bluuebunny/crossref_metadata_2025_split huggingface dataset:
Results from a CURE course project reveal novel insights into the structure of bacterial Toll/interleukin-1 receptor (TIR) domain proteins
Authors: Snyder Michelle , Vallejo-Schmidt Tamara E , Palm Cheyenne , Obiorah Trinity E , Koudjra Rachida E , Snyder Greg A 
Published: August 2023

Abstract
Toll-like receptors (TLRs) are critical for innate immune signaling, and various bacteria express Toll/interleukin-1 receptor (TIR) domains to evade this signaling. Bacterial TIRs exhibit NADase activity, although it is unclear what role NADase activity plays in evasion of innate responses. To better characterize the NADase activity of the TIR from Acenitobacter baumanii (AbTIR), we used site directed mutagenesis to identify amino acids predicted to be important for NADase activity. Studies were carried out in a Course-Based Undergraduate Research Experience (CURE) involving 16–20 undergraduate research students per year. Students used Chimera to analyze the AbTIR structure and hypothesize amino acids they predicted would play a role in enzymatic activity. They then performed site-directed mutagenesis to create plasmids for expression of mutant AbTIR. Mutant TIRs were expressed, purified and tested for NADase activity. So far, our mutagenesis results suggest a potential conformational change in the NAD binding site upon NAD binding. Identifying amino acids critical for enzymatic activity may inform drug development for diseases associated with dysregulated TIR activity. In addition, the application of the CURE class to carry out these studies has provided project-based research training experiences for approximately 80 undergraduate students, about half of whom are members of groups traditionally underrepresented in science. Several students, including some who had limited research experience before completing the course, are currently pursuing graduate studies and research-based careers, highlighting the value of CURE-based projects for facilitating the inclusion of students from diverse groups in research.
TVS and TO were supported by the NIH Bridges to Baccalaureate Grant R25GM058264; CP was supported by the NIH Bridges to the Doctorate Grant R25GM119970, and the CURE course was supported by an HHMI Inclusive Excellence Grant
Teaching Methods and Instructional Efficacy of Virtual Course Presentation in a Pre-Clerkship Medical Immunology Module
Authors: Robinson Isabelle L , Ryan Madison R , Edwards Penny S , Chosed Renee J , Grier Jennifer T 
Published: August 2023

Abstract
In March of 2020, education across the world transitioned to virtual delivery and medical education was not exempt. This study explored the efficacy of virtual medical education in a pre-clerkship immunology course during the COVID-19 pandemic at the University of South Carolina School of Medicine Greenville. Course delivery methods and educational resources provided to students during either 2019 or 2020 were evaluated to generate a picture of the learning environment by year. Deidentified summative exam performance data from 204 students was analyzed across 61 matched questions that did not differ between the 2019 and 2020 exams. Statistical significance was determined by unpaired t-test using GraphPad Prism. As expected, in 2020, the content delivery shifted towards remote and asynchronous sessions. The total number of integrated clinical cases decreased in 2020 compared to 2019, although a greater percentage of the clinical cases directly mapped to the USMLE Content Outline in 2020. Formative multi-attempt online quizzes replaced in-person practice questions and the overall number of available review questions increased from 2019 to 2020. There was no significant difference in student performance between the two years on matched exam questions or on exam performance as a whole. While the switch to virtual learning offered fewer synchronous sessions for student-faculty interactions, modifications to course materials appears to have counteracted any potential learning loss. This suggests that virtual medical immunology education with adequate resources can be as effective as in-person delivery, which has implications for undergraduate and medical immunology instruction and for the entirety of medical education.
Reverse Mentoring in STEM: Changing Perspectives to Foster Dialogue
Authors: Banda France-Elvie , Seger Yvette , Brute Maribet 
Published: August 2023

Abstract
Reverse mentoring provides organizations with key insights into the personal and professional needs and preferences of staff members from a variety of backgrounds and experiences. Although pairing a senior-level mentee with a junior-level mentor has been successfully utilized by the corporate world for over a decade to help employees gain appreciation of colleagues’ lived experiences, traditional mentoring remains the norm in academic research settings. Given the well-documented barriers to participation and attrition of individuals from historically excluded groups from the professoriate, FASEB established a reverse mentoring program for scientists and staff of scientific societies to assess the ability of this strategy to foster a greater understanding and appreciation of differences between individuals at different career stages and lived experiences. Through participation in reverse mentoring, pairs learn to develop inclusive spaces and foster a culture of inclusive decision-making in their spheres of influence. This engagement will enable participants to better assess existing procedures and policies that create institutional barriers for historically excluded groups. Our presentation will summarize key findings from our pilot cohort of four reverse mentoring pairs, including applicant statistics, reverse mentoring curriculum, and preliminary participant feedback.
Chronic opioid use is associated with higher flu vaccine induced Ab response in HIV+ and HIV− individuals
Authors: Dang Christine Minh , Iyer Akshay , Nelson C Mindy , Feaster Daniel , Forrest David , Ghanta Priya , Pahwa Rajendra , Jayaweera Dushyantha , Rodriguez Allan , Tookes Hansel , Pallikkuth Suresh , Pahwa Savita 
Published: August 2023

Abstract
Opioid dependence is frequent in people with HIV (PWH), but impact of opioids on immune system of PWH is unknown. This study tested the hypothesis that chronic opioid use exacerbates known impairment of flu vaccine responses in PWH. Chronic opioid users were compared to non-opioid users in HIV+ and HIV− groups termed HIV+OP+ (n=28), HIV-OP+ (n=55), HIV+OP− (n=53), HIV-OP− (controls, n=58). HIV+ individuals were on ART with plasma virus load &lt;200 copies/mL. Flu antibody titers were determined at pre- (T0) and 4 weeks post (T2) seasonal quadrivalent influenza vaccinations (2020–2022) by hemagglutination inhibition (HAI) along with a plasma cytokine score at T0. Vaccine response (VR) was defined as a 4-fold change (FC) in titer from T0 to T2. In Kruskal Wallis analysis, all 4 groups had increases in HAI titer from T0 to T2 (p&lt;0.01 to p&lt;0.0001) for antigens (H1N1, H3N2, B Victoria) and whole vaccine. HIV+ status was associated with decreased responses to all antigens but OP+ status in HIV+ and HIV− had higher response than HIV+OP−, both in T2/T0 FC and T2 titer for one or more vaccine antigens (p&lt;0.05 to p&lt;0.001). In regression analysis controlling for demographics, opioid use was associated with increased response to H1N1 and B Yamagata antigens. In a random forest model, predicted probability of VR was lower for HIV+ status and higher for OP+ status. Cytokine score was highest in HIV+OP+. Results of quantitative Ab response to flu vaccine refute our hypothesis that flu vaccine response would be impaired in PWH who chronically use opioids. Qualitative assessments of Ab and mechanism of opioid intersection with B cell function are ongoing to understand if opioid use overcomes humoral immune deficits in PWH.
Role of IgG glycans in severe COVID-19 inflammatory cytokine responses
Authors: Scherer Erin M , Sherman Jacob D , Karmali Vinit , Kumar Bhoj , Simon Trevor W , Bechnak Sarah , Ciric Caroline R , Gray Natalie , Huerta Christopher , Hussaini Laila , Johnson Brandi , Lee Matthew , Panganiban Bernadine , Panjwani Anusha , Samaha Hady , Wang Dongli , Anderson Evan J , Azadi Parastoo , Collins Matthew H , Rostad Christina A , Rouphael Nadine 
Published: August 2023

Abstract
Severe COVID-19 and multisystem inflammatory syndrome in children (MIS-C) are characterized by hyperinflammation. Severe COVID in adults is associated with development of afucosylated SARS-CoV-2 IgG that induce monocytes to secrete inflammatory cytokines. This study aimed to analyze the relationship between spike IgG Fc glycosylation and inflammatory markers during acute pediatric COVID or MIS-C or following COVID vaccination in adults.
We assessed cross-sectional and longitudinal samples from 195 participants: Adult vaccinees, acute COVID patients, and healthy controls, as well as pediatric acute COVID patients, MIS-C patients, and healthy controls. We developed capillary electrophoresis (CE) methods to analyze bulk and spike IgG Fc glycans and measured ten relevant cytokines/chemokines by multiplexed assay.
Analysis of healthy control bulk IgG by CE found 95.3–99.6% fucosylated, 2.7–6.9% bisected, 55.8–66.8% galactosylated, and 4.6–15.6% sialylated Fc glycans, compared to 99.8%, 6.7%, 45.4%, and 8.2% by mass spectrometry, respectively. IFN-γ, IL-10, CXCL10, and CCL3 were significantly increased in MIS-C compared to pediatric healthy controls; IL-10, CCL2, and TNF were only increased in severe adult COVID compared to healthy controls or less severe COVID. Initial results confirmed significantly more afucosylated spike IgG in adult severe COVID than afucosylated bulk IgG in controls.
Our high-throughput methods for IgG Fc glycan profiling generated comparable results to mass spectrometry. Continued assessment will determine if COVID- or vaccine-induced spike IgG glycoforms correlate with inflammatory markers. Future mechanistic work will examine how modified glycosylation induces inflammatory cytokines.
Supported by an IDCRC New Investigator Pilot Award to E.M.S. (NIH UM1AI148684), by the Centers for Disease Control and Prevention through a cooperative agreement with the Georgia Emerging Infection Program (grant no. U50CK000485), and by grant from NIH to P.A. (R24GM137782).
Streptococcus pneumoniae coinfection results in increased disease in a murine model of influenza B virus infection
Authors: Lorton Ana T , Beaver Emily , Ray Sean , Page Caroline , Shepard Justin , Tompkins Stephen Mark 
Published: August 2023

Abstract
Secondary Streptococcus pneumoniae (Spn) infection is commonly associated with increased hospitalization and death following seasonal influenza virus infection. The animal models of influenza A virus (IAV) and secondary Spn coinfection have described the increased morbidity and mortality leading to several proposed mechanisms of enhanced disease. Influenza B virus (IBV) causes 25% of seasonal influenza cases and is associated with bacterial coinfection, yet little is known about the immune response to IBV and Spn coinfection. Here, we established a mouse model of IBV and Spn coinfection to study disease in singly and coinfected animals and define mechanism(s) of enhanced disease. Like IAV and Spn coinfection, infection with IBV followed by Spn inoculation resulted in increased morbidity and mortality compared to IBV or Spn infection alone. In IBV primed mice, Spn titers were increased early following bacterial inoculation and Spn rapidly migrated to the lung. Analysis of cytokines in the lung showed increased pro-inflammatory cytokine (TNF-α, IL-1β, and IL-6) responses with secondary Spn infection, which may be associated with enhanced disease. These data suggest that IBV infection initiates innate immune responses that are rapidly enhanced with Spn infection, leading to worsening disease. Ongoing studies include analysis of infiltrating cells during coinfection and histopathologic analysis to complement the innate response data to define mediators of clinical disease.
NIH CIDER Contract #75N93021C00018
Development of a novel serology assay for the detection of IgG antibodies to identify exposures to orthopoxviruses
Authors: Lee William Thomas , Hunt Danielle T , Kulas Karen E , Howard Kelly M , Carson Kyle , Lamson Theresa , Chakravarthi Sandhya , Chave Karen , Bievenue Rachel , Rock Jean M , Styer Linda M , Noone Philip , Hogrefe Wayne , Cai Gianna Y , Bermúdez-González Maria C , Kleiner Giulio , Srivastava Komal , Simon Viviana , Feihel Dennis , McGowan Joseph P , Slifka Mark K , Yates Jennifer L 
Published: August 2023

Abstract
The World Health Organization has declared a Public Health Emergency for an ongoing mpox (formerly monkeypox) outbreak, due to multiple cases of mpox in regions of the world in which infections have not been previously seen. The outbreak is notable for the high degree of human-human transmissions spreading primarily through sexual contacts. Currently, as a prophylactic (and post-exposure) measure, high-risk individuals are being offered immunization with cross-reactive vaccinia virus (VV) vaccines, a strategy previously used for the closely related variola (smallpox) virus. Commercial tests to measure binding antibodies to mpox for diagnostics or surveillance are not (yet) available. We present here a novel IgG detection assay to identify or measure binding antibodies to mpox and VV. This multiplexed, Luminex-based serology assay contains multiple recombinant proteins or peptides derived from VV and mpox. Because the assay contains antigens present on orthopoxviruses including VV, but also contain antigens present on orthopoxviruses except VV, the test allows for serological distinction between infection with virulent orthopoxviruses and attenuated strains of VV. We present data on detection of antibodies elicited by childhood (smallpox) or recent (mpox) vaccination, mpox infection, and breakthrough infection in previously vaccinated individuals. We demonstrate that the baseline antibody profiles of high-risk populations are distinct and must be considered when assessing mpox serology information. Overall, these data support the use of Luminex-based serology as a diagnostic tool for mpox infections and as a surveillance tool to detect the prevalence of mpox infections, even within a vaccinated population.
This work was supported by Cooperative Agreement Number NU50CK000516 funded by the Centers for Disease Control and Prevention. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the Centers for Disease Control and Prevention or the Department of Health and Human Services.
Characterizing the effect of type I interferon signaling within regulatory T cells in HSV-2 infection
Authors: Warrier Lakshmi , Graham Jessica B , Swarts Jessica , Lund Jennifer M 
Published: August 2023

Abstract
Herpes simplex virus type-2 (HSV-2) is a lifelong, recurrent sexually transmitted infection that affects about 13% of the world population between ages 13–49. There is currently no available vaccine for HSV-2 and understanding the nuances of cell-mediated immunity is crucial to developing improved therapeutics and vaccines. HSV-2 infection induces a potent, localized type I interferon (TIIFN) signaling cascade that can stimulate both innate and adaptive antiviral effector responses. Interestingly, regulatory T cells (Tregs), an immunosuppressive CD4+ T cell subset, also respond to TIIFNs. TIIFNs have previously been shown to transiently inhibit Tregs early during a systemic lymphocytic choriomeningitis virus (LCMV) infection to promote the development of antiviral CD4+ and CD8+ T cell responses, and Tregs that lack the interferon alpha receptor (IFNAR) have enhanced suppressive function. The role of TIIFNs in a mucosal infection such as HSV-2 is unclear and studying the tissue-specific effects of TIIFNs warrants further study. To investigate the role of TIIFN signaling within Tregs in the context of HSV-2, we infected IFNARfl/fl x FoxP3Cre mice intravaginally with attenuated or WT HSV-2 and assessed phenotypic Treg markers and viral titers within the vaginal tract by flow cytometry and plaque assays, respectively. Preliminary data indicates that WT, and not attenuated HSV-2 infected mice show enhanced disease progression and increased viral titers. This suggests that TIIFN signaling in Tregs could be aiding in mediating protection early in HSV-2 infection.
NIH R01AI141435
Imaging of lung resident memory CD8+ T cells reveals antigen-specific early protective responses during secondary infection
Authors: van de Wall Stephanie , Anthony Scott M , Varga Steven M , Langlois Ryan A , Zehn Dietmar , Badovinac Vladimir P , Harty John T 
Published: August 2023

Abstract
Tissue-resident memory T (TRM) cells are poised at mucosal sites to confer robust protection against secondary infection. In contrast to skin and intestine, lung TRM cells are short lived with a decline in number coinciding with a decline in protective immunity against subsequent heterosubtypic influenza infections. Despite this causal relationship, the localization, dynamics and function of lung Trm during secondary responses remains poorly defined. To address this, we used two-photon intravital microscopy of lungs to assess in real time the spatiotemporal dynamics of lung TRM cells during recall infection. At a memory time point, lung TRM cells (representing both CD103+ and CD103− subsets) exhibit significantly lower motility and less displacement compared to an CD8+ effector T cells at an earlier time point post infection. Depletion of circulating CD8+ T cells using a low dose antibody administered systemically led to enrichment of lung TRM cells that exhibited slow to moderate speed scanning patterns. A fate mapping model for CD103+ TRM cells specifically revealed limited speed (&lt;5 um/min) and patrolling behavior compared to CD103− TRM cells. Within 24 hr of rechallenge, CD103+ TRM cells co-localized with infected cells and by 48 hr, lung-resident CD8+ T cells exhibit significantly lower motility with recruitment and clustering in infected regions in an antigen-specific manner. Protective immune responses by lung TRM cells is further accompanied by CXCR3-dependent migration of dendritic cells to draining lymph nodes as well as recruitment of inflammatory monocytes. Overall, these data unravel the spatiotemporal dynamics of lung TRM cells and interactions with recruited immune cells for early protective immunity.
Supported by grants from NIH (GM134880, AI42767, AI114543, AI151183, AI167249, the Holden Cancer Center at the University of Iowa and its National Cancer Institute Award P30CA086862
Double negative T cell-NK cell axis orchestrates antitumor immunity against triple negative breast cancer
Authors: Zhang Hui , Little Alex A 
Published: August 2023

Abstract
Triple-negative breast cancer (TNBC) is hard to treat because of the development of chemoresistance and metastasis. It is recently found that the combination of chemotherapy and immune checkpoint blockade can significantly improve TNBC treatment. However, the low response rate to immunotherapy greatly hampers the application of immunotherapy to TNBC. It is urgent to find new paradigms to improve immunotherapy for TNBC. We previously found that transferring thymocytes into Ja281 KO mice could inhibit E0771 TNBC formation. Herein, we further determine the underlying mechanism. We found that transferring splenocytes, CD4+, or CD8+ T cells into the Ja281 KO mice could not induce antitumor immunity. Transferring thymic double-negative T (DN T) cells into the Ja281 KO mice could inhibit tumor formation. Treating the cell-transferred Ja281 mice with anti-TCRβ or anti-TCRγδ antibodies two days before tumor inoculation, E0771 TNBC grew in the anti-TCRβ treated mice but not anti-TCRγδ-treated mice. These results indicated that the cell transfer-induced antitumor immunity in Ja281 KO mice was mediated by TCRαβ+ DN T cells. Transferring iNKT or MAIT KO DN T cells did not affect cell-transfer-induced antitumor immunity. Only NK cell depletion abrogated DN T transfer-mediated antitumor immunity by using an antibody to deplete NK or macrophage, suggesting that NK cells are essential for DN T-mediated antitumor immunity against E0771. Antibody depletion of the circulating transferred cells did not affect cell transfer-induced antitumor immunity, suggesting that the DN T cells mediating antitumor immunity are tissue-resident cells. Collectively, these data indicate that the DN T/NK cell axis orchestrates antitumor immunity against TNBC.
Supported by Washington State University College of Pharmacy and Pharmaceutical Sciences Start-up funds
Tumor infiltrating natural killer cells in soft tissue sarcoma are predominantly CD56dim: Implications for outcomes
Authors: Sholevar Cyrus , Cruz Sylvia M , Iranpur Khurshid , Judge Sean J , Gingrich Alicia , Farley Lauren E , Razmara Aryana , Lammers Marshall , Thorpe Steven , Monjazeb Arta M , Murphy William J , Canter Robert J 
Published: August 2023

Abstract
Natural killer (NK) cells have been shown to be important mediators of anti-tumor responses, including in soft tissue sarcomas (STS). NK cells show significant heterogeneity, depending on maturation, tissue residency, and inflammatory environment. As previous studies have suggested that tumor-infiltrating NK cells (TiNKs) acquire a less cytotoxic CD56bright tissue resident phenotype, we sought to compare blood versus tumor NK cell phenotype in STS and evaluate any association with survival. Blood and tumor from 27 STS patients undergoing surgery from 2018–2022 were collected prospectively for flow cytometry and retrospectively analyzed. 52% were female, the mean age was 59, and 74% were AJCC stage 3. Increasing absolute number of NK cells in the blood correlated with longer survival (P&lt;0.005, r=0.6). As expected, CD56dim NK cells predominated in the blood (91.5±5.8% of CD3-CD56+ lymphocytes compared to 80.1±13% in tumor, P&lt;0.005). In contrast, CD56bright cells were enriched in tumor representing 18.5±13% compared to 8.4±5.8% in blood (P&lt;0.005). Although CD56bright NK cells were approximately 2.4±3-fold higher in tumor compared to blood, CD56dim NK cells still represented the majority of TiNKs. Higher proportions of both overall NK cells and CD56dim NK cells in STS tumors were associated with better metastasis-free survival on Kaplan-Meier analysis (P&lt;0.05). CD56dim TiNKs had higher NKp46 expression than CD56brights. In conclusion, both blood and intra-tumoral NK cells appear prognostic in STS. Although the proportion of CD56bright TiNKs in STS is higher than blood the majority of TiNKs are CD56dim consistent with a cytotoxic phenotype. Better characterization of NK subsets in STS is likely to have translational significance.