bluuebunny/crossref_metadata_2025_split huggingface dataset:
Implementing participatory immunology education in a diverse Latin American student population
Authors: Ferreira Leonardo , Carosso Giovanni A. , Duran Natalia Montellano , Bohorquez-Massud Soad , Vaca-Diez Gustavo , Rivera-Betancourt Laura Ines , Rodriguez Yara , Ordonez Dalila G. , Alatriste-Gonzalez Diana K. , Vacaflores Aldo Fabian , Auza Lilian Gonzalez , Schuetz Christian , Alvarado-Arnez Lucia Elena , Alexander-Savino Carolina , Gandarilla Omar , Mostajo-Radji Mohammed A. 
Published: January 2023

Abstract
Immunology is typically first encountered by students in advanced college or graduate courses. Additional challenges to teaching immunology exist in the developing world, where limited resources require curricula balancing state-of-the-art knowledge with region-specific practical considerations. Project-based teaching is especially difficult to execute due to lack of infrastructure and teacher training. Here, we report the results of implementing short hands-on low-cost microbiology and immunology courses for high school and college students in Bolivia. This initiative, “Clubes de Ciencia Bolivia”, brings graduate students and postdoctoral fellows from research institutions in developed nations to teach weeklong intensive courses in Bolivia. These courses (“clubes”) feature project-based and active learning components and are designed and taught with local Bolivian science instructors. Our approach was effective at transmitting concepts in microbiology and immunology, as measured by subject-specific test scores before and after course completion. Importantly, student learning was not significantly affected by their background, education level, socioeconomic status, or initial interest in the specific course taken. Moreover, participants reported a heightened interest in scientific careers after course completion. Strikingly, participants scored higher in subject-specific tests than Bolivian college students who had taken related semester-long college courses. Altogether, these data indicate that participatory learning for immunology in a developing nation can be used to effectively transmit knowledge. Similar approaches are likely to drive scientific engagement in other developing economies.
In for the long haul: teaching immunology in a longitudinal medical curriculum
Authors: Bottaro Andrea 
Published: January 2023

Abstract
In recent years, undergraduate medical education has undergone significant transformations, including stronger integration of clinical and basic science content in both early and late phases of the curriculum; frequent replacement of traditional topic-specific courses with system-specific blocks in which all basic sciences can be weaved in longitudinally; and an increasing utilization of active learning approaches to replace traditional lectures. At the same time, immunology has greatly expanded its relevance to clinical care, impacting an increasing number of medical subspecialties. As a consequence, medical graduates now require a higher degree of immunology literacy than ever before.
Here, I will discuss the experience of building (and sustaining) a longitudinally integrated immunology curriculum at a new (now 7-year-old) medical school. I will first highlight a number of key challenges, including the ballooning amount of information that medical students are required to learn in increasingly time-constrained curricula; the balancing of the competing needs of updating content for scientific advancements vs maximizing student success in the slower-moving but ever-important medical board exams; collaborating with faculty colleagues whose training in immunology often happened decades in the past; and dealing with content gaps and redundancies in shifting course curricula. Lastly, I will provide some examples of how we have addressed some of these challenges locally, while others may require more concerted, global solutions.
Open Discovery in an Undergraduate Immunology Course Lab: Testing Anti-Inflammatory Drugs
Authors: Brennan Catherine , Roshandell Mobina 
Published: January 2023

Abstract
I will present the design and outcomes from a course laboratory module in which undergraduates apply techniques learned earlier in the semester (cell culture, spectrophotometry, standard curves, ELISA) towards the testing of an anti-inflammatory drug of their choice. Students rationalize their dose and assay selections, calculate all dilutions, perform the assays, and analyse and interpret all results. Student-directed course research experiences have been shown to solidify student understanding of advanced scientific concepts. Additionally, we observed that this exercise, in which students analyse the function of medications used by themselves or loved ones, leads to enthusiastic engagement with and mastery of complex concepts.
Role of GILT and MHC class II in melanoma cells on regulating the anti-tumor immune response
Authors: Macy Anne , Adams Anngela , Hastings Karen Taraszka 
Published: January 2023

Abstract
The MHC class I antigen presentation pathway in melanoma cells has a well-established role in immune-mediated destruction of melanoma. However, the role of the MHC class II pathway in melanoma cells is not fully understood. Gamma-interferon-inducible lysosomal thiol reductase (GILT) is critical for MHC class II-restricted presentation of multiple melanoma antigens by antigen presenting cells. While GILT and MHC class II expression is typically limited to antigen presenting cells, GILT and MHC class II can be expressed constitutively or induced by IFN-γ in melanoma cells. In human melanoma specimens, high GILT expression and an active and intact MHC class II pathway are associated with improved survival. The goal of this project is to investigate a causal role for GILT and MHC class II in melanoma cells, using Yale University Mouse Melanoma (YUMM) lines YUMM2.1, YUMMER1.7, and YUMMERG. These cell lines constitutively expressed GILT, and MHC class II expression was IFN-γ-inducible in a subset of cells under serum-free conditions. We have genetically engineered GILT−/− YUMM2.1 cells and MHC class II (H2Ab1)−/− YUMM2.1 cells via CRISPR/Cas9 and verified the deletion. We have generated lentiviral constructs expressing GILT alone, CIITA alone, and dual expression of H2Ab1 and CIITA, to allow for the generation of melanoma cell lines with and without GILT and MHC class II. Transduction of GILT−/− YUMM2.1 cells with GILT resulted in high transduction efficiency and overexpression. Thus, we have developed models to determine the effect of GILT and MHC class II expression in melanoma cells on regulating in vivo tumor growth and the anti-tumor immune response.
This work was supported by a Basic Science Award from the Harry J. Lloyd Charitable Trust and a Merit Review Award I01-BX005336 from the US Department of Veterans Affairs (VA), Biomedical Laboratory Research and Development Service (K.T.H.). The contents do not represent the views of the VA or the US Government.
Distinct myeloid antigen presenting cells dictate differential fates of tumor-specific CD8 T cells in pancreatic cancer
Authors: Burrack Adam L , Schmiechen Zoe , Patterson Michael , Miller Ebony , Spartz Ellen J , Rollins Meagan , Raynor Jackson , Mitchell Jason , Kaisho Tsuneyasu , Fife Brian T , Stromnes Ingunn 
Published: January 2023

Abstract
Pancreatic ductal adenocarcinoma (PDA) contains numerous protumor myeloid cells at the expense of antitumor dendritic cells (cDC1s). We identify that tumor antigenicity is a major determinant of myeloid composition and functionality. Neoantigen-tumors contained pro-tumor macrophages and a paucity of cDC1s whereas neoantigen+ tumors accumulate cDC1s via Xcr1 signaling and macrophages exhibit a diminished role. Effective immunotherapies increased splenic cDC1s, which were required for endogenous T cell expansion following α-PD-L1 and transferred effector and memory T cells. Batf3−/− mice, which lack cDC1s, failed to spontaneously generate tumor-specific CD8 T cells and were resistant to T cell therapy and α-PD-L1. In contrast, agonistic α-CD40 exhibited partial benefit in Batf3−/− mice and expanded atypical tumor-specific CD8 T cells. Monocyte depletion abrogated atypical tumor-specific CD8 T cell priming yet enhanced α-CD40-mediated antitumor activity in Batf3−/− mice. In contrast, α-Gr1 abrogated the therapeutic benefit of CD40 agonist in Batf3−/− mice. In sum, our study supports that CD40 agonist promotes a cDC1-dependent antitumor immunity and a monocyte-dependent protumor arm of immune system. These results further underscore an essential antigen presenting role for cDC1s in the expansion and differentiation of naïve, effector, and memory T cells into Klrg1+ potent cytotoxic effector T cells capable of targeting pancreatic cancer.
M.R. is supported by National Institutes of Health (NIH) T32 AI 007313 E.J.S. is supported by NIH T35 AI118620 I.M.S. is supported by NIH R01 CA249393, R01 CA255039, Department of Defense #PA200286, an American Association for Cancer Research (AACR) Pancreatic Cancer Action Network Career Development Award (17-20-25-STRO), an AACR Pancreatic Cancer Action Network Catalyst Award (19-35-STRO), American Cancer Society Institutional Research Grant (124166-IRG-58-001-55-IRG65).
The impact of GSK3β on the metabolism of IFNγ activated macrophages
Authors: Ankley Laurisa M , Olive Andrew J 
Published: January 2023

Abstract
The activation of macrophages by IFNγ induces transcriptional and metabolic changes that enable the direct control of pathogens and improves interactions with T cells. In particular, IFNγ induces the expression of antigen presentation machinery like MHCII. Despite its central role in host immunity, the regulation of IFNγ-induced MHCII is poorly understood. Using CRISPR Cas9 screens we examined regulators of distinct IFNγ-mediated mechanisms in macrophages including MHCII. One key regulator found was the multifunctional kinase, GSK3β. Not only was GSK3β required to induce transcriptional activation of MHCII, but RNAseq analysis suggests broad changes to the IFNγ response in the absence of GSK3β. Recent work in our group suggests that changes in metabolism are essential for macrophages to respond to IFNγ. While GSK3β is known to directly modulate metabolism, its role in controlling metabolic changes following IFNγ activation remains unknown. Given that GSK3β interfaces with metabolic pathways like mTORC1 and glycolysis we hypothesize that the loss of GSK3β inhibits important metabolic shifts required for macrophages to effectively respond to IFNγ. Using a Seahorse Analyzer we are now determining how genetic or chemical inhibition of GSK3β changes the rate of glycolysis or oxidative phosphorylation following IFNγ stimulation. We will then directly target metabolic networks in the presence and absence of GSK3β to determine how these impact IFNγ responses including the expression of MHCII. Given the broad impact of GSK3β on several disease states, such as inflammatory disease and tumor progression, understanding the regulation of GSK3β has the potential to contribute to several aspects of human health.
Dissecting the role of H-2Db class I molecule in the development of brain atrophy during Theiler’s murine encephalomyelitis virus (TMEV) infection
Authors: Wininger Katheryn M , Goddery Emma , Khadka Roman , Tritz Zachariah P , Fain Cori , Ayasoufi Katayoun , Jin Fang , Hansen Michael , Johnson Aaron J 
Published: January 2023

Abstract
Brain atrophy is a common feature of many neurological diseases as diverse as Alzheimer’s disease, cerebral palsy, Huntington’s disease, Krabbe disease, multiple sclerosis, Pick’s disease, epilepsy, encephalitis, neurosyphilis, neuroAIDS, and Covid-19 infection. We have developed a murine model of brain atrophy using the Theiler’s murine encephalomyelitis virus (TMEV) infection mouse model of multiple sclerosis. In this study, we investigated the contribution of the MHC class I molecule, H-2Db, in generating an immune response associated with the onset of brain atrophy. To define the roll of the MHC class I molecule in the cellular and molecular mechanisms of brain atrophy, we developed a novel bi-transgenic mouse lines with tamoxifen induced conditional ablation of the H-2Db class I molecule in Cx3CR1+ brain resident myeloid cells on the C57BL/6 background. Interestingly, both CX3CR1cre/Db LoxP mice and their cre- littermate controls experienced short-term memory loss using novel object recognition tests. However, Cx3CR1cre+/Db LoxP mice presented with significantly less brain atrophy as assessed by analysis of lateral ventricular volume at 45 d.p.i. using T2-weighted MRI, compared to cre- littermates. This change in ventricular volume was sustained at 6 months post infection. These results strongly imply a requirement for antigen presentation by H-2Db on myeloid cells for the development of brain atrophy and provided insight into the molecular mechanism of this poorly understood neuropathology.
Supported by NIH (RFI NS122174)
Mechanism of Peptide Loading as Revealed by Structure of tapasin/MHC-I Complex
Authors: Jiang Jiansheng , Taylor Daniel K , Kim Ellen , Boyd Lisa F , Cresswell Peter , Mage Michael G , Margulies David H , Natarajan Kannan 
Published: January 2023

Abstract
MHC-I molecules loaded with peptide for subsequent cell surface expression are critical for adaptive immunity. Tapasin, a component in the Peptide Loading Complex (PLC), serves as an MHC-I chaperone, and it facilitates peptide loading onto MHC-I in the endoplasmic reticulum (ER). Although the structure of TAPBPR (a homolog of tapasin) complexed with MHC-I was reported several years ago, the detailed mechanism of peptide loading by tapasin remains unclear. Here, we report the structure of a complex of tapasin with the MHC-I molecule, HLA-B* 44:05. The model of tapasin/HLA-B*44:05 compared with that of unliganded HLA-B*44:05 and to the structure of tapasin bound to ERp57 reveals dramatic changes that refect both chaperone and catalytic activities. Tapasin cradles the MHC-I molecule through contacts with the N-terminal alpha1 and alpha2 domains, as well as with alpha3 and beta-2 microglobulin interaction via its C-terminal IgC domain. Thus the MHC-I molecule is stabilized in a peptide accessible form. We compare this tapasin/HLA-B*44:05 structure with our previously determined structure of TAPBPR/H2-D(d). Although the general dispositions of tapasin and TAPBPR are the same in their complexes with MHC-I, the structural details of their interactions differ. Analyses of tapasin mutants that disrupt the interaction with MHC-I are consistent with the X-ray crystal structure. Thus, the two MHC-I dedicated chaperones, tapasin and TAPBPR, although acting at distinct stages of the MHC-I loading pathway, show broadly conserved binding modes and similar mechanisms of peptide loading in antigen presentation.
Supported by the intramural research program of the NIAID, NIH
Investigating the potential binding partners of H2-O to further elucidate its role in MHCII antigen presentation
Authors: Dobkin Julie , Golovkina Tatyana , Denzin Lisa K 
Published: January 2023

Abstract
The production of neutralizing antibodies in response to viral pathogens requires CD4+ T cell help. For this to occur, pathogen-derived peptides must be loaded onto class II major histocompatibility complex (MHC II) molecules in endosomal/lysosomal compartments and trafficked to the surface of antigen presenting cells. The αβ heterodimeric nonclassical MHCII molecules, H2-M and H2-O, modulate peptide loading. While H2-M directly catalyzes MHCII peptide loading, H2-O is a negative regulator of MHCII antigen presentation. Current dogma supports that H2-O is an MHCII structural mimic that exerts control over peptide loading via direct competition with MHCII for H2-M-binding. Work done in our labs, however, suggests this current model is incomplete and suggests the involvement of other, yet undefined, factors. I/LnJ mice, which are resistant to retroviruses through the production of potent neutralizing antibodies, inherit an H2-Ob allele that encodes nonfunctional H2-O. I/LnJ H2-O binds to but does not inhibit H2-M, directly challenging current dogma. Compared with H2-Oβ from retrovirus-susceptible mice, I/LnJ H2-Oβ contains three unique amino acid (AA) substitutions in the conserved immunoglobulin domain which we have found to be critical for the H2-O inhibitory activity of H2-M. We hypothesize that altered H2-O function in I/LnJ mice is due to an inability to interact with essential binding partners. We recently generated B6J mice in which we have knocked-in these three AA substitutions. This novel tool will be exploited to discover potential binding partners of variant H2-Oβ and elucidate the mechanism by which H2-O activity is modulated.
T Cell Recognition of Disulfide Modified Antigens
Authors: Dai Shaodong , Li Wei , Wang Yang 
Published: January 2023

Abstract
Disulfide formation is one of the most common post-translational modifications and plays important in protein structures and functions. We discovered that the disulfide bridge was the epitope of a highly diabetogenic CD4 T cells. This is a shift from the paradigm that T cells commonly recognize the linear peptide antigens and showed a novel to create neo-epitopes. We identified in vivo that the different subsets of CD4 T cells could be stimulated by both reduced and oxidized form of the same antigens, IAPP. This suggested an imbalance in redox homeostasis may be important for T cell mediated pathogenesis. A disulfide bridge specific monoclonal antibody was created and showed extraordinary ability in preventing type 1 diabetes. The disulfide bridge specific T cells were able to be generalized another protein, CGRP. The lysosomal thiol reductase knock-out experiments showed that the reductase may be important for disulfide modified antigen generation during antigen processing. Elevating GSH levels through glutamate cysteine ligase pathway inhibited CD4 T cell response in a dose dependent manner. Although NOX attributed to the importance of hydrogen peroxide production in pancreatic beta cells, a Nox inhibitor effectively attenuated the BDC5.2.9 T cell activation. Our results shed light on how thiol-disulfide exchange reaction impacts the T cell responses.
Supported by NIH R56 AI153488 NIH R21 AI149655
Lupus Susceptibility Gene Pbx1 Regulates STAT3 in T cells via JAK2/STAT3 Signaling Pathway
Authors: Roach Tracoyia A , Choi Seung-Chul , Park Yuk Pheel , Morel Laurence 
Published: January 2023

Abstract
Systemic Lupus Erythematosus (SLE) is an autoimmune disease in which poorly characterized genetic factors lead to the production of autoreactive or inflammatory T cells. Pre-B cell leukemia homeobox 1 (Pbx1) is a transcription factor whose dominant negative splice isoform (Pbx1-D) is overexpressed in CD4 T cells of lupus patients and lupus-prone mice as compared to the normal isoform (Pbx1-B). Pbx1-D overexpression impairs Treg cell development and function while favoring the production of follicular helper T cells. Based on previous studies showing that Pbx1 promotes cell proliferation via JAK2/STAT3 signaling pathway in a renal cell carcinoma, we hypothesized that Pbx1-D decreases T cell proliferation and viability by attenuating the JAK2/STAT3 signaling pathway. U3A STAT3 reporter cells transfected with Pbx1-B plasmid have a greater luciferase expression, and hence pSTAT3 transcriptional activity in comparison to cells transfected with Pbx1-D or control plasmids. We also observed that Jurkat T cells stably overexpressing Pbx1-B increases STAT3 gene expression as compared to Pbx1-D. Furthermore, Pbx1 silencing in Jurkat T cells leads to the attenuation of STAT3 message. These results suggest a mechanism by which Pbx1 contributes to impaired T cells in lupus pathogenesis is via the JAK2/STAT3 pathway. Current studies are conducted to dissect the mechanism by which Pbx1 regulates this pathway in primary T cells.
Supported by a grant from the NIH (R01 AI045050) to LM.
PKCq modulates LKB1 isoforms downstream of IL-6 signaling to mediate iTreg-Th17 cell plasticity
Authors: Mohan Deeksha , Minter Lisa 
Published: January 2023

Abstract
Following activation, CD4 T cells undergo metabolic and transcriptional changes responding to external cues and differentiate into T helper (Th) cells. This differentiation process was perceived to be terminal. However, it has been shown that T cells exhibit plasticity between subsets in highly inflammatory environments. This phenomenon is prevalent in autoimmune conditions, such as colitis, in which high levels of IL-6 promote plasticity between regulatory T (Treg) cells and Th17 cells. Protein Kinase C theta (PKCq) is a serine/threonine kinase that promotes Th17 differentiation while negatively regulating Treg polarization. The tumor suppressor liver kinase B1 (LKB1) encoded by Stk11, a serine/threonine kinase, enforces energy-preserving mechanisms such as fatty acid oxidation, a major energy source for Tregs. Recent studies demonstrate loss of LKB1 disrupts Treg survival and function. Stk11 can be alternatively spliced into a short variant by the inclusion of a cryptic exon. However, specificity of LKB1 isoforms influencing Th plasticity has not been elucidated. Our study shows that the expression of the Stk11 short splice variant (Stk11S) is upregulated in Th17 compared to induced (i)Tregs. We demonstrate that, in Th17 cells heterogeneous ribonucleoprotein L-like (hnRNPLL), a splicing factor promoting cryptic exon inclusion, binds to transcripts of Stk11 and correlates with increased expression of Stk11S. We further show in iTregs exposed to IL-6, Prkcq, Stk11S, and Rorc are upregulated, suggesting iTreg-Th17 plasticity is induced in response to IL-6 resulting in Stk11 splicing downstream of PKCq. Thus, PKCq may destabilize iTreg programming through its regulation of Stk11 splicing under pro-inflammatory conditions.
Supported by grants from the department of defense.
USP19 Suppresses Th17 Pathogenicity in Autoimmunity
Authors: Bouch Ronald 
Published: January 2023

Abstract
Th17 cells play an essential role in the pathogenesis of murine autoimmune models and human autoimmune diseases. However, Th17 cells exhibit notable plasticity in vivo, adopting pro- or anti-inflammatory effector states in a distinct milieu. The underlying mechanisms that regulate pathogenicity of Th17 cells in health and disease remain unclear. In this study, we investigate the role of the deubiquitinase USP19 in the differentiation and function of Th17 cells. We found that lineage-specific transcription factor RORγt directly binds to ubiquitination-related gene promoters, upregulating the ubiquitination pathway in pathogenic Th17 cells (pTh17). The enhanced ubiquitination activity in pTh17 cells directly correlates with autoimmune disease severity in murine models. The deubiquitinase USP19 suppresses pTh17 differentiation in vitro and Th17-mediated autoimmune disease in vivo. Mechanistically, we found that USP19 removes the K63-linked ubiquitin chain from RORγt lysine 313, resulting in the blocking of the RORγt-SRC3 interaction and ultimately, RORγt activation. Thus, these findings suggest that USP19 selectively suppresses Th17 pathogenicity in murine autoimmune disease.
Supported by P30 Award from NIH, National Cancer Institute of Health (P30CA012197) and NIH T32 Training Grant (5T32A1007401-28)