bluuebunny/crossref_metadata_2025_split huggingface dataset:
CCR10 regulates intestinal immune homeostasis through regulating both Treg and Teff cells in the intestine (MUC2P.832)
Authors: Hu Shaomin , Yang Jie , Xiong Na 
Published: January 2023

Abstract
Intestinal immune homeostasis is an active process of balanced tolerant responses towards the gut-resident commensals and other food-born stimulations. Failure of the balanced response could lead to inflammatory diseases and cancers. Understanding molecular mechanisms that differentially regulate the homeostatic vs. inflammatory activation of T cells are fundamentally important in the intestinal immunity. CCR10, through interaction with its mucosa-specific ligand CCL28, was previously found important in regulation of IgA antibody-producing B cells in intestines. On the other hand, CCR10 was suggested to be a skin- but not gut- homing molecule for T cells. Using a novel strain of CCR10-knockout/EGFP-knockin mouse, we found that CCR10 is specifically induced on activating T cells in healthy intestines in situ and is critically important in balanced maintenance and functions of Treg and Teff cells to promote the intestinal homeostasis. On the other hand, during intestinal immune activations associated with infection or other inflammatory stimulations, the induction of CCR10 on activated intestinal T cells were suppressed that allows the T cell activation along different lineages. Our findings represent the first demonstration that a chemokine receptor is specifically associated with the homeostatic status of the intestine and reveal that there exists an auto-promoting feedback circuit of intestinal homeostasis in which the CCR10/ligand axis are a critical molecular link.
Janus-faced immunity to an enteric pathogen: how Campylobacter mediates colitis and autoimmunity (MUC2P.820)
Authors: Malik Ankit , Gadsden Barbie , Sharma Deepika , A. Dybas Leslie , Mansfield Linda 
Published: January 2023

Abstract
Campylobacter jejuni is a leading cause of foodborne enteritis and has also been linked to the autoimmune neuropathy, Guillain Barré syndrome. IL-10−/− mice serve as C. jejuni colitis model, but a mouse model for GBS is lacking. Immune mechanisms responsible for C. jejuni mediated colitis, protection from colitis or initiation of autoimmune sequelae are also largely unknown. We demonstrate that IL-10−/− mice infected with colitogenic human isolates of C. jejuni had significantly upregulated Type1/17 but not Type2 cytokines in the colon coincident with infiltration of phagocytes, T cells and Innate Lymphoid Cells. Both ILCs and T cells participated in IFN-γ, IL-17 and IL-22 upregulation but in a time- and organ-specific manner. However, T cells were necessary for colitis. Furthermore, depleting IFN-γ, IL-17 or both significantly ameliorated colitis and drove the colonic responses towards Type2 cytokine and antibody induction. In contrast, C. jejuni GBS patient strains induced mild colitis associated with blunted Type 1/17 but enhanced Type2 responses. Moreover, IL-4 and CD4+ T cell dependent Type2 antibodies (but not the Type1/17 antibodies) elicited exclusively by the GBS strains cross-reacted with peripheral nerve antigens. This lead to macrophage mediated demyelination of sciatic nerves that manifested with gait ataxia and hind limb paraparesis in the IL-10-/- mice. Therefore, this is the first mouse model of an autoimmune disease induced directly by an enteric bacterium.
A next-gen pipeline for generation, error correction and annotation of high-throughput immunosequencing data (TECH1P.842)
Authors: Emerson Ryan , Sherwood Anna , DeWitt William , Howie Bryan , Rieder Mark , Robins Harlan 
Published: January 2023

Abstract
Immunosequencing is emerging as a new technology with substantial scientific and clinical relevance in immunology, hematology and oncology. Thanks to the highly-randomized somatic rearrangement of antigen receptor genes in lymphocytes, each clonal population of lymphocytes bears a receptor rearrangement that is essentially unique. Thus, sequencing of rearranged antigen receptors in B and T cells can provide a thorough characterization of the diversity and clonal structure of the adaptive immune system. Despite the significant promise of immunosequencing, accurate and consistent methods for data generation and cleaning have proven elusive in practice. Outstanding issues include biased amplification of rearranged antigen receptor genes; computational correction of PCR and sequencing errors in raw sequencing data; alignment of receptor sequences to germline gene receptor loci for accurate reconstruction of rearrangement events; and methods for extrapolation from sequencing reads attributed to each unique rearrangement to absolute input cell numbers. Using a set of robust experiments in which carefully-controlled conditions and synthetic molecules are utilized to precisely determine correct output, we have developed an experimental paradigm and data analysis pipeline that address these concerns and set a new standard for accuracy and data quality in the field of immunosequencing.
Annotation of pseudogenous gene segments by high-throughput sequencing of rearranged lymphocyte receptor loci (TECH1P.844)
Authors: Sherwood Anna , Emerson Ryan , Williamson David , Chung Moon-Wook , Robins Harlan , Rieder Mark , Carlson Christopher 
Published: January 2023

Abstract
The adaptive immune system generates a huge breadth of antigen-specific T (TCRs) and B cell receptors (BCRs) by combinatoric shuffling of gene segments. Lymphocytes undergo somatic recombination of variable (V), diversity (D), and joining (J) region gene segments, with deletions and non-templated insertions at the junctions, collectively encoding the CDR3 region. Many of the gene segments are annotated as non-functional; however, full annotation of genes as functional, pseudogene or ORF has proven elusive. We used high-throughput sequencing to identify 100K’s of rearranged TCRB and IGH receptors from a large cohort of healthy adults. We then annotated V and J gene segments as functional or pseudogenes based on selection. Random chance predicts that less than one-third of somatic rearrangements will lead to in-frame transcripts, but only lymphocytes with a productive receptor are selected. Non-functional rearrangements (out-of-frame or pseudogene) persist only as a second allele in mature T and B cells. Based on these data we annotated the functional status of each gene segment based on the realized ratio of productive to non-productive rearrangements. Some gene segments are currently mis-annotated: in TCRB 2 presumed-functional gene segments are pseudogenes, in IgH 3 presumed-functional genes are pseudogenes and 1 annotated pseudogene is functional. Additionally, in both loci we identified cases of gene segment functional/pseudogene status segregating among healthy subjects.
Improved GAA activity in tregitope-treated pompe disease: preclinical studies (TECH1P.864)
Authors: Cousens Leslie , Koeberl Dwight , Mingozzi Federico , Terry Frances , Martin Bill , De Groot Anne 
Published: January 2023

Abstract
“Tregitopes” are immunoglobulin-derived regulatory T-cell (Treg) epitopes that lead to suppression of inflammation and, when administered with a target antigen, adaptive tolerance. One potential application of Tregitopes is mitigating immune responses to therapeutic proteins, manifest when a protein appears foreign either because it is derived from a different species or because patients express a genetic deletion or modification. In Pompe disease, genetic deficiencies in acid alpha-glucosidase (GAA) result in lysosomal glycogen accumulation throughout the body. Life-saving treatment with fully human recombinant GAA can trigger high titer anti-drug-antibodies (ADA) that interfere with drug efficacy. We report here our efforts to apply Tregitopes to provide GAA-specific tolerance induction. Therapeutic GAA protein was expressed in GAA-deficient mice by administering AAV-GAA under conditions known to induce ADA +/- Tregitopes, also delivered by AAV expression vectors. Tregitope administration inhibited ADA formation compared to vehicle-treated controls. In addition, a significant increase in GAA activity was observed for Tregitope, consistent with Tregitope inhibition of glycogen buildup in muscle. Current experiments are designed to refine the Tregitope treatment regimen to achieve clinically-relevant ADA inhibition and to evaluate cellular immune responses towards the goal of providing effective and lasting Tregitope-mediated tolerance to GAA for Pompe patients.
Deletion of both Id2 and Id3 in developing T cell promotes NKT lymphoma formation in mice (TUM2P.887)
Authors: Li Jia , Jima Dereje , Dave Sandeep , Zhuang Yuan 
Published: January 2023

Abstract
NKT lymphoma is a rare form of tumor, the etiology and pathogenesis of which is still largely unknown. There are also no animal models to study its course. The Id2 and Id3 genes and their targets, E proteins, have been demonstrated to be involved in multiple lymphoma and cancer subtypes, but their role in the development of NKT lymphomas is unexplored. Here we show that combined deletion of Id2 and Id3 by LckCre in early T cell development results in NKT cell expansion, which rapidly progresses into malignancy. Adoptive transfer experiments indicate that the malignant cells are able to invade healthy tissue. cDNA Microarray analysis of NKT lymphoma and pre-malignant NKT cells showed several cytokine signaling pathways are altered during tumor progression. Therefore, regulation of E proteins by Id2 and Id3 may play a role in NKT lymphoma development. Furthermore, our double knockout mice may provide a model to study mechanisms involved in human NKT lymphoma progression.
The change of T cell repertoire due to central tolerance determines the antigenic requirement and strategy for inducing effective anti-tumor CD8 T cell response (TUM2P.888)
Authors: Leng Qibin , Guo Nining , Zhu Kai , Zhang Na , Yang Chunhui , Wang Jianrong , Yang Juhao , Luo Kaimin , Wang Yan , Shen Hao , Zehn Dietmar , Leng Qibin 
Published: January 2023

Abstract
Tumor antigens are mainly low-avidity antigens to T cells and unable to elicit effective anti-tumor immunity. Little knowledge on how low-avidity antigens elicit T cell response in vivo further hinders designing strategies to improve anti-tumor immune response. Using a unique mouse tumor model, we found that neither native tumor antigen nor majority of altered peptide ligands (APLs) derived from tumor antigen but only intermediate avidity APL was able to elicit robust cross-reactive tumor-specific CD8 T cell response. Central tolerance shifted T cell repertoire to allow host T cells only respond to low avidity APLs and spare the lower cross-reactivity to self antigens (tumor antigens). In addition, the functional avidity of cross-reactive tumor-specific CD8 T cells was over 1000 times less than that of pathogen-type antigen-specific CD8 T cells. These results indicate that the extremely low functional avidity and low cross-reactivity to native tumor antigen are two key reasons why APLs are ineffective for eliciting cytotoxic activity of CD8 T cells to kill tumor cells. Our further studies revealed that two distinct mechanisms control the functionality and responsiveness of tumor-specific CD8 T cells. This study also discovered a novel immunization strategy to enhance the functionality and responsiveness of tumor-specific CD8 T cells. Altogether, our study will shed light on strategies to enhance the efficacy of both immunotherapy and vaccination against cancers.
Neuropilin-1 enforces Treg stability and function in the tumor microenvironment. (TUM2P.889)
Authors: Overacre Abigail , Delgoffe Greg , Vignali Dario 
Published: January 2023

Abstract
Regulatory T cells (Tregs) play an integral role in the adaptive immune system by suppressing detrimental self-reactive responses. However, Tregs have deleterious effects in cancer through suppression of the anti-tumor immune response. Therefore, it is advantageous to understand the role of Tregs in the tumor microenvironment in order to create targeted cancer therapies. Our lab has uncovered a novel function for neuropilin-1 (Nrp1) on Tregs in tumors. In the absence of Nrp1 ligation by its ligand, Sema4a, Tregs lose stability and suppressive capabilities, leading to enhanced anti-tumor immunity and reduced tumor growth or tumor clearance. We hypothesize that the loss of Nrp1 expression on Tregs reduces intratumoral stability through altered intracellular pathways and diminished Foxp3 expression, transforming Tregs from an anti-inflammatory to a pro-inflammatory T cell population. Nrp1-deficient Tregs have a hypermethylated Foxp3 CNS2/TSDR locus in the tumor microenvironment, comparable to Teff, and display TH1-like characteristics, with upregulation of Tbet, CXCR3, and IFNγ, and the reduction of suppressive molecules, IL-10 and CD73. Furthermore, in competitive environments Nrp1-deficient Tregs undermine and limit the function of wild-type Tregs. Further mechanistic analysis and fate mapping studies will elucidate the cell-intrinsic and cell-extrinsic impact of Nrp1 loss on Treg function and tumor clearance.
Gene related to anergy in lymphocytes deficiency confers spontaneous tumor regression (TUM2P.897)
Authors: Wang Junmei , Zou Qiang , Sahoo Anupama , Alekseev Andrei , Sun Shao-cong , Nurieva Roza 
Published: January 2023

Abstract
T cell tolerance is the main obstacle to successful cancer immunotherapy; thus, today it is a high priority to develop strategies to break T cell tolerance and thereafter enhance T-cell cytotoxicity towards tumor cells. We, as well as other groups, have acknowledged the E3 ubiquitin ligase, Gene related to anergy in lymphocytes (Grail), as an essential component of T-cell tolerance phenotype. Here we report that Grail expression is highly upregulated in CD8+ T cells infiltrated into the EG-7 tumors compared to T cells in lymphoid tissues, and Grail deficiency confers spontaneous protection against inoculated EG-7 tumors. Importantly, therapeutic transfer of naïve tumor-antigen-specific Grail deficient CD8+ T cells into the wild-type hosts was sufficient to reject established tumors. Mechanistically, Grail deficient CD8+ T cells exhibit augmented effector cytokine expression and enhanced cytolytic function compared to wild-type CD8+ T cells, and are resistant to the suppression mediated by regulatory T cells. Moreover, the elevated cytotoxic function of Grail-deficient CD8+ T cells essentially depends on TCR and IL-21-signaling pathway; Grail promotes IL21R ubiquitination and degradation. Altogether, our data demonstrate that Grail is a crucial factor that controls anti-tumor activity of cytotoxic T cells, thus, inhibition of Grail may be a beneficial immunotherapeutic approach to induce potent immune responses against established tumors.
TCR transgenic and patient-derived CTL killing of CTL-resistant melanoma: modulation of TRAIL apoptotic pathway and immune sensitization by epigenetic modifier SAHA (TUM2P.901)
Authors: Jazirehi Ali , Kurdistani Siavash , Economou James 
Published: January 2023

Abstract
Resistance to apoptotic signals delivered by cytotoxic T cells is likely to become an important limiting factor with increasingly effective immune therapies. We report that TCR transgenic CTLs primarily use the TRAIL apoptotic pathway, via DR5 receptor, in killing sensitive melanomas. CTL-resistant melanomas had both DR5 down-regulation and aberrant expression of apoptotic molecules. Clinically achievable concentration of FDA approved chromatin remodeling drug SAHA increased surface DR5 expression, favorably inverted the expression of apoptotic genes, and restored the sensitivity of R melanomas to MART CTL. TCR transgenic MART CTLs were used as a reliable and reproducible source of highly potent melanoma-reactive CTLs. Acquisition of CTL-resistance and its reversal by SAHA was also confirmed using naturally occurring T cells expressing endogenous MART-1 TCR derived from melanoma patients. Further, using rhTRAIL and Drozitumab explicitly verified the contribution of DR5/TRAIL pathway. Apoptosis negative (c-IAP-2 and Bcl-xL) and positive (DR5) regulators were potential candidates partly regulating CTL sensitivity. Combination of SAHA and rhTRAIL or Drozitumab destabilized mitochondria and facilitated cytosolic release of apoptotic molecules. These preclinical findings suggest that exposure to chromatin remodeling drug of immune-resistant melanomas can skew towards an intracellular pro-apoptotic milieu, increase death receptor expression, and overcome acquired immune-resistance.
IP3 3-kinase B controls hematopoietic stem cell homeostasis and prevents lethal hematopoietic failure in mice (HEM5P.228)
Authors: Sauer Karsten , Conche Claire , Broaten Blake , Schaffer Lana , Rigaud Stephanie , Westernberg Luise , Head Steven , Siegemund Sabine 
Published: January 2023

Abstract
Tight regulation of HSC homeostasis ensures life-long hematopoiesis and prevents blood cancers. The mechanisms balancing HSC quiescence with expansion and differentiation into hematopoietic progenitors are incompletely understood. Here, we identify inositoltrisphosphate (IP3) 3-kinase B (Itpkb) as a novel essential regulator of HSC quiescence and function. Young Itpkb-/- mice accumulated phenotypic HSC which were less quiescent and proliferated more than wildtype controls. Itpkb-/- HSC downregulated quiescence associated mRNAs, but upregulated activation, oxidative metabolism, protein synthesis and lineage associated transcripts. Although they showed no significant homing defects and had normal to elevated viability, Itpkb-/- HSC had a severely reduced competitive long-term repopulating potential. Aging Itpkb-/- mice lost hematopoietic stem and progenitor cells and died with severe anemia. Wildtype HSC normally repopulated Itpkb-/- hosts, indicating a HSC-intrinsic Itpkb requirement. In vitro, Itpkb-/- HSC had reduced cobblestone-area forming cell activity and showed increased stem cell factor activation of the phosphoinositide 3-kinase (PI3K) effector Akt. This was reversed by exogenous provision of the Itpkb product IP4, a known PI3K/Akt antagonist. Itpkb-/- HSC also showed transcriptome changes consistent with hyperactive Akt/mTOR signaling. Thus, we propose that Itpkb ensures HSC quiescence and function in part by limiting cytokine-induced PI3K signaling in HSC.
Blockade of PD-L1 accelerates the development of Sjögren’s syndrome in non-obese diabetic mice (HUM3P.256)
Authors: Zhou Jing , Jin Jun-O , Du Juan , Yu Qing 
Published: January 2023

Abstract
Programmed death-ligand 1 (PD-L1) is a B7-family member and plays a negative role in T cell -mediated immunity by binding to its receptors such as programmed death-1 (PD-1). Previous reports have shown enhanced expression of PD-L1 and PD-1 in the salivary gland of patients with Sjögren’s syndrome (SS), a prevalent systemic autoimmune disease characterized by progressive lymphocyte infiltration of exocrine glands and generation of autoantibodies, which together lead to impaired secretory function of salivary and lacrimal glands. However, the particular function of PD-L1 in SS disease development still remains poorly understood. Here we employed non-obese diabetic (NOD) mice, a model of SS-like disease to define the role of endogenous PD-L1 in the pathogenesis of SS. Administration of an anti-PD-L1 antibody to NOD mice enhanced leukocyte infiltration in salivary glands and elevated the levels of serum antinuclear antibodies. Importantly, PD-L1 blockade also has a crucial effect on the clinical phenotype of SS. We demonstrated blockade of PD-L1 markedly reduced salivary secretion, indicating an exacerbated secretory dysfunction. These results suggest that blockade of endogenous PD-L1 promotes the SS-characteristic pathological and clinical changes and accelerates SS development in NOD mice. Further understanding of the molecular mechanisms underlying the function of PD-L1 in SS could identify critical molecular targets for development of alternative therapeutic strategies.
Differentially expressed microRNAs decrease regulatory T cell development in multiple sclerosis (HUM3P.266)
Authors: Severin Mary , Guerau-De-Arellano Mireia , Lee Priscilla , Racke Michael , Lovett-Racke Amy 
Published: January 2023

Abstract
Multiple sclerosis (MS) is an immune-mediated demyelinating disease of the CNS. The disease is thought to be an autoimmune reaction mediated by autoreactive T cells. Interestingly, regulatory T cells (Tregs), protectors against autoimmune responses, are defective in MS patients. To better understand defects in the CD4 T cells of MS patients, a miRNA profiling study was performed on naïve CD4+ T cells of MS patients and healthy donors, identifying 85 differentially expressed miRNAs. By performing bioinformatics analyses, 23 miRNAs were predicted to target the TGFβ signaling pathway. TGFβ is critical for the development and function of Tregs. We hypothesized that dysregulated miRNAs in the naïve CD4+ T cells of MS patients target the TGFβ signaling pathway, resulting in defective Tregs and enhanced susceptibility to developing MS. To test this hypothesis, we conducted quantitative real-time PCR, flow cytometry, and luciferase assays to analyze TGFβ-associated genes and Treg markers in cells overexpressing TGFβ-targeting miRNAs. Our data indicate that 1) TGFβ signaling genes are decreased in MS patients’ naïve CD4 T cells, 2) several of the identified miRNAs directly target these TGFβ associated genes, and 3) overexpression of the TGFβ-targeting miRNAs cause a decrease in iTreg development in vitro and in vivo. Our data suggest that when overexpressed these identified miRNAs decrease TGFβ signaling and disrupt Treg development in multiple sclerosis.
Soluble mediators and clinical features predict transition to systemic lupus erythematosus classification in previously unaffected relatives of SLE patients (HUM3P.243)
Authors: Munroe Melissa , Young Kendra , Kamen Diane , Guthridge Joel , Niewold Timothy , Weisman Michael , Ishimori Mariko , Wallace Daniel , Gilkeson Gary , Olsen Nancy , Karp David , Harley John , Norris Jill , James Judith 
Published: January 2023

Abstract
Healthy relatives of lupus patients have an increased risk of developing SLE, but predictors and mechanisms of disease transition are unknown. Unaffected relatives of SLE patients (n=409) were enrolled in this follow-up study to identify factors associated with transition to SLE. Participants provided detailed demographic and clinical information, including the Connective Tissue Disease Screening Questionnaire (CSQ). Medical records were reviewed for ACR classification criteria. Plasma samples were tested for the presence of autoantibodies and soluble mediators. Forty-five relatives (11%) transitioned to classified SLE during follow-up (mean time = 6.4 years) who had more autoantibodies (p&lt;0.0001) and higher pre-classification (baseline) CSQ scores (p&lt;0.0001). Relatives who transitioned also had elevated baseline levels of inflammatory mediators, including BLyS, SCF, and IFN-associated chemokines (p&lt;0.05), with concurrent decreased regulatory mediators, including TGF-β and IL-10 (p&lt;0.05). Logistic regression analysis revealed that baseline ACR and CSQ scores (but not autoantibodies), and baseline plasma levels of SCF and TGF-β were significant and independent predictors of SLE transition. Thus, perturbations in immune-mediated processes precede SLE clinical classification and can help identify unaffected relatives at highest risk of future SLE classification for early intervention studies.