bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunosurveillance of cancer stem cells (P2217)
Authors: Gross Emilie , Peinado Carlos , Han Semi , Washington Allen , Bui Jack 
Published: January 2023

Abstract
The cancer stem cell (CSC) hypothesis stipulates that tumors follow a hierarchical organization where a subpopulation of cells endowed with stem cell characteristics initiates and maintains the malignant progression. Using the murine MCA sarcoma model system, a widely recognized model for studying cancer immunosurveillance and rejection mechanisms, we sought to explore how hierarchical tumor progression occurs during cancer immunosurveillance. The first step was to define CSC in the MCA sarcoma model. We identified an interesting heterogeneity of the tumor population based on Sca1 and CD90 expression. Purification of the Sca1+ and Sca1+/CD90+ in various cell lines and subsequent in vitro culture demonstrated that the Sca1+/CD90-population is able to regenerate the initial tumor heterogeneity. Transplantation of those two populations in RAGxγc-/- hosts demonstrated either a more rapid onset of sarcoma in mice injected with Sca1+ cells. Interestingly, phenotypic analysis of escaping tumors revealed an increase in Sca1+ cells. Finally, in vitro treatment with IFNg led to a substantial increase in Sca1 expression that correlated with an increase in cells' stemness properties as suggested by the elevated sphere forming capacity of IFNg treated cells. This data suggest that cancer stemness may be modulated by anti-cancer immune responses and may represent an additional mechanism of tumor immune escape.
The role of macrophages and T regulatory cells in capsaicin mediated regression of colon cancer (P3217)
Authors: Kadaveru Krishna , Hodio Jennifer , Basu Sreyashi 
Published: January 2023

Abstract
Colorectal cancer (CRC) is the third most commonly diagnosed cancer in both men and women. In humans genetic changes in intestinal epithelia commencing with inactivation of the APC tumor suppressor gene along with mucosal inflammation, facilitate the development of intestinal adenomas. The role of host immune system to detect and eliminate these altered cells is debatable based on its failure to prevent tumor growth. A physiological approach to prevent CRC formation is to use dietary compounds, which can inhibit inflammation and induce apoptosis to transformed cells. Capsaicin (CP), the principal pungent ingredient of red chili peppers is highly consumed spice throughout the world. It exhibits a wide range of immunological and pharmacological properties. We have previously reported that orally administered CP enhances a discreet population of CD11b+/F4/80+ macrophages and a CD4+ T cells population in the gut associated lymphoid tissue (GALT) expressing anti-inflammatory factors such as IL-10 and PD-L1. Here we demonstrate that orally administered CP can suppress spontaneous intestinal polyposis in the Apc(Min/+) mouse model, a widely used spontaneous colorectal cancer model for human familial adenomatous polyposis (FAP) in a time and dose dependent manner. We further demonstrate the role of different immune cells (myeloid and lymphoid) in CP mediated prevention of intestinal tumorigenesis. The use of dietary CP will be a novel approach for chemoprevention of human CRC.
The meninges: A staging site for immune cell interactions in early experimental autoimmune encephalomyelitis (P4166)
Authors: Hatfield Julianne , Brown Melissa 
Published: January 2023

Abstract
Multiple sclerosis (MS) is a demyelinating disease affecting the central nervous system in which immune mediate damage to white matter myelin is well characterized. However, improved imaging has revealed that inflammation in the meninges, tissues that are proximal to the CNS, also occurs and corresponds to cortical grey matter lesion formation in early disease. Together with evidence of activated T cells and tertiary lymphoid follicle formation within these tissues, these data suggest that the meninges are critical sites of immune cell activity. Yet little is known about the basal and disease-associated immune status of these tissues. Using a murine model of MS, we evaluated the immune cell makeup of the meninges in naïve mice and asked how this changes during the course of disease. CD4+ and CD8+ T cells, macrophages, dendritic cells, mast cells and neutrophils are detected in naïve mice and increased numbers of neutrophils and macrophages are observed in preclinical disease. A population of innate lymphoid cells (ILCs: CD3-, CD4+/-, RORγt+, IL-7Rα+) also normally resides in the meninges and this population is reduced in mast cell deficient mice. These data confirm that the meninges are an active site of immune response in early disease. We speculate that meningeal ILCs may provide a mechanism for the development of tertiary lymphoid follicle formation as well as contribute to Th17 cell activation and thereby amplify meningeal inflammation.
HMGB1-LPS complex promotes the transformation of SFs to a RASF-like phenotype (P4168)
Authors: Qin Yanghua , Chen Yan , Shen Qian 
Published: January 2023

Abstract
The specific pathogenesis of rheumatoid arthritis (RA) remains unclear. Current views hold that some inflammatory antigens recognize the Toll-like receptors on the synovial fibroblasts (SFs) and then activate downstream signals, leading to the formation of RASFs and the induction of RA. Our previous studies showed that intra-articular injection of the complex of HMGB1 and lipopolysaccharide (LPS) directly induced experimental arthritis in DBA/1 mice; histopathological analysis further revealed synovial thickening with excessive proliferation of SFs. In the present study, osteoarthritis synovial fibroblasts (OASFs) were co-cultured with HMGB1-LPS complex in vitro for 5-8 generations to induce the transformation of human SFs to RA-like SFs (tOASFs). Then the pathogenicity of tOASFs was evaluated, both in vitro and in vivo. Our results showed that tOASFs attached to, invaded and degraded co-implanted cartilage in a SCID mouse model. In addition, histochemistry showed excessive proliferation of tOASFs and the expression of MMPs. In vitro, cell cycle analysis showed more tOASFs passing through the G1/S checkpoint and moving to S or G2 phase. Flow cytometry and confocal microscopy showed significantly reduced apoptosis and enhanced autophagy in tOASFs compared to OASFs, respectively. Finally, we showed upregulated expression of certain receptors and adhesion molecules in tOASFs. This accumulated evidence led to the conclusion that HMGB1-LPS complex promoted the formation of RASFs.
Cell intrinsic role for MAVS in regulating CD8 T cell fitness during acute virus infection (P6113)
Authors: Thomas Sunil , Huang Albert , Loo Yueh-Ming , Ramos Hilario , McMillan Aimee 
Published: January 2023

Abstract
Mitochondrial antiviral Signaling molecule (MAVS) has been described in it’s role as an adaptor molecule that links viral RNA sensing by RIG-I-like receptors (RLR)s to type I IFN and inflammatory cytokine production. However it’s role in adaptive immune responses to viruses is not well defined. Using an adoptive cell transfer system we examined the requirement for MAVS directly in CD8 T cells during adaptive responses following acute Lymphocytic choriomeningitis virus (LCMV) infection. Here, we found that MAVS plays an RLR-independent role in determining CD8 effector T cell fitness. MAVS-deficient cells were deficient in virus-induced expansion and cytokine production. This deficiency correlated with altered cell size and granularity, differential mitochondrial membrane potential and reduced levels of steady-state intracellular calcium. We found cytokine production deficiencies were acute at earlier stages of CD8 cell expansion, suggesting that the requirement for MAVS is most critical when cellular need for mitochondrial metabolism is maximal. These results suggest that MAVS is required for metabolic support of CD8 effector cell expansion during acute virus infection. Our study therefore assigns a novel metabolic role to MAVS in adaptive CD8 T cell energetics. Defining the function of MAVS in enhancing T cell metabolism therefore has implications for vaccine strategies that aim to increase the quality and quality of the CD8 T cell response to virus infection.
An in vitro culture system for studying the human B cell repertoire (P1459)
Authors: Su Kuei-Ying , Nojima Takuya , Kuraoka Masayuki , Moody M. Anthony , Kelsoe Garnett 
Published: January 2023

Abstract
Defining the human B-cell repertoire is important for understanding human B-cell development and autoimmunity. Currently, re-expression of immunoglobulins from single B cells is the standard method to study human B-cell repertoire; however, this method is labor-intensive and costly as it requires large numbers of recombinant antibodies to be generated by single-cell PCR and expression of recombinant heavy and light chain pairs. We have developed an alternative method to characterize the B-cell repertoire by culture that supports the proliferation and differentiation of human pre-B, immature/transitional, and mature B cells recovered from bone marrow, fetal liver, or peripheral blood. Briefly, single B cells are cultured in IL-2, IL-4, IL-21, and BAFF on CD154+ stromal cells. This culture supports extensive cell proliferation and differentiation into IgG-secreting plasmacytes. The monoclonal IgG antibodies in each cell culture can be readily screened by ELISA, Luminex, or immunofluorescence against endogenous and exogenous antigens. We find that autoantibodies from pre-B cell cultures have higher avidities for self-antigens than do those from immature/transitional and mature B cell cultures, suggesting the loss of high avidity cells at the first tolerance checkpoint. V(D)J rearrangements are easily recovered from selected clones for sequencing and expression. This culture method offers a pathway to determine B-cell repertoires with high throughput and reduced cost.
CD28 promotes long lived plasma cell survival, sustained antibody responses and upregulation of BLIMP-1 expression through its distal proline motif. (P1475)
Authors: Lee Kelvin , Utley Adam , Murray Megan , Carlson Louise , Farren Matthew , Russell Lisa , Nair Jayakumar , Garrett-Sinha Lee Ann , Green Jonathan , Boise Lawrence , Rozanski Cheryl 
Published: January 2023

Abstract
Durable humoral immunity is dependent upon the long term maintenance of serum antibody titers. Evidence now suggests that antibody levels are maintained by a distinct subset of bone marrow resident long lived plasma cells (LLPC). However, the molecular basis for the longevity of LLPC is largely uncharacterized. We have recently shown that CD28 signaling supports the survival and function of LLPC but not short lived splenic PC (SLPC). We have now extended these studies by characterizing the signaling pathways and molecular targets downstream of CD28 in PCs using CD28 knock-in mice in which the CD28 cytoplasmic tail is mutated at either the Y170MNM (CD28-Y170F) or P187YAP190 (CD28-AYAA) regions, resulting in loss of downstream PI3K or Grb2-Vav signaling, respectively. Interestingly, only the CD28-AYAA animals had significantly diminished LLPC numbers with no effect on SLPC numbers in vivo, consistent with the inability of CD28 activation to rescue CD28-AYAA LLPC from serum starvation-induced death in vitro. Further characterization of the CD28 (PYAP)-Vav-NFκB pathway in PC uncovered a previously undescribed regulation of the central transcription regulator BLIMP-1 through a CD28 responsive element in the prdm1 promoter. These results demonstrate that CD28 (PYAP)-Vav-NFκB activation in LLPC but not SLPC induces BLIMP-1 expression, which then can regulate the broader transcriptional reprogramming allowing for LLPC function and survival.
B cell development and IL-10 secretion (P1476)
Authors: Basu Uttiya , Sun Jianbo , Wang Jiguang 
Published: January 2023

Abstract
Accumulating evidence confirms the existence of regulatory B cells (Bregs). Bregs are hypothesized to negatively regulate immune responses in autoimmunity and inflammation through the production of regulatory cytokines such as IL-10 and TGF-β. The precise cellular origin and function of Bregs remains unknown. IL-10 is known to exert broad anti-inflammatory effects which may negatively influence the activation of T cells, macrophages, and dendritic cells (DCs) and thereby influence both cellular and humoral immunity. Many questions exist regarding the function and biogenesis of Bregs. Do IL-10-generating Bregs only transiently express IL-10 during particular developmental stages? And what is the functional consequence of B cell mediated IL-10 secretion in vivo? We observe that in a population of naïve splenic B cells, a significant fraction is competent to secrete IL10. Even in B cells stimulated for class switch recombination (CSR), the unswitched population shows a high proportion of IL-10 secreting Bregs. Finally, we discuss the expansion of IL-10 secreting Bregs and their ex vivo matured progenitors in activation-induced cytidine deaminase (AID) deficient mice which are incompetent to undergo CSR and somatic hypermutation, two processes that generate antibody diversity. Put together, these observations point to a correlation between Breg biogenesis and antibody diversification processes.
Endothelin-1 but not endothelial nitric oxide synthase polymorphisms are implicated in sickle cell disease (P3311)
Authors: Thomas Bolaji , Thakur Tanya , Guindo Aldiouma , Ott Jurg , Diallo Dapa 
Published: January 2023

Abstract
Sickle cell disease (SCD) shows marked variability in severity among individuals; this variability probably linked to differential expression of adhesion molecules. Polymorphisms of the endothelial nitric oxide synthase (eNOS) have been implicated in disease severity, potentially delineating vascular complications. There is paucity of information on endothelin-1 (ET-1) diversity and disease severity or ethnic stratification. We examined genomic diversity and estimated haplotype frequency of eNOS and ET-1 polymorphisms in children with SCD. Thirteen SCD children (HbSS) and 176 adult and pediatric controls (HbAA) from Mali were recruited. Genotypic analysis of eNOS variants (T786C, Glu298Asp and intron 4) and ET-1 (G5665T) was carried out with a PCR-RFLP assay. Our results show the frequency distribution and allelic diversity of eNOS and ET-1 variants. Endothelin-1 (G5665T) mutant form was the most frequent, and probably most significant among children with SCD, though the distribution is out of Hardy-Weinberg equilibrium (P=0.0008). eNOS variants showed no difference between SCD and controls. We show that eNOS variants are less frequent, and may have no functional significance in SCD. Additionally, eNOS allele and genotype frequencies are not significantly different in cases and controls. Association between ET-1 and eNOS variants in African versus African American SCD children will be explored and the reason(s) for the HWE deviation will be investigated.
The IRF5-TNPO3 association has two components in systemic lupus erythematosus, which are shared with other autoimmune disorders (P3315)
Authors: Kottyan Leah , Zoller Erin , Kaufman Kenneth , Harley John 
Published: January 2023

Abstract
Improved technical and bioinformatics genetic methods now allows us to capture more complete relevant genetic variation for candidate causal disease risk variant identification. Exploiting genotyping, DNA sequencing, imputation, and trans-ancestral mapping, we modeled the IRF5-TNPO3 genetic association on chromosome 7, now implicated in two immunotherapies and seven autoimmune diseases. Specifically, in systemic lupus erythematosus (SLE) we resolve separate associations in the IRF5 promoter (all ancestries) and an extended European haplotype. We capture 7,650 IRF5-TNPO3 genetic variants across five ethnicities in 8,395 SLE cases and 7,367 controls. The six plausible causative IRF5 promoter variants are confined to 5.7 kb (meta-analysis p-value=6x10^-49; Odds Ratio=1.38-1.97). The 22 potentially causal variants of an 85.5 kb haplotype of IRF5 and TNPO3 (haplotypic p-value in European Americans=2.4x10^-45, Odds Ratio=0.58) (individual p-value in European Americans=10^-27 - 10^-32, Odds Ratio=1.7-1.81). The possible models from this sample virtually eliminate the previously purported IRF5 functional variants as causal. Finally, this model also appears to operate in Sjögren’s syndrome and systemic sclerosis (both components) and primary biliary cirrhosis (haplotype only), demonstrating the nuances of similarity and difference in autoimmune disease risk mechanisms at IRF5-TNPO3.
Functional characterization of the MECP2/IRAK1 lupus risk haplotype in human T cells and a human MECP2 transgenic mouse (P3319)
Authors: Sawalha Amr , Koelsch Kristi , Webb Ryan , Jeffries Matlock , Dozmorov Mikhail , Frank Mark , Guthridge Joel , James Judith , Wren Jonathan 
Published: January 2023

Abstract
Genetic polymorphism in MECP2/IRAK1 is a confirmed susceptibility locus for lupus. DNA methylation changes in T cells play a central role in the pathogenesis of lupus, and MeCp-2 is a master regulator of gene expression and is known to recruit DNA methyltransferase 1 (DNMT1) during DNA synthesis. Using human T cells from normal individuals, we demonstrate that the lupus risk variant in MECP2/IRAK1 increases MECP2 mRNA expression in stimulated T cells. By assessing DNA methylation levels at ~ 485,000 methylation sites across the genome, we further show that the lupus risk variant in this locus is associated with significant DNA methylation changes, including in the HLA-DR and HLA-DQ loci, as well as interferon-related genes such as IFI6, IRF6, and BST2. Further, using a human MECP2 transgenic mouse, we show that overexpression of MECP2 alters gene expression in stimulated T cells. This includes overexpression of Eif2c2 that regulates the expression of multiple microRNAs (such as miR-21), and the histone demethylase Jhdm1d. We also show that MECP2 transgenic mice develop antinuclear antibodies. Our data suggest that the lupus associated variant in the MECP2/IRAK1 locus has the potential to affect all 3 epigenetic mechanisms. Importantly, we demonstrate that variants within the MECP2 gene can alter DNA methylation in other genetic loci including the HLA and interferon-regulated genes, thereby providing evidence for genetic-epigenetic interaction in lupus.
rhIL-15 and rhIL-2 together enhances growth and function of TCR transduced T cells for adoptive immunotherapy (P4358)
Authors: Scurti Gina , Moxley Kelly , Mehrotra Shikhar , Nishimura Michael 
Published: January 2023

Abstract
The use of TCR transduced T cells for the treatment of melanoma has shown promise in recent years. However, there is a need to improve the function and growth of transduced T cells to allow for better clinical outcomes. We wanted to evaluate the impact of rhIL-15 on TCR transduced T cells. T cells from multiple donors were transduced with a retrovirus encoding the TIL 1383I T cell receptor. The transduced T cells were cultured and expanded for approximately 21 days under several different cytokine conditions including 300IU/mL of rhIL-2, 100ng/mL of rhIL-15 or both rhIL-2 and rhIL-15. For each culture condition, data was collected throughout the process for growth, function, phenotype and TCR expression. We found that the T cells cultured in rhIL-15 had a greater TCR expression and produced more IFN-γ in response to tumor than T cells cultured only in rhIL-2. rhIL-15 alone did not increase the growth of TCR transduced T cells when compared to rhIL-2 whereas the use of both rhIL-2 and rhIL-15 not only enhanced the TCR expression and anti-tumor activity but also improved the growth of the T cells. We also noticed that T cells cultured with rhIL-15 increased resistance to apoptosis during oxidative stress. Based on these results, we have initiated our TCR gene transfer clinical trial using rhIL-15 and rhIL-2 to transduce and expand our T cells.
Mutant HSP70i prevents effector T cell responses in autoimmune vitiligo (P4352)
Authors: Mosenson Jeffrey , Zloza Andrew , Nieland John , Garrett-Mayer Elizabeth , Eby Jonathan , Huelsmann Erica , Kumar Previn , Denman Cecele , Lacek Andrew , Kohlhapp Frederick , Alamiri Ahmad , Hughes Tasha , Bines Steven , Kaufman Howard , Overbeck Andreas , Mehrotra Shikhar , Hernandez Claudia , Nishimura Michael , Guevara-Patino Jose , Le Poole I. Caroline 
Published: January 2023

Abstract
A single amino acid change (Q435A) within HSP70i renders an isoform with treatment potential for vitiligo, as demonstrated by ablated depigmentation in gene gun vaccinated, vitiligo-prone TCR transgenic mice. We hypothesized that HSP70iQ435A inhibits T cell mediated immune responses. We probed skin from gene gun vaccinated P-mel1 and h3TA2 mice for T cell infiltrates and analyzed splenocytes for T cell activation markers by FACS, comparing findings to those in tissues from vitiligo patients and among cells collected from human skin explants exposed to HSP70iQ435A. Decreased skin infiltration was observed for T cells responsible for melanocyte killing in both mouse strains. Eomes/T-bet ratios were significantly decreased by wild-type HSP70i driving an effector phenotype, whereas vaccination with HSP70iQ435A maintains a quiescent phenotype. This ratio was similarly decreased in circulating and skin-infiltrating T cells from vitiligo patients where we also observed an increase in CXCR3 expression, supportive of skin homing. In cells from skin explants exposed to wild-type or mutant HSP70i the Eomes/T-bet ratio is upregulated by HSP70iQ435A whereas upregulation of skin homing receptor CLA followed exposure to wildtype HSP70. In conclusion, the wild-type HSP70i-induced T cell effector phenotype accompanied by elevated CXCR3 and CLA expression observed in vitiligo is countered by HSP70iQ435A exposure, providing a mechanism for HSP70iQ435A induced inhibition of autoimmunity.
cAMP-based adjuvants (LT, mLT, dmLT) promote Th17 and germinal center responses through dendritic cells, while direct CD4 T-cell contact induces a T-suppressor phenotype. (P4512)
Authors: Norton Elizabeth , Clements John 
Published: January 2023

Abstract
Enterotoxin-based adjuvants with the ability to induce cAMP (LT, mLT and dmLT) elicit multifaceted antigen-specific response after vaccination, including Th1, Th2, Th17 and T-regulatory (Treg) responses unlike related non-cAMP inducing proteins (LT-E112K and B-subunit). While cAMP induction is linked to enterotoxin immunologic activity, many studies have now demonstrated a key regulatory role for cAMP in directing Treg responses. We investigated these paradoxical abilities to promote both suppressive and stimulatory T-cell subset using a panel of LT enterotoxin based proteins for cAMP induction, adjuvanticity, and in vitro effects on immune cells. We found that direct stimulation with cAMP-inducing adjuvants (LT, mLT or dmLT) induced a T-suppressive phenotype linked to cAMP activity but not Foxp3 or IL-10 expression. However, cAMP-inducing adjuvants promoted both proliferation and secretion of all classical Th-defining cytokines, especially IL-17, via indirect stimulation of T-cells through dendritic cells (DCs). Neutralization of IL-17 during vaccination reduced germinal center development and antibody responses. These results indicate cAMP-based adjuvants strongly promote Th17 cells via stimulated DCs, while direct contact mediates T-suppressive phenotype. In addition, DCs play a key role in directing these adjuvant signals away from suppression and towards activation.