bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inherited human PD-1 deficiency underlies tuberculosis and autoimmunity
Authors: Ogishi Masato , Yang Rui , Aytekin Caner , Boisson-Dupuis Stéphanie , Casanova Jean-Laurent 
Published: February 2023

Abstract
The pathogenesis of adverse reactions to PD-1 blockade immunotherapy, including tuberculosis (TB) and autoimmunity, remains poorly characterized. Here, we study a patient with inherited PD-1 deficiency who suffered from TB and died of pulmonary autoimmunity. The patient’s leukocytes displayed no PD-1 expression and did not respond to PD-1-mediated suppression. Similar to various inborn errors of immunity impairing interferon (IFN)-γ production and underlying TB, the patient’s lymphocytes poorly produced IFN-γ in response to mycobacterial stimuli owing to both the depletion of multiple IFN-γ-producing T and NK lymphocyte subsets, including Vδ2+ γδ T, MAIT, and CD56bright NK lymphocytes, and dysfunction of residual T lymphocytes. Another set of inborn errors, including STAT3 gain-of-function (GOF) mutations, trigger an expansion of CD4−CD8− double-negative (DN) αβ T cells and underlie lymphoproliferative autoimmunity. Remarkably, the patient displayed hepatosplenomegaly and expansion of total, activated, and RORγT+ DN αβ T cells, as observed in PD-1-deficient mice and cancer patients with PD-1 blockade. Mechanistically, the patient’s myeloid cells produced excessive amounts of STAT3-activating cytokines IL-6 and IL-23 upon stimulation with lipopolysaccharide. Overall, inherited human PD-1 deficiency underlies TB through undermining IFN-γ production by T and NK lymphocytes and lymphoproliferative autoimmunity through over-activating STAT3.
RGC-32 Facilitates Reactive Astrocytosis in Experimental Autoimmune Encephalomyelitis (EAE) by Maintaining a Mature Phenotype
Authors: Rus Horea G , Tatomir Alexandru , Beltrand Austin , Cudrici Cornelia , Courneya Jean Paul , Badea Tudor Constantin , Rus Violeta 
Published: February 2023

Abstract
We previously showed that lack of Response Gene to Complement 32 (RGC-32) decreases the severity of clinical scores of mice with EAE and renders spinal cord astrocytes a progenitor phenotype reminiscent of radial glia. In this study, we used RNA-sequencing to investigate whether RGC-32 deferentially regulates biological processes related to astrocyte maturation and activation. We purified neonatal astrocytes from WT and RGC-32 knock-out (KO) mice, stimulated them with TGF-β and then performed RNA-seq (Genewiz, S. Plainfield, NJ). Spinal cords from WT and RGC-32 KO mice with EAE were harvested at day 0 and at the peak of disease (day 14) and subjected to immunohistochemistry for astrocytes lineage marker GFAP, as well as radial glial marker HOPX, neural stem cell marker CD133, and proliferation marker Ki-67. Pathway enrichment analysis revealed that the genes deferentially expressed in WT but not RGC-32 KO astrocytes belonged to GO categories related to brain development, axonal guidance, cell migration and extracellular matrix (ECM) regulation. Real-Time PCR confirmed that Ephrin A7, bone morphogenetic protein 1 and polycystin 1, other genes related to brain development and ECM were significantly up-regulated in WT astrocytes after TGF-β stimulation. We found that spinal cords of RGC-32 KO mice displayed a higher number of HOPX+ cells at day 14. Similarly, the number of CD133+ cells with a radial glial morphology was higher in RGC-32 KO mice. In addition, we found that RGC-32 KO astrocytes had a higher proliferative index at the peak of EAE, as shown by their increased expression of Ki-67. These data suggest that RGC-32 is an important regulator of astrocyte maturation and reactive astrocyte formation during EAE
A survey of current research trends and potential future directions in reproductive immunology clinical trials
Authors: Rossavik Eevar B , Rossavik Carolin Haeusler , Evans Michael Blake 
Published: February 2023

Abstract
Reproductive immunology is a relatively recent field with a wide spectrum of research opportunities. Within the reproductive system, immune system dysfunction has been suggested to contribute to a variety of conditions - such as endometriosis, infertility, recurrent miscarriage, preeclampsia, and preterm delivery. Although there is preliminary research supporting these possible connections, further studies are necessary. A survey was conducted of current clinical trials to show the current state of reproductive immunology research to assess needs in less researched conditions.
The survey was performed by accessing the ClinicalTrials.gov registry. “Reproductive immunology” was entered into the database, and the following filters were applied: “recruiting,” “active, not recruiting,” and “enrolling by invitation.” 153 studies were obtained, and the associated conditions were counted for how many times they were listed. Similar conditions were combined into one category – e.g. different types of leukemias were categorized as “Leukemia.”
Although the rate of new female breast cancer cases is significantly higher than the rate of new leukemias in the general population, this survey suggests reproductive immunology research is more focused on leukemia than breast cancer. Male and female reproductive/breast cancer/endometrial cancer/endometriosis (6.17%), autoimmune diseases (4.43%), and pregnancy/contraception (0.58%) are less frequently listed compared to leukemia (45.28%) and hematologic diseases (15.80%) in the clinical trials registry as associated conditions. This information can potentially guide future research towards less studied conditions in the relatively new field of reproductive immunology.
DOCK8-expressing Tfh cells cause SLE: autoimmunity via repeated TCR stimulation by antigen to levels that surpass system’s self-organized criticality
Authors: Shiozawa Shunichi , Tsumiyama Ken , Miyazaki Yumi , Uto Kenichi , Sakurai Keiichi , Nakashima Toshie , Matsuyama Hiroko , Doi Ai , Tarui Miho , Izumikawa Manabu , Kimura Mai , Satonaka Chisato , Fujita Yoko , Horiuchi Takahiko , Matsubara Tsukasa , Oribe Motohiro , Yamane Takashi , Kagawa Hidetoshi , Li Quan-Zhen , Mizuno Keiko , Mukai Yohei , Murakami Kazuhiro , Enya Takuji , Tsukimoto Shota , Hakata Yoshiyuki , Miyazawa Masaaki , Shiozawa Kazuko 
Published: February 2023

Abstract
INTRODUCTION: The cause of autoimmunity remains unclear. Candidate pathogens including autoantigens all failed to induce, exist in, or heal upon their removal the prototypical autoimmune disease, systemic lupus erythematosus (SLE). We here investigated, instead of antigen, the immune response of host that recognized antigen.

METHODS:
We test the integrity of host’s immune response by stimulating TCR maximally to levels that surpass the limit of host’s steady-state immune response, self-organized criticality, by an antigen.

RESULTS:
After repeated stimulation by any immunogenic antigen, SLE was induced in mice normally not prone to autoimmune disease, wherein a novel T follicular helper (Tfh) cell type expressing the guanine nucleotide exchange factor DOCK8 on the cell surface was newly generated via resuscitation from antigen-induced anergy. DOCK8+ Tfh cells passed through TCR revision at the periphery, and induced a variety of autoantibodies and the lesion characteristic of SLE. They existed in splenic red pulp and in the circulation of active patients with SLE, which subsequently declined after conventional therapy. Lupus lesions of mice and the classical (NZBxNZW) F1 model mice were mostly cured by anti-DOCK8 antibody.

CONCLUSION:
Thus, disruption of host’s steady-state immune response by repeated TCR stimulation by immunogenic pathogen, either exogenous or endogenous, results in generation of DOCK8-expressing Tfh cells that cause autoantibodies and SLE.
GPR65, a critical regulator of Th17 pathogenicity, is regulated by the CREB/CRTC2 pathway
Authors: Hernandez Jeniffer B , Cave Si , Yang Qiyuan , Decker Carl , Swanson Jack , Wiater Ezra , Zheng Ye , Montminy Marc 
Published: February 2023

Abstract
GPR65 has been shown to be a critical regulator of Th17 pathogenicity. Loss of GPR65 in mice results in a decrease in Th17 cells and reduced susceptibility to a mouse model of multiple sclerosis. The CREB/CRTC2 pathway has emerged as an important regulator of immune function. We have previously shown that the CREB/CRTC2 pathway modulates autoimmune disease by promoting differentiation of Th17 cells. In this study we performed RNA-seq to identify Th17 genes regulated by the CREB/CRTC2 pathway. Our RNA-seq analysis led us to uncover the first mechanism of regulation of the orphan receptor GPR65 by the CREB/CRTC2 pathway. We show that GPR65 is a target of the CREB/CRTC2 pathway through expression studies and chromatin immunoprecipitation. In addition, we show that targeting of GPR65 with small molecules alters the expression of IL-17A. Understanding the regulation of GPR65 will be crucial in developing small molecules to treat patients with Th17-mediated diseases.
Emerging role of the mitochondrial branched-chain aminotransferase (BCATm) as an immunosuppressive enzyme during CD4+ T cell activation
Authors: Ananieva-Stoyanova Elitsa A , Adam Christie , Betar Rebekah , Martin Alexander , Hutson Susan , Boyer Michael 
Published: February 2023

Abstract
The mitochondrial branched-chain aminotransferase (BCATm) catalyzes the transamination of leucine, an amino acid that is essential for the upregulation of complex 1 of the mammalian target of rapamycin (mTORC1) during T cell activation. Because BCATm is responsible for the mitochondrial degradation of leucine, we hypothesized that BCATm limits leucine availability for mTORC1 signaling and subsequently suppresses T cell activation. To test the hypothesis, we isolated CD4+T cells from the global BCATmKO mouse and measured the rates of leucine transamination/oxidation, the activity of the mTORC1 downstream targets S6 and 4EBP-1, and the release of IFNγ after 24–72h of T cell activation. To elucidate the role of BCATm in T cells in vivo, we used the Cre-LoxP strategy to create a T cell specific deletion of BCATm in C57BL/6 mice. We completed the initial characterization of the mice, which included a validation of absence of BCATm in T cells by using qRT-PCR and Western blotting. Results demonstrated that BCATm-deficient T cells experienced significant reductions in the transamination and oxidation of leucine, which led to increased intracellular leucine concentrations. This correlated with an increased phosphorylation of S6, but not 4EBP-1, suggesting that BCATm regulates mTORC1 signaling in S6 dependent manner. The T cells released more IFNγ in the absence of BCATm, which was highest after 72h of T cell activation. Thus, BCATm appears to play an immunosuppressive role by reducing CD4+ T cell activation. The newly designed T cell conditional BCATmKO mice, which do not express BCATm in CD4+ and CD8+ T cells will be engaged in preclinical studies aiming at deciphering the therapeutic potential of BCATm in T cell-driven immunological responses.
Exploring Mechanisms of Human PD-1 Gene Regulation in T Follicular Helper Cells
Authors: Powell Michael Duane , Neeld Dennis Kyle , Rose James , Scharer Christopher , Boss Jeremy M 
Published: February 2023

Abstract
Programed Cell death (PD-1), encoded by Pdcd1, is an immune inhibitory receptor expressed on the surface of lymphocytes. Surface PD-1 expression is amongst its highest in a subset of CD4+ T cells called T follicular helper cells (TFH). TFH cells are pivotal for optimal germinal center formation and subsequent generation of pathogen- and vaccine-induced antibodies by B cells. Despite the clear connection to human health, the majority of previous work has centered on elucidating the mechanisms surrounding murine Pdcd1 regulation. Surprisingly, there is almost nothing known about how human Pdcd1 (hPdcd1) is regulated! Here we seek to close this gap in knowledge by identifying the cis-, trans-, and epigenetic pathways that regulate hPdcd1 in TFH cells. To this end, human TFH cells were sourced from both discarded tonsil tissue as well as from a recently described model to generate ex vivo human TFH cells through treatment with the cytokines Activin A, IL-12 and TGFβ. RNA-sequencing demonstrated that ex vivo TFH cells exhibit characteristics of bona-fide tonsillar TFH cells, including increased expression of PD-1. Furthermore, ATAC-seq and H3K27Ac Hi-ChIP experiments revealed the presence of 7 conserved accessible regions within the hPdcd1 locus that exhibit unique looping characteristics and potential enhancer properties. Functionality of each regulatory element was further defined through promoter reporter assays. Additionally, PAGE-RANK analysis elucidated transcription factors that may play a role in the induction of hPdcd1 expression. Collectively, these findings begin to unearth the mechanisms that control hPdcd1, informing the design of future therapeutics aimed at precisely manipulating human PD-1 expression.
A genome-wide CRISPR screen reveals a role for the BRD9-containing non-canonical BAF complex in Foxp3 expression and regulatory T cell function
Authors: Zheng Ye , Loo Chin-San , Gatchalian Jovylyn , Hargreaves Diana 
Published: February 2023

Abstract
Regulatory T cells (Tregs) play a pivotal role in suppressing auto-reactive T cells and maintaining immune homeostasis. Treg development and function are dependent on the transcription factor Foxp3. Although a large body of work has been devoted to dissecting the molecular mechanisms regulating Foxp3 expression, a systematic, genome-wide approach has not been reported. Here we performed a genome-wide CRISPR/Cas9 knockout screen to identify the regulators of Foxp3 in mouse primary natural Tregs. The screen results not only confirmed a number of known Foxp3 regulators, but also revealed many novel factors that control Foxp3 expression. Gene ontology analysis showed that Foxp3 regulators are highly enriched in genes encoding SWI/SNF and SAGA complex subunits. Among the three SWI/SNF-related complexes, the non-canonical or ncBAF (also called GBAF or BRD9-containing BAF) complex promoted the expression of Foxp3, whereas the PBAF complex repressed its expression. Gene ablation of BRD9 led to reduced Foxp3 expression and compromised Treg function in inflammatory disease and anti-tumor immunity. BRD9 co-localized with Foxp3 in genome-wide binding studies, including at the CNS0 and CNS2 enhancers at the Foxp3 locus. Functional genomics revealed that BRD9 is required for Foxp3 binding and expression of a subset of Foxp3 target genes. Thus, we provide an unbiased analysis of genes and networks regulating Foxp3, and reveal the ncBAF complex as a novel target that could be exploited to manipulate Treg function.
Factor VIII-Fc activates natural killer cells via Fc-mediated interactions with CD16
Authors: Lagasse H.A. Daniel , Hopkins Louis B. , Jankowski Wojciech , Sauna Zuben E. , Golding Basil 
Published: February 2023

Abstract
FactorVIII Fc-fusion (rFVIIIFc) is an enhanced half-life antihemophilic factor usedin replacement therapy of hemophilia A. Fc-fusion is a successful protein bio-engineering platform technology. In addition to enhancement of plasmahalf-life via neonatal Fc receptor (FcRn) binding, other Fc-mediatedinteractions, including engagement with Fc gamma receptors (FcγR), may haveimmunological consequences. Previouslywe and others demonstrated rFVIIIFc interactions with activatingFcγRIIIA/CD16. Here, we investigated ifrFVIIIFc activates natural killer (NK) cells via CD16. We demonstrated rFVIIIFc signaling via CD16independent of Von Willebrand Factor (VWF):FVIII complex formation. We established that rFVIIIFc potentlyactivated NK cells in a CD16-dependent fashion resulting in IFNγ secretion andcytolytic perforin and granzyme B release. We also demonstrated an association between rFVIIIFc-mediated NK cellIFNγ secretion levels and the high affinity (158V) CD16 genotype. Furthermore, we show that rFVIIIFc-activatedCD16+ NK cells were able to lyse a B-cell clone bearing ananti-FVIII BCR (BO2C11) in an antibody-dependent cellular cytotoxicity (ADCC)assay. These findings provide anunderlying molecular mechanism that may help explain clinical case reports andretrospective studies that suggest rFVIIIFc may be more effective in tolerizinghemophilia A patients with anti-FVIII inhibitory antibodies compared to FVIIInot linked to Fc. Our findings suggest apotential use of Fc-fusion proteins acting via NK cells to target B-cells, inthe management of unwanted immune responses directed against immunogenicself-antigens or therapeutic protein products.
Curricular Framing of the Undergraduate Immunology Classroom
Authors: Bruns Heather A. , Taylor Rebekah , Wisenden Brian , Kleinschmit Adam , Liepkalns Justine , Lal Archana , Vanniasinkam Thiru , Mixter Philip F , Paustian Timothy , Sletten Sarah , Sparks-Thissen Rebecca , Condry Danielle , Pritchard Rachel E , Elliott Samantha L , Justement Louis B , Pandey Sumali 
Published: February 2023

Abstract
Immunology education has historically been delivered at the graduate level. Calls for reform in biology education and increased workforce demand for individuals trained in immunology have spurred efforts to examine immunology education at the undergraduate level. The Undergraduate Immunology Education Consortium, a newly formed international group of immunology educators representing diverse institutions, is contributing to this effort through the development of curricular resources and student learning outcomes. We have identified a set of fundamental concepts that are critical to the understanding of immunology for undergraduate students and align with the 2011 AAAS Vision and Change Report. To identify consensus among immunology educators this group distributed an initial online survey to immunology educators to evaluate the identified fundamental concepts. Following survey feedback, the concepts were revised, submitted for further evaluation by immunology educators in focus groups, and then distributed in a final survey. Analyses of the data from the initial survey demonstrated that the respondent pool represented a varied and diverse range of expertise. Although teaching experience and current role in teaching influenced the proportion of time allotted to some immunology subtopics, topics of greatest emphasis included innate immunity, adaptive immunity, host-pathogen interactions, and molecular mechanisms. An understanding of current curricular practices by immunology educators will further aid in the refinement of the fundamental concepts, their interdisciplinary use in a diverse range of undergraduate science curricula, and future collection of assessment data to analyze effective best practices.
Learning outcomes with flipped and traditional classrooms in medical immunology.
Authors: Jafferjee Mufaddal , Abdullah Ikram , Brown Teresa , Geis Abby L 
Published: February 2023

Abstract
Education literature reports the benefits of flipped classrooms (FC), but medical educators still typically rely on traditional didactic lectures (TL) as a primary teaching modality. In fact, when given a choice of delivery modalities, learners often prefer to engage in the easier and comfortable route to knowledge, including listening to lectures. However, learning through passive techniques is less effective because it fails to facilitate application and recall of concepts. High order learning and retention is significantly improved when the learning process is challenging and thought-provoking.
We used immunology within a medical school curriculum to compare the effectiveness of and student preferences for two distinct teaching modalities: FC and TL. Data was collected from 170 first year medical students at our institution. In the FC modality, prerecorded lectures were provided to students prior to class. In class, students completed a quiz with low-level questions, the quiz was reviewed, and the remaining class time was spent doing high-level application problems. For the TL, students were provided with the low-level quiz and high-level application problems to do at their own discretion. No aspect of the learning activities was required or graded. Our data show that students who performed poorly on exam items linked to the traditional lecture benefitted disproportionately from the flipped classroom modality, though there was no difference in learning outcomes between the two modalities overall. Finally, students who provided feedback about their learning preferences overwhelmingly preferred traditional lectures and denied learning gains from the flipped modality.
Enhancing engagement of graduate students in class via use of an interactive teaching platform improves learning and long-term retention
Authors: Alard Pascale , Kosiewicz Michele M 
Published: February 2023

Abstract
Engagement of students in the classroom has been shown to improve understanding and retention of material. However, the most common teaching format in Graduate, Medical or Dental School is lecturing via powerpoint, which minimally engages students, an issue amplified during the pandemic due to remote teaching. Assessing previous knowledge, predicting/retrieving information, and using academic games are teaching strategies that facilitate more effective learning. We used TopHat, an interactive teaching platform allowing integration of these strategies, to teach a graduate Immunology course. Two cohorts of first-year graduate students were provided with the opportunity to anonymously discuss topics learned during the current or previous sessions, hear different perspectives, and correct what they had not understood during face-to-face or virtual sessions. Only some instructors in the course used the platform. Students who perform overall at a lower level obtained higher exam scores with instructors who used the platform. However, all students received higher scores on comprehensive final exam questions provided by instructors who used the platform, suggesting that engaging graduate students in class helped with long-term retention. Lastly, a student survey about the utility of the platform indicated that a large majority of students felt more engaged, and believed that they were better able to understand the material and retrieve the information both short-term and long-term. This study suggests that using an interactive teaching platform with graduate students resulted in better engagement, improved learning and long-term material retention, and could be easily implemented at any level of Immunology education.
Generation and implications of oral resident memory T cells
Authors: Stolley J. Michael , Masopust David 
Published: February 2023

Abstract
CD8+ tissue resident memory T cells (TRM) represent the predominant memory T cell population in many nonlymphoid tissues (NLT) where they are major contributors to localized immunosurveillance. TRM accelerate protection against reinfection, may be associated with tumor control, and may also facilitate the persistence of certain allergic and autoimmune diseases. While extensively studied in other mucosal sites, there is presently a fundamental void in our understanding of the ontogeny, function, and therapeutic implications of oral-mucosal TRM. Given their well-documented and critical functions in mediating barrier immunosurveillance in other NLT, oral TRM are likely to play a major role in antiviral immunity and oral immune homeostasis. TRM may also perpetuate chronic immune responses observed in periodontal disease and oral lichen planus. However, addressing their role in these clinically relevant settings has been mired by a lack of animal models for generating sufficient oral TRM to manipulate and study. Here we introduce a novel oral ‘prime-pull’ strategy for generating large quantities of tractable TRM of a defined antigen specificity within the oral mucosa. Leveraging this approach, we demonstrate that oral TRM 1) are abundant within mouse gingiva, 2) are intimately associated with salivary ducts of the tongue and buccal mucosa, and 3) can be locally reactivated to potentiate oral immunity. Ongoing work will further define the tissue-specific functions of oral TRM and describe new mechanisms by which oral immunity can be attenuated or augmented to achieve the desired clinical outcome.