An automated and closed system for patient specific CAR-T cell therapies

Autologous cell therapies, particularly chimeric antigen receptor T-cell (CAR-T) immunotherapies, are becoming a promising treatment option for difficult diseases. Immunotherapies for blood cancers have dominated the pipeline, while treatments for solid tumors have started to become more successful. However, as the market continues to grow and more clinical trials begin globally, the challenge of manufacturing autologous cell therapies remains significant. A greater number of patients will lead to an increase in cost, labor, and the complexity of logistics for scaling out the commercial production of patient specific therapies. To enable clinical and commercial success, novel manufacturing platforms, such as closed and automated systems, will be required to produce cost effective and robust therapies. This abstract highlights a successful CAR-T process translation from a manual process to an automated patient scale system. To accomplish a CAR-T process translation, we utilized a platform that automates cell seeding, activation, transduction, real time process monitoring, feeding, washing and concentration, and harvesting. In order to mimic a therapeutic CAR-T cell process, manual research scale processes were optimized, scaled up, and then programmed to run automatically without manual intervention. In these processes, 100 million peripheral blood mononuclear cells (PBMC) were first inoculated with CD3/CD28 activation beads. The following day, cells were transduced with HER-2 lentivirus vector. Cells were then expanded with a defined feeding strategy and IL-2 supplements until harvested when target yields were reached. After harvest, cells were analyzed for cell yield, viability, transduction efficiency, and an array of cell phenotype, potency and functionality via FACS and killing assays. Specifically, CAR-T cells were analyzed for the presence of naïve T cells, T stem cell memory, T central memory, T effector memory, and T effector cells. We show here how we optimized, scaled up, and automated manual processes to reach clinical requirements. Automated runs using the above process with cells transduced by HER-2 virus yielded an average of 2 x 109 cells post harvest with a viability \u3e 90%. Automated runs and associated controls were able to support the expansion of both CD4+ and CD8+ T cells with 73% CD4+ T cells and 20% CD8+ T cells. Harvested cells yielded approximately 80% NGFR+ cells with a higher detection of NGFR in the CD4+ fraction than in the CD8+ fraction for all samples. Both CD4+ and CD8+ subsets demonstrated T cell phenotype such as naïve T cells, T stem cell memory, T central memory, T effector memory, and T effector cells. Both subsets also only expressed between 15-20% of immunosuppressive regulatory T cells. Cell health was evaluated by the levels of exhaustion marker, PD-1, which was 19% in CD4+ T cells and \u3c 1% in CD8+ T cells. Furthermore, there was a negligible amount of senescent T cells and anergic cells and \u3c 10% expression of the apoptotic marker, Caspase-3. Subsequently, cells from multiple automated runs showed the specific killing of NGFR+ tumor line were correlated with high levels of effector cytokines: TNF-alpha (~34%) and IFN-gamma (20-25%) as compared to a manual control. In summary, automated CAR-T process in the Cocoon system yields a healthy populations of T cell subsets. This system is a viable solution to translate labor-intensive CAR-T process into a fully automated system, thus allowing scalability, high yield, reduction of manufacturing cost, and better process control to yield high quality CAR-T cells

Changes in Body Mass Index and Atherosclerotic Disease Risk Score After Switching From Tenofovir Disoproxil Fumarate to Tenofovir Alafenamide.

Background: Switching from tenofovir disoproxil fumarate (TDF) to tenofovir alafenamide (TAF)-containing antiretroviral therapy (ART) can improve renal function and bone mineral density in people with human immunodeficiency virus (PWH). The switch can also negatively influence cholesterol, but changes in body mass index (BMI) and atherosclerotic cardiovascular disease (ASCVD) risk are unknown. Methods: This retrospective observational study evaluated BMI and ASCVD risk score changes in virologically suppressed PWH who switched from TDF to TAF without switching other ART regimen components. Adults on TDF for ≥1 year with 2 consecutive HIV ribonucleic acid values/mL before a TAF switch were included. Body weight, BMI, cholesterol, and ASCVD risk score were collected for the year before and after the switch. Pre- and postswitch values were compared with the Wilcoxon signed-rank test. Changes in BMI and ASCVD scores were modeled using generalized estimating equations regression. Results: One hundred ten patients were included. In unadjusted analyses, there were significant increases in weight, BMI, total cholesterol, LDL, HDL, and ASCVD risk score in the year after switching from TDF to TAF (each P ≤ .01). In regression models, switching from TDF to TAF was associated with a 0.45 kg/m2 increase in BMI (95% confidence interval [CI], 0.14-0.76) and a 13% increase in ASCVD risk score (95% CI, 4%-23%). Conclusions: We observed significant BMI and ASCVD score increases in PWH 1 year after switching from TDF to TAF. The mechanism of changes is unclear and requires additional study

1961

My Ideal Course, Underwater, U.S.A. (page 1) From the Editor (3) Turf Management Club News (3) Quotes from 1961 Seniors (4) The United States Most Western Owned Golf Course: Armed Forces Golf Course, Guam (5) Turf Majors Participate in Horticultural Show (7) Picture - G.C.S.A Scholarships Awarded to Three Turf Seniors (8) Picture - Stockbridge - Majors in turf Management (9) Opportunity and Education (10) The Most Outstanding Turf Senior for the Year - 1961 (11) How We Prepare Our Greens Before Topdressing (12) An Inexpensive Cure for Weeds and Poa Annua (13) Watering (14) Picture - Honorary Members of the Turf Management Club (16) Picture - Graduates of Winter School for Turf Managers - 1961 (17) Welcome Speech by Narry Sperandio (A-1) Handle with Care by Dr. Ellsworth H. Wheeler (A-2) Current Ideas on Green Construction - Panel Discussion (A-4) Automatic Systems for Watering by Robert F. Harper (A-14) History of Golf Course Architecture by Geoffrey S. Cornish (A-22) Effect of Nutrition on Turf Diseases by Dr. Houston B. Couch Turf Disease Control and Use of Fungicides by Dr. R. J. Lukens Trees and Tree Care by Gordon S. King (A-38) Arsenical Toxicity by Dr. C. R. Skogley (A-41) Soil Reaction to Arsenical Compounds by Joseph E. Steckel Brush Control For the Golf Course by Dr. William I. Boyd (A-51) Massachusetts Highway Herbicide Program by Joseph L. Beasley (A-54) General Turf (Alternate Session): Observations on Highway Turf Establishment & Maintenance by E.F. Button (A-62) Pre-emerge Chemicals for the Control of Crabgrass by Dr. John R. Havis, John M. Zak & Joseph Troll (A-70) Root Growth of Turf Grasses as Affected by Different heights of Cut and Nutrient Levels by Evangel J. Bredakis (A-71) The Use of Sod by Daniel Pellegrino (A-72

LAB/NTAL Facilitates Fungal/PAMP-induced IL-12 and IFN-γ Production by Repressing β-Catenin Activation in Dendritic Cells.

Fungal pathogens elicit cytokine responses downstream of immunoreceptor tyrosine-based activation motif (ITAM)-coupled or hemiITAM-containing receptors and TLRs. The Linker for Activation of B cells/Non-T cell Activating Linker (LAB/NTAL) encoded by Lat2, is a known regulator of ITAM-coupled receptors and TLR-associated cytokine responses. Here we demonstrate that LAB is involved in anti-fungal immunity. We show that Lat2−/− mice are more susceptible to C. albicans infection than wild type (WT) mice. Dendritic cells (DCs) express LAB and we show that it is basally phosphorylated by the growth factor M-CSF or following engagement of Dectin-2, but not Dectin-1. Our data revealed a unique mechanism whereby LAB controls basal and fungal/pathogen-associated molecular patterns (PAMP)-induced nuclear β-catenin levels. This in turn is important for controlling fungal/PAMP-induced cytokine production in DCs. C. albicans- and LPS-induced IL-12 and IL-23 production was blunted inLat2−/− DCs. Accordingly, Lat2−/− DCs directed reduced Th1 polarization in vitro and Lat2−/−mice displayed reduced Natural Killer (NK) and T cell-mediated IFN-γ production in vivo/ex vivo. Thus our data define a novel link between LAB and β-catenin nuclear accumulation in DCs that facilitates IFN-γ responses during anti-fungal immunity. In addition, these findings are likely to be relevant to other infectious diseases that require IL-12 family cytokines and an IFN-γ response for pathogen clearance

mRNA Coronavirus Disease 2019 Vaccine-Associated Myopericarditis in Adolescents: A Survey Study

In this survey study of institutions across the US, marked variability in evaluation, treatment, and follow-up of adolescents 12 through 18 years of age with mRNA coronavirus disease 2019 (COVID-19) vaccine-associated myopericarditis was noted. Only one adolescent with life-threatening complications was reported, with no deaths at any of the participating institutions

A Revision of the Upper Cretaceous Lepidosirenid Lungfishes from the Quseir Formation, Western Desert, Central Egypt

We evaluate new lungfish remains from the Upper Cretaceous Quseir Formation in the Western Dessert of southern Egypt. Taxa include Lavocatodus protopteroides, L. humei, L. giganteus, and Protopterus nigeriensis. We treat Lavocatodus as members of Lepidosirenidae based on the presence of a well-defined medial articular surface, in the absence of a symphyseal surface of corresponding prearticular bones. Material of L. protopteroides represents the first example of an adult specimen, supporting the retention of the species as valid, and not a junior synonym of L. humei. Specimens of L. humei are the most abundant in the assemblage, and all exhibit the medial articular surface, a feature that is absent from specimens of Ceratodus; therefore, we reassign additional specimens of C. humei to Lavocatodus. New specimens of L. giganteus extend the temporal and spatial ranges of the species (the type material of L. giganteus is from Paleocene deposits of In Farghas, Mali). Finally, a new specimen of Protopterus represents the oldest record of the genus in Egypt. The prearticular is confidently assigned to Protopterus based on the presence of only three ridges on the tooth plate, a medial symphyseal suture, a coronoid process, and a shallow tooth crown plus prearticular depth. We further assign it to P. nigeriensis based the length and merger of tooth ridges. Furthermore, the oldest Cenomanian specimens reported from the Sudan Wadi Milk Formation are assigned to P. nigeriensis, which would extend the temporal range of P. nigeriensis by roughly 14 Ma. © 2014 by the Society of Vertebrate Paleontology

Stratigraphy and Vertebrate Paleoecology of Upper Cretaceous? Lowest Paleogene Strata on Vega Island, Antarctica

The Upper Cretaceous (Maastrichtian) Sandwich Bluff Member of the López de Bertodano Formation is well exposed on Vega Island in the James Ross Basin off the northeastern coast of the Antarctic Peninsula. Although this unit is one of the richest sources of end-Cretaceous vertebrate fossils in Antarctica, it is also one of the least sedimentologically and stratigraphically characterized units in the basin. New facies and stratigraphic analyses of the Sandwich Bluff Member and the underlying Cape Lamb Member of the Snow Hill Island Formation were performed in tandem with intensive prospecting for fossil vertebrates and stratigraphic assessment of historic paleontological localities on Vega Island. This effort has led to a revised stratigraphy for the Sandwich Bluff Member and the precise stratigraphic placement of important terrestrial and marine vertebrate fossil localities. Facies analysis reveals a fining and shallowing upward trend through the section that culminates in a newly recognized sequence boundary near the top of the Sandwich Bluff Member, followed by the deposition of a previously unrecognized, 6 m-thick, matrix-supported pebble–cobble conglomerate of probable alluvial origin. Immediately overlying this unit, well-developed Thalassinoides burrow networks in fine-grained transgressive sandstones and siltstones indicate a rapid return to marine conditions. A similar stratigraphic pattern is well documented at the top of the López de Bertodano Formation and the base of the overlying (Paleocene) Sobral Formation on Seymour Island in the southern part of the basin. Although no fossils were recovered to constrain the age of the upper 10–15 m of the succession on Vega Island that preserves the newly recognized upper sequence boundary, strata below this level can be confidently placed within the Manumiella bertodano interval zone, which extends to a short distance below the K–Pg boundary on Seymour Island. Hence, based on sequence stratigraphic and lithostratigraphic evidence, the uppermost 10–15 m of the succession on Vega Island may encompass the Cretaceous–Paleogene boundary together with a few meters of the Paleocene Sobral Formation

Hypertension: An Important But Reversible Cause of Systolic Dysfunction in a Cohort of Pediatric Patients.

Cardiac dysfunction due to hypertension (CDHTN) in pediatrics is not well described. We aimed to describe the presentation and outcomes of pediatric CDHTN and identify clinical features associated with resolution of dysfunction. A single-center retrospective cohort study of patients ≤ 21 years with CDHTN from January 2005-September 2020 was performed. Patients with systolic dysfunction without another cause, blood pressure \u3e 95th percentile, and physician judgment that dysfunction was secondary to hypertension were included. Demographics, clinical characteristics, echocardiographic findings, and outcomes were examined using Fisher\u27s exact and Mann-Whitney U tests. Multiple correspondence analysis was used to explore the relationship of resolution of dysfunction to clinical features. Thirty-four patients were analyzed at a median age of 10.9 (IQR 0.3-16.9) years. Patients were divided into groups \u3c 1 year (n = 12) and ≥ 1 year (n = 22). Causes of hypertension were varied by age, with renovascular disease most common in infants (42%) and medical renal disease most common in older patients (77%). Echocardiography demonstrated mild LV dilation (median LV end-diastolic z-score 2.6) and mild LV hypertrophy (median LV mass z-score 2.4). Most patients (81%) had resolution of dysfunction, particularly infants (92%). One patient died and one patient was listed for heart transplant. None required mechanical circulatory support (MCS). No clinical features were statistically associated with resolution of dysfunction. Hypertension is an important but reversible cause of systolic dysfunction in children. Patients are likely to recover with low mortality and low utilization of MCS or transplantation. Further studies are needed to confirm features associated with resolution of dysfunction

EIRSAT-1 - The Educational Irish Research Satellite

The 2nd Symposium on Space Educational Activities (SSEA), Budapest, Hungary, 11-13 April 2018The Educational Irish Research Satellite, "EIRSAT-1", is a collaborative space project that aims to build, launch and operate the first ever Irish satellite. The EIRSAT-1 spacecraft is a 2U CubeSat incorporating three novel experiment payloads: GMOD, a gamma-ray detector; EMOD, a thermal management coating demonstration; and WBC, an attitude control algorithm. The spacecraft is currently under construction at University College Dublin and will be delivered to ESA in late 2019.Enterprise IrelandEuropean Space AgencyIrish Research CouncilUniversity College DublinENIBOIDEASSensL.Department of Business, Enterprise and Innovatio

mRNA Coronavirus Disease 2019 Vaccine-Associated Myopericarditis in Adolescents: A Survey Study

In this survey study of institutions across the US, marked variability in evaluation, treatment, and follow-up of adolescents 12 through 18 years of age with mRNA coronavirus disease 2019 (COVID-19) vaccine-associated myopericarditis was noted. Only one adolescent with life-threatening complications was reported, with no deaths at any of the participating institutions