bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human-derived raw materials: Controlled, consistent collections enable successful manufacturing of cell-based regenerative medicine products (TECH1P.847)
Authors: Ramos Thomas , Wang Wenshi , Okhovat Jacob , McPherson Gaytha , Burger Scott 
Published: January 2023

Abstract
Human cells are critical raw materials for manufacturing cell therapy products, but introduce significant variability. HemaCare controls apheresis procedures and collection sites under a formal quality system, with GMP-compliant, validated procedures and equipment, and GTP-compliant donor screening and tracking. HemaCare performed 86,799 cellular apheresis collections in the last 7 years for research, clinical trials, and commercial products. HemaCare’s unmobilized apheresis products showed consistently high MNC purity, with 93.8% of products containing ≥75% MNC, and an average of 85.66% MNC±7.1% (mean±1 SD). RBC contamination was low, with hematocrit averaging 1.78%±0.7%. Approximately 85% of HemaCare donors have donated apheresis products 5 or more times. HemaCare’s laboratory is equipped with Miltenyi Biotec technology for isolation of raw material into purified cellular subpopulations. The consistency and viability of the purified end products are measured with flow cytometry. Using BioLife Solutions’ serum-free and protein-free cGMP CryoStor™ cryopreservation media with purified cells, post-thaw recovery rates of cell fractions have been above 95%; dendritic cell and macrophage have demonstrated recovery rates of ≥90%. CryoStor™, in combination with freezing in the BioCision CoolCell™ freezing container, has enabled HemaCare to standardize the cryopreservation process, reducing variability while optimizing post-thaw viable cell recovery of its research products.
Automation of immune repertoire amplification on the iC-Processor (TECH1P.851)
Authors: Byrne-Steele Miranda , Pan Wenjing , Brown Brittany , Alleyne Jessica , Yang Qunying , Wang Chunlin , Han Jian 
Published: January 2023

Abstract
Immune repertoire amplification of T and B cell receptors coupled to Next Generation Sequencing (NGS) provides detailed sequence-level insight into the immune system. Previously, we developed a patented, nested multiplex PCR method (arm-PCR) to amplify an immune repertoire, inclusively and semi-quantitatively. Here, we describe the automation of this process in cassette format on the iC-system (iCubate, Inc.) and compare the sequencing results from repeat amplification experiments of the TCRβ and human Ig heavy chains from the same individual. The iC-system is an open, nested multiplex PCR diagnostic development platform, which allows for the development of automated diagnostic panels in a closed cassette format. In addition to the automation, the closed format helps to reduce contamination of the laboratory environment with multiplex primers. Template RNA is added to the cassette at the appropriate concentration with RT enzyme. After two sets of PCR, residual primer-dimers are removed by magnetic selection of the primary product in the cassette. The cassette is then removed from the iC-Processor, and the amplified library is eluted from the magnetic beads. The amplified immune library is directly ready for pooling, QC, and sequencing using the Illumina MiSeq NGS platform. Repeat amplification experiments from the same individual using this automated approach demonstrated high repeatability among shared CDR3 sequences for both the TCRβ and Ig heavy chains.
Predicting conversion to multiple sclerosis using antibody genetics: comparing Sanger and next generation sequencing methods. (TECH1P.858)
Authors: Rounds William , Ligocki Ann , Levin Mikhail , Greenberg Benjamin , Bigwood Douglas , Eastman Eric , Cowell Lindsay , Monson Nancy 
Published: January 2023

Abstract
We have previously identified a distinct mutation pattern in the B cell receptors of cerebrospinal fluid B cells that can predict conversion to multiple sclerosis. This antibody gene signature, developed using Sanger sequencing on singly-sorted B cells, is limited in its clinical application by the time and effort required to obtain such single-cell repertoires. In order to increase the throughput of antibody gene signature evaluation for patients, we used 454 sequencing on bulk-sorted cerebrospinal fluid B cell pellets. We focused our analysis on identifying the mutation and distribution profiles of the repertoires obtained via both sequencing methods. Here we outline the additional filtering and data processing steps used to correct for 454 platform bias and compare the 454 and Sanger sequencing-derived repertoires which include multiple sclerosis samples sequenced by both methods in parallel. Although we observe variable amounts of background mutation and uneven sequence amplification noise by 454 sequencing for our matched samples, most of the antibody gene signature scores remain above the multiple sclerosis conversion threshold established by Sanger sequencing. We expect that additional improvements directed at the sample preparation, data filtering and sequence clustering methods presented here will help further improve the reliability of next generation sequencing-based multiple sclerosis conversion diagnosis.
Over-expression of microRNA-494 is involved in repression of genes but independent of antigen receptor-mediated apoptosis in a T cell leukemia cell line (TECH1P.867)
Authors: Jayaraman Sundararajan 
Published: January 2023

Abstract
Treatment of the human T cell leukemia cell line, Jurkat with the histone deacetylase inhibitor, Trichostatin A (TSA) resulted in histone hyperacetylation, dissipation of mitochondrial transmembrane potential, and augmentation of T cell receptor mediated apoptosis. Global gene expression profiling of TSA-treated Jurkat cells revealed the repression of several genes involved in T cell receptor signaling and PVRIG, a non-coding member of the paired immune-like receptor family genes. Analysis of total RNA by qRT-PCR validated these findings. Genome wide microRNA expression profiling uncovered drug-mediated down-regulation of microRNAs, including the miR-17-92 cluster, critical for T cell biology. Notably, TSA treatment induced the up-regulation of miR-494, which correlated with Jurkat cell death. This is in stark contrast to previous reports indicating that the up-regulation of miR-494 is essential for the survival of carcinomas. Interestingly, transfection of Jurkat cells with a miR-494 inhibitor partially blocked TSA-mediated down-regulation of PVRIG expression, without abrogating the antigen receptor mediated apoptosis. These data demonstrate that the over-expression of miR-494 as a consequence of histone hyperacetylation results in tumor cell death, which is unrelated to antigen receptor-mediated apoptosis. Thus, up-regulation of miR-494 via epigenetic mechanisms appears to be a novel treatment strategy for T cell leukemia.
Mild cold stress resulting from standard housing conditions for laboratory mice influences baseline dendritic cell properties (TUM2P.896)
Authors: Kokolus Kathleen , Spangler Haley , Povinelli Benjamin , Farren Matthew , Lee Kelvin , Repasky Elizabeth 
Published: January 2023

Abstract
The ability of dendritic cells (DC) to stimulate and regulate T cells is critical for effective anti-tumor immunity. Therefore, it is important to recognize any inherent factors which may influence DC function under experimental conditions, especially in laboratory mice which are commonly used to model anti-tumor immunity. Recent data shows that CD8+ T cell frequency within the tumor microenvironment and anti-tumor function is dependent upon the ambient temperature used to house laboratory mice. As DC function is crucial for CD8+ T cell activation, we hypothesized that DC function is also dependent upon housing temperature. We observed increased numbers of splenic DCs (CD11c+ ) in tumor-bearing mice at housed at standard temperature (ST; 22°C) vs. mice housed at a thermoneutral temperature (TT; 30°C) which alleviates the mild cold stress experienced at ST. Despite increased frequency, DCs from mice at ST primarily display immature phenotypes (MHC II+CD86- ), likely rendering them tolerogenic and unable to activate T cells. Further, splenocytes from tumor-bearing mice at ST were unable to induce T cell proliferation following tumor inoculation while DCs from mice at TT elicited T cell activation. These findings likely contribute to faster tumor growth observed in mice at ST vs. TT. This data strongly suggests that the housing temperature can affect fundamental properties of DC function which in turn influence the ability of DCs to regulate the anti-tumor immune response.
IL-33 precedes IL-5 in regulating early commitment to eosinophil development (HEM5P.229)
Authors: Johnston Laura , Berdnikovs Sergejs , Krier-Burris Rebecca , Hsu Chia-Lin , Chien Karen , Bryce Paul 
Published: January 2023

Abstract
IL-33 is strongly associated with allergic disease, in part because of the potent effects it exerts on Th2 lymphocytes, ILC2s, mast cells, and eosinophils, which express its receptor ST2. However, ST2 has also recently been observed on hematopoietic progenitor cells. The effect of IL-33 on these cells is unclear. We observed that naïve ST2KO and IL-33KO mice had significantly less peripheral blood eosinophils (PBEs), whereas exogenous IL-33 was sufficient to elevate PBEs in WT and IL-33KO mice, but not in ST2KO mice. This was associated with increased myeloid/erythroid cell ratios in the bone marrow itself. IL-33 treatment expanded a Sca1-/Linint/SiglecF+ population, characteristic of a committed eosinophil progenitor. Using in vitro eosinophilopoesis conditions, IL-33 promoted mature eosinophil commitment (Sca1-/Lin+/SiglecF+) that was significantly diminished in ST2KO cells. Instead, ST2KO cells accumulated a Sca1-/Lin-/SiglecF+ population, characteristic of an early committed precursor. In contrast, IL-5, which has been shown to drive eosinophil development, expanded mature eosinophils. Finally, we assessed the hierarchy of IL-5 and IL-33 by crossing IL-5 overexpressing transgenic mice (NJ1638) with ST2KO mice. While IL-5 transgenic mice exhibited a robust age-dependent increase in PBEs, ST2 deficiency significantly prevented this IL-5-driven eosinophilia. Our findings demonstrate a novel role for IL-33 and ST2 in supporting eosinophil development that precedes IL-5.
Reduced kinase inhibitory region containing suppressor of cytokine signaling molecule levels in SLE patients is correlated to enhanced STAT1 activation (HUM3P.242)
Authors: Sukka-Ganesh BhagyaLaxmi , Wilson Tenisha , Westley Reeves , Johnson Howard , Larkin Joseph 
Published: January 2023

Abstract
Systemic Lupus erythematosus (SLE) is a chronic inflammatory autoimmune disorder with unknown etiology. Given the importance of the suppressor of cytokine signaling (SOCS) family of intracellular proteins in the regulation of lupus promoting cytokine signaling, we tested the hypothesis that SOCS deficiencies would be present in SLE patients. We therefore compared a group of 34 SLE patients, by disease activity (SLEDIA) and prednisone treatment, to 11 healthy controls for the presence of the kinase inhibitory region (KIR) bearing SOCS1 and SOCS3. We found that protein and mRNA levels of SOCS1 and SOCS3 were significantly reduced in PMBC’s isolated from SLE patients compared to controls. Further, decreased SOCS1, but not SOCS3 protein levels in the SLE patients was inversely correlated to pSTAT1 activation, but not Erk 1/2 or Akt. Notably, the inverted SOCS1/pSTAT1 ratio correlated to significantly enhanced MHC class II levels amongst SLE patients. Together these data suggest that SOCS1 deficiencies may play an essential role in the onset/progression of SLE in a subset of SLE patients, possibly through dysregulated STAT1 mediated antigen presentation. These studies represent a critical first step in implicating a role of SOCS1 and SOCS3 deficiencies in the progression of human SLE. Moreover, these studies point to the possibility that peptides, which partially mimic SOCS signaling and inhibit rodent autoimmune disease, may have efficacy in the treatment of human SLE.
The potential involvement of plasmablast antibodies in clinically isolated syndrome and multiple sclerosis (HUM3P.268)
Authors: Monson Nancy , Rivas Jacqueline , Ligocki Ann , Rounds William , Ireland Sara , Ma Lisha , Frohman Elliot , Greenberg Benjamin , Cowell Lindsay , Stowe Ann 
Published: January 2023

Abstract
Plasmablasts are a transient B cell subtype that secretes large amounts of antibody and arises in response to antigen exposure and T cell help. Once plasmablasts form they die off in a few days or differentiate into a long lived plasma cells, and therefore are only present during an active immune response. In our laboratory a subset of Clinically Isolated Syndrome (CIS) patients presenting with Transverse Myelitis (TM) symptoms at high risk to develop Multiple Sclerosis (MS) also have an elevated level of plasmablasts in the peripheral blood and/or the cerebrospinal fluid. Interestingly, only those patients in our cohort who have elevated plasmablast levels continue to accumulate brain MRI activity. The antibody genetics of plasmablasts from CIS TM patients are altered in comparison to those of plasmablasts responding to influenza from otherwise healthy individuals. There is increased evidence of receptor editing in CIS patients, as well as enrichment of the Antibody Gene Signature (AGS), which predicts whether CIS patients will convert to MS later in life. Antibodies from CIS plasmablasts that are enriched in overall mutations and AGS mutations recognize mouse brain by ELISA and histology, binding to multiple different cell types. By flow cytometry we have shown that many of these antibodies recognize mouse astrocyte and neuron cell lines. If these antibodies are able to bind brain tissue in vivo, they could exacerbate brain damage and contribute to disease in CIS patients.
A second wave of monocyte expansion may contribute to the severe dengue in 2014 outbreak in China (HUM4P.272)
Authors: Yu Lei , Hong Wenxin , Zhao Lingzhai , Zhang Ling , Wang Qianqian , Lei Yang , Qiu Shuang , Yin Chibiao , Feng Hanping , Guan Yongjun , Zhang Fuchun 
Published: January 2023

Abstract
The 2014 dengue outbreak, the largest one in the last ten years in China, resulted in 13,800 hospitalized patients and more than 300 severe dengue cases in Guangzhou. Both of the viral strain and the host immune background may contribute to severe dengue. Here we report the monocyte response in dengue fever (DF) and severe dengue (SD) during the acute infection of seven dengue patients (DF n=4 and SD n=3) using multiparameter flow-cytometry with a panel of 9-surface markers. We also compared dengue specific IgG, IgM and NS1 in serum at the same time points for each patient. Six of the seven patients were diagnosed as secondary DENV1 infection, while one DF diagnosed as primary DENV1 infection. NS1 was nearly cleared in all patients when the serum IgG and IgM reached the high level at day 8-10 from the onset of fever. Obvious monocyte expansion was observed in both of DF and SD groups, frequently companied with the expansion of CD14++CD16+ subset. But the pattern of the response in the two groups was different at the recovery phase. The monocyte expansion in DF group maintained at a high level at day 7-10. Interestingly, the SD group underwent a second wave of monocyte expansion, in parallel with clinical course, starting from day 7 to day 10, especially the CD16+CD64+ monocyte subset. The characteristics of monocyte responses in DF and SD groups will advance our understanding of human dengue pathogenesis and may implicate the potential targets for early therapy.
Severity of infectious mononucleosis correlates with the frequency of crossreactive influenza A/Epstein Barr virus-specific CD8+ T cells (HUM4P.301)
Authors: Selin Liisa , Watkin Levi , Gil Anna , Mishra Rabinarayan , Luzuriaga Katherine , Aslan Nuray 
Published: January 2023

Abstract
During EBV-associated infectious mononucleosis (IM) IAV-specific crossreactive memory T cells are activated and play a role in disease severity. In HLA-A2+ IM patients, influenza M158 (IAV-M1)-specific CD8 memory T cell responses crossreacted with two different EBV lytic epitopes, BMLF1280 (17/29) and BRLF1109 (19/20). Disease severity of IM directly correlated with significantly increased frequencies of crossreactive IAV-M1/EBV-BMLF1, IAV-M1, and EBV-BMLF1 specific CD8 cells, and with mean viral load over the first 5 weeks of infection. Disease severity did not correlate with BRLF1 or M1/BRLF1 crossreactive responses. When severity of IM was scored and patients were assigned to either mild or severe groups, disease severity correlated with specific TCR Vb usage in IAV-M1 population suggesting that TcR selection is driving disease outcome. Consistent with crossreactive responses driving increased immunopathology was the observation that tetramer-sorted crossreactive responses had completely altered immune response signatures than cognate responses by gene array. These results suggest that T cell crossreactivity impacts T cell selection and function and ultimately disease outcome. Insights on these issues are important for the intelligent design of vaccines and to develop therapeutic interventions for virally induced disease (NIHAI49320).
Basophil surface expressions of CD203c and CD63 following peanut allergen ex vivo stimulation correlate with severity of allergic reactions during food challenge (HYP4P.307)
Authors: Rosa Jaime , Chinthrajah Rebecca , Dominguez Tina , Brock Emily , Nadeau Kari 
Published: January 2023

Abstract
Introduction: Food allergies are a potentially fatal disorder. However, reliable prognostic markers for predicting the severity of allergic reactions during exposures have not yet been established. Recent studies have found that basophil activation tests (BATs) may be a useful adjunct for the diagnoses of peanut and other food allergies. We hypothesize that BAT can also be a useful biomarker for predicting reaction severity during food challenges. Methods: 19 (5-25 years old) peanut-allergic participants (PA) and 5 age-matched healthy controls (AC) who enrolled in our Stanford IRB and FDA approved double blind placebo controlled food challenge (DBPCFC) protocol were given escalating oral doses (5, 20, 50, 100, 100, 100, 125 mg) of peanut and placebo (oat) on separate occasions. The DBPCFC was stopped at the first sign of an objective symptom, which was graded on the modified Bock’s scale. Prior to the DBPCFC, basophils gated via flow cytometry from 200 μl of whole blood obtained from all subjects were stimulated with peanut ex vivo for 20 minutes. Results: For PAs, 8 had grade 1, 7 had grade 2, and 4 had grade 3 reactions to peanut; the ACs had no symptom. Using linear regression, high correlation coefficients were found between Bock’s reaction grades and basophil surface expressions of CD203c (r2 =0.89) and CD63 (r2 =0.85), both p&lt;0.001. Conclusions: Our findings suggest that suitable BATs can be a useful biomarker for predicting clinical reactivity during peanut challenges.
Peanut allergy: a mouse model comparison (HYP4P.309)
Authors: Jhaveri Devi , Bonfield Tracey 
Published: January 2023

Abstract
Peanut induced anaphylaxis often results in a bi-phasic response, associated with a second anaphylaxis once current treatment interventions have been removed. Previous reports have suggested that this response is associated with elevated levels of platelet activating factor. Our studies have focused on refining the murine model of peanut anaphylaxis with the hypothesis that we can generate a model that recapitulates the anaphylactic response to peanuts. We used BALB/c and C3H/HeJ mice (N=20 for each study) mice were sensitized weekly for 6 weeks with crude peanut extract in cholera toxin. Mice were then grouped for crude peanut extract or saline challenge by intra-peritoneal injection. In all mice, temperatures and clinical scores were accessed both early post injection as well as 2 hours later. Serum, lungs, spleen, and liver were stored from all animals for future studies. The BALB/c strain had a mean temperature change from baseline of 0.2 C⁰ for the controls initial post IP versus the 1.89 C⁰ for the challenged mice. The C3H/HeJ strain had a mean temperature change from baseline of 0.1 C⁰ for the controls at the initial post IP versus the 4.2 C⁰ change for the challenged mice. Similar results were noted at the 2 hour mark with the C3H/HeJ strain with the greater temperature change. Both strains had notable changes in clinical scores between the control and challenge groups. This was more pronounced in the C3H/HeJ mice both clinically and by temperature measurement.
Existing allergic inflammation alleviates influenza infection (HYP5P.319)
Authors: Lee Debbie , Prabhu Nayana , Kemeny David 
Published: January 2023

Abstract
Asthma is a disease suffered by millions throughout the world and its prevalence continues to increase. During the 2009 influenza pandemic asthma was found to be the commonest comorbid illness of patients admitted to hospital. It was also observed that asthmatics admitted to hospital with influenza infection were less likely to die or require admission to intensive care compared with non-asthmatics. Using an in vivo model of asthma and influenza infection we demonstrate that prior exposure to Blomia tropicalis extract (BtE) led to a reduction in weight loss and faster recovery following influenza infection compared to PBS/influenza infected mice. This was accompanied by an earlier increase in both CD4 and CD8 T cells in the lungs, which produced more INF-γ, TNF-α and expressed higher levels of CD107a at day 8 post infection. At day 14 post infection levels of T cells in the lungs of the BtE/influenza group were significantly reduced, which parallels the faster recovery from weight loss seen in this group. In summary, we have shown that prior exposure to BtE leads to an altered immune response to influenza infection, comprised of less severe weight loss and the faster recruitment of more highly activated T cells in to the lungs. The earlier recruitment of T cells led to faster resolution of disease and a reduction in T cell numbers. We believe this data provides a clue as to why asthmatic patients present with less severe disease when infected with influenza.