bluuebunny/crossref_metadata_2025_split huggingface dataset:
Reduced T cell priming to vaccination in microbially-experienced (‘dirty’) mice
Authors: Sjaastad Frances V , Huggins Matthew A , Swanson Whitney , Kucaba Tamara A , Hamilton Sara E. , Griffith Thomas S 
Published: January 2023

Abstract
Successful vaccination strategies can provide lifelong immunity against infectious diseases and cancer. Vaccines are widely considered one of the greatest advancements in public health; however, recent failures in vaccine development indicate our inadequate understanding of the immune response to vaccination. There has been increased attention recently regarding the limited translation of preclinical findings generated using specific pathogen free (SPF) laboratory mice to humans. One potential reason for the difference between laboratory mice and humans lies in maturation status of the immune system at the time of challenge. We have been using a ‘dirty’ mouse model, where SPF laboratory mice are cohoused with pet store mice to permit pathogen transfer and immune system maturation, to investigate the naive T cell response after peptide vaccination with Trivax (polyI:C and anti-CD40 mAb) or Complete Freund’s Adjuvant (CFA). Despite having no differences in the number of antigen-specific naïve CD4 and CD8 T cell precursors, the expansion of these CD4 and CD8 T cell populations in cohoused (CoH) B6 mice was significantly reduced (compared to SPF B6 mice) following vaccination with either Trivax or CFA. Furthermore, memory recall responses to cognate antigen – as measured by proliferative capacity or cytokine production – were reduced in the CoH mice. Evaluation of the dendritic cell compartment revealed reduced expression of costimulatory molecules (e.g., CD80/CD86) on DC from CoH mice shortly after vaccination. One consequence of the muted T cell priming in CoH mice was reduced antitumor immunity after vaccination. Collectively, our data show the potential limitation of exclusive use of SPF mice when testing vaccine efficacy.
Effect of CRISPR/Cas9-mediated PD-1-disrupted primary human third-generation CAR-T cells targeting EGFRvIII on human glioblastoma cell growth
Authors: Nakazawa Tsutomu , Natsume Atsushi , Nishimura Fumihiko , Matsuda Ryosuke , Nakamura Mitsutoshi , Nakagawa Ichiro , Motoyama Yasushi , Park Young Soo , Tsujimura Takahiro , Wakabayashi Toshihiko , Nakase Hiroyuki 
Published: January 2023

Abstract
Glioblastoma (GBM) is the most common and malignant brain tumor. As GBM is resistant to standard treatment, immunotherapy might be a promising alternative. Chimeric antigen receptor (CAR) is an artificially modified fusion protein that can be engineered to direct the specificity and function of T cells against tumor antigens. The antitumor effects of EGFRvIII-targeting CAR-T (EvCAR-T) cells are limited in GBM, on account of the interaction between programmed cell death protein 1 (PD-1) on activated EvCAR-T cells and its ligands on target tumor cells. Antibodies against PD-1 have shown limitations, so permanent deletion of the PD-1 gene in CAR-T cells might be a promising solution. Therefore, in the present study, PD-1-disrupted EvCAR-T cells were established for the first time with the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) gene-editing tool. We determined the disruption efficiency of the human PD-1 gene in EvCAR-T cells and evaluated the antitumor effect of PD-1-disrupted EvCAR-T cells against human GBM cells. The results showed that our designed-sgRNA/Cas9 expression vectors precisely disrupted the target region of PD-1 and inhibited the expression of PD-1 in EvCAR-T cells. Further, the PD-1-disrupted EvCAR-T cells also had a growth inhibitory effect on EGFRvIII-expressing GBM cells. Thus, this study provides a simple, easy, and clinical applicable strategy for inducing PD-1-disrupted EvCAR-T cells, which could potentially be useful for improving the efficacy of EvCAR-T cell-based adoptive immunotherapy for GBM, and even other cancers.
Macrophages genetically engineered to secrete IL-12 induce tumor killing and immune activation in human tumor slice cultures
Authors: Kreuser Shannon A , Brempelis Katherine J , Ladabie Kevin P , Cowan Courtney M , Chinn Harrison K , Davis Amira , Pillarisetty Venu G , Crane Courtney A 
Published: January 2023

Abstract
Cell-based immunotherapies for cancer, such as chimeric antigen receptor (CAR) T-cells, are effective against certain hematologic malignancies, but less reliably overcome the immunosuppressive tumor microenvironment (TME) of solid tumors to effectively cause cancer remission. Preclinical studies testing interleukin-12 (IL-12), an immune stimulating cytokine that activates T-cells and NK cells, as an anti-tumor agent have been promising. However, clinical trials have demonstrated that IL-12 is toxic if delivered systemically. To achieve local, low-dose delivery of IL-12, we have made genetically engineered macrophages (GEMs) to produce IL-12. In vitro, IL-12 GEMs stably produce IL-12 for at least one month. To evaluate their ability to stimulate an immune response and induce tumor cell death in an intact immune environment, we co-cultured them with tumor slices from both human colorectal cancer liver metastases and pancreatic ductal adenocarcinoma. In both tumor subtypes, the addition of IL-12 GEMs increased tumor cell death and generated an inflammatory response within the tumor slice, as evidenced by an increase in interferon-gamma and C-X-C Motif Chemokine Ligand 9 and 10, when compared to control GEMs. We demonstrate our GEM platform as a promising way to locally and sustainably deliver IL-12 to solid tumors to overcome the immunosuppressive TME.
Interferon gamma signaling regulates the generation of influenza-specific memory B cells
Authors: Risley Chris A , Stone Sara L , Scharer Christopher D. , Boss Jeremy M , Lund Frances E 
Published: January 2023

Abstract
Memory B cells(Bmem), which can rapidly differentiate into antibody secreting cells (ASC) upon secondary challenge, are key mediators of humoral immunity. However, the signals that govern the formation and function of these cells remain poorly understood. To date, most Bmem studies have been performed with alum-adjuvanted proteins that induce type 2 immune responses. This differs significantly from the type I and II interferon (IFN) response generated after viral infection. Using a murine model of influenza, we showed that Bmem cell intrinsic expression of the IFNγ-inducible transcription factor (TF) T-bet was required for Bmem secondary ASC responses following viral challenge. Here, we demonstrate that B cell-specific deletion of the IFNγR and the IFNg-induced TF STAT1 also affects the flu-specific Bmem compartment and greatly diminishes Bmem recall responses. We demonstrate that T-bet is expressed by a subset of all flu-specific Bmem, including those expressing IgG1 and IgM receptors. Therefore, we hypothesized that T-bet expression could be used to subset the Bmem cells into those that did or did not receive prior IFNg and STAT1 programming signals. Sorting Bmem from flu-infected mice, we show that Bmem cells with the highest levels of T-bet differentiate quickly and robustly following ex vivo stimulation. By contrast, those Bmem expressing lower levels of T-bet differentiate with delayed kinetics. Taken together, these data support the notion that B cell intrinsic IFNγR signaling, mediated by STAT1, drives formation of a distinct population of T-bet expressing effector memory B cells that are programmed to rapidly differentiate following re-exposure to antigen.
Human Lymph: Formation Transport and Immunological role
Authors: Santambrogio Laura , Clement Cristina C , Stern Lawrence J. 
Published: January 2023

Abstract
The pre-nodal afferent lymph is the fluid which directly derives from the extracellular milieu from every parenchymal organ and, as it continues to circulate between the cells, it collects products deriving from the organ metabolism/catabolism. A comprehensive qualitative and quantitative investigation of the self-antigenic repertoire transported by the human lymph and its role in immunological recognition is still missing and is the focus of this investigation.
Using advanced proteomics and biochemical approaches; we sequenced the proteome/degradome/peptidome of the human lymph and compared it with the MHC II peptidome displayed by HLA-DR1+ dendritic cells.
Analysis of more than 3000 sequences identified self-peptides derived from both intracellular and extracellular proteins revealing the variety of catabolic products transported by human lymph. Quantitative analysis established that at least some of these peptides are present in the circulating lymph in nanomolar concentration. Processing enzymes were identified by reference to a database of experimentally determined cleavage sites. We found that self-antigens processed by tissue-specific proteases, including matrix metalloproteases, caspases, and granzymes, and carried by lymph contribute to the MHC II self-peptidome presented by conventional dendritic cells in vivo. Our work brings new attention to the role of “immunological self-recognition” as a dynamic interaction between dendritic cells and the lymph-transported metabolic/catabolic products from parenchymal organ derived from tissue growth, remodeling, and physiological apoptosis.
Oral feeding as well as intravenous administration of biodegradable nanoparticles containing antigen and dexamethasone induces antigen-specific immune tolerance
Authors: Kim Sang-Hyun , Park Ha-Eun , Kim Kyungjae , Lee Chong-Kil 
Published: January 2023

Abstract
Polymeric nanoparticles containing both antigens and drugs have remarkable advantages because they can specifically deliver the antigens and the drugs to phagocytes such as dendritic cells and macrophages, thereby reducing the potential systemic side effects of the drugs. In the present study, we fabricated polymeric nanoparticles (NP) containing both ovalbumin (OVA) and dexamethasone (Dex) (NP[OVA+Dex]) using biodegradable and biocompatible polymer poly(lactic-co-glycolic acid) (PLGA), after which their immunomodulatory effects were examined. Immature dendritic cells treated with NP[OVA+Dex] were profoundly suppressed in both MHC class II- and class I-restricted OVA peptide presentation. In addition, mice injected with NP[OVA+Dex] were profoundly suppressed in generating OVA-specific cytotoxic T cells and also in producing OVA-specific IgG with concomitant increase of regulatory T cells. Furthermore, feeding of mice with NP[OVA+Dex] induced OVA-specific immune tolerance, as demonstrated by significant decrease in generating OVA-specific immune responses upon subsequent intravenous challenge with the nanoparticles containing OVA only. The present study demonstrates that oral feeding as well as intravenous injection of PLGA nanoparticles encapsulated with both antigen and Dex is a useful means for inducing antigen-specific immune tolerance.
Podocyte-targeting autoimmunity promotes diabetic nephropathy progression
Authors: Li Limin , Li Shan , Liu Ying , Zhang Mingchao , Zen Ke 
Published: January 2023

Abstract
Diabetic nephropathy (DN) is the leading cause of kidney failure worldwide. High levels of advanced glycation end products (AGEs) derived from chronic hyperglycemia occur in diabetes. As AGEs are highly antigenic, it promotes us to hypothesize that AGEs may exert an autoimmune response in DN development. First, we demonstrated that podocytes have the ability to present antigens under inflammatory condition. Second, we confirmed the presence of AGEs-recognizing autoantibodies in the serum of DN patients, not nondiabetic individuals or diabetic patients without nephropathy. Third, we showed that the autoantobodies in sera of DN patients specifically identify kidney biopsies of DN patients, not nondiabetic individuals or diabetic patients without nephropathy. In particular, the autoantobodies recognize signal specificity in podocytes from DN patients. Finally, employing the kidney tissue sections from DN patients as antigen to screen serum samples from diabetic or DN patients, we found that significant DN patients (48%) but not diabetic patients without nephropathy (&lt;5%) exhibited podocyte labeling with autoantibodies. These results suggest that podocyte-targeting autoimmunity may plays an important role in promoting immune-mediated DN progression.
Effects of FcμR expression on B cells activation
Authors: Blandino Rebecca A , Lou Zheng , Cook Sarah , Sciammas Roger , Baumgarth Nicole 
Published: January 2023

Abstract
FcμR is present on the cell surface of B cells as well as other antigen presenting cells where they bind the secreted form of IgM (sIgM). Our previous studies have shown that follicular B cells internalize sIgM continuously in vivo and that lack of sIgM or the FcμR on B cells results in diminished IgG responses following influenza virus infection. We aim to determine the mechanisms underlying the need for sIgM in the development of IgG responses. For this we generated HELB-FcmR−/− and HELB-FcmR+/+ mice. In vitro stimulation of purified B cells from these mice with HEL-OVA conjugate in the presence of OVA-specific OTII CD4 T cells showed an antigen-dose-dependent stimulation of both types of B cells. However, FcmR−/− B cells proliferated less at intermediate and low doses of antigen compared to controls, while they proliferated similar as wild type B cells following high-dose antigen stimulation. Intravenous injection of CD45.1-expressing C57BL/6 mice with B cells from either FcmR-deficient or -sufficient CD45.2+ HELB mice together with sheep red blood cells conjugated to HEL showed no significant differences in HEL-specific serum IgM or IgG, or frequencies of HEL-specific B cells between mice receiving either type of B cell, as assessed by ELISA and FACS, respectively. Ongoing studies are aimed at determining whether antigen-dose affects the need for FcmR expression in vivo. For that we have created mixed bone marrow (BM) irradiation chimeras with 2% BM from CD45.2 HELB or HELB-FcmR−/− mice, and 98% CD45.1 wild type BM. To-date our studies suggest that FcmR-IgM interaction enhances B cell proliferation when antigen is limiting. Future studies are aimed at testing this conclusion and explore how sIgM-FcmR interaction enhanced B cell responses.
The E3 Ubiquitin Ligase Itch Restricts the Magnitude of Antigen-Driven B Cell responses
Authors: Moser Emily K , Roof Jennifer , Dybas Joseph , Spruce Lynn , Seeholzer Stephen , Cancro Michael P , Oliver Paula M 
Published: January 2023

Abstract
More than 23 million Americans suffer from autoimmune disease, driven in large part by germinal center (GC)-derived autoantibodies. In GC B cells, somatic hypermutation of immunoglobulin (Ig) genes can render autoreactivity, but cells bearing autoreactive Ig receptors are eliminated due to strict selection of GC B cells by mechanisms that are still poorly defined. The E3 ubiquitin ligase Itch prevents the emergence of autoimmune disease and autoantibodies in humans and mice, and patients lacking Itch develop debilitating, multi-faceted, potentially fatal, autoimmune disease; yet how Itch regulates GC B cell fate or function has not previously been explored. By studying spontaneous and immunization-induced GC B cell responses in Itch deficient mice, we now show that Itch directly limits B cell activity to shape antibody responses. While Itch-deficient mice displayed normal numbers of pre-immune B cell populations, they showed elevated numbers of antigen-experienced B cells. Mixed bone marrow chimeras revealed that Itch acts within B cells to limit follicular and GC B cell numbers. Proteomic profiling of acutely activated B cells uncovered that Itch limited abundance of a subset of proteins involved in cell cycle and mTORC1 activity. We next found that B cells lacking Itch showed increased proliferation, glycolytic capacity, and mTORC1 activation in vitro and in vivo. Moreover, stimulation of these cells by in vivo immunization resulted in elevated numbers of GC B cells, plasma cells, and serum IgG. These results support a novel role for Itch in limiting B cell metabolism and proliferation to suppress GC B cell responses, supporting the idea that overly-exuberant GC B cell responses may contribute to autoimmunity in Itch deficiency.
Genetic contribution to loss of B cell anergy during development of autoimmunity
Authors: Smith Mia J , Rihanek Marynette , Stensland Zachary , Sarapura Virginia , Gottlieb Peter , Cambier John C 
Published: January 2023

Abstract

Background
Development of autoimmunity requires the activation of autoantigen-reactive T and B cells that are normally silenced by immune tolerance mechanisms. Many potentially dangerous autoreactive B cells are silenced by anergy, wherein cells persist in the periphery in an unresponsive state.

Goals of Study
Determine whether perturbation of self-reactive anergic B cells occurs during development of autoimmunity, and whether this is associated with autoimmunity risk alleles that may act by compromising B cell anergy.

Results
Using a novel enrichment method, we assessed the status of self-reactive B cells in peripheral blood of type 1 diabetic (T1D) and autoimmune thyroid disease (AITD) patients, as well as healthy controls. We observed a &gt;90% reduction in anergic autoreactive B cells in new-onset autoimmune patients, as well as in pre-diabetic subjects and some first degree relatives (FDRs). In new-onset T1D and mice, absence of anergic insulin-reactive B cells from peripheral blood corresponded with their activation and migration to the pancreas and pancreatic lymph node. To assess any genetic contribution to subversion of B cell tolerance, we genotyped FDRs of T1D patients with normal or low levels of anergic B cells. We found that the high risk HLA:DR4-DQ8 haplotype and certain non-HLA risk alleles (PTPN2, PTPN22, IKZF3, and INS) relevant to B and T-cell development and signaling are associated with loss of anergy.

Conclusions
Findings reveal a breach in B cell anergy occurs early in development in multiple autoimmune disorders in humans, and demonstrate an association between loss of anergy and autoimmunity risk alleles.
Ikaros regulates gene expression programs required for proper BCR-activation in mature B cells
Authors: Rodriguez Princess D , Howell William , Boyd Joseph , Frietze Seth , Schjerven Hilde 
Published: January 2023

Abstract
The zinc-finger (ZnF) transcription factor Ikaros (encoded by Ikzf1) plays an essential role in regulating transcriptional programs required for B cell development, as Ikzf1null/null mice do not develop B cells. Ikaros contains two ZnF domains which function in DNA-binding and protein-protein interactions, respectively. Recently, mutations within the central DNA-binding ZnF domain of the IKZF1 gene has been linked to autoimmunity. Likewise, our murine model containing a germ-line deletion of the fourth ZnF (Ikzf1DF4/DF4) displays a B cell hyper-reactive (HR) phenotype, with B cells activated by anti-IgM without the requirement for a second co-stimulatory signal. Although Ikaros has been well-studied in early developing B cells, how loss-of-function mutations in Ikzf1 impacts downstream effector functions of mature B cells, remains largely unknown. Here, we investigated the role of Ikaros in regulating an epigenetic program required for co-stimulation-restricted activation of B cells. We utilized transcriptomic and epigenomic approaches (RNA-seq and ATAC-seq) to define transcriptional programs which dynamically change upon ex vivo stimulation and investigated their Ikaros-dependency. Interestingly, we found that the largest subset of genes deregulated between wt and Ikzf1DF4/DF4 B cells, occur prior to stimulation. In addition, CUT&RUN performed in wt B cells shows Ikaros binding to a large subset of genes expressed upon stimulation with anti-IgM and CD40L. Together, these results demonstrate that Ikaros regulates the transcriptional landscape of BCR-activated mature B cells. In the future, we will be integrating single cell RNA-seq analysis to investigate B cell heterogeneity in wt and Ikzf1DF4/DF4 B cells.
Autoantibody Prevalence in Cerebrospinal Fluid in Autoimmune Encephalitis and Related Disorders: The Need for a More Targeted Screening Approach
Authors: Klonoski Josh , Haven Thomas R , Peterson Lisa K 
Published: January 2023

Abstract
Detection of a neural autoantibody can confirm diagnosis and guide patient management and treatment. Recent recommendations suggest concurrent serum and cerebrospinal fluid (CSF) testing. However, most studies on prevalence of these autoantibodies have focused on serum testing. Herein we performed a retrospective review of CSF specimens submitted to Mayo Clinic Laboratories for CSF panel (Encephalopathy, Dementia, Paraneoplastic or Epilepsy) testing. Autoimmune encephalitis-associated and other autoimmune neurologic disorder-associated autoantibody prevalence was evaluated in conjunction with ordering patterns for CSF testing only or paired with serum testing to evaluate the clinical relevance of autoantibody testing in CSF.
An overall positivity rate of 4.63% (112/2420) was observed, with 95.54% (107/112) CSF specimens positive for a single autoantibody. The CSF positivity rates were highest for antibodies associated with autoimmune encephalitis (GAD65&gt;NMDA&gt;GABA-B&gt;LGI1&gt;AMPA&gt;VGKC) except CASPR2 which was not detected, while antibodies associated with paraneoplastic syndrome were less prevalent (ANNA-1/Hu&gt;PCA-1/Yo&gt;CRMP-5/CV2&gt;AGNA-1/SOX-1&gt;PCA-2) or not detected (amphiphysin, ANNA-2/Ri, ANNA-3 and PCA-Tr). Most CSF specimens (62.56%; n=1514) did not have a paired serum submitted for testing. Of 906 CSF/serum pairs, only 49 were CSF autoantibody positive. Rarely were antibodies identified in CSF with negative serum results while antibodies were more frequently identified in serum with negative CSF results. These data indicate that test utilization for neural autoantibodies in CSF could be substantially improved by basing ordering practices on clinical manifestations and autoantibody prevalence.
The tetraspanin CD53 impacts splenic B cell migration and function
Authors: Greenberg Zev Joshua , Schuettpelz Laura Graves 
Published: January 2023

Abstract
Tetraspanins are a family of transmembrane proteins important for membrane organization and regulation of cellular migration, adhesion, and activation (Hemler, M.E., Nat. Rev. Mol. Cell Biol., 2005). The tetraspanin CD53 has been implicated in B cell development and function, and human deficiency of CD53 results in recurrent infections and reduced serum immunoglobulin (Mollinedo, F. et al., Clin. Diagn. Lab. Immunol., 1997). We have previously reported a novel function for CD53 in supporting early B cell development by maintaining IL-7 signaling (Greenberg, Z.J., et al., J. Immunol., (in press)). However, characterization of CD53 in peripheral B cell subsets has not been described. We found that Cd53−/− mice have significantly fewer splenic transitional 1 (T1, 48% reduced, p&lt;0.01) and follicular B (Fo, 30% reduced, p&lt;0.05) cells than wild type (WT) littermate controls. Congruent with human data, Cd53−/− mice have reduced resting serum IgG and IgM (40% reduced, p&lt;0.01) compared to WT. CD53 has been shown to interact with integrin α4 and we found in vitro chemotaxis of both cultured primary B cells and B220-enriched splenocytes towards CXCL12 was significantly impaired in the absence of CD53 (45% reduced, p&lt;0.005). The lymph nodes of naïve Cd53−/− mice were smaller (35% reduced, p&lt;0.05) with significantly fewer B cells (65% reduced, p&lt;0.0001) than WT, furthermore suggesting that CD53 may be important for B cell localization. Interestingly, surface expression of CXCR4 and VLA-4 was significantly upregulated on Cd53−/− T1, T2, and marginal zone B cells (15% increased, p&lt;0.05). In summary, we hypothesize that CD53 maintains naïve chemotactic signaling in splenic B cell populations and is necessary for proper B cell localization.