bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Young Scientist Program: successful use of a volunteer based outreach program created by graduate and medical students to improve science education in the St Louis Public School system (51.10)
Authors: Lynch Jennifer , Moss Britney , Chiappinelli Katherine , Mosher Jennifer , Woolsey Thomas 
Published: January 2023

Abstract
YSP aims to supplement science curricula and attract young students to scientific careers through a series of programs designed for underserved middle and high school students and their teachers. Teaching Teams develop and lead inquiry-based science modules in classrooms, after school programs, and to community organizations throughout the year. In the summer, high school students and teachers perform independent research at Washington University under the guidance of graduate student or post doctoral mentors through our Summer Focus (SF) and Teacher Researcher Partnership (TRP) programs. SF culminates in written and oral presentations of laboratory findings. TRP participants develop tools to bring bench work into their classrooms. Analysis of the past five years elucidates the immense success YSP has had within the St Louis community. Students report more interest and aptitude in science due to YSP. Applications for SF and TRP have reached another record high and come from an expanding demographic. Participants of both programs often pursue advanced degrees in science or education. Analyses of SF writing samples show remarkable improvement in YSP’s ability to teach scientific writing and critical thinking skills. YSP continues to increase the number of volunteer hours and reach new community venues. We present YSP as a model for the successful use of a volunteer based approach to improve science education and inspire future generations of science professionals.
Integrating immunology into a case based medical curriculum (51.3)
Authors: Moticka Edward 
Published: January 2023

Abstract
Several innovations in the delivery of the pre-clinical medical curriculum have focused on non-discipline approaches. These include Problem Based Learning, Team Based Learning and the Clinical Presentation (CP) Curriculum. All these approaches require the integration of various science disciplines into clinical scenarios organized by organ systems. The CP Curriculum involves integrating didactic material in the context of approximately 125 clinical presentations (i.e. chest pain, hematuria, shortness of breath). During the pre-clinical years, a clinician provides an overview of the clinical problems which might be seen followed by a series of basic science presentations relevant to the clinical presentation. Incorporation of immunology into a CP curriculum requires coverage of both basic concepts and pathophysiology of immunologic diseases in each organ system. Our approach is to use a small number of clinical presentations (sore throat, recurrent infections) to cover basic concepts. This is followed by presentations in each organ system during which disease mechanisms and additional immunological concepts are introduced. This presentation will review the ability to include topics identified by USMLE as well as those identified by ASM in such a curriculum along with data on the performance of students enrolled in a medical school using this approach.
Using differentiated instruction to teach immunological concepts to a diverse group of learners. (51.6)
Authors: Rodriguez Victor , Havran Wendy 
Published: January 2023

Abstract
Last summer, I was a teacher intern in the Havran laboratory at The Scripps Research Institute where the research focused on how to define the mechanisms by which γδ T cells interact with neighboring epithelial cells, how they recognize and respond to self produced antigen, and ultimately how they accelerate the repair of damaged tissue. Upon completion of my internship, I developed a curriculum unit with a Biology Action Model (BAM) and a double diffusion test that demonstrates the interaction between antibodies and antigen. Students will be introduced to the role of γδ T cells by interpreting pictures of wild type and γδ knock out epidermal mouse tissue that have received identical wounds. Students will observe delayed wound closure and healing in the tissue that comes from the γδ knock out mouse. In addition to the content, students will apply laboratory skills such as microscopy and tissue staining. This unit is aligned with the California Content Standards for the Immune System and is designed to be implemented in a college prep biology class. This unit will incorporate a variety of teaching strategies that engage students in active learning and will foster a better understanding of immunology.
Real-time tracking ATAK cells in vivo (52.1)
Authors: Lin Lin , Ibrahim Ashraf S. , Baquir Beverlie , Palosaari Andrew , Spellberg Brad 
Published: January 2023

Abstract
Background: Activated Targeted Killer (ATAK) cells are a cellular immunotherapy which protects neutropenic mice from invasive candidiasis and aspergillosis. However,the lifespan and distribution of ATAK cells into organs has not been determined in vivo. Methods: ATAK cells were stably transfected with a luminescence reporter system. Neutropenic mice were imaged by IVIS after administration of coelentrazine substrate. RT-PCR was used to confirm the luminescence results. Results: On day 1 after ATAK cell administration, livers, kidneys, and spleens were infiltrated by the ATAK cells. By day 3, cells had infiltrated more significantly into the spleen, and also spread into the lungs. By day 5, cell signal began to wane. By day 8, the day after neutrophil recovery, there was no detectable signal. By RT-PCR, viable cells were detected at 6 hours and 3 days after treatment. By day 5, no mRNA was detected. Mouse immunity to ATAK cells was detectable by ELISA and flow cytometry post-neutrophil recovery. Discussion: ATAK cells are viable and circulate widely into visceral organs in neutropenic mice, commensurate with their ability to protect mice from tissue invasive fungal infections. Hence the cells can provide protection during the critical neutropenic period yet the risk of engraftment is minimized by host-versus-graft affect after marrow recovery.
Memory CD8+ T cell proliferation after anti-CD40/IL-2 treatment is independent of CD4+ T cells (52.17)
Authors: Tietze Julia , Hsiao Hui-Hua , Koehn Brent , Wilkins Danice , Sckisel Gail , Blazar Bruce , Murphy William 
Published: January 2023

Abstract
Systemic cancer immunotherapy (IT) with anti-CD40/IL-2 in mice leads to marked antigen-independent proliferation of CD44high CD8+ cells, which are NKG2D+/CD25- and do not upregulate PD1 significantly. Here we investigated the role of CD4+ T cells on the memory CD8+ expansion in vivo using CD4 knockout mice and in mice whose CD4+ T cells were depleted with antibody. We observed that CD4 depletion significantly increased total numbers of CD44highCD8+ cells in untreated mice due to homeostatic proliferation. After IT there was no difference in the number of CD44highCD8+ cells or in the expression of NKG2D compared to IT treated control mice. However, we observed upregulation of PD1 on the CD8+ T cells in CD4-depleted mice after IT compared to control mice. Treatment of IL21 RKO mice with IT also did not affect the expansion of the CD44highCD8+ T cells compared to WT mice. We conclude that the effect of anti-CD40/IL-2 treatment on CD44highCD8+ T cell proliferation is independent of CD4 help or IL21. However, the absence of CD4+ T cells appears to affect the phenotype of the expanded CD8+ T cell population resulting in the upregulation of PD-1.
Targeting and manipulating mIgD-expressing B cells by anti-migis-δ monoclonal antibodies (52.18)
Authors: Chen Nien-Yi , Hung Alfur , Yu Hui-Ming , Chang Hwan-You , Chang Tse-Wen 
Published: January 2023

Abstract
Membrane IgD and IgM double positive cells are the major population of B cells in the peripheral circulation. The δ chain of mIgD (mδ) differs from the δ chain of IgD in that mδ contains an extra C-terminal transmembrane-anchor peptide, of which the N-terminal extracellular segment of 26 a. a. residues, also referred as migis-δ segment, potentially provides as a unique antigenic site for isotype-specific targeting mIgD+ B cells. Here we report the preparation of mouse monoclonal antibodies (mAbs) specific for migis-δ using hybridoma methodology with ovalbumin conjugated with synthetic migis-δ peptide as the immunogen. The mAbs bound to recombinant IgD.Fc-migis-δ fusion protein in ELISA and to mIgD-expressing transfectants of a CHO cell line in fluorescence flow cytometric analysis. Only mAb 20E6, which binds to an epitope toward the N-terminal of migis-δ, was found to stain human B cell line MC116, which expressed surface IgD and IgM. MC116 cells could be induced to undergo apoptosis by the treatment of 20E6, in the absence or presence of crosslinking second antibody. These results encourage further investigation on the potential of anti-migis-δ mAbs to target or control mIgD+ B cells.
Evaluation of the human CD8 T cell response to exogenous antigen using MIMIC® technology (52.5)
Authors: Byers Anthony , Haupfear Kelly , Wittman Vaughan 
Published: January 2023

Abstract
Vaccines typically achieve immune protection by induction of humoral immunity. Recently, interest has increased in vaccines capable of eliciting both humoral and cellular immunity. Since the majority of vaccines are subunit or protein-based and CD8 T cells are typically primed by endogenous proteins expressed within the target cell, effective induction of CD8 T cell (CTL) responses by vaccines has been erratic. Exogenous antigen can be presented on MHC class I to CD8 T cells via an inefficient and poorly defined process termed cross-priming. We have used our in vitro MIMIC® technology to optimize CTL cross-priming immune responses to subunit vaccines. We have shown in vitro CTL responses to soluble influenza nucleoprotein (NP) by activating, or licensing, of dendritic cells (DC) by CD4 T cells. Further optimization has involved modification of various assay parameters: cell ratios/kinetics, antigen load, DC types used, addition of DC maturation/activation factors. Generation of the cross-priming response has been evaluated by the ability of CTL to make combinations of multiple cell factors (CD107a, IFN-γ, IL-2 and TNF-α) via intracellular cytokine staining and the ability of the CTL to eliminate targets in a fluorescent-based cytotoxic assay. The MIMIC® system can be used to evaluate CTL response to various protein/adjuvant combinations and identify corresponding vaccine candidates capable of efficiently inducing cellular immunity.
Promotion of the neutralization-competent structure of the HIV-1 gp41 membrane proximal external region when tethered to its native transmembrane domain, and expressed in the context of the plasma membrane: implications for vaccine design (53.24)
Authors: Gulzar Naveed , Montero Marinieve , Klaric Kristina-Ana , Donald Jason , Wang Shixia , Lepik Christa , Tsai Sue , Wu Sampson , Julien Jean-Philippe , Hessell Ann , Lu Shan , Burton Dennis , Pai Emil , DeGrado William , Scott Jamie 
Published: January 2023

Abstract
The limited success of vaccines targeting the MPER, a target of three broadly neutralizing (bNt) monoclonal antibodies (MAbs), we hypothesize, reflects the difficulty of mimicking the neutralization-competent structure (NCS) of the MPER. We determined the contribution of the amino-acid sequence and the transmembrane domain (TM), to the antigenicity of the MPER in the context of the plasma membrane. DNA constructs encoding various gp41 ectodomain fragments, and the TM of either the platelet-derived growth factor receptor (PGDFR), or that of gp41, were produced and transiently expressed in COS-7 cells. Constructs expressing the MPER tethered to the gp41 TM followed by a 27-residue cytoplasmic tail fragment, MPER-TM1, produced optimal binding of MAbs 2F5, 4E10 and Z13e1. A series of 24 single amino-acid substitutions in the MPER-TM1 revealed critical binding residues for the three MAbs; similar substitutions were previously shown to ablate Ab-mediated viral neutralization. Neutralization-incompetent 2F5 Fab and 4E10 IgG mutant Abs failed to bind MPER-TM1, yet retained the ability to bind to peptide epitopes, indicating the plasma-membrane expressed MPER-TM1 closely approaches the NCS of the MPER. Substitution of the TM of gp41 with that from the PGDFR reduced binding by MAb 4E10, but not MAbs 2F5 or Z13e1. Our studies reveal that the gp41 TM appears to play a pivotal role both in orienting the 4E10 epitope, and affecting exposure of the MPER epitopes for all three bNtMAbs.
Induction of memory responses following transcutaneous immunization requires presence of immunoadjuvantative cholera holotoxin (53.4)
Authors: Kalsy Anuj , Bhuiyan Md , Charles Richelle , Arifuzzaman Mohammad , Clemens John D. , Larocque Regina C. , Harris Jason B. , Calderwood Stephen B. , Czerkinsky Cecil , Qadri Firdausi , Ryan Edward 
Published: January 2023

Abstract
Vaccination induces both mucosal & systemic immune response. Polysaccharide antigen (PAg) which is T cell independent antigen does not stimulate production of memory responses. Cholera holotoxin (CT) is shown to be a potent immunoadjuvant. Transcutaneous immunization (TCI) is recently shown as an important alternative route of immunization to induce both systemic & mucosal immune responses. Induction of memory responses upon TCI with PAg is unkown. We evaluated if TCI with Vi & Vi-conjugate vaccine (Vi-DT)could induce anti-Vi specific memory responses in mice. Parenteral & TCI of Vi can not produce memory response. Splenic memory B cell IgG responses were induced by parenteral immunization with Vi-DT but not Vi, but were induced following TCI with Vi-DT/CT and Vi/CT. TC & parenteral immunization with Vi & Vi-DT induced serum anti-Vi IgG & IgA responses but more prominently along with CT & induced T cell independent PAg specific memory response in mice. Stool & lamina proprial lymphocyte (LPL) anti-Vi IgA responses were highest following TCI. Splenic antibody-secreting-cell (ASC) anti-Vi IgG responses were induced by parenteral immunization with Vi or Vi-DT, and by TCI with Vi/CT or Vi-DT/CT. Our results show that TCI induces both systemic & mucosal immune responses with PAg & use of CT as adjuvant induces this response more prominently. Memory responses only found when CT was used TC. This observation could assist in developing of vaccines providing long-term protection.
Virus induced airway hyperreactivity in allergen sensitized animals is mediated by immunological memory and is ablated by dexamethasone (55.14)
Authors: Skappak Christopher , Illarraza Ramses , Wu Ying-qi , Adamko Darryl 
Published: January 2023

Abstract
Common cold viruses are the leading cause of asthma exacerbations. Why people with asthma develop such severe reactions is unclear. We hypothesize that it is not the severity of virus infection but the activation of memory T-cells and eosinophils, which cause the airway hyperreactivity (AHR). We have a model with virus memory T cells in allergen-sensitized guinea pigs (GP). GP are sensitized (i.p.) to ovalbumin (OVA) or not. All receive an inhaled OVA challenge. Some of each group are exposed once to parainfluenza virus (PIV) then, 6 weeks later, re-exposed to either live virus or UV-inactivated virus. Measurements of AHR (histamine i.v.), inflammation (BAL/histology), and T cell memory (BrdU incorporation ex vivo) are measured. Additionally some animals from each group were pre-treated with dexamethasone (DEX) (i.p.) before re-exposure to PIV. Both non-sensitized and OVA-sensitized GPs develop T cell memory to virus based on BrdU incorporation data from lymph node T cells. Unlike non-sensitized GP, OVA-sensitized GP show elevated numbers of eosinophils in the airways, except those pre-treated with DEX. Unlike non-sensitized GP, OVA-sensitized GP develop AHR upon re-exposure to UV-inactivated virus, which is prevented by DEX. GP can develop T cell memory to PIV. Upon re-exposure to PIV antigens, this T cell memory can cause AHR but only sensitized animals, which we believe is mediated by eosinophils. DEX prevents AHR in sensitized animals re-exposed to viral antigens.
A novel composition from herbal extract (SCAP) alleviates skin severity and serum IgE level in NA/Nga mice, an atopic dermatitis model (55.17)
Authors: Lee Geum Seon , Oh Se Koon , Cheong Jae Hoon , Kim Kyungjae , Kang Tae Jin 
Published: January 2023

Abstract
Pimecrolimus (Elidel cream®) is an immunomodulating agent and is used for the treatment of atopic dermatitis (AD). However, it is reported that this treatment has side effects regarding the potential increased risk of lymph node or skin malignancy and is not sufficiently effective. SCAP is a composition of herb extracts from Sonchus oleraceus, Chelidonium majus var. asiaticum, Arctii Fructus, Portulacae Herba. which were traditionally used in oriental medicine as an analgesic, diuretic, and anti-inflammatory agents. The anti-atopic effect of SCAP was investigated in an atopic dermatitis animal model, NC/Nga mice. AD was induced by repeated application of 1-chloro 2, 4-dinitrobenzene (DNCB) in NC/Nga mice. And then mice were treated with SCAP or Pimecrolimus cream by dermal application. Scratching behavior, clinical skin severity, and the levels of total serum IgE were measured. After AD induction, scores of scratching behavior and clinical skin severity, and the levels of total serum IgE were increased. Treatment with SCAP significantly attenuated the scratching behavior and improved skin severity in a dose dependent manner in NC/Nga mice. Especially, treatment of SCAP 0.5 % reduced more rapidly scratching behavior, clinical skin severity, and the level of total serum IgE, compared to that of pimecrolimus cream. These results suggest that the SCAP may be an alternative substance for the treatment of AD.
Non-apoptotic Fas (CD95) signaling on T cells regulate the resolution of airway inflammation in a murine model of asthma (55.9)
Authors: Williams Jesse , Ferreira Caroline , Blaine Kelly , Rayon Crystal , Velazquez Francisco , Peter Marcus , Sperling Anne 
Published: January 2023

Abstract
Asthma is a Th2-mediated airway disease associated with chronic lung inflammation and mucus production. The mechanism by which inflammation resolves in the lung remains unidentified. We have previously shown that Fas deficiency on T cells in an adoptive transfer model of experimental asthma, results in a delay in resolution of inflammation compared with control. Surprisingly, using T cells with survival advantages independent of Fas on T cells (Bim-/- and BcLxL-tg), we found that inflammation resolution was not delayed, thus suggesting a dependency of T cells on a Fas-specific mechanism for normal resolution of inflammation. Fas has long been known to function as a death initiator by activating caspase-mediated apoptosis. However, recently it has also become clear that Fas has non-apoptotic functions including activation of NF-κB via MAPK pathways. Using an adoptive transfer model, Fas mutant T cells capable of non-apoptotic Fas signaling, but not apoptotic Fas signaling, were sufficient to produce normal resolution. Further, we have found that in vitro skewed Th2 cells that are resistant to Fas-mediated apoptosis, still signal through the non-apoptotic Fas pathway. Together these findings demonstrate that non-apoptotic Fas signaling, rather than apoptotic signaling, in T cells is important for normal resolution of airway inflammation in a murine model of experimental asthma.
A Toll/IL-1R/resistance domain-containing thioredoxin regulates phagocytosis in Entamoeba histolytica (56.23)
Authors: Mancilla Ismael , Mendez Alfonso , Wong Isabel , Alcántara Marcela , Arriaga Lourdes , López Constantino , González Jorge , Isibasi Armando 
Published: January 2023

Abstract
Entamoeba histolytica is a parasite protozoan that infects humans and causes amebiasis in developing countries. Phagocytosis of epithelial cells, erythrocytes, leucocytes, and commensal microbiota bacteria is a major pathogenic mechanism used by this parasite. A Toll/IL-1R/Resistance (TIR) domain-containing protein is required for bacterial phagocytosis in the social ameba Dyctiostelium discoideum, both for nutrition and for protection against infection. In insects and vertebrates, TIR domain-containing proteins regulate phagocytic cell activation. Therefore, we investigated whether E. hystolytica expresses TIR domain-containing molecules that may be involved in the phagocytosis of erythrocytes and bacteria. Using Hidden Markov models (HMM), we identified an E. histolytica thioredoxin containing a TIR-like domain. The primary structure of this putative TIR domain exhibited low similitude with the TIR domain of human Toll-like Receptor (TLR)-2; however, the predicted tertiary structures of these two TIR domains were remarkably similar. Interestingly, the phagocytosis of erythrocytes and E. coli by E. histolytica trophozoites increased after the inhibition of the expression of thioredoxin using a specific siRNA. This suggests that TIR domain-containing thioredoxin is a negative regulator of phagocytosis in E. histolytica.
Toxoplasma gondii infection of human plasmacytoid dendritic cells inactivates TLR7 and -9 mediated induction of IFN-α and TNF-α in response to HIV and HSV (56.26)
Authors: Pierog Piotr , Zhao Yanlin , Singh Sukhwinder , Yap George , Fitzgerald-Bocarsly Patricia 
Published: January 2023

Abstract
pDC are the major producers of IFN-α, an antiviral cytokine involved in control of HIV replication, while T.gondii is a life-threatening opportunistic infection in AIDS patients. In this study, we investigated pDC responses in the context of virus and T.gondii co-infection. T.gondii invaded but did not induce IFN-α or TNF-α in human pDC. However, Class I, II and III strains of T.gondii inhibited IFN-α and TNF-α in response to HSV and HIV. Within the T. gondii exposed pDC population, IFN-α production was inhibited only in cells infected by T. gondii, whereas TNF-α was inhibited in both the infected and to a lesser extent, uninfected pDC. T. gondii inhibited neither uptake of GFP-HSV nor recruitment of TLR9 to EEA1+ endosomes. Using imaging flow cytometry we found that virus-induced nuclear translocation of IRF-7 was abolished by the parasite. Taken together, these data indicate that the block of the intracellular signaling cascade by T.gondii occurs downstream of TLR9 recruitment but upstream of IRF-7 translocation. Unexpectedly, T.gondii did not inhibit HSV-induced nuclear translocation of NFκB. We identified that Rop16, a parasite-derived kinase, is partially responsible for IFN-α and TNF-α inhibition. Additionally, T.gondii altered the virus-induced maturation profile of pDC. These findings suggest a novel mechanism of inhibition of TLR signaling by T.gondii and suggest potential negative consequences of HIV/T. gondii co-infection.