bluuebunny/crossref_metadata_2025_split huggingface dataset:
Adjuvant effects of peptidoglycan on CD4+ T cell activation by MHC-II+ fibroblast-like synoviocytes
Authors: Rouse Joe , Danner Rebecca , Wahhab Amanda , Jutras Brandon , Lochhead Robert B 
Published: August 2023

Abstract
Lyme arthritis (LA) is the predominant late-stage manifestation of Lyme disease, caused by tick-borne spirochete Borrelia burgdorferi. LA can resolve with antibiotic therapy or persist after treatment in a subset of patients, known as post-antibiotic LA. Post-antibiotic LA is characterized by proliferative synovitis, dysregulated Th1/ IFNɣ responses and MHC-II+ fibroblast-like synoviocytes (FLS), a common cell type in inflamed joints. High levels of B. burgdorferi peptidoglycan (PG) have been found in LA patient synovia, yet the effects of PG on host immune responses are not well characterized. We hypothesize that PG acts as an adjuvant to enhance antigen presentation and CD4+ T cell activation in the joint. To test this, primary murine macrophages and FLS, both abundant in LA synovia, were treated in vitro with IFNɣ, PG, or both. Stimulation of macrophages led to upregulation of MHC-II in an IFNɣ-dependent manner, as expected. FLS stimulated with IFNɣ led to upregulation of MHC-II and production of proinflammatory cytokines, which was enhanced by PG as well as the muramyl-dipeptide subunit of PG, a known NOD2 ligand. To examine T cell responses, IFNɣ-primed MHC-II+ FLS were co-cultured with OT-II mouse T cells (all with identical antigen-specific T cell receptors) and supplemented with the cognate OVA323–339 peptide. MHC-II+ FLS activated naïve CD4+ T cell proliferation, which was enhanced by PG stimulation. Co-culture supernatants showed upregulated IFNɣ and IL-12 production, indicating a Th1 response. Negative controls using an alternative OVA257–264 peptide or NOD2-deficient FLS failed to induce T cell responses. These data show that PG acts as an immune adjuvant enhancing peptide-specific CD4+ T cell activation by MHC-II+ FLS.
Heterogeneous Specificities of Nickel Presentation to Nickel Specific CD4 T Cells
Authors: Dai Shaodong , Zhang Yan , Chen Lan , Wang Yang , Li Wei 
Published: August 2023

Abstract
Sensitization to nickel (Ni2+) is the most common T cell mediated type IV hypersensitivity, although the mechanisms by which Ni2+ alters antigen presentation are not understood. To study how the TCR then recognizes Ni2+ bound to the MHC molecule, we performed paired single cell RNA sequencing of PBMCs from two highly Ni2+ sensitized subjects to orthopedic implants and characterized three Ni2+ specific CD4+ T cell clones. One Ni2+ specific CD4+ T cell is highly restricted to HLA-DR7. But the other highly cross-reactive CD4+ T cell clones was restricted to multiple HLA alleles, including HLA-DR, HLA-DP, and HLA-DQ expressing antigen presenting cells. We identified a HLA-DQ8 restricted Ni2+ dependent mimotope reactive to the highly cross-reactive T cell clone. This mimotope appeared to present Ni2+ ions with the Glu at P2 position, involving βH81. The mutational studies of HLA-DP2 molecule showed that the same T cell recognized the Ni2+ ion bound to a completely different location in the P4-P7 pocket. Ni2+ ion was also can be presented efficiently by HLA-DR53 to the same T cells. mutational and structural study of HLA-DR53 molecules suggested that HLA-DR53 presented Ni2+ in the P4-P7 pocket. This is the first evidence showing that a metal ion can be presented by HLA using the different sites. The metal binding sites are HLA and peptide dependent. The heterogeneous specificities of Ni2+ specific TCRs are determined by different alleles of HLAs together with particular self-peptides.
NEI supplementation improves alum-based immunity in CFTR KO mice
Authors: Yoo Sungjun , Kim Eunsoo , Mazik Logan , Steiner Haley E , Cormet-Boyaka Estelle , Boyaka Prosper N 
Published: August 2023

Abstract
Cystic fibrosis (CF) is a genetic disorder that causes severe damage to the lungs and digestive system. CF affects the cells that produce mucus, sweat, and digestive enzymes and is caused by mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR). Expression and function of CFTR have been extensively studied on epithelial cells and several recent publications have shown that CFTR also regulates the functions of myeloid cells. We addressed whether CFTR is expressed by B cells and the role of CFTR in B cell functions. We found that CFTR is expressed on murine and human B cells. Compared to control wild-type (WT) mice, CFTR KO exhibited reduced frequency of B cells in lymphoid tissues and lower levels of serum immunoglobulin isotypes. Upon systemic immunization with an alum-based vaccine, CFTR KO mice developed lower antibody responses than control mice. Interestingly, vaccine supplementation with an inhibitor of the serine protease elastase (NEI) enhanced the titers of vaccine-specific serum antibody responses in CFTR KO mice and CFTR heterozygote (CFTR Het) mice. We have previously reported that NEI supplementation could promote mucosal immunity in mice immunized with the alum-based injected vaccine. In this regard, CFTR KO and CFTR Het mice immunized with NEI-supplemented alum-based injected developed higher IgA and IgG responses in saliva than control WT mice. These results provide new insights on the regulatory role of CFTR for B cell differentiation and function. They also suggest that vaccine supplementation with NEI could improve protection of individuals leaving with CF against respiratory pathogens.
NIH R01 DK101323
Investigating the immunomodulatory effects of histotripsy ablation for osteosarcoma
Authors: Patwardhan Manali Nitin , Hay Alayna , Vickers Elliana , Hixson Haleigh , Ruger Lauren , Allen Irving C , Vlaisavljevich Eli , Tuohy Joanne 
Published: August 2023

Abstract
Osteosarcoma (OS) is the most common primary bone cancer in humans and dogs. Canine OS shares many biological similarities with human OS and is a valuable comparative oncology research model. In both species, metastasis remains a major hurdle against improved survival, thus warranting a novel therapeutic approach such as histotripsy, a non-thermal, non-invasive, focused ultrasound ablation modality. Histotripsy can mechanically disintegrate gross tumor and potentially induces an anti-tumor immune response to inhibit metastatic development. The objective of this in vitro study was to investigate the immunomodulatory effects of histotripsy ablation of canine OS. We hypothesized that histotripsy ablation of OS promotes a pro-inflammatory immune signature in primary monocytes and macrophages. We isolated CD14+ primary monocytes using positive selection immunomagnetic cell separation from peripheral blood mononuclear cells of healthy dogs, and differentiated primary macrophages by culturing primary monocytes with canine GM-CSF for 72 hours. We co-cultured primary monocytes and macrophages with histotripsy-ablated canine D17 OS cells for 24 hours, then characterized their cell surface markers using flow cytometry and performed multiplex supernatant cytokine analysis. We observed upregulation of CD80 and CD62L on primary monocytes and a downregulation of CD206 on macrophages exposed to histotripsy-ablated D17 cells compared to controls, suggesting immune activation. We observed decreased levels of the pro-tumor cytokine, TGF-β, from primary monocytes exposed to histotripsy-ablated D17s compared to controls. Our results indicate that histotripsy ablation of OS can induce a pro-inflammatory immune signature.
This work is supported by grants from the American Kennel Club Canine Health Foundation, the Focused Ultrasound Foundation.
TGFβ1 and BMP signals modulate CD8+ T cell functionality and responsiveness to immune checkpoint blockade therapy
Authors: Saadey Abbey , Yousif Amir , Osborne Nicole , Shahinfar Roya , Castillo Chelsea , Laster Brooke , Rajeev Meera , Kumar Shrea , Webb Amy , Ghoneim Hazem 
Published: August 2023

Abstract
T cell dysfunctionality is a major challenge during chronic infections and cancer, limiting durable responses to T-cell based immunotherapies including immune checkpoint blockade (ICB). However, the role of upstream environmental signals in modulating the development of terminal dysfunction in CD8+ T cells remains to be explored. We have recently established that persistent TGFβ1 exposure drives an epigenetically stabilized, terminal dysfunction program in chronically stimulated human and mouse CD8+ T cells. In contrast, boosting BMP signaling induces a striking ability of T cells to maintain effector functionality and survival under chronic stimulation. Here, we show how rebalancing TGFβ1/BMP-signals in dysfunctional CD8+ T cells retains functionality in chronically stimulated human cord blood or tumor-infiltrating CD8+ T cells in vitro, while enhancing chronic virus or tumor control in mice. We also explore the therapeutic potential of BMP-signals in promoting durability of CD8+ T cell responses. Altogether, these findings provide an exciting new approach to enhance T cell responses to immunotherapy.
Sulfides in the gut mediate protection against gastrointestinal infection via alterations to local immunity and the microbiome
Authors: Band Victor I , Stacy Apollo , Chau Joanna , Belkaid Yasmine 
Published: August 2023

Abstract
Sulfide is a gaseous molecule, which has toxic effects at high concentrations yet plays key roles in homeostasis throughout the body. Sulfides are produced endogenously by both host tissues and the bacterial cells of the gut microbiome, which results in the gut containing the highest concentrations of sulfide in the body. We sought to assess the role of the highly abundant sulfide molecule in gut immunity, microbiome homeostasis and resistance to enteric infections. Local sulfides can be depleted by the compound bismuth subsalicylate (BSS), a common anti-diarrheal medication, which acts locally by sequestering sulfides in the gut. Key gut commensals such as Lactobacillus and segmented filamentous bacteria, major mediators of gut immunity and resistance to pathogen colonization, were profoundly depleted following sulfide sequestration. Additionally, we observed significant downstream immune effects, specifically within the local immunity of the small intestine. Depletion of gut sulfides resulted in profound collapse of CD4 T cells, especially among Th1 cells in the lamina propria of the small intestine. Using a mouse model of Salmonella Typhimurium, mice treated with BSS were extremely susceptible to infection, with a 5-log increase in fecal bacterial load at 24 hours post infection. These data reveal a central role for sulfides in gut homeostasis and prevention of enteric infection. Strategies to manage gut sulfide levels, including diet supplementation and microbiome engineering, could be a possible intervention to promote gut health.
NIH Office of Dietary Supplements (Research Scholar Grant 2022)
Jedi Mindset: Discussing Effective Learning Strategies with Students
Authors: Alard Pascale 
Published: August 2023

Abstract
Students’ ability to retrieve acquired knowledge long-term is critical for every discipline, especially Immunology. The research on learning shows undergraduate students use ineffective learning strategies, misjudging passive vs active ways of learning. Moreover, first generation graduates may be at a disadvantage as they have less opportunity to learn from family member experience. Sixty percent of students who attend medical, dental or graduate school continue to use ineffective strategies, e.g., re-reading. Our goal was to develop a bookclub for our incoming graduate students to discuss effective learning strategies, and assess student’s willingness to switch to more effective learning strategies as well as evaluate the benefit of the bookclub.
We held three consecutive weekly sessions before courses started to discuss the book by Dr. Saundra McGuire entitled, “Teach yourself how to learn”. Incoming graduate students read a few chapters for each session and answered questions about their reading. A discussion was facilitated with senior students, who shared their experience about the program and provided advice. Short podcasts from the UCLA Lasting Learning website were viewed to revisit the important points.
Students recognized that re-reading is not effective and were willing to modify their study habits and continued to use effective learning strategies the following semester. In addition, students agreed that the bookclub was beneficial and provided tools to help them succeed in their coursework, and emphasized the positive impact of senior students’ participation. Our study suggested that a bookclub about effective learning strategies is beneficial and can help incoming graduate students reach their full potential.
Polyomavirus ependyma infection induces region-specific and Stat1-dependent expression of chemokines in the CNS
Authors: Spencer Samantha A , Lukacher Aron 
Published: August 2023

Abstract
Immunity of the central nervous system to pathogens is partly established by critical barrier sites. One barrier within the four ventricles, the blood-cerebrospinal fluid (CSF)-brain barrier, is comprised of the choroid plexus epithelial cells separating the vasculature from the CSF, and the CSF from the brain parenchyma by the ependymal cells lining the ventricles. Intracerebroventricular delivery of mouse polyomavirus (MuPyV) in wild-type mice leads to infection of ependymal cells. In Stat1 null mice, viral infection is increased, and ependyma effacement is associated with hydrocephalus. Unknown is whether innate immunity and immune cell recruitment via Stat1 signaling by the ependyma limits MuPyV infection of the parenchyma. Dcdc2α-Cre mice, which show recombination in the ependyma and choroid plexus, were crossed with Stat1fl/fl mice to yield ependyma-specific Stat1 knockout. MuPyV infection was greater in Dcdc2αCre+ x Stat1fl/fl mice than in wild-type (WT) controls at 4 days post infection (dpi), but by 7 dpi the Dcdc2α-Cre+ x Stat1fl/fl mice exhibited higher levels of MuPyV infection in the periventricular region. Prior studies using the MuPyV-infected Stat1 null mice showed increased monocyte and T cell recruitment. Several chemokines (Ccl2, Cxcl10, Cxcl11, and Cxcl16) were elevated at 7 dpi in Dcdc2αCre+ x Stat1fl/fl mice relative to WT mice, while Cxcl13 was reduced at 4 and 7 dpi. Of these, Ccl2, Cxcl10 and Cxcl11 showed greater levels near the ventricles in both wild-type and Dcdc2αCre+ x Stat1fl/fl mice. Our data highlight the ependyma as a critical barrier protecting the brain from MuPyV infection that is dependent on Stat1 signaling.
This work was supported by grants from the NIH (R35NS127217-01, T32CA60395-25).
Immunological and genetic analysis of vaccine strains and regional variants of infectious bronchitis virus is examined
Authors: Lee Rangyeon , Jung Ji Seung , Kwon Hyuk Moo , Sung Haan Woo , Park Jeongho 
Published: August 2023

Abstract
Infectious bronchitis virus (IBV) is an acute and highly contagious coronavirus characterized by respiratory and systemic pathology in chickens. Frequent genetic mutation in the spike (S) glycoprotein repeatedly induces regional variants. Although IBV vaccines are extensively utilized in chicken flocks, their efficacy is diminished by variant serotypes and poor cross-protection. To provide restored strategy for the IBV protection, we analyzed the genetic distance between local IBV variants and other coronaviruses, including SARS-CoV-2. We also compared the genetic sequence of the S1 glycoprotein in three commercial vaccine strains. Then, we examined immune responses by vaccine and regional strains in primary chicken embryonic kidney cells and splenocytes. Featured pathways for innate immunity and gene expressions for innate immune response was observed. Our study is expected to provide valid information for vaccine development against emerging variants.
Supported by funding from the office of Research Support, Kangwon National University, a 2020 Research Grant from Kangwon National University (no. 520200065), and a National Research Foundation of Korean (NRF) grant funded by the Korea government (MSIT) (no. 2020R1G1A1099715)
Amniotic fluid derived extracellular vesicles as novel anti-inflammatory therapeutics against SARS-CoV-2 infection
Authors: Chanda Debarati , Bellio Michael A , del Rivero Tania , Mitrani Maria Ines , Channappanavar Rudragouda 
Published: August 2023

Abstract
The ongoing Covid-19 pandemic caused by SARS-CoV-2 is associated with acute respiratory distress syndrome (ARDS) and fatal pneumonia. Excessive inflammation caused by SARS-CoV-2 is the key driver of ARDS and lethal disease. Despite strong evidence for the role of virus induced inflammation in severe COVID19, no effective anti-inflammatory drug is available to control robust inflammation. Although corticosteroids are in clinical use to suppress excessive inflammation, global immunosuppressive effects of these drugs have limited their clinical success. Moreover, cytotoxicity caused by several chemically derived drugs is a cause of concern. Therefore, there is an urgent need to identify biologically derived immunomodulators that suppress inflammation without affecting antiviral immunity. In this study, we evaluated human amniotic fluid derived extracellular vesicles (hAF-EVs) as a potential safe biologic for immunomodulation during Covid-19. Our results showed that hAF-EVs significantly reduced inflammatory cytokine production in TLR2/4/7 and CoV structural protein stimulated mouse macrophages, in-vitro. Importantly, we also found that intraperitoneal administration of hAF-EVs in mice infected with a mouse-adapted SARS-CoV-2 reduced morbidity and mortality. A detailed examination of infected lungs showed that the increased protection in hAF-EV-treated mice was associated with reduced myeloid cell accumulation and inflammatory cytokine production. Collectively, we identify a novel biologic that has potential to suppress excessive inflammation and promote survival following SARS-CoV-2 infection, highlighting the translational potential of hAF-EVs against Covid-19 and other viral lung infections.
This study was supported in part by institutional research funds from Oklahoma State University—Stillwater and NIGMS 5P20GM103648-10.
Fc properties of SARS-CoV-2 spike-specific antibodies elicited by vaccines and infection
Authors: Hioe Catarina E , Klingler Jeromine , Bandres Juan , Emami-Gorizi Rozita , Amanat Fatima , Kowdle Shreya , Gleason Charles , Srivastava Komal , Simon Viviana , Perandones Claudia , Lee Benhur 
Published: August 2023

Abstract
Since its emergence in 2019, SARS-CoV-2 has infected millions of people and several vaccines have been approved for clinical use worldwide. Antibodies against the spike protein of SARS-CoV-2 are a critical immune determinant for protection against this virus. While virus neutralization is a key function of spike-specific antibodies, antibodies also mediate Fc-dependent activities that can play a role in protection or pathogenesis. This study compared the Fc properties and functions of anti-spike antibodies elicited by Pfizer/BioNtech or Moderna mRNA vaccines, Sputnik rAd26+rAd5 vaccines, or natural SARS-CoV-2 infection. We first examined immunoglobulin (Ig) isotypes in sera or plasma from the three groups. IgG1 antibodies against the spike protein were the predominant isotype, produced to high levels by the vast majority of the subjects in the three groups. In addition to IgG1, the mRNA vaccines also elicited IgM, IgG2-4, and IgA1-2, whereas the Sputnik vaccines induced only IgG4. Convalescent COVID-19 patients had yet a distinct set of isotypes, displaying mainly IgM and IgA1 alongside IgG1. When we examined antibody functions, spike-specific antibodies from both mRNA and Sputnik vaccine groups showed greater capacities to bind and activate complement than those from the convalescent COVID-19 patients. However, the Sputnik vaccine-induced antibodies had a greater potency of binding the ADCC-activating FcγRIIIa receptor, although the Sputnik vaccines induced lower Ig levels against spike than the mRNA vaccines. These data demonstrate that antibodies with distinct Ig isotypes/subtypes and Fc functions were elicited by different COVID-19 vaccines and by natural SARS-CoV-2 infection.
This work was supported in part by the Department of Veterans Affairs [Merit Review Grant I01BX005794] (C.E.H.) and [Research Career Scientist Award 1IK6BX004607] (C.E.H.); the National Institutes of Health [grant AI139290] to C.E.H., [grants R01 AI123449, R21 AI1498033] to B.L, the Department of Medicine of the Icahn School of Medicine at Mount Sinai (C.E.H.); the Department of Microbiology and the Ward-Coleman estate for endowing the Ward-Coleman Chairs at the Icahn School of Medicine at Mount Sinai (B.L.), and institutional funding as well as anonymous philanthropic donations (V.S.).
Biomarkers of Neurological Injury for Emerging Viral Threats and Post Viral Disease
Authors: Bartlett Maggie , Poeck-Goux Heather , Johnson Linwood Johnson , Scully Kevin , Vittor Amy , Hayes Ronald , Carrera Jean-Paul , Smith Darci R 
Published: August 2023

Abstract
Neurotropic viral infection and the ensuant immune response are a significant cause of morbidity and mortality worldwide, which can range in severity from mild to permanent central nervous system (CNS) damage and death. Encephalitic alphaviruses include Venezuelan and eastern equine encephalitis viruses (VEEV and EEEV; Alphavirus; Togaviridae). Injury to the CNS is an important determinant of poor outcome and tools to predict this outcome are lacking. Neurons are the primary target cells of encephalitic alphaviruses where cytopathology plays a major role in CNS dysfunction. Proteins are released following cell death, such as ubiquitin carboxy-terminal hydrolase L1 (UCH-L1) and glial fibrillary acidic protein (GFAP) are neuromarkers of CNS tissue injury which could serve as biomarkers to assess injury severity, monitor disease progression, direct treatment, and as reliable endpoints to help develop novel medical countermeasures. Recent advances in the use of blood-based biomarkers for diagnosis of traumatic brain injury (TBI) which have FDA approved assays have provided a scientific foundation for expanding the biomarker technology to brain damage caused by other CNS pathologies like viral encephalitis. Here we evaluated the ability to detect these biomarkers for encephalitic alphaviruses, encephalitis of unknown origin, and hospitalized patients with severe coronavirus disease (COVID-19). Higher levels of specific biomarkers were detected in patients diagnosed with alphavirus or unknown encephalitis which may be useful in prognosis and treatment guidance of post viral disease. Collectively, our results suggest that these blood-based biomarkers may be a good indicator for brain injury resulting from viral infection.