13 results on '"Rolf O. Ehrhardt"'
Search Results
2. Process automation helps standardize and de-risk thawing of cellular therapy products
- Author
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Maria L. Thompson, Brian Schryver, and Rolf O. Ehrhardt
- Subjects
Cell therapy ,Cancer Research ,Transplantation ,Oncology ,Risk analysis (engineering) ,Computer science ,Immunology ,Immunology and Allergy ,Cell Biology ,Process automation system ,Genetics (clinical) - Published
- 2017
3. Technological developments for small-scale downstream processing of cell therapies
- Author
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Maria L. Thompson, Lee Buckler, Eric J. Kunkel, and Rolf O. Ehrhardt
- Subjects
0301 basic medicine ,Cancer Research ,Computer science ,Immunology ,Cytological Techniques ,Cell- and Tissue-Based Therapy ,Specimen Handling ,03 medical and health sciences ,0302 clinical medicine ,Bioreactors ,Downstream (manufacturing) ,Immunology and Allergy ,Humans ,Genetics (clinical) ,Cryopreservation ,Transplantation ,Downstream processing ,Scale (chemistry) ,Cell Biology ,030104 developmental biology ,Oncology ,Risk analysis (engineering) ,Batch Cell Culture Techniques ,030220 oncology & carcinogenesis ,Microtechnology - Abstract
Despite considerable regulatory and clinical hurdles, the development and use of cell-based therapies are gaining momentum. As more of these therapies move toward commercial approval and larger-scale distribution, associated manufacturing and processing technologies are being advanced. Modern technologies directed at downstream processing seek to distribute such therapies from the manufacturing site to the patient more efficiently and reliably. Novel small-scale downstream solutions boost the transformation of cell therapies from abstraction to reality.
- Published
- 2015
4. Blockade of Experimental Atopic Dermatitis via Topical NF-κB Decoy Oligonucleotide
- Author
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Kim Mai, Tony Muchamuel, Susan Hudak, Christopher G. De Vry, Debra Mutnick, Jie Zhang, Christi Parham, Leslie M. McEvoy, Brian Schryver, Maya Dajee, Timothy Colby, Srinivasa Prasad, Tina Le, László G. Kömüves, Aung Oo, Donald Ruhrmund, Jennifer Alleman-Sposeto, Radha Shyamsundar, and Rolf O. Ehrhardt
- Subjects
Male ,Allergy ,Administration, Topical ,Apoptosis ,Mice, Inbred Strains ,Inflammation ,Dermatology ,Biochemistry ,Permeability ,Dermatitis, Atopic ,Ointments ,Mice ,chemistry.chemical_compound ,Immune system ,Antigen ,medicine ,Animals ,Edema ,Ear, External ,Molecular Biology ,Skin ,integumentary system ,business.industry ,NF-κB ,Atopic dermatitis ,Cell Biology ,medicine.disease ,Betamethasone valerate ,Oligodeoxyribonucleotides ,chemistry ,Immunology ,Atrophy ,medicine.symptom ,business ,Decoy ,Cell Division ,Immunosuppressive Agents - Abstract
Atopic dermatitis (AD) is a common chronic skin inflammatory disease. Long-term use of topical corticosteroids in skin inflammation poses risks of systemic and local side effects. The NF-kappaB transcription factor family plays a central role in the progression and maintenance of AD. This study explores the possibility of using topical NF-kappaB Decoy as a novel therapeutic alternative for targeting Th1/Th2-driven skin inflammation in experimental AD. A high-affinity, topical NF-kappaB Decoy developed for human efficacy demonstrates: (i) efficient NF-kappaB Decoy penetration in pig skin, (ii) NF-kappaB Decoy nuclear localization in keratinocytes and key immune cells, and (iii) potent "steroid-like" efficacy in a chronic dust-mite antigen skin inflammation treatment model. NF-kappaB Decoy exerts its anti-inflammatory action through the effective inhibition of essential regulators of inflammation and by induction of apoptosis of key immune cells. Unlike betamethasone valerate (BMV), long-term NF-kappaB Decoy treatment does not induce skin atrophy. Moreover, topical NF-kappaB Decoy, in contrast to BMV, restores compromised stratum corneum integrity and barrier function. Steroid withdrawal causes rapid rebound of inflammation, while the NF-kappaB Decoy therapeutic benefit was maintained for weeks. Thus, topical NF-kappaB Decoy provides a novel mechanism of reducing chronic skin inflammation with improved skin homeostasis and minimal side effects.
- Published
- 2006
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5. The ThawSTARTM Automated Cell Thawing Platform: De-Risking Thawing From Research to the Clinic
- Author
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Maria L. Thompson, Rolf O. Ehrhardt, Brian Schryver, and Eric J. Kunkel
- Subjects
Andrology ,Cancer Research ,Transplantation ,Oncology ,business.industry ,Immunology ,Immunology and Allergy ,Medicine ,Cell Biology ,business ,Genetics (clinical) - Published
- 2016
6. The ThawSTARTM automated cell thawing system: standardizing the last step in the cryopreservation work flow
- Author
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Eric J. Kunkel, Rolf O. Ehrhardt, Brian Schryver, and Maria L. Thompson
- Subjects
Cancer Research ,Transplantation ,Oncology ,Computer science ,business.industry ,Immunology ,Immunology and Allergy ,Work flow ,Cell Biology ,Process engineering ,business ,Genetics (clinical) ,Cryopreservation - Published
- 2015
7. BioT™ cryopod carrier: standardized cryogenic temperature handling of cellular therapies
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Scott Comiso, John Fink, Maria L. Thompson, Rolf O. Ehrhardt, and Eric J. Kunkel
- Subjects
Cancer Research ,Transplantation ,Biot number ,Computer science ,Immunology ,Airlock ,Critical zone ,Process (computing) ,Sampling (statistics) ,Classification scheme ,Cell Biology ,Reliability engineering ,Oncology ,Immunology and Allergy ,Work site ,Cryogenic temperature ,Genetics (clinical) - Abstract
personnel and process flows, and resulting air flow patterns. Within the BSC critical zone, sites should be in close proximity to operations such as centrally located settle plates and particle sensors not more than 1 foot from the work site. Surface viable sites should be in close proximity to processing activity, at locations contacted by operator gloves, areas where materials transfer from lower classification, and other facility surfaces (e.g. floors, walls) to assess disinfectant efficacy. Suggested sampling frequencies are shown in Tables 1 and 2. EM Levels: Cell therapy areas typically will have ISO 5 BSCs placed within an ISO 7 room with ISO 8 support such as a gowning airlock. This classification scheme is outlined in USP Cellular and Tissue-Based Products. Suggested EM levels for cell therapy aseptic processing are shown in Table 3. Conclusion: Considerations are offered to help cell therapy manufacturers to set-up and an appropriate risk-based EM sampling program that meets criteria of being meaningful, manageable, and defendable.
- Published
- 2015
8. Non-viral delivery of nuclear factor-kappaB decoy ameliorates murine inflammatory bowel disease and restores tissue homeostasis
- Author
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Christopher G. De Vry, Christi Parham, Sarvesh Adda, Leslie M. McEvoy, Kim Mai, Srinivasa Prasad, Nicole Kahoud, Tony Muchamuel, Jennifer Hoffman, Radhika Garlapati, Jie Zhang, Brian Schryver, Rolf O. Ehrhardt, Radha Shyamsundar, László G. Kömüves, Tina Le, Carlos Lorenzana, and Maya Dajee
- Subjects
Budesonide ,Male ,Colon ,Inflammatory bowel disease ,Oxazolone ,chemistry.chemical_compound ,Mice ,Medicine ,Animals ,Homeostasis ,Colitis ,Interleukin 6 ,Tissue homeostasis ,Mice, Inbred BALB C ,Microscopy, Confocal ,biology ,business.industry ,Dextran Sulfate ,Inflammatory Bowel Disease ,Gastroenterology ,Gene Transfer Techniques ,NF-kappa B ,T helper cell ,Genetic Therapy ,medicine.disease ,Inflammatory Bowel Diseases ,Ulcerative colitis ,Mice, Inbred C57BL ,Disease Models, Animal ,medicine.anatomical_structure ,chemistry ,Oligodeoxyribonucleotides ,Trinitrobenzenesulfonic Acid ,Immunology ,Cancer research ,biology.protein ,Female ,Inflammation Mediators ,business ,medicine.drug - Abstract
Background: Nuclear factor-κB (NF-κB) is a key transcriptional regulator of inflammatory bowel disease (IBD). Aim: To investigate the therapeutic potential of a locally administered “non-viral” nuclear factor-κB decoy (NFκBD) in multiple experimental models of IBD. Methods: A fully phosphorothioated decoy oligonucleotide with improved stability that specifically binds NF-κB and blocks inflammatory mediators regulated by this transcription factor without the help of viral envelope-assisted delivery was developed. The therapeutic effects of NFκBD were studied in the trinitrobenzene sulphonic acid, oxazolone and dextran sodium sulphate induced colitis models. Results: Intracolonic administration of NFκBD results in the delivery of NFκBD to inflammatory cells and a reduction of NF-κB heterodimers. In the T helper cell 1-driven trinitrobenzene sulphonic acid-induced colitis model, mice receiving NFκBD treatment exhibit a dose-dependent reduction in disease severity and a more rapid recovery to normal body weight, similar to a clinically relevant dose of budesonide. Clinical efficacy was corroborated by considerable reductions in colitis pathology and tissue levels of several pro-inflammatory markers, including tumour necrosis factor α, interleukin 6, interleukin 1β and monocyte chemotactic protein 1. NFκBD also mitigates disease activity in the T helper cell 2-like oxazolone colitis and epithelial injury-related acute dextran sodium sulphate colitis models. Interestingly, restoration of tissue homeostasis is observed in NFκBD-treated animals with the rapid re-emergence of functional goblet cells and a return to normal patterns of cell proliferation in the mucosal epithelium and smooth muscle cell layers. Conclusions: These data support the potential use of “naked” NFκBD as a cross-functional therapeutic in IBD, and show for the first time that it can facilitate the restoration of colon homeostasis and function.
- Published
- 2006
9. Role of IFN-gamma in Th1 differentiation: IFN-gamma regulates IL-18R alpha expression by preventing the negative effects of IL-4 and by inducing/maintaining IL-12 receptor beta 2 expression
- Author
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Ronald B. Smeltz, June Chen, Rolf O. Ehrhardt, and Ethan M. Shevach
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Mice, Inbred A ,Immunology ,Mice, Transgenic ,Biology ,Neutralization ,Interferon-gamma ,Mice ,Adjuvants, Immunologic ,Immunology and Allergy ,Animals ,Receptor ,Interleukin 4 ,Cells, Cultured ,Mice, Inbred BALB C ,Receptors, Interleukin-18 ,Interleukin-18 ,Receptors, Interleukin-12 ,Cell Differentiation ,Receptors, Interleukin ,Th1 Cells ,Interleukin-12 ,Cell biology ,DNA-Binding Proteins ,Mice, Inbred C57BL ,Interleukin 12 ,Female ,Interleukin-4 ,Signal transduction ,Interleukin-18 Receptor alpha Subunit - Abstract
Two key events occur during the differentiation of IFN-gamma-secreting Th1 cells: up-regulation of IL-12Rbeta2 and IL-12-driven up-regulation of IL-18Ralpha. We previously demonstrated that IL-12-driven up-regulation of IL-18Ralpha expression is severely impaired in IFN-gamma(-/-) mice. However, it was unclear from these studies how IFN-gamma influenced IL-18Ralpha since IFN-gamma alone had no direct effect on IL-18Ralpha expression. In the absence of IL-4, IL-12-dependent up-regulation of IL-18Ralpha/IL-12Rbeta2 was independent of IFN-gamma. However, in the presence of IL-4, IFN-gamma functions to limit the negative effects of IL-4 on both IL-18Ralpha and IL-12Rbeta2. Neutralization of IL-4 restored IL-12-driven up-regulation of IL-18Ralpha/IL-12Rbeta2 in an IFN-gamma-independent fashion. In the absence of both IL-12 and IL-4, IFN-gamma up-regulates IL-12beta2 expression and primes IFN-gamma-producing Th1 cells. When T cells were primed in the presence of IL-4, no correlation was found between the levels of expression of the IL-18Ralpha or the IL-12Rbeta2 and the capacity of these cells to produce IFN-gamma, suggesting that IL-4 may also negatively affect IL-12-mediated signal transduction and thus Th1 differentiation. These data clarify the role of IFN-gamma in regulation of IL-18Ralpha/IL-12Rbeta2 during both IL-12-dependent and IL-12-independent Th1 differentiation.
- Published
- 2002
10. Validation of a novel portable freezing device in the optimal freezing of peripheral blood mononuclear cells for potential cell therapy use
- Author
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Qizhi Tang, Maria L. Thompson, Rolf O. Ehrhardt, and Brian Schryver
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Cancer Research ,Transplantation ,business.industry ,Immunology ,Cell Biology ,Peripheral blood mononuclear cell ,Cryopreservation ,Cell therapy ,Clinical study ,Oncology ,Immunology and Allergy ,Medicine ,Drug product ,business ,Genetics (clinical) ,Biomedical engineering - Abstract
In order to ensure a standardized and consistent Drug Product freezing and to minimize batch-to-batch differences in cell recovery and viability postthaw, TxCell scientists tested the CoolCell container, a passive freezing device, as an alternative to the classical controlled rate freezer. Results showed that by using a CoolCell freezing device, Ag-Tregs can be successfully cryopreserved and recovered in a standardized way acceptable to the processing and manufacturing of cell therapies. TxCell now intends to use the CoolCell device in its phase IIb clinical study with its lead AgTreg cell product candidate, Ovasave . Use of the CoolCell passive freezing device for cell therapy manufacturing of Ag-Treg represents a new standardized method of cell therapy product cryopreservation. The ability to develop and store functional Treg in a cost-effective and reproducible manner is an important milestone in the ultimate use of these cells in the clinic.
- Published
- 2014
11. Persistence of pathogenic CD4+ Th1-like cells in vivo in the absence of IL-12 but in the presence of autoantigen
- Author
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Kenneth Hong, Rolf O. Ehrhardt, and Ellen L. Berg
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CD4-Positive T-Lymphocytes ,Adoptive cell transfer ,Cell Survival ,Immunology ,Inflammation ,Mice, SCID ,Biology ,Autoantigens ,Lymphocyte Depletion ,Mice ,In vivo ,Recurrence ,T-Lymphocyte Subsets ,medicine ,Immunology and Allergy ,Animals ,Psoriasis ,Lymph node ,Immunization Schedule ,Autoimmune disease ,Mice, Inbred BALB C ,Anti-Inflammatory Agents, Non-Steroidal ,Antibodies, Monoclonal ,Th1 Cells ,medicine.disease ,Adoptive Transfer ,Interleukin-12 ,Disease Models, Animal ,medicine.anatomical_structure ,Chronic Disease ,biology.protein ,Interleukin 12 ,Female ,Lymph ,Lymph Nodes ,medicine.symptom ,Antibody ,Injections, Intraperitoneal - Abstract
Despite recent successful treatment of murine autoimmune disease with anti-IL-12 mAb, it has not yet been addressed whether anti-IL-12 mAb can also be effective in late stages of disease and whether it can provide lasting protection against recurrence, especially during continued presence of autoantigen. We used a newly developed psoriasis model in scid/scid mice, which allows easy tracking of pathogenic T cells, to show that when anti-IL-12 mAb is given for 2 wk (1 mg/wk) in the late stage of severe disease, inflammation is greatly reduced, as measured by ear thickness and histology (scores, 1.1 ± 0.1 vs 2.0 ± 0.4). Moreover, prolonged treatment (4 wk) of chronic psoriatic mice with high doses of mAb (1 mg/wk; prolonged active anti-inflammatory treatment (PAAIT)) results in the almost complete resolution of lesions (scores, 0.3 ± 0.1 vs 2.7 ± 0.2). Surprisingly, however, despite these significant treatment results, the psoriasis-like lesions return soon after the anti-IL-12 mAb treatment is discontinued. This rapid relapse of disease may be attributed to large populations of activated CD4+ T cells present in the lymph nodes of PAAIT animals still expressing an effector/memory phenotype (CD45RBlow, L-selectinlow). Upon stimulation in vitro such PAAIT lymph node cells secrete high amounts of IFN-γ (129 ng/ml); when transferred into naive scid/scid animals they are able to rapidly induce disease without costimulation. Our data indicates an alternative IL-12-independent pathway for pathogenic Th-1-like cells in vivo during the chronic phase of disease that allows these cells to persist and maintain their pathogenicity in the draining lymph tissue of the autoimmune site.
- Published
- 2001
12. T helper type 2 cell differentiation occurs in the presence of interleukin 12 receptor beta2 chain expression and signaling
- Author
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Rolf O. Ehrhardt, Ryuta Nishikomori, and Warren Strober
- Subjects
CD4-Positive T-Lymphocytes ,Immunology ,Mice, Transgenic ,Lymphocyte Activation ,interleukin 4 ,Interleukin 21 ,Interferon-gamma ,Mice ,Th2 Cells ,reversibility ,cytokine ,Immunology and Allergy ,Cytotoxic T cell ,Animals ,IL-2 receptor ,Interleukin 4 ,Interleukin 3 ,CD40 ,biology ,Receptors, Interleukin-12 ,Cell Differentiation ,Receptors, Interleukin ,STAT4 Transcription Factor ,Th1 Cells ,Molecular biology ,DNA-Binding Proteins ,Interleukin 15 ,signal transducer and activator of transcription 4 ,biology.protein ,Interleukin 12 ,Trans-Activators ,Original Article ,Interleukin-4 ,T helper type 1 ,Signal Transduction - Abstract
The differentiation of CD4(+) T cells into T helper type 1 (Th1) cells is driven by interleukin (IL)-12 through the IL-12 receptor beta2 (IL-12Rbeta2) chain, whereas differentiation into Th2 cells is driven by IL-4, which downregulates IL-12Rbeta2 chain. We reexamined such differentiation using IL-12Rbeta2 chain transgenic mice. We found that CD4(+) T cells from such mice were able to differentiate into Th2 cells when primed with IL-4 or IL-4 plus IL-12. In the latter case, the presence of IL-4 suppressed interferon (IFN)-gamma production 10-100-fold compared with cells cultured in IL-12 alone. Finally, in studies of the ability of IL-12 to convert Th2 cells bearing a competent IL-12R to the Th1 cells, we showed that: (a) T cells bearing the IL-12Rbeta2 chain transgene and primed under Th2 conditions could not be converted to Th1 cells by repeated restimulation under Th1 conditions; and (b) established Th2 clones transfected with the IL-12Rbeta2 chain construct continued to produce IL-4 when cultured with IL-12. These studies show that IL-4-driven Th2 differentiation can occur in the presence of persistent IL-12 signaling and that IL-4 inhibits IFN-gamma production under these circumstances. They also show that established Th2 cells cannot be converted to Th1 cells via IL-12 signaling.
- Published
- 2000
13. Induction and Regulation of Colitis in IL-2-/- Mice
- Author
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Belinda Gray, Bjorn R. Ludviksson, Warren Strober, and Rolf O. Ehrhardt
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business.industry ,Immunology ,Emergency Medicine ,Medicine ,Colitis ,Critical Care and Intensive Care Medicine ,business ,medicine.disease - Published
- 1997
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