminosugars has been described in several publications with a subset of these demonstrating in vivo activity against flaviviruses, herpesviruses, retroviruses and filoviruses. Although there is compelling non-clinical in vivo evidence of antiviral efficacy, the efficacy of iminosugars as antivirals has yet to be demonstrated in humans. In the current study, we report a novel iminosugar, UV-12, which has efficacy against dengue and influenza in mouse models. UV-12 exhibits drug-like properties including oral bioavailability and good safety profile in mice and guinea pigs. UV-12 is an example of an iminosugar with activity against multiple virus families that should be investigated in further safety and efficacy studies and demonstrates potential value of this drug class as antiviral therapeutics.
- Published
- 2015
- Full Text
- View/download PDF
34. Dengue Virus Evolution under a Host-Targeted Antiviral.
- Author
-
Plummer E, Buck MD, Sanchez M, Greenbaum JA, Turner J, Grewal R, Klose B, Sampath A, Warfield KL, Peters B, Ramstedt U, and Shresta S
- Subjects
- Animals, Dengue Virus physiology, Disease Models, Animal, Evolution, Molecular, Genetic Variation, Host-Pathogen Interactions drug effects, Humans, Imino Sugars pharmacology, Mice, Mice, 129 Strain, Mice, Knockout, Mutation, Polymorphism, Single Nucleotide, Selection, Genetic, Viral Proteins genetics, Virus Replication drug effects, Antiviral Agents pharmacology, Dengue drug therapy, Dengue virology, Dengue Virus drug effects, Dengue Virus genetics
- Abstract
Unlabelled: The host-targeted antiviral drug UV-4B reduces viral replication and promotes survival in a mouse model of experimental dengue virus (DENV) infection. UV-4B is an iminosugar that inhibits the α-glucosidase family of enzymes and subsequently the folding of glycosylated proteins, both viral and host. Here, we utilized next-generation sequencing to investigate evolution of a flavivirus under selective pressure by a host-targeted antiviral in vivo. In viral populations recovered from UV-4B-treated mice, there was a significant increase in the number of single-nucleotide polymorphisms (SNPs) and the ratio of nonsynonymous to synonymous SNPs compared to findings in viral populations from vehicle-treated mice. The strongest evidence of positive selection was in the glycosylated membrane protein, thereby providing in vivo validation of the mechanism of action of an iminosugar. In addition, mutations in glycosylated proteins were present only in drug-treated mice after a single passage. However, the bulk of the other mutations were present in both populations, indicating nonspecific selective pressure. Together with the continued control of viremia by UV-4B, these findings are consistent with the previously predicted high genetic barrier to escape mutations in host-targeted antivirals., Importance: Although hundreds of millions of people are infected with DENV every year, there is currently no approved vaccine or antiviral therapy. UV-4B has demonstrated antiviral activity against DENV and is expected to enter clinical trials soon. Therefore, it is important to understand the mechanisms of DENV resistance to UV-4B. Host-targeted antivirals are thought to have a higher genetic barrier to escape mutants than directly acting antivirals, yet there are very few published studies of viral evolution under host-targeted antivirals. No study to date has described flavivirus evolution in vivo under selective pressure by a host-based antiviral drug. We present the first in vivo study of the sequential progression of viral evolution under selective pressure by a host-targeted antiviral compound. This study bolsters support for the clinical development of UV-4B as an antiviral drug against DENV, and it provides a framework to compare how treatment with other host-targeted antiflaviviral drugs in humans and different animal models influence viral genetic diversity., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)
- Published
- 2015
- Full Text
- View/download PDF
35. In vivo therapeutic protection against influenza A (H1N1) oseltamivir-sensitive and resistant viruses by the iminosugar UV-4.
- Author
-
Stavale EJ, Vu H, Sampath A, Ramstedt U, and Warfield KL
- Subjects
- 1-Deoxynojirimycin adverse effects, 1-Deoxynojirimycin pharmacology, Animals, Antibodies, Viral biosynthesis, Antiviral Agents adverse effects, Female, Influenza A Virus, H1N1 Subtype immunology, Influenza A Virus, H1N1 Subtype physiology, Lung drug effects, Male, Mice, Mice, Inbred BALB C, Safety, Time Factors, Viral Load drug effects, 1-Deoxynojirimycin analogs & derivatives, Antiviral Agents pharmacology, Drug Resistance, Viral drug effects, Influenza A Virus, H1N1 Subtype drug effects, Oseltamivir pharmacology
- Abstract
Our lead iminosugar analog called UV-4 or N-(9-methoxynonyl)-1-deoxynojirimycin inhibits activity of endoplasmic reticulum (ER) α-glucosidases I and II and is a potent, host-targeted antiviral candidate. The mechanism of action for the antiviral activity of iminosugars is proposed to be inhibition of ER α-glucosidases leading to misfolding of critical viral glycoproteins. These misfolded glycoproteins would then be incorporated into defective virus particles or targeted for degradation resulting in a reduction of infectious progeny virions. UV-4, and its hydrochloride salt known as UV-4B, is highly potent against dengue virus in vitro and promotes complete survival in a lethal dengue virus mouse model. In the current studies, UV-4 was shown to be highly efficacious via oral gavage against both oseltamivir-sensitive and -resistant influenza A (H1N1) infections in mice even if treatment was initiated as late as 48-72 hours after infection. The minimal effective dose was found to be 80-100 mg/kg when administered orally thrice daily. UV-4 treatment did not affect the development of protective antibody responses after either influenza infection or vaccination. Therefore, UV-4 is a promising candidate for further development as a therapeutic intervention against influenza.
- Published
- 2015
- Full Text
- View/download PDF
36. An iminosugar with potent inhibition of dengue virus infection in vivo.
- Author
-
Perry ST, Buck MD, Plummer EM, Penmasta RA, Batra H, Stavale EJ, Warfield KL, Dwek RA, Butters TD, Alonzi DS, Lada SM, King K, Klose B, Ramstedt U, and Shresta S
- Subjects
- Animals, Antiviral Agents chemistry, Cytokines, Dengue immunology, Dengue Virus physiology, Humans, Imino Sugars chemistry, Mice, Mice, Inbred Strains, Structure-Activity Relationship, Antiviral Agents pharmacology, Dengue drug therapy, Dengue virology, Dengue Virus drug effects, Imino Sugars pharmacology
- Abstract
The aim of the present study was to evaluate the ability of the iminosugar drug UV-4 to provide in vivo protection from lethal dengue virus (DENV) challenge. This study utilized a well-described model of dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS)-like lethal disease in AG129 mice lacking the type I and II interferon receptors. Herein, we present UV-4 as a potent iminosugar for controlling DENV infection and disease in this mouse model. Specifically, administration of UV-4 reduced mortality, as well as viremia and viral RNA in key tissues, and cytokine storm. In addition, UV-4 treatment can be delayed, and it does not alter the anti-DENV antibody response. These results have set the foundation for development of UV-4 as a DENV-specific antiviral in phase I human clinical trials., (Copyright © 2013 Elsevier B.V. All rights reserved.)
- Published
- 2013
- Full Text
- View/download PDF
37. In vivo electroporation enhances the potency of poly-lactide co-glycolide (PLG) plasmid DNA immunization.
- Author
-
Barbon CM, Baker L, Lajoie C, Ramstedt U, Hedley ML, and Luby TM
- Subjects
- Adjuvants, Immunologic pharmacology, Animals, Antigen-Presenting Cells immunology, Aryl Hydrocarbon Hydroxylases immunology, Cell Line, Tumor, Cytochrome P-450 CYP1B1, Cytokines immunology, Female, Immunity, Innate, Injections, Intramuscular, Mice, Mice, Inbred BALB C, Mice, Inbred C3H, Mice, Transgenic, Microspheres, Phagocytes immunology, Plasmids immunology, Polylactic Acid-Polyglycolic Acid Copolymer, T-Lymphocytes immunology, Electroporation, Lactic Acid pharmacology, Polyglycolic Acid pharmacology, Vaccines, DNA immunology
- Abstract
Immunization with plasmid DNA that has been encapsulated in poly lactide-co-glycolide (PLG) microparticles targets the plasmid DNA to antigen presenting cells and elicits immune responses to the encoded antigen(s). Application of a series of electrical pulses (EPT) immediately following unformulated DNA injection enhances expression of the encoded antigen and increases immune responses. The combination of using EPT before or after PLG-encapsulated plasmid DNA immunization was tested to determine if enhanced immune responses would be generated. The results show that the combination lead to both enhanced expression of antigen and more robust T cell responses, even if EPT was applied prior to immunization. The data also demonstrate that recruitment of phagocytes to the injection site was markedly enhanced by EPT, and this resulted in an increase of the antigen expression levels in these cells. Co-administration of microparticles and EPT also effected localized necrosis of muscle fibers, caused persistent Th-1-modulated cytokine production, and lead to the release of two endogenous adjuvants, uric acid and HMGB1. In all, we describe that increased immunogenicity observed with the combination of PLG-encapsulated plasmid DNA microparticle with EPT was caused by an increase in the recruitment of antigen presenting cells which mediated a more robust T cell response than observed with immunization alone., (Copyright © 2010 Elsevier Ltd. All rights reserved.)
- Published
- 2010
- Full Text
- View/download PDF
38. Repeated immunization with plasmid DNA formulated in poly(lactide-co-glycolide) microparticles is well tolerated and stimulates durable T cell responses to the tumor-associated antigen cytochrome P450 1B1.
- Author
-
Luby TM, Cole G, Baker L, Kornher JS, Ramstedt U, and Hedley ML
- Subjects
- Animals, Aryl Hydrocarbon Hydroxylases, Cancer Vaccines adverse effects, Cancer Vaccines genetics, Cancer Vaccines immunology, Cytochrome P-450 CYP1B1, Cytochrome P-450 Enzyme System genetics, Female, Glycolates pharmacology, HLA-A2 Antigen immunology, Lactic Acid, Lymphocyte Activation immunology, Mice, Mice, Inbred C57BL, Mice, Transgenic, Microspheres, Polyglycolic Acid, Polylactic Acid-Polyglycolic Acid Copolymer, Rats, Rats, Sprague-Dawley, Toxicity Tests, Vaccines, DNA adverse effects, Vaccines, DNA genetics, Vaccines, DNA immunology, CD8-Positive T-Lymphocytes immunology, Cancer Vaccines administration & dosage, Cytochrome P-450 Enzyme System immunology, Immunization methods, Vaccines, DNA administration & dosage
- Abstract
Injection of microparticle-encapsulated DNA elicits immune responses to plasmid-encoded antigens in mice and humans. Cytochrome P450 CYP1B1 (CYP1B1) is a member of the CYP1 P450 enzyme family that is overexpressed in a variety of solid tumors. The work described herein was performed to study the kinetics of stimulating T cell responsiveness with an encapsulated DNA encoding CYP1B1 and provides support for the clinical development of this formulation. Immunization of HLA-A2/Kb transgenic mice with human CYP1B1 encoding plasmid DNA formulated in poly(lactide-co-glycolide) (PLG) microparticles elicits CD8+ T cells that respond to human CYP1B1-positive target cells. The duration of the immune response, the effect on the immune response of multiple injections, and the safety of repeated injections were studied. These results show that the PLG-encapsulated DNA therapeutic elicits durable immune responses to CYP1B1, the responses are dependent on repeat immunization, and that the formulation is well tolerated.
- Published
- 2004
- Full Text
- View/download PDF
39. The shared tumor-associated antigen cytochrome P450 1B1 is recognized by specific cytotoxic T cells.
- Author
-
Maecker B, Sherr DH, Vonderheide RH, von Bergwelt-Baildon MS, Hirano N, Anderson KS, Xia Z, Butler MO, Wucherpfennig KW, O'Hara C, Cole G, Kwak SS, Ramstedt U, Tomlinson AJ, Chicz RM, Nadler LM, and Schultze JL
- Subjects
- Animals, Blood Cells, Cell Line, Tumor, Cytochrome P-450 CYP1B1, Cytotoxicity, Immunologic, Epitopes, T-Lymphocyte immunology, HLA-A Antigens, HLA-A2 Antigen genetics, HLA-A2 Antigen immunology, Humans, Mice, Mice, Transgenic, Neoplasms blood, Neoplasms pathology, Peptides immunology, Antigens, Neoplasm immunology, Aryl Hydrocarbon Hydroxylases immunology, Neoplasms immunology, T-Lymphocytes, Cytotoxic immunology
- Abstract
Cytochrome P450 1B1 (CYP1B1), a drug-metabolizing extrahepatic enzyme, was recently shown to be overexpressed in multiple types of cancer. Such tumor-associated genes may be useful targets for anticancer therapy, particularly cancer immunotherapeutics. We identified HLA-A*0201-binding peptides and a naturally processed and presented T-cell epitope capable of inducing CYP1B1-specific cytotoxic T lymphocytes (CTLs) in HLA-A2 transgenic mice. Furthermore, the induction of CYP1B1-specific T cells was demonstrated in healthy donors and cancer patients. These T cells efficiently lysed target cells pulsed with the cognate peptide. More important, HLA-A2-matched tumor cell lines and primary malignant cells were also recognized by CYP1B1-specific CTLs. These findings form the basis of a phase 1 clinical trial exploring a DNA-based vector encoding CYP1B1 for widely applicable cancer immunotherapy conducted at the Dana-Farber Cancer Institute.
- Published
- 2003
- Full Text
- View/download PDF
40. Inhibition of human immunodeficiency virus replication and growth advantage of CD4+ T cells and monocytes derived from CD34+ cells transduced with an intracellular antibody directed against human immunodeficiency virus type 1 Tat.
- Author
-
Poznansky MC, La Vecchio J, Silva-Arietta S, Porter-Brooks J, Brody K, Olszak IT, Adams GB, Ramstedt U, Marasco WA, and Scadden DT
- Subjects
- Adult, Base Sequence, CD4-Positive T-Lymphocytes immunology, Cell Division immunology, Cell Lineage, DNA Primers, Humans, Immunophenotyping, Monocytes immunology, Reverse Transcriptase Polymerase Chain Reaction, Transduction, Genetic, Transgenes, tat Gene Products, Human Immunodeficiency Virus, Antigens, CD34 analysis, CD4-Positive T-Lymphocytes cytology, Gene Products, tat immunology, HIV-1 physiology, Monocytes cytology, Virus Replication immunology
- Abstract
Current clinical gene therapy protocols for the treatment of human immunodeficiency virus type 1 (HIV-1) infection involve the ex vivo transduction and expansion of CD4+ T cells derived from HIV-positive patients at a late stage in their disease (CD4+ cell count <400 cells/mm3). We examined the efficiency of transduction and transgene expression in adult bone marrow (BM)- and umbilical cord blood (UCB)-derived CD34+ cells induced to differentiate into T cells and monocytes in vitro with an MuLV-based vector encoding the neomycin resistance gene and an intracellular antibody directed against the Tat protein of HIV-1 (sFvtat1-Ckappa). The expression of the marker gene and the effects of antiviral construct on subsequent challenge with monocytotropic and T cell-tropic HIV-1 isolates were monitored in vitro in purified T cells and monocytes generated in culture from the transduced CD34+ cells. Transduction efficiencies of CD34+ cells ranged between 22 and 27%. Differentiation of CD34+ cells into T cells or monocytes was not significantly altered by the transduction process. HIV-1 replication in monocytes and CD4+ T cells derived from CD34+ cells transduced with the intracellular antibody gene was significantly reduced in comparison with the degree of HIV replication seen in monocytes and CD4+ T cells derived from CD34+ cells transduced with the neomycin resistance gene alone. Further, T cells and monocytes derived from CD34+ cells transduced with the intracellular antibody gene were demonstrated to express the sFvtat1-Ckappa transgene by RT-PCR and had a selective growth advantage in cultures that had been challenged with HIV-1. These data demonstrate that sFvtat1-Ckappa inhibits HIV-1 replication in T cells and monocytes developing from CD34+ cells and supports the continuing development of a stem cell gene therapy for the treatment of HIV-1 infection.
- Published
- 1999
- Full Text
- View/download PDF
41. Inhibition of human immunodeficiency virus type 1 replication in vitro in acutely and persistently infected human CD4+ mononuclear cells expressing murine and humanized anti-human immunodeficiency virus type 1 Tat single-chain variable fragment intrabodies.
- Author
-
Mhashilkar AM, LaVecchio J, Eberhardt B, Porter-Brooks J, Boisot S, Dove JH, Pumphrey C, Li X, Weissmahr RN, Ring DB, Ramstedt U, and Marasco WA
- Subjects
- Amino Acid Sequence, Animals, CD4-Positive T-Lymphocytes virology, Giant Cells, HIV Antibodies genetics, HIV-1 physiology, Humans, Immunoglobulin Fragments genetics, Immunoglobulin Variable Region genetics, Leukocytes, Mononuclear cytology, Leukocytes, Mononuclear virology, Mice, Molecular Sequence Data, Monocytes cytology, Monocytes virology, Transfection, Tumor Cells, Cultured, tat Gene Products, Human Immunodeficiency Virus, CD4-Positive T-Lymphocytes immunology, Gene Products, tat immunology, Gene Transfer Techniques, HIV Antibodies immunology, HIV-1 immunology, Immunoglobulin Fragments immunology, Immunoglobulin Variable Region immunology, Virus Replication
- Abstract
We have previously reported that a murine anti-Tat sFv intrabody, termed sFvtat1Ck, directed against the proline-rich N-terminal activation domain of HIV-1, is a potent inhibitor of HIV-1 replication [Mhashilkar, A. M., et al. (1995). EMBO J. 14, 1542-1551]. In this study, the protective effect of sFvtat1Ck expression on HIV-1 replication in both acutely infected and persistently infected CD4+ cells was examined. Stably transfected CD4+ SupT1 cells were resistant to HIV-1 infection at high MOI with both the laboratory isolate HxB2 and six syncytium-inducing (SI) primary isolates. Persistently infected U1 cells, which can be induced to increase HIV-1 mRNA synthesis on addition of PMA or TNF-alpha, showed decreased production of HIV-1 in the presence of sFvtat1Ck. In transduced CD4+-selected, CD8+-depleted, and total PMBCs, the sFvtat1Ck-expressing cells showed marked inhibition of HIV-1 replication. The anti-Tat sFv was subsequently humanized by substituting compatible human framework regions that were chosen from a large database of human V(H) and V(L) sequences on the basis of high overall framework matching, similar CDR length, and minimal mismatching of canonical and V(H)/V(L) contact residues. One humanized anti-Tat sFv intrabody, termed sFvhutat2, demonstrated a level of anti-HIV-1 activity that was comparable to the parental murine sFv when transduced PBMCs expressing the murine or humanized sFv intrabodies were challenged with HxB2 and two SI primary isolates. Because Tat is likely to have both direct and indirect effects in the pathogenesis of AIDS through its multiple roles in the HIV-1 life cycle and through its effects on the immune system, the strategy of genetically blocking Tat protein function with a humanized anti-Tat sFv intrabody may prove useful for the treatment of HIV-1 infection and AIDS, particularly when used as an adjuvant gene therapy together with highly active antiretroviral therapies that are currently available.
- Published
- 1999
- Full Text
- View/download PDF
42. Intracellular antibodies against HIV-1 envelope protein for AIDS gene therapy.
- Author
-
Marasco WA, Chen S, Richardson JH, Ramstedt U, and Jones SD
- Subjects
- Adverse Drug Reaction Reporting Systems, Anti-HIV Agents immunology, Antibodies immunology, Gene Transfer Techniques, Genetic Therapy adverse effects, Genetic Vectors, HIV Antigens immunology, HIV Envelope Protein gp120 genetics, HIV-1 genetics, Humans, Patient Selection, Pilot Projects, T-Lymphocytes, Transduction, Genetic, Antibodies genetics, Clinical Protocols, Genetic Therapy methods, HIV Envelope Protein gp120 immunology, HIV Infections therapy, HIV-1 immunology
- Published
- 1998
- Full Text
- View/download PDF
43. Inhibition of human immunodeficiency virus replication and growth advantage of CD4+ T cells from HIV-infected individuals that express intracellular antibodies against HIV-1 gp120 or Tat.
- Author
-
Poznansky MC, Foxall R, Mhashilkar A, Coker R, Jones S, Ramstedt U, and Marasco W
- Subjects
- Animals, CD4-Positive T-Lymphocytes cytology, Cell Division, Gene Transfer Techniques, Genetic Vectors genetics, HIV Infections immunology, HIV-1 genetics, HIV-1 immunology, Humans, Leukemia Virus, Murine genetics, Mice, tat Gene Products, Human Immunodeficiency Virus, CD4-Positive T-Lymphocytes virology, Gene Products, tat immunology, HIV Antibodies immunology, HIV Envelope Protein gp120 immunology, HIV-1 physiology, Virus Replication immunology
- Abstract
Current clinical gene therapy protocols for the treatment of human immunodeficiency virus type 1 (HIV-1) infection often involve the ex vivo transduction and expansion of CD4+ T cells derived from HIV-positive patients at a late stage in their disease (CD4 count <400). These protocols involve the transduction of T cells by murine leukemia virus (MLV)-based vectors encoding antiviral constructs such as the rev m10 dominant negative mutant or a ribozyme directed against the CAP site of HIV-1 RNA. We examined the efficiency and stability of transduction of CD4+ T cells derived from HIV-infected patients at different stages in the progression of their disease, from seroconversion to AIDS. CD4+ T cells from HIV-positive patients and uninfected donors were transduced with MLV-based vectors encoding beta-galactosidase and an intracellular antibody directed against gp120 (sFv 105) or Tat. (sFvtat1-Ckappa). The expression of marker genes and the effects of the antiviral constructs were monitored in vitro in unselected transduced CD4+ T cells. Efficiency and stability of transduction varied during the course of HIV infection; CD4+ T cells derived from asymptomatic patients were transducible at higher efficiencies and stabilities than CD4+ T cells from patients with acquired immunodeficiency syndrome (AIDS). Expression of the anti-tat intracellular antibody was more effective at stably inhibiting HIV-1 replication in transduced cells from HIV-infected individuals than was sFv 105. The results of this study have important implications for the development of a clinically relevant gene therapy for the treatment of HIV-1 infection.
- Published
- 1998
- Full Text
- View/download PDF
44. Cyclic AMP-mediated growth arrest is associated with increased expression of human T cell leukemia virus type I structural and transforming genes.
- Author
-
Poteat HT, Ramstedt U, Yoon S, Dardik M, Terwilliger E, and Sodroski JG
- Subjects
- Bucladesine pharmacology, Caffeine pharmacology, Cyclic AMP biosynthesis, Gene Expression Regulation, Viral, HTLV-I Infections genetics, HTLV-I Infections immunology, Human T-lymphotropic virus 1 genetics, Human T-lymphotropic virus 1 metabolism, Humans, Prostaglandins E pharmacology, Proviruses genetics, RNA, Viral biosynthesis, Receptors, Interleukin-2 biosynthesis, Retroviridae Proteins, Oncogenic metabolism, T-Lymphocytes drug effects, T-Lymphocytes metabolism, Viral Proteins biosynthesis, Cyclic AMP metabolism, HTLV-I Infections metabolism, Human T-lymphotropic virus 1 growth & development, T-Lymphocytes virology
- Abstract
The effect of increased intracellular cyclic AMP levels on gene expression of the human T cell leukemia virus type I (HTLV-I) provirus was examined. Induction of infected cells to produce elevated levels of cyclic AMP was associated with specific increases in viral surface antigen expression, protein synthesis, p24 release into the supernatant, and RNA levels. The patterns of HTLV-I proviral gene expression observed support results from transfection experiments regarding the function of Tax, Rex, and cyclic AMP in HTLV-I gene regulation. As evidenced by thymidine incorporation, treatment of the infected cells to produce cyclic AMP caused reversible growth arrest. The data indicate that HTLV-I RNA and protein synthesis can proceed at an elevated level in the absence of cell growth. Sustained increases in the intracellular level of cyclic AMP may represent a method for enriching cell cultures in HTLV-I proteins.
- Published
- 1996
- Full Text
- View/download PDF
45. In human monocytes a strong correlation exists between expression of the M3 antigen, Fc-mediated phagocytic activity and failure to participate in extracellular antibody-dependent cytotoxicity.
- Author
-
Gidlund M, Rossi P, Cotran P, Ramstedt U, and Wigzell H
- Subjects
- Cell Separation, Flow Cytometry, Humans, Leukocytes, Mononuclear immunology, Leukocytes, Mononuclear physiology, Lymphoma, Large B-Cell, Diffuse pathology, Tumor Cells, Cultured immunology, Tumor Cells, Cultured physiology, Antibody-Dependent Cell Cytotoxicity, Antigens, Surface analysis, Leukocytes, Mononuclear classification, Phagocytosis
- Abstract
Human fresh blood monocytes can phagocytize and lyse antibody-coated target cells by contact with membrane Fc receptors. Recently, the monocyte differentiation antigen Leu M3 has been described to be associated with monocyte/macrophage maturation pathway and to be linked to functionally distinct monocyte subsets. In the present study peripheral blood monocytes were separated into M3+ and M3- subsets, and evaluated for their ability to mediate antibody-driven effector functions. A clear cut functional difference could be demonstrated. M3+ monocytes phagocytize antibody-coated sheep red cells but do not carry out contact-mediated extracellular lysis. In contrast, M3- monocytes have exactly the opposite functional features and they mediate cytolysis without exhibiting any phagocytic activity. By using phenotypically defined clones of the U937 histiocytic cell line, the segregation of different lytic abilities and their linkage of the M3 phenotype have been confirmed.
- Published
- 1988
- Full Text
- View/download PDF
46. Immunologic lesions during Toxoplasma gondii infection.
- Author
-
Goidl EA, Ramstedt U, and Jones TC
- Subjects
- Animals, Brain pathology, Mice, Toxoplasma immunology, Brain immunology, Toxoplasma parasitology, Toxoplasmosis, Animal pathology
- Published
- 1983
- Full Text
- View/download PDF
47. Cyclosporin a permits the distinction between specific T and NK activity generated in a human MLC.
- Author
-
Landegren U, Ramstedt U, Axberg I, Orn A, and Wigzell H
- Subjects
- Cyclosporins administration & dosage, Cytotoxicity, Immunologic drug effects, Drug Interactions, Humans, Killer Cells, Natural drug effects, Kinetics, Lymphocyte Culture Test, Mixed, Cyclosporins pharmacology, Interferons administration & dosage, T-Lymphocytes drug effects
- Abstract
Human lymphocytes, activated in mixed lymphocyte culture (MLC), express cytolytic activity against a range of target cell types. Cyclosporin A (CyA) efficiently prevents the development of cytotoxic T cells directed against target cells expressing the phenotype of the stimulator cells whilst not preventing an increase in natural killing (NK) activity measured against tumor targets. The drug was further shown not to influence the short-term induction of increased NK activity by interferon. Already established T killer cells are not sensitive to CyA. The drug thus seems to be selectively involved in the induction of immune responses requiring T-cell proliferation.
- Published
- 1981
- Full Text
- View/download PDF
48. NK-cell stimulating properties of a membrane proteoglycane from non-capsulated Klebsiella pneumoniae biotype a.
- Author
-
Normier G, Pinel AM, Dussourd D'Hinterland L, Ramstedt U, and Wigzell H
- Subjects
- Adjuvants, Immunologic immunology, Animals, Chromatography, Gel, Cytotoxicity, Immunologic drug effects, Dose-Response Relationship, Drug, Female, Injections, Intraperitoneal, Interferon Type I metabolism, Male, Mice, Mice, Inbred Strains, Spleen immunology, Time Factors, Bacterial Outer Membrane Proteins immunology, Killer Cells, Natural immunology, Klebsiella pneumoniae, Proteoglycans immunology
- Abstract
Upon testing the individual fractions of a composite bacterial vaccine for biological activities, a potent immuno-stimulatory capacity could be demonstrated within a crude membrane proteoglycan preparation from Klebsiella pneumoniae. One of its characteristic features was the capacity to induce alpha-type interferon and increased NK activity in vivo in mice, following intraperitoneal or oral administration. A highly purified fraction from the crude preparation was obtained using alkaline hydrolysis, delipidation and size fractionation. This fraction was shown to be a very potent inducer of NK cells in vivo or in vitro, where in the latter systems concentrations as low as 0.1 microgramme per ml were highly efficient.
- Published
- 1985
- Full Text
- View/download PDF
49. Lipoxins: stereochemistry, biosynthesis and biological activities.
- Author
-
Serhan CN, Hamberg M, Ramstedt U, and Samuelsson B
- Subjects
- Animals, Arachidonic Acids metabolism, Humans, Hydroxyeicosatetraenoic Acids chemical synthesis, Hydroxyeicosatetraenoic Acids pharmacology, In Vitro Techniques, Killer Cells, Natural drug effects, Leukocytes drug effects, Leukocytes metabolism, Lipid Peroxides metabolism, Masoprocol pharmacology, Neutrophils drug effects, Protein Kinase C metabolism, Stereoisomerism, Hydroxyeicosatetraenoic Acids biosynthesis, Leukotrienes, Lipoxins
- Published
- 1986
50. Natural killer activity in (C57BL/6 X DBA/2)F1 hybrids undergoing acute and chronic graft-vs.-host reaction.
- Author
-
Pattengale PK, Ramstedt U, Gidlund M, Orn A, Axberg I, and Wigzell H
- Subjects
- Acute Disease, Animals, Antibody-Dependent Cell Cytotoxicity, Chronic Disease, Crosses, Genetic, Graft vs Host Disease diagnosis, Graft vs Host Disease genetics, Hematopoiesis, Leukemia, Experimental immunology, Lymphocyte Activation, Lymphoma immunology, Male, Mice, Mice, Inbred C57BL, Mice, Inbred DBA, Spleen cytology, Spleen immunology, Spleen pathology, Cytotoxicity, Immunologic, Graft vs Host Disease immunology, Killer Cells, Natural immunology
- Abstract
The present findings demonstrate that a total i.v. transfer of 100 X 10(6) C57BL/6 (B6) parental spleen cells into untreated (C57BL/6 X DBA/2)F1 hybrids (B6D2F1) resulted in acute runting, which was associated with a significantly elevated graft-vs.-host (GVH) index over a one-month period following GVH induction. Furthermore, this B6-induced acute GVH disease was associated with a marked depression of natural killer (NK) cell activity (spleen and peripheral blood) (with or without addition of mouse fibroblast interferon), which correlated with lymphoid cell hypocellularity, prominent splenic extramedullary hematopoiesis (EMH), and parallel depressions of both concanavalin A- and lipopolysaccharide-induced mitogenesis. Significantly increased killing by antibody-dependent cellular cytotoxicity of antibody-coated chicken red blood cells, as well as increased T cell killing of the NK-insensitive cell line P815 (as compared to the significantly decreased killing of the NK-sensitive cell line YAC-1) was also observed in the spleens of this 100 X 10(6) B6-injected F1 group. In marked contrast to this 100 X 10(6) B6-injected acute GVH group, untreated mice injected i.v. with the same or greater numbers of parental DBA/2 spleen cells (100 X 10(6)-150 X 10(6) DBA/2 spleen cells) exhibited a milder and more chronic form of GVH disease, which was not associated with a significant decrease of NK activity. It was of considerable interest that a total i.v. transfer of 50 X 10(6) B6 spleen cells (i.e. one-half of that required to produce acute GVH, markedly depressed NK, and prominent splenic EMH) into B6D2F1 hybrids also resulted in a more chronic form of GVH disease, but was associated with significantly increased levels of NK activity at two weeks post GVH induction.
- Published
- 1983
- Full Text
- View/download PDF
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