bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison of antibody gene mutation patterns in first attack optic neuritis and transverse myelitis leading to Multiple Sclerosis (47.18)
Authors: Ligocki Ann , Rounds William , Cowell Lindsay , Chaudhary Parul , Xiang Diane , Guidry Paula , Scheuermann Richard , Racke Michael , Courtney Ardith , Frohman Elliot , Greenberg Benjamin , Monson Nancy 
Published: January 2023

Abstract
Multiple Sclerosis (MS) is the leading disease of the central nervous system and a significant, costly cause of disability in young adults. Patients at risk for MS often present with a single demyelinating event, typically called a “clinically isolated syndrome” (CIS). A CIS can occur in the brain, spinal cord (Acute Partial Transverse Myelitis, APTM), or optic nerve (Optic Neuritis, ON) at similar rates and present a challenging diagnostic and therapeutic dilemma. Stratifying CIS patients most likely to develop MS is a complicated process, but desirable since treatment with the appropriate immunomodulatory agents early in the disease course can delay long-term disability. Thus, a primary focus of our laboratory has been to develop a completely novel type of biomarker that can identify CIS patients that will develop MS. This unique biomarker is based on a pattern of antibody gene mutations (i.e. antibody gene signature or “AGS”) that is exclusive to B cells from the cerebrospinal fluid and brain lesions of MS patients that initially presented with either TM or ON. In fact, the AGS is consistently elevated in patients with one attack of ON that go on to develop definite MS, and can predict conversion to MS with 91% accuracy. More recently, we are testing the hypothesis that the AGS serves as a predictor for conversion to MS in CIS patients that present with TM. Preliminary data demonstrate that the AGS is also prevalent in CIS patients presenting with TM.
Overexpression of short CYLD leads to autoimmunity and production of autoantibodies (47.20)
Authors: Reissig Sonja , Hövelmeyer Nadine , Weih Debra , Weih Falk , Nikolaev Alexei , Waisman Ari 
Published: January 2023

Abstract
CYLD is a tumour suppressor gene which plays an important role in NF-kB signaling. To analyze the function of CYLD in vivo we used the CYLDex7/8 mouse strain, which is characterized by loss of the full-length transcript and overexpression of a naturally occurring short splice variant of the cyld gene (sCYLD). This overexpression resulted in splenomegaly and lymphadenopathy. Further, the B cell numbers in spleen and lymph nodes were increased at the expense of T cells. CYLDex7/8 mice showed a strong reduction of CD4 single positive (SP) and CD8 SP T cells in the thymus and periphery. In in vitro and in vivo studies we could show that CD4+ T cells display a hyperactive phenotype accompanied with an increased production of inflammatory cytokines. We could also demonstrate a defect in negative selection of CYLDex7/8 thymocytes by using the HY-TCR transgenic system. mTECs in the thymus, which are important for the negative selection of autoreactive T cells, were drastically reduced in number. Since abnormal T cell responses are often associated with chronic inflammations, we examined if CYLDex7/8 mice develop immunological abnormalities. Importantly, we could observe that CYLDex7/8 mice crossed onto TCR transgenic background developed severe colonic inflammation accompanied with high production of nuclear autoantibodies.
Impact of TLR9 on hyper-responsive ITAM signaling in a mouse model of SLE (47.24)
Authors: Mills Robyn , Lam Viola , Tobler Diana , Oksenberg Nir , Cresalia Nicole , Anderson Mark , Mao Jian-Hua , Weiss Arthur , Hermiston Michelle 
Published: January 2023

Abstract
Anti-nuclear autoantibody (ANA) production by autoreactive B cells is a hallmark symptom of the autoimmune disease Systemic Lupus Erythematosus (SLE), which has a complex genetic etiology with environmental influences. In the CD45E613R knock-in mouse model of SLE, mice homozygous for an activating point mutation in CD45 develop high titer anti-dsDNA autoantibodies on the BALB/c (BA) and not the C57BL6/J (B6) or 129/Sv genetic backgrounds despite hyper-responsive ITAM signaling in several immune lineages in each genetic background. A screen for genetic modifiers of autoantibody production identified the candidate gene Tlr9. Sequence analysis of B6 and BA alleles of Tlr9 indicate several point mutations, resulting in five amino acid substitutions in TLR9. Splenic CD11b+ myeloid cells from BA express lower levels of TLR9 and BA-derived BMDCs are less responsive to synthetic TLR9 ligand CpG DNA. BA B cells are also less responsive to TLR9-mediated signaling than B6 B cells despite similar levels of TLR9 protein, indicating the BA TLR9 allele is hypo-responsive. Genetic ablation of TLR9 in CD45E613R mice on the non-autoreactive B6 genetic background augments disease, with mice developing splenomegaly and increased frequencies of memory T cells by 3 months of age. Some aged animals also developed ANAs, supporting the hypothesis that reduced TLR9 responsiveness permits increased autoreactivity in the context of hyper-responsive ITAM signaling.
Regulation of the NKG2D ligand Mult1 by shedding (48.22)
Authors: Deng Weiwen , Nice Timothy , Raulet David 
Published: January 2023

Abstract
Natural Killer group 2D (NKG2D) is an activating receptor expressed by NK cells and some T cells that recognizes a diverse family of ligands, including the Rae-1 family, H60, and Mult1 in the mouse, and the MICA and ULBP families in humans. The binding of NKG2D to its ligands plays an important role in immunosurveillance of tumors and infectious pathogens, but disregulation of this system has been tied to autoimmunity and chronic inflammatory disease. Intriguingly, some of human NKG2D ligands such as MICA and MICB can be shed from tumor cells and have been detected in sera from patients with various types of cancer. Here we report that the mouse NKG2D ligand Mult1 protein is shed from the surface of cells and that the resulting soluble Mult1 can still bind to NKG2D. While cell-bound Mult1 causes downregulation of surface NKG2D, soluble Mult1 did not alter NKG2D expression on NK cells in vitro. Furthermore, shedding of Mult1 could be blocked by the general matrix metalloproteinase (MMP) inhibitor BB94. Finally, soluble Mult1 is elevated in the sera from Eu-Myc transgenic mice, a strain that develops B lymphomas. This study represents the first instance of NKG2D ligand shedding in the mouse, which provides an important animal model to probe the mechanisms underlying shedding and its impact on immune recognition.
Effect of complement C5a in tumor progression and the local tumor microenvironment (48.27)
Authors: Gunn Lacey , Ding Chuanlin , Hansen Richard , Hu Xiaoling , Cai Yihua , Aggarwal Deep , Qi Chunjian , Ma Yungfeng , Yan Jun 
Published: January 2023

Abstract
Promoting complement activation may enhance immunological mechanisms of anti-tumor antibodies for tumor destruction. However, complement activation components, such as C5a, trigger inflammation which can promote tumor growth. It has been demonstrated, in one tumor model, that C5a in the tumor can inhibit anti-tumor CD8 T cell responses through effects on myeloid-derived suppressor cells. We address the role of C5a on tumor growth by transfecting tumor cells with mouse C5a: SKOV-3 human ovarian carcinoma and RMA murine lymphoma. The two models allow us to study C5a in immunocompromised and immunocompetent hosts. In vitro growth kinetics of C5a, control vector (CV), or wild-type cells revealed no significant differences. In both models, tumor-bearing mice with C5a-transfected tumor cells have significantly less tumor burden as compared to CV tumors. In the immunocompromised model, DX5 CD11b NK cells and F4/80 macrophages infiltrated C5a expressing tumors with significantly greater frequency while VEGF and arginase production were significantly less. The syngeneic model revealed C5a leads to a greater tumor influx of CD11c and CD4 T cells. A greater percentage of CD4 T cells infiltrating RMA C5a tumors demonstrated an activated status, CD44hi CD62L low. Thus our studies demonstrate tumor C5a chemoattracts innate NK cells and macrophages while downregulating negative mediators, and promotes infiltration of important antigen-presenting cells and activated CD4 T effector cells.
Tumors adapt rapidly to thwart vaccine-induced T cell attack but this adaptive response can be overcome by manipulation of costimulation and immune checkpoints (48.30)
Authors: McGray Aj , Bernard Dannie , Kelly Ryan , Jha Mayank , Gregory Caitlin , Bassett Jennifer , Wan Yonghong , Bramson Jonathan 
Published: January 2023

Abstract
We have developed a prototypic recombinant adenovirus vaccine encoding dopachrome tautomerase (rHuAd5-hDCT) that leads to moderate suppression of B16F10 melanoma growth through a CD8+ T cell- and IFN-γ-dependent mechanism. The immunogenicity of rHuAd5-hDCT was improved by concomitant 4-1BB stimulation leading to enhanced control of tumor growth; however, most tumors ultimately relapsed. Gene expression analysis revealed a rapid adaptation of the tumor environment within 5 days of vaccination. Intratumoral IFN-γ and TNF-α expression increased rapidly following vaccination but, paradoxically, expression was almost completely attenuated at the time of highest infiltration by vaccine-induced CD8+ T cells. We also noted robust, yet transient, induction of PD-L1 and PD-L2 in the tumor following vaccination, which correlated with the attenuation of IFN-γ and TNF-α expression. Blockade of PD-1 signaling in combination with rHuAd5-hDCT enhanced effector cytokine expression in the tumor and improved growth control, but the tumors ultimately relapsed. Strikingly, the combination of PD-1 blockade and 4-1BB stimulation with rHuAd5-hDCT produced a synergistic enhancement in intratumoral T cell activity, resulting in complete regression despite no increase in the functionality or number of tumor infiltrating CD8+ T cells. These results punctuate the need to understand the dynamic changes within the tumor to optimize cancer immunotherapy.
NKT cell mediated responses to mantle cell lymphoma (48.35)
Authors: Sun Wenji , Kimball Amy , Hester Danubia , Webb Tonya 
Published: January 2023

Abstract
Mantle cell lymphoma (MCL) is an aggressive subtype of Non-Hodgkin’s lymphoma that is characterized by the abnormal accumulation of CD20+CD5+ B cells in the lymph nodes, spleen, gastrointestinal tract, bone marrow and blood. Although treatment with combination chemotherapy can be effective, most patients relapse, and the outcome for MCL remains poor. Natural killer T (NKT) cells are important in regulating immune responses to tumors. The observation that lymphoma patients have a reduction in both NKT cell number and function suggests that the anti-tumor effects mediated by these cells may be compromised. In these studies, we have investigated NKT cell activation in response to MCL. We found that CD1d cell surface expression was highly variable in a panel of well-characterized MCL cell lines. Importantly, cell surface expression of CD1d did not directly correlate with the ability of the MCL lines to activate primary NKT cells. Circulating NKT cells in the peripheral blood of MCL patients were not stimulated by autologous B cells, even in the presence of the potent antigen, α-GalCer. However, when NKT cells from healthy donors were co-cultured with purified B cells from MCL patients, the NKT cells specifically recognized the malignant B cells and produced IFN-γ, but not inflammatory or regulatory cytokines. These data suggest that B cell lymphomas may condition autologous NKT cells not to recognize them in order to evade NKT cell mediated anti-tumor responses.
BALB/c mouse model of RSV clinical isolate A2001/2-20 pathogenesis (49.13)
Authors: Stokes Kate , Chi Michael , Sakamoto Kaori , Newcomb Dawn , Huckabee Matthew , Lee Sujin , Pretto Carla , Williams John , Peebles R. , Moore Martin 
Published: January 2023

Abstract
Airway mucus is a hallmark of respiratory syncytial virus (RSV) lower respiratory tract illness. Laboratory RSV strains (A2, Long, and Line 19) differentially induce airway mucus production in mice. We tested whether RSV strains differ in pathogenesis by screening low passage RSV clinical isolates for mucogenicity and virulence in BALB/c mice. RSV clinical isolates induced variable disease severity. Infection of BALB/c mice with clinical isolate RSV A2001/2-20 (2-20) resulted in greater disease severity and higher lung IL-13 levels than infection with RSV laboratory strains and other clinical isolates. Like the line 19 strain, 2-20 induced airway mucin expression in BALB/c mice and resulted in higher viral antigen levels in the bronchiolar epithelium than A2. 2-20 caused epithelial desquamation, airway hyperresponsiveness, and increased breathing effort in BALB/c mice. We hypothesize that the fusion (F) protein of 2-20 contributes to this phenotype. We generated a chimeric RSV by replacing the F gene of A2 with the F gene of 2-20. A2-2-20F grew to similar levels as 2-20 in mouse and human cells in vitro. In BALB/c mice, the A2-2-20F exhibited viral loads similar to 2-20 over a time course and showed comparable mucin levels to 2-20. We established a mouse model of clinical isolate strain-dependent RSV pathogenesis that recapitulates key features of RSV disease. Identification of virulence factors of RSV 2-20 may contribute to our understanding of RSV disease mechanisms.
P-selectin regulation of T cell immunity during influenza virus infection (49.14)
Authors: Tinoco Roberto , Deiro Mia , Lin Melissa , Bradley Linda 
Published: January 2023

Abstract
Influenza viruses represent a major global health concern as they have the potential to generate worldwide pandemics characterized by increased morbidity and mortality. In response to infection of lung epithelial cells, an immune response is generated that will clear the virus and prevent re-infection with the same virus. T cells are critical for eliminating virus-infected cells and their localization to sites of viral replication is crucial to resolving infection. However, the underlying mechanisms regulating effector and memory T cell responses after influenza virus infection remain unclear. Using a mouse model, we demonstrated that the adhesion molecule P-selectin, is induced on lung endothelium after influenza virus infection. More importantly, virus-specific T cells acquire the ability to bind P-selectin through expression and glycosylation of its ligand PSGL1, which coincided with effector function. P-selectin binding antigen-specific CD4+ T cells produced increased levels of IFN-γ. Similarly, P-selectin binding antigen-specific CD8+ T cells also had increased IFN-γ levels in the lung when compared to non-binding CD8+ T cells. Our findings highlight an important role for the adhesion molecule P-selectin in regulating T cell responses by inducing a differential effector program, which may be important for T cell memory and immunity to influenza virus.
The C3a anaphylatoxin receptor is critical in the pathophysiology of RSV infection (49.2)
Authors: Bera Monali , Lu Bao , Martin Thomas , Cui Shun , Gerard Craig , Gerard Norma 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) is linked to severe lung disease and may lead to wheezing in infants. RSV infection in mice demonstrates infiltration of inflammatory cells, mucus cell metaplasia, and airway hyperresonsiveness (AHR) to methacholine. Previous studies demonstrated activation of complement by RSV. Other studies showed a requirement for C3 or C3a in AHR associated with antigen sensitization as well as inhalation of particulate matter. We hypothesized, therefore, that C3a is a critical component for the AHR associated with RSV infection. We found that mice lacking the C3aR (C3aR1-/-) do not develop AHR secondary to acute RSV infection in contrast to wild type mice. Further, viral clearance was accelerated in C3aR1-/-mice. The AHR phenotype was restored to wild type when C3aR1-/- mice were transplanted with wild type bone marrow cells,supporting a role for a hematopoetic cell type. T cells were the dominant cell type increased in the lungs of both wild type and C3aR1-/-mice following RSV infection, however, the increases in lung IL-23 and IL-17 were significantly lower in C3aR1-/-mice compared with wild type animals. Additionally the increased levels of the transcription factor RORγT, responsible for regulating IL17 synthesis, seen in wild type mice were greatly reduced in C3aR1-/- mice. Our findings identify a key role for activation of complement anaphylatoxin C3aR in the pathophysiology of RSV disease, and may help identify a therapeutic target in humans.
MHV68 latency modulates host immune responses to H1N1 influenza virus (49.9)
Authors: Saito Fumitake , Ito Toshihiro , Schaller Matthew , Carson William , Hogaboam Cory , Lukacs Nicholas , Standiford Theodore , Rochford Rosemary , Kunkel Steven 
Published: January 2023

Abstract
Human gammaherpesviruses such as Epstein-Barr virus (EBV) are clinically important pathogens, as diseases caused by gammaherpesvirus infection are a significant human health concern. Additionally, latent gammaherpesvirus infection is widespread, and can have a profound effect on subsequent immune responses. Murine gammaherpesvirus 68 (MHV68) is a natural rodent pathogen that has been used as a model to study the pathogenesis of human gammaherpesviruses. Like other herpesviruses, MHV68 causes acute infection and establishes life-long latency in the host. Recently, it has been shown that mice latently infected with MHV68 have resistance to unrelated pathogens in secondary infection models. We therefore hypothesized that latent MHV68 infection can modulate host response to H1N1 influenza virus, a serotype with high associated mortality rates in humans. To test this hypothesis, mice were infected intranasally with influenza virus following the establishment of MHV68 latency. Mice latently infected with MHV68 showed significantly higher survival of H1N1 influenza virus infection than mock-infected mice. Latent MHV68 infection led to lower influenza viral loads in the lungs, as well as decreased inflammatory pathology in the lungs. Collectively, our findings indicate that latent MHV68 infection has a protective effect against H1N1 influenza virus infection.
Regulatory T cell conversion by migratory but not lymphoid tissue resident dendritic cells (50.11)
Authors: Broggi Achille , Vitali Caterina , Mingozzi Francesca , Raimondi Giorgio , Zanoni Ivan , Granucci Francesca 
Published: January 2023

Abstract
There is evidence that dendritic cells (DCs) induce peripheral tolerance. Although studies on specific DC subsets have proven their ability to induce regulatory T cell conversion (iTreg cells), little is know about the DC subtypes actually able to promote peripheral CD4+ T cell tolerance to autoantigens in vivo. Here, we show that, when autoantigen presentation is not confined to specific DC subsets but is extended to all conventional DCs, steady state migratory DCs (ssmDCs) possess unique ability to induce antigen-specific iTreg cells in cutaneous lymph nodes (CLNs). Diversely, lymphoid tissue resident DCs do not show this capacity. Moreover, DC-derived CCL22 contributes to the retention of iTreg cells in CLNs and skin. Therefore migratory DCs represent the DC population devoted to the maintenance of peripheral self-tolerance.
Host defense against extracellular microbes-integrating the parts into a big picture in a clinical presentation-based medical school curriculum (51.1)
Authors: Piskurich Janet 
Published: January 2023

Abstract
The 2009 Scientific Foundations for Future Physicians Report of the Association of American Medical Colleges (AAMC)-Howard Hughes Medical Institute (HHMI) Committee recommends a list of specific competencies that all medical students should demonstrate before receiving the M.D. degree. Competency M4, most related to immunology, recommends students should demonstrate the ability to “Apply the principles of the cellular and molecular basis of immune and nonimmune host defense mechanisms in health and disease to determine the etiology of disease, identify preventative measures, and predict response to therapies”. An associated learning objective states that students should be able to “Apply knowledge of the mechanisms utilized to defend against intracellular or extracellular microbes to the development of immunological prevention or treatment”. Yet medical students, many learning the various parts of the immune system for the first time, may express frustration that they cannot assemble the parts into a comprehensive big picture. One integrated approach for presenting host defense mechanisms against extracellular microbes will be illustrated.
Successful integration of practical flow cytometric experience into undergraduate education. (51.4)
Authors: Rogers Clare , Gubbels Bupp Melanie , Kim Unyoung , DeKrey Gregory , Stranford Sharon , Medh Rheem , Tomich Gloria 
Published: January 2023

Abstract
Flow cytometry is an essential tool in almost every discipline of cell biology, and is increasingly utilized in a variety of other associated yet diverse fields, including molecular biology, bioengineering, microbiology and marine biology. For many years, access to the use of flow cytometers as teaching tools, even in large universities, was limited to graduate programs, due to the cost of instrument purchase and maintenance, and to the complexity of operation. Recently, several affordable, bench-top flow cytometers have appeared on the market. The affordability and user-friendly nature of these instruments have made them attractive to the faculty of various departments at medium- and small-sized universities and colleges for use both as teaching tools and in faculty research projects. We have compiled information here on the funding sources for both purchase and maintenance of these instruments at our various institutions, and share information on applications, protocols and lab exercises that have worked well with our students. Example protocols described include applications such as immunophenotyping, apoptosis (annexin/propidium iodide), monocytic phagocytosis, intracellular and mitochondrial reactive oxygen species detection, and detection of fluorescent protein expression in bacteria.
A forensic analysis of food poisoning using a dip-stick immunoassay (51.9)
Authors: Goudie John 
Published: January 2023

Abstract
Since bacteria are ubiquitous, they are likely to be present on most foods that you purchase. Proper cooking or processing of food destroys bacteria but poor personal hygiene, improper cleaning of storage and preparation areas and unclean utensils cause contamination of raw and cooked foods and allow bacteria to grow. Although cooking destroys the bacteria, the toxins produced by some are heat stable and may not be destroyed and remain in or on food when it is eaten. Students will examine food samples from a simulated (non-toxic) food poisoning outbreak for the presence of a specific antigen by utilizing a Dip-stick immunoassay. Dip-stick immunoassays are a less expensive technology for high schools and produce excellent results.