bluuebunny/crossref_metadata_2025_split huggingface dataset:
Chemerin aggravates dextran sulfate sodium-induced colitis by impairing the protective innate responses (P3274)
Authors: He Rui , Lin Yuli , Yang Xuguang , Zou Linlin , Li Bingji , Yue Wenjie 
Published: January 2023

Abstract
Chemerin is characterized by a chemoattractant protein present in various inflammatory sites. Several studies have suggested that chemerin plays an anti-inflammatory role in acute tissue inflammation by inhibiting the recruitment of inflammatory cells. It has been recently reported that chemerin is elevated in patients with inflammatory bowel disease (IBD), however, the role of chemerin in IBD remains unknown. In the present study, we examined a role for chemerin in the development DSS-induced experimental colitis. Chemerin expression was up-regulated on days 5 and 8 after continuous DSS exposure for 5 days. Administration of exogenous chemerin aggravated the severity of DSS-induced colitis, which was characterized by more severe clinical disease scores, extensive epithelial damage and significantly increased expression of inflammatory cytokines in the colon on days 5 and 8. The exacerbated colitis was associated with decreased early colonic infiltration of neutrophils and decreased colonic expression of M2 macrophage-associated genes. In vitro study further demonstrated that chemerin negatively regulated IL-4-induced M2 macrophage polarization from the peritoneal macrophages. Our results suggest that chemerin plays an inflammatory role in DSS-induced colitis by impairing the protective innate responses and the targeting of chemerin for anti-inflammatory treatment should be cautious.
Spontaneous and persistent activation of NLRP3/NALP3 inflammasome plays a critical pathological role in colitis of IL-10 deficient (IL-10-/-) mice (P3240)
Authors: Li Xuhang , Liu Ling , Ye Mei , Yang Jianbo , Joosse Maria , Marohn Michael 
Published: January 2023

Abstract
IL-10-/- mice spontaneously develop chronic colitis. Our study was to evaluate the role and regulation of NLRP3 inflammasome in colitis of IL-10-/- mice and Crohn’s disease (CD). Two major NLRP3 complex proteins, NLRP3 and ASC, were upregulated in colonic mucosa of both IL-10-/- mice and CD. Importantly, NLRP3 inflammasome in IL-10-/- mice was activated prior to the onset of colitis; it progressively increased as disease worsened, peaking as macroscopic disease emerged. NOD2, a major IBD susceptible gene product, was upregulated only at a later stage of colitis when NLRP3 inflammasome was fully activated. The NLRP3 complex, including NLRP3, ASC and caspase-1, was demonstrated in vivo in colonic mucosa and increased in IL-10-/- mice, using 3 independent approaches (density fractionation, crosslinking, and co-IP). NLRP3 inflammasome activation occurred spontaneously in macrophages of IL-10-/- but not WT mice, and was effectively inhibited by IL-10. Glyburide, an inflammasome inhibitor, suppressed NLRP3 inflammasome expression/activation in IL-10-/- mice, resulting in not only alleviation of preexisting colitis, but also prevention of colitis development when given before disease onset. Thus, spontaneous and persistent activation of NLRP3 inflammasome, which is negatively regulated by IL-10, plays a major pathogenic role in colitis, it functions as a disease initiator whereas NOD2 may act as an accelerator. Inhibitors of the NLRP3 inflammasome may have utility in IBD therapy.
TRAF5 negatively regulates Toll-like receptor signaling (P1226)
Authors: Buchta Claire , Bishop Gail 
Published: January 2023

Abstract
Dysregulation in Toll-like receptor (TLR) signaling has been implicated in various inflammatory and autoimmune disorders. The cytoplasmic adaptor proteins TNF receptor associated factor (TRAF)3 and TRAF6 are known to mediate TLR signaling. Data from our lab suggest for the first time that another family member, TRAF5, is a negative regulator of TLR signaling. B lymphocytes, macrophages, and dendritic cells from TRAF5-/- mice have enhanced early signaling pathways including increased phosphorylation of the kinases ERK1/2, JNK, and Akt . They also produce more cytokines after TLR ligation. Consistent with these results, overexpression of TRAF5 in B cells inhibits TLR-induced cytokine production. Pursuing the mechanism of TRAF5-mediated regulation, we discovered that TRAF5 negatively regulates the transcription and protein levels of TRAF6. Additionally, TRAF5 associates in a complex with TRAF3, TRAF6, and the TLR adaptor protein MyD88 after TLR stimulation in B cells. Results suggest multiple mechanisms by which TRAF5 negatively regulates TLR signaling. TRAF5 deficiency has been reported to accelerate atherogenesis in mice. Elucidating the role of TRAF5 in TLR signaling may thus provide a new target for therapies against a variety of inflammatory conditions.
CD4+FoxP3+ regulatory T cell-mediated suppression of dendritic cells (P1049)
Authors: Liu Sang , Joly Anne-Laure , Mailer Reiner , Andersson John 
Published: January 2023

Abstract
CD4+FoxP3+ regulatory T cells are a subset of T cells that play central role in regulating inflammatory responses during many different immunological diseases. However the mechanisms of CD4+FoxP3+ regulatory T cells-mediated suppression remain controversial and poorly understood. Several studies have shown that CD4+FoxP3+ regulatory T cells can interact with and suppress a large number of cell types including CD4+ T cells and antigen presenting cells. For instance in vitro studies have shown that CD4+FoxP3+ regulatory T cells can inhibit cytokine production especially IL-2 production on CD4+ T cells hence the inhibition of CD4+ T cell proliferation; CD4+FoxP3+ regulatory T cells can also confer infectious tolerance by converting conventional T cells into functional FoxP3+ T cells in a cell contact TGF-β-dependent manner.In addition, CD4+FoxP3+ regulatory T cells suppress dendritic cell maturation and function by down-regulating the expression of co-stimulatory molecules CD80/CD86 via the interaction with CTLA-4.In our study, we have observed that CD4+FoxP3+ regulatory T cells may also modulate dendritic cell function via the PD-1:PD-L pathway. Dendritic cells that have been previously cultured with CD4+FoxP3+ regulatory T cells have decreased ability to support CD4+CD25- T cell proliferation and they can also induce higher level of FoxP3 expression in CD4+CD25- T cells.
Potentiation of regulatory T cell stability and function via a neuropilin-1:semaphorin-4a axis (P1057)
Authors: Delgoffe Greg , Woo Seng , Turnis Meghan , Gravano David , Guy Cliff , Bettini Matthew , Finkelstein David , Vogel Peter , Bonnevier Jody , Workman Creg , Vignali Dario 
Published: January 2023

Abstract
Regulatory T cells represent a critical immunoregulatory component of the immune system. The signals that maintain Treg stability and potentiate their function remain obscure. Here we show that the immune cell surface ligand semaphorin-4a on conventional murine and human T cells and dendritic cells interacts with the Treg-restricted receptor neuropilin-1 to potentiate Treg function and enhance survival. Mice with a Treg-restricted deletion of Nrp1 exhibit extremely limited tumor-induced tolerance, and thus substantial resistance to certain tumors, yet do not develop any autoimmune or inflammatory manifestations. Sema4a or Nrp1 blockade also had therapeutic efficacy against pre-existing tumors. In addition, Nrp1-deficient Treg cells fail to suppress inflammatory colitis. Upon Sema4a ligation, Nrp1 was recruited to the immunological synapse and repressed Akt activity via PTEN, which facilitated Foxo nuclear translocation. This induced a transcriptional program that promoted Treg stability, survival and function while repressing the induction of lineage-specific transcription factors. Thus, Nrp1 ligation enforces Treg stability and function in highly inflammatory sites but is dispensable for the maintenance of immune homeostasis, highlighting Nrp1 as a potential immunotherapeutic target in cancer.
Virus-specific Tregs are terminally differentiated upon peripheral activation and dependent on thymic output after both primary and secondary infection (P1035)
Authors: Liu Jie , Kim Sung-Han , Cao Shirley , Peppers Gretchen , Graham Barney 
Published: January 2023

Abstract
Regulatory CD4 T cells (Tregs) are defined as FoxP3+ cells and are categorized as natural (nTreg) or induced (iTreg). iTregs can be further classified as pathogen or epitope specific (pTreg). pTregs can share epitope specificity with effector cells (Teffs) and expand in response to infection. Although pTregs reduce infection-induced immunopathology, over-expansion and persistence can have a negative impact on pathogen clearance. Studying CD4 T cell responses to respiratory syncytial virus (RSV) infection, we found that the pTreg response had different dynamics than Teffs, which promoted efficient RSV clearance with minimal immunopathology and effectively established adaptive memory responses. The IA(b)M209-specific pTreg response was subdominant to IA(b)M209-specific Teffs. It peaked early to suppress responses in lymphoid organs after primary infection, but had limited regulation on CD8 T cell responses at the site of infection. The IA(b)M209-specific pTreg response contracted when the specific Teff responses peaked, and was diminished after secondary infection when an adequate adaptive Teff responses were established. The dynamics and differentiation pattern of the IA(b)M209-specific pTreg response was TCR clonotype-specific and enriched for recent thymic immigrants even on secondary infection, which suggests that pTreg responses are recalibrated with each infection to provide an optimal balance of Teff-mediated virus clearance and immunoregulation to limit tissue damage.
Cellular sentinels for heat shock protein-peptide complexes in vivo. (P4270)
Authors: Messmer Michelle , Binder Robert 
Published: January 2023

Abstract
Heat shock proteins (HSPs), such as gp96, harvested from cancer cells elicit potent anti-tumor immunity in mice and in patients with cancer. HSPs in the extracellular environment, released by cells or introduced through vaccination, are taken up by antigen presenting cells of the immune system, which allows for cross-presentation of the HSP-chaperoned peptides to T-cells. The goal of this study is to identify the subset(s) of cells that sample HSPs in vivo prior to initiating T cell responses. By vaccinating mice with fluorescent-labeled gp96, cells in the draining lymph node (dLN) that engage extracellular HSPs have been explored in a time and dose dependent manner. Our results show that CD4+CD11b+ cells are the predominant cells that internalized the HSP in the subcapsular region of the dLN while CD11c+CD8+ cells, which have been shown by other groups to be the major cross-priming dendritic cells, did not significantly internalize the HSP at any time points. Other CD11c+ cells did not appreciably internalize HSP until 24+ hours after. Internalization corresponded to higher CD91 expression by the CD4+CD11b+ cell subset. The functional capabilities of the CD4+CD11b+ and CD11c+ cells in T cell priming are examined. These studies illustrate how HSPs act to alert the immune system to cellular damage and will be useful for optimizing HSP-based vaccination regimes by identifying possible cellular targets.
Elucidating the role of bovine heavy- and light-chain variable domains in polyspecific antigen-binding (P6104)
Authors: Pasman Yfke , Kaushik Azad 
Published: January 2023

Abstract
The bovine antibody repertoire is generated from a limited germline sequence divergence as well as combinatorial diversity. Generation of an exceptionally long CDR3H (up to 61 amino acids) in the heavy chain variable region (VH) provides an additional mechanism to generate antibody diversity, not found in other species to date. These long CDR3H loops, present in polyspecific IgM, originate from VDJ recombination encoded by a specific VH gene, unusually long single DH-gene (capable of encoding up to 51 codons) and insertion of 15-18 base long conserved short nucleotide sequences (CSNS) specifically at VH-DH junction. In cattle, it is thought that antigen-binding is mainly a function of the variable-region of the heavy chain where light chain provides only structural support. To test this hypothesis, single chain variable fragment (scFv) and single domains (Fd) from polyspecific IgM were constructed. Both purified scFv as well as FdVH showed polyspecific binding to structurally dissimilar antigens in an ELISA. Whether scFv and FdVH bind to multiple epitopes on an antigen or if they recognize the same epitope can now be determined. The structural-functional complexities of these antibody fragments and the role of the heavy- and light-chains in antigen-binding will be discussed. [Supported by NSERC Canada]
Reactivity of bovine WC1+ γδ T cells to viral dsRNA (P6107)
Authors: Toka Felix , Krukowska Ewelina , Golde William 
Published: January 2023

Abstract
Bovine γδ T cells have properties of both innate and adaptive immune cells. However, detailed studies of how bovine γδ T cells execute their functions as innate or adaptive immune cells are lacking. In addition, their precise role in viral infections of cattle is not known. We have examined the reactivity of bovine γδ T cells to viral dsRNA isolated from bovine respiratory syncytial virus. Viral dsRNA was added to sorted peripheral blood WC1+ γδ T cells or directly delivered into the cells with DOTAP. Cell surface expression of CD80, CD335, MHC I and II, and synthesis of TNFα and IFNγ were investigated following stimulation with viral dsRNA. Addition of viral dsRNA to the culture of γδ T cells does not lead to changes in expression of CD335, MHC I and II but induces increased expression of CD80, although not statistically significant compared to non-stimulated cells. Direct delivery of viral dsRNA into WC1+ γδ T cells by DOTAP induced a twofold increase in expression of CD335 and MHC I with smaller increases in expression of MHC II and CD80. Although WC1+ γδ T cells synthesized low levels of TNFα and IFNγ in the presence of viral dsRNA or not, they significantly increased synthesis of IFNγ upon direct delivery of viral dsRNA into the cells. Results suggest that viral dsRNA may induce expression of molecules that promote innate function of γδ T cells and are discussed in context of pathways for enhancing bovine γδ T cell function against viral infections.
Polymorphisms and tissue expression of the Feline Leukocyte Antigen class I loci FLA I-E, -H and -K (P6086)
Authors: Hess Paul , Holmes Jennifer , Holmer Savannah , Gojanovich Gregory , Ross Peter 
Published: January 2023

Abstract
The domestic cat is a popular model of human retroviral diseases, but surprisingly, knowledge of the MHC class I complex (FLA I) in this species is limited. Only a few full-length coding sequences with uncertain locus assignments are known, and allelic variability, genotypic frequency and expression patterns remain undetermined. Of 19 class I genes, 3 loci - FLA I-E, -H and -K - are predicted to encode classical molecules. Our objective was to confirm their presumptive class Ia status by investigating polymorphisms and tissue expression of these genes. To accomplish this task, we developed a PCR-based, locus-specific, genotyping protocol, and then amplified 33 different FLA I-E, -H, and -K alleles from the gDNA of 12 cats of various breeds. We identified, for the first time, allele genotypes across all three loci in a feline species. We also amplified alleles from cDNAs from eight different tissues using an FLA I multi-loci-permissive primer set; only FLA I-E, -H, and -K transcripts were detected, and all three were widely expressed. The putative proteins encoded by our FLA I-E, -H, and -K alleles are highly variable and fit the class Ia model of homologous domains. Previously reported full-length FLA I sequences were confirmed, as were the predictions of classical status from the feline genome; FLA I-E, -H, and -K meet two major criteria of class Ia genes. These data lay the important groundwork for studying epitope-specific cytotoxic T-lymphocyte responses in the cat.
Fowlpox-based Survivin vaccination for malignant mesothelioma therapy (P2117)
Authors: Bertino Pietro , Panigada Maddalena , Soprana Elisa , Rose Aaron , Gaudino Giovanni , Carbone Michele , Siccardi Antonio , Hoffmann Peter 
Published: January 2023

Abstract
Survivin is an attractive candidate for cancer immunotherapy since it is abundantly expressed in most human cancers and absent in normal adult tissues. Malignant mesothelioma (MM) is a deadly cancer associated with asbestos exposure with no successful therapies currently available. We evaluated a Survivin-based vaccine as therapeutic tool for malignant mesothelioma in different MM mouse models. BALB/c mice, injected subcutaneously or intraperitoneally with murine fiber-induced MM tumor cells, were vaccinated with recombinant Fowlpox virus replicons encoding Survivin. Vaccination generated immune responses in both models, which led to delayed tumor growth and improved survival. Flow cytometric and immunofluorescent analyses of tumors from vaccinated mice showed CD8+ T cells infiltration, and real-time PCR demonstrated increased mRNA and protein levels of immunostimulatory cytokines. ELISPOT and intracellular cytokine staining analyses of Survivin peptide pulsed spleen cells from vaccinated mice confirmed antigen-specific, interferon-γ-producing CD8+ T cell responses. In addition pentamer-based flow cytometry showed vaccination generated Survivin specific CD8+ T cells. Importantly, vaccination did not affect fertility or induce autoimmune abnormalities in mice. Our results demonstrate that vaccination with recombinant Fowlpox expressing Survivin improves targeting of aggressive MM cells by the immune system and may form the basis for promising clinical applications.
Tumorigenesis of NIH3T3 cells induced by the exogenously over-expression of the novel tumor antigen OVA66 (P2121)
Authors: Ge Hailiang , Rao Wei , Xie Guohua , Li Weiyi , Xi Yibin , Wang Ying 
Published: January 2023

Abstract
The tumor associated antigen OVA66 which is firstly defined by serological analysis of human ovary cancer of recombinant cDNA expression library, has been demonstrated to be highly expressed in the majority of malignant tumors. We constructed a eukaryotic expression vector pFlag-OVA66 to establish an OVA66 stably over-expressed mouse fibroblast NIH3T3 cell line. The OVA66 over-expressed NIH3T3 cells exhibited several significantly malignant changes, the S and G2/M phage was markedly increased whereas the G1/G0 phage was decreased in the total cell cycle, indicating that the over-expression of OVA66 is able to promote cell cycling and proliferation. MTT and colony formation assay validated that OVA66 promotes the cell growth and colony formation in vitro. NIH3T3 cells with the over-expression of tumor antigen OVA66 displayed more resistance to the cell apoptosis induced by 5-FU. In vivo assay of the tumor xenograft studies in nude mice revealed the OVA66 over-expressed NIH3T3 cells were capable of forming tumors in the nude mice compared to the NIH3T3-mock cells with low expression of OVA66. Analysis of the phosphorylation of AKT and ERK1/2 stimulated with serum indicated a hyper activation of ERK1/2 MAPK and PI3K/AKT pathway in OVA66 stably over-expressed NIH3T3 cells. The results suggesting that OVA66 as a novel tumor antigen with its strong tumorigenic ability might be a novel target for the early detection, prevention and treatment of tumor in the future.
Indoleamine 2,3-dioxygenase-1 (IDO1) expression by childhood acute myeloid leukemia restrains IFN-γ production by T cells and may portend an unfavorable prognosis (P2157)
Authors: Rutella Sergio , Folgiero Valentina , Masetti Riccardo , Pagliara Daria , Caruso Roberta , Vinti Luciana , Moretta Francesca , Pession Andrea , Locatelli Franco 
Published: January 2023

Abstract
Background: Indoleamine 2,3-dioxygenase 1 (IDO1) degrades tryptophan (TRP) into kynurenine (KYN), driving the development, stabilization and activation of regulatory T cells, and suppressing anti-tumor T-cell responses. Patients and methods: We investigated IDO1 expression and function in 22 children with acute myeloid leukemia (AML; median age at diagnosis: 8 years, range 1-16). IDO expression was assessed with Western blotting; TRP and KYN levels were measured with RP-HPLC. Results: Leukemia blasts from 10 out of 21 evaluable AML cases up-regulated IDO1 expression and function after in vitro challenge with IFN-γ, as indicated by higher KYN levels in supernatants of stimulated (24.3 μM/L, range 10.8-37.7) compared with unstimulated AML cells (1.3 μM/L, 0.9-2.7; p=0.0022). IDO induction was abrogated by STAT3 inhibitors; furthermore, co-immuno-precipitation assays suggested the occurrence of physical interactions between IDO and STAT3 in leukemia cells. IDO-expressing AML blasts restrained Th1 cytokine production by allogeneic CD8+ T cells, an effect that was partially abrogated by D,L-1-methyl-tryptophan, an IDO inhibitor. A Kaplan-Meier estimate of the 3-year overall survival indicated a clear benefit for IDO- AML cases compared with IDO+ ones (68% versus 33%). Conclusions: Blast cells from childhood AML express functional IDO1. From a therapeutic standpoint, interference with IDO1 activity may restore anti-leukemia Th1 immune responses and improve disease outcome.
Activated B cells infiltrate lung cancer (P2201)
Authors: Bruno Tullia , Kern Jeffrey , Slansky Jill 
Published: January 2023

Abstract
Lung cancer is the leading cause of cancer death, and although some novel therapies are emerging, we are interested in boosting antigen-specific responses. In a comprehensive survey of the immune infiltrate in non-small cell lung cancer (NSCLC) patients, we found that the total number of B cells (TIL-Bs) in the tumor versus the tumor-adjacent tissue was increased in comparison to other immune subsets, specifically T cells. NSCLC TIL-Bs are activated (CD21lo and CD69+), proliferate, and have a memory phenotype (CD27+). Preliminary functional studies indicate that these cells generate activation cytokines (IFN-γ and IL-12), which suggest that the TIL-Bs have an effector phenotype that could aid in targeting the tumor. Further, these cells secrete total IgG and may present antigen because of their juxtaposition to T cells at the site of the tumor. In addition, we are analyzing the surface expression of Fas-ligand and TRAIL on the TIL-Bs to determine if these cells could be involved in the direct killing of tumor cells. Because we are interested in uncovering the unique potential of these TIL-Bs, we are currently collecting cells from tumor and tumor-adjacent tissue for gene-expression profiling. By comparing these two populations to normal lung B cells, we aim to discover a unique gene or pathway in the TIL-Bs that can be targeted for an immunotherapy clinical trial in lung cancer patients.