bluuebunny/crossref_metadata_2025_split huggingface dataset:
Neutrophils Disrupt B-1a Cell Homeostasis by Targeting Siglec-G to Exacerbate Sepsis
Authors: Tan Chuyi , Aziz Monowar , Reilly Bridgette , Ma Gaifeng , Wang Ping 
Published: November 2024

Abstract
In sepsis, B-1a cells diminish in the peritoneal cavity (PerC) as they relocate to the spleen via CXCL12/CXCR4. In the spleen, these cells differentiate into plasma cells, skewing from innate to adaptive phenotype. The mechanism governing the PerC B-1a cell relocation in sepsis remains elusive. Sialic-acid-binding immunoglobulin-like lectin (Siglec)-G is a B-1a cell-specific receptor. We found that Siglec-G-/- PerC B-1a cells had a higher migration rate than WT B-1a cells. Siglec-G negatively regulated B-1a cell migration by interacting with CXCR4, as Siglec-G-/- B-1a cells showed increased activation of the CXCR4’s downstream pathway. LPS-activated neutrophils decreased Siglec-G expression on B-1a cells and promoted B-1a cell migration, while NE inhibitor reversed these. Recombinant NE treatment decreased Siglec-G expression by cleavage on B-1a cells. A small Siglec-G-derived decoy peptide (SDP) was developed to prevent Siglec-G cleavage based on the interaction site of NE at the Siglec-G protein in silico. Treatment of SDP significantly reduced the decrease in PerC B-1a cell numbers and Siglec-G expression on B-1a cells in septic mice. SDP treatment significantly reduced serum IL-6 and TNFα levels and improved survival rates in septic mice. In conclusion, we discover Siglec-G to be a new regulator of B-1a cell migration. Preventing NE-mediated Siglec-G cleavage upholds a promising strategy for maintaining peritoneal B-1a cell homeostasis to mitigate sepsis.
The Role of Textile Dyes in the Immune System Dysregulation
Authors: Lorenz -Cortes Lizette , Vinueza-Benitez Nelson 
Published: November 2024

Abstract
There has been an increase in the incidence of atopic diseases and cancer in the last decades. There is an urgent need to identify the environmental triggers that fuel these conditions. In recent years, the use of synthetic textile dyes has increased. The effects of these chemicals or their metabolites on our skin and our immune system have not been well studied. There have been reports on the effects of irritants to the skin on their ability to break the dermal barrier to start a chain of immune reactions that dysregulate the immune system. The purpose of this study was to investigate the effects of the most common textile dyes in the viability, IFNg production and mitochondrial function of epidermal skin, and intestinal epithelial cells using swine as an animal model. Cultures of MPEK-BL6 and IPEC cells were cultured with disperse textile dyes Red 11, Orange 37, Blue 1, Blue 124, and Brown 1, at low and high concentrations for 3-6 hours and 1-3 days. Viability was measured with LUNA cell counter and by flow cytometry. IFNg production by flow cytometry and ELISPOT and Oxygen consumption rate (OCR) by Seahorse Agilent Technology kit. Disperse Blue 124 and Brown 1 induced IFNg production, and impaired cell viability and mitochondrial function as early as 3 hours after exposure with IPEC and MPEK cells. These data suggest that textile dyes can influence cell viability and possibly cause inflammation as a starting point to dysregulate the immune system. Supported by NCSU-DSA
Non-tumor cell IDO1 non-enzymatically decreases radiation + PD-1 mAb treatment efficacy in older adults with glioblastoma
Authors: Zhai Lijie , Bell April , Asimakidou Evridiki , Lauing Kristen , Wainwright Derek 
Published: November 2024

Abstract
To understand the role of indoleamine 2,3 dioxygenase 1 (IDO1) in the age-dependent immunotherapeutic effect, we compared 20-26 month-old C57BL/6 background mice that were IDO1-T2A-GFP [(WT); n=16], IDO1 enzyme null [H350A; (n=16)], IDO1KO (n=4), Ido1fl/fl(n=14), Lysm-Cre+→Ido1fl/fl (n=20), CD11c-Cre+→Ido1fl/fl (n=23), Tie2-Cre+→Ido1fl/fl (n=14) treated with RT + PD-1 mAb. Mice were intracranially engrafted (ic.) with GL261 cells followed by RT + PD-1 mAb beginning at 14-days post-ic. IDO1 WT and IDO1 enzyme null (H350A) mice had a survival rate of 12.5% and 18.75% at 90-days post ic., respectively, which was significantly decreased as compared to IDO1KO mice with a 75% survival rate (p&lt;0.05). Tie2-Cre+→Ido1fl/fl mice, CD11c-Cre+→Ido1fl/fl, and Lysm-Cre+→Ido1fl/fl mice had a 100%, 96%, and 70% survival rate at 90-days post ic., respectively, as compared to uncrossed Ido1fl/fl mice with a 42% survival rate (p&lt;0.05). Significantly reduced kynurenine (KYN) levels were found in IDO1 enzyme null (H350A) and IDO1KO mice as compared to WT mice (p&lt;0.001). Unexpectedly, KYN levels were also decreased in Tie2-Cre+→Ido1fl/fl but not in CD11c-Cre+→Ido1fl/fl, Lysm-Cre+→Ido1fl/fl or Ido1fl/fl mice (p&lt;0.05). Tryptophan (TRP) levels did not change between mouse groups. These data support the hypothesis that non-tumor cell IDO1 reduces immunotherapeutic efficacy in older adults with GBM through a TRP-KYN pathway-independent mechanism.
Targeting RAS/RAF/PI3K pathway for CD40 induction in melanoma cells to overcome resistance to anti-PD1 immunotherapies
Authors: Yan Chi , Luo Weifeng , Chen Sheau-Chiann , Johnson Douglas , Ayers Gregory , Richmond Ann 
Published: November 2024

Abstract
The first-line treatment for metastatic melanoma is immune checkpoint blockade (ICB), but it fails in many patients and may have serious adverse events. We showed that rigosertib (RGS), a RAS-pathway inhibitor, promotes CD40 upregulation on melanoma cells and synergizes with ICB in preclinical melanoma models. However, residual disease still occurred with uncontrolled ERK activation. Our present study explores the efficacy of RGS plus trametinib (T), a MEK1/2 inhibitor, to overcome ICB resistance. The concurrent RGS+T+αPD1 resulted in an additive effect to suppress ICB-resistant NRASmut 1014 tumor growth, and 44.4% (4/9) tumors completely regressed after 2 weeks of treatment. The RGS+T regimen promoted CD8+ T cell responses in the tumor microenvironment (TME) and extended the time to αPD1 resistance (p&lt;0.0001). Combining RGS+T with agonist CD40 (aCD40) resulted in increased CD8+ T cells, natural killer cells, and M1 macrophages in the TME, with a reduction of myeloid-like CD11b+PD-L1+ regulatory B cells in the tumors (~70%, p&lt;0.0001) and tumor-draining lymph nodes (~40%, p&lt;0.0001). CRISPR/dCas9-based overexpression of CD40 (CD40-OE) in 1014 melanoma cells reduced in vivo tumor growth and successfully turned the ICB-resistant tumors into responders to αPD1 (p=0.025), which further regressed with aCD40+αPD1 (p&lt;0.001). Our preclinical data support the therapeutic use of RAS/RAF/PI3K inhibition plus CD40 agonism for metastatic melanoma patients who do not respond to ICB.
T cell immune landscape in melanoma patients before and post immune checkpoint blockade treatment
Authors: Luo Ying , Yao Chen , Wu Tuoqi 
Published: November 2024

Abstract
Immunotherapy based on immune checkpoint blockade (ICB) has revolutionized cancer treatment. Immune cells, especially T cells, play a crucial role in ICB. However, ICB-induced T cell response remains incompletely investigated. Revealing T cell landscape changes caused by ICB could promote the discovery of novel biomarkers with predictive efficacy and toxicity and the potential strategy to improve the therapeutic efficacy while reducing side effects. We used single cell-based sequencing approaches to analyze transcriptome and surface protein of T cells in peripheral blood collected before and post ICB treatment from melanoma patients with immune-related adverse events. A total of 17 cell clusters were identified and annotated according to the expression of RNA and protein. Collectively, variation existed among the recruited participants. ICB treatment decreased the proportion of memory CD4 T cells while increasing the proportion of T follicular helper cells. In summary, these findings provide insight into how ICB modulates T cell responses in melanoma patients.
Investigating MAIT cell phenotype using MR1 and TCR Dextramer™ reagents
Authors: Blicher Thomas , Hansen Bjarke , Jacobsen Kivin , Brix Liselotte 
Published: November 2024

Abstract
Objective
Here, we demonstrate a workflow, which enables the study of mucosal-associated invariant T (MAIT) cells using MR1 reagents. The workflow covers initial detection of MAIT cells with TCR sequencing as well as cloning, production, and validation of selected MAIT TCR molecules.
Methods
Human PBMC samples were stained with relevant antibodies and MR1/5-OP-RU dCODE Dextramer™ reagents. Following staining, sorting, and partitioning on a BD RhapsodyTM Single-Cell Analysis System, the cells underwent sequencing for characterization of gene and surface marker expression as well as paired alpha-beta TCR sequences. Selected TCR sequences were produced as TCR molecules and subsequently used to detect MR1/5-OP-RU on the surface of antigen-presenting cells.
Results
The majority (70%) of the MAIT cells express typical TCRs containing TRAV1-2 and TRAJ33 gene segments. The remaining MR1/5-OP-RU-reactive MAIT cells express highly diverse, non-classical MAIT TCR sequences. The three most frequent TCR clonotypes were produced and their specificity validated using in vitro assays and staining of MR1/5-OP-RU-presenting cells.
Summary
Our workflow allows identification of MAIT cells and elucidation of their TCR sequences. The high sensitivity of MR1 dCODE Dextramer reagents enables detection of both weak interactions and rare cell types. Subsequent production of MAIT TCR molecules allows deeper characterization of individual MAIT cell specificity and its role in health and disease.
Divergent coactivator usage and CREB1 activity controls tuft lineage commitment
Authors: Eskiocak Onur , Subhash Santhilal , Shah Vyom , Moresco Phill , Beyaz Semir 
Published: November 2024

Abstract
The precise coordination of co-activators and transcription factors in shaping distinct intestinal epithelial cell types during intestinal stem cell (ISC) differentiation, especially the roles of CBP and P300, remains poorly understood despite extensive gene regulation studies. Here using conditional and inducible genetic models, we identify P300 but not CBP as a specific repressor of tuft and goblet lineages. Both ISC- and IEC-specific deletion of P300 elicited selective expansion of tuft and goblet cells with upregulation of gene modules that orchestrate their differentiation without affecting Paneth and enteroendocrine linages. Intestinal organoids that lack P300 more robustly upregulated tuft cell marker genes in response to IL13, a cytokine that drives type-II immunity and tuft cell expansion. Because P300 often acts as co-activator of TFs that regulate lineage specific gene expression, we hypothesized that lack of P300 repressed a repressor of tuft cell lineage. Among TFs that associate with P300, we tested whether CREB1 regulates P300 mediated restriction of tuft lineage. Surprisingly, we found that CREB1 is necessary for tuft cell commitment in vivo and expansion in response to IL13 treatment in organoids. Overall, these results contribute to our understanding of the complex regulatory mechanisms underlying the ISC differentiation to form specialized cell types in the intestinal epithelium.
IL-22 is required for preserving tissue integrity and host-microbe homeostasis in the peridontium
Authors: Saha Gundappa , Li Xiaofei , Li Lu , Diaz Patricia , Hajishengallis George 
Published: November 2024

Abstract
Periodontal Disease (PD) is a dysbiotic inflammatory condition of the periodontium (tooth-supporting tissues) that exerts an adverse impact on systemic health. IL-22 mediates unidirectional communication from immune cells to tissue stromal cells and plays a key role in promoting homeostatic immunity with some implications in certain inflammatory disorders. In this study, we investigated whether IL-22 is required for periodontal tissue homeostasis at steady state using IL-22-deficient (IL-22–/–) mice and IL-22+/+ littermate controls. The mice were analyzed in terms of disease phenotype, gingival transcriptomic profile (bulk RNA sequencing and quantitative real-time PCR) and microbiota composition (16S rRNA sequencing). Analysis of the bone levels in 10-week-old IL-22–/– mice revealed significantly increased naturally occurring bone loss vs controls. The IL-22 deficiency-induced bone loss was associated with significantly increased gingival tissue expression of the pro-inflammatorycytokine IL-17 and several molecules with pro-inflammatory and antimicrobial properties (e.g., S100 and Reg3 proteins). Moreover, IL-22 deficiency was associated with a significant increase in the periodontal microbial burden and alterations to the microbiota composition consistent with dysbiosis. Therefore, IL-22 appears to be required in setting the homeostatic tone of the periodontal tissue, suggesting that loss-of-function genetic polymorphisms in IL-22 may affect periodontal health.
Elucidating inflammation-induced cell fate decisions in primary human Tregs
Authors: Ho Patrick , Bluestone Jeffrey , Tang Qizhi 
Published: November 2024

Abstract
Adoptive regulatory T-cell (Treg) therapy has remarkable efficacy in promoting immune tolerance in preclinical models of transplantation and autoimmune disease. However, lineage-tracing studies have revealed that Tregs can undergo epigenetic reprogramming in chronically inflamed tissue environments, resulting in the acquisition of proinflammatory functions and the capacity to exacerbate autoimmunity. Despite intense interests in developing Treg therapy for humans, inflammation-induced human Treg lineage-decommitment remains poorly understood. Here, we present a robust in vitro model of IL6, IL1β, and IL23-driven human Treg instability characterized by progressive FOXP3 and HELIOS downregulation, FOXP3 conserved non-coding sequence (CNS)-2 enhancer re-methylation, diminished in vitro suppressive function, and elevated proinflammatory cytokine expression. Single-cell transcriptomic and chromatin accessibility analyses of human Tregs undergoing destabilization revealed a gradual closing of a Treg-specific element (A) and a reciprocal opening of a competing element (B) at the IRF4 locus. CRISPR-mediated excision of the B element (IRF4ΔB) preserved Treg identity and enhanced in vitro suppressive activity following prolonged inflammatory exposure. Ongoing experiments aim to evaluate the in vivo function and stability of IRF4ΔB Tregs in a xenogeneic graft-versus-host model. These findings may facilitate the design of Treg therapeutics with greater potency and improved safety.
Bi-functional IL2/CD25/IL10 fusion protein promotes Tregs, reduces inflammation, and is highly effective in limiting diabetes in NOD mice
Authors: Santos Savio Alicia , Welford Sarah , Malek Thomas 
Published: November 2024

Abstract
Clinical trials indicate that low-dose IL-2 is a promising approach for patients with autoimmunity. To improve upon IL-2, the mouse (m) IL-2/CD25 fusion protein was developed, which has a long half-life, is selective to the high-affinity IL-2R and expands Tregs to limit diabetes in NOD mice. However, after an extended time off-therapy, many NOD mice exhibited T1D. To improve efficacy, we developed a bifunctional fusion protein, consisting of IL-2/CD25 covalently linked to IL-10 to increase Tregs and limit inflammation. With respect to IL-2 activity, IL2/CD25/IL10 supported IL-2-dependent proliferation of CTLL and STAT5 activation by Tregs. With respect to IL-10 activity, it supported IL-10-dependent STAT3 activation by RAW264.7 cells and limited IFNγ and IL-6 production by activated T and B cells, respectively. In vivo, IL2/CD25/IL10 supported expansion of Tregs as well as activation of pSTAT3 in macrophages and reduced LPS-dependent production of IL-6 and TNFα, indicative of bifunctional activity. When 12-week-old female NOD mice received IL2/CD25/IL10 for 5-weeks, IL2/CD25/IL10 (mean survival &gt;50 week) was more effective than IL-2/CD25 (mean survival 26 weeks) in preventing progressive diabetes. Collectively, these data support the notion that a combination approach, where Tregs are increased and autoreactive T cells are limited, in this case through IL-10, will enhance the efficacy of a Treg-targeted immunotherapy of autoimmunity.
Comparative assessment of selective Janus Kinase inhibitors in rheumatoid arthritis mouse model: Insights into immune modulation and therapeutic implications
Authors: Xue Chengfeng , Wang Jingjing , Xu Na , Pei Yaqiong , Chen Donghai , Sun Jiaping , Gu Qingyang , Zhang Qiyao 
Published: November 2024

Abstract
Janus kinase inhibitors (JAKi) are a class of orally available drugs for treating rheumatoid arthritis (RA). With a focus on enhancing safety profiles and therapeutic efficacy, the development of JAKi has transitioned from first-generation non-selective inhibitors to second-generation selective inhibitors. To better understand the roles of distinct JAK isoforms in the pathogenesis and progression of RA, we employed the Collagen-Induced Arthritis (CIA) mouse model to evaluate the effects of four selective JAK inhibitors: Upadacitinib (JAK1 inhibitor), CEP-33779 (JAK2 inhibitor), Ritlecitinib (JAK3 inhibitor), and Deucravacitinib (TYK2 inhibitor).
All four JAK inhibitors showed varying degrees of alleviation in paw swelling and histological severity. We utilized flow cytometry to characterize the lymphoid and myeloid compartments within the spleen and Olink technology to analyze cytokines in the plasma.
Our findings suggested that JAK1 and JAK3 play an important role in the differentiation and proliferation of T cells, while JAK2 is crucial in the development of myeloid cells. Notably, the TYK2 inhibitor exhibited significant effect in suppressing the expression of inflammatory cytokines.
In summary, this study represents a comprehensive comparison of four selective JAKi in the CIA mouse model, offering valuable insights into immune modulation and potential implications for optimizing the benefit-risk profile of selective JAK inhibitors in the treatment of RA.
Hydroxychloroquine and its derivative enhance efferocytosis and modulate the gut microbiome in pristane-induced lupus mice
Authors: Tsay Gregory , Xue Ting-Yin , Hsieh Fei-Hung , Tsay Jun-Chieh , Peng Hsin-Yi , Yeh Yung-Ju , Tsao Yen-Chi , Yin Mei-Chin , Chao Che-Yi , Tsai Shu-Yao , Lin Chung-Ping , Chen Jiunn-Horng , Huang Chung-Ming , Chang Hen-Hong 
Published: November 2024

Abstract
Background Impaired clearance of apoptotic cells (efferocytosis) and microbiota dysbiosis is related to pathogenesis of systemic lupus erythematosus (SLE). Hydroxychloroquine (HCQ) is frequently prescribed for the treatment of SLE. Here, we evaluate changes in efferocytosis and the gut microbiome in mice with pristane-induced lupus (PIL) by HCQ administration.
Methods PIL mice were studied with or without HCQ, the HCQ derivatives were modified from HCQ by molecular modification. Efferocytosis was determined in RAW 264.7 cells and peritoneal macrophages from mouse ascites fluid. The gut microbiome was analyzed using Illumina sequencing.
Results HCQ enhanced efferocytosis in RAW 264.7 cells, the HCQ derivative AL127 also much enhanced efferocytosis. HCQ also significantly suppressed ascites production of proinflammatory cytokines interleukin (IL)-6 and tumor necrosis factor alpha (TNF-α) in PIL mice. The Firmicutes/Bacteroidetes (F/B) ratio was notably decreased in comparison to untreated controls (p&lt;0.05). Furthermore, administration of HCQ in PIL mice resulted in an increased F/B ratio.
Conclusions In this study, both HCQ and its derivative exhibited a capacity to enhance efferocytosis in RAW 264.7 cells and peritoneal macrophages of PIL mice. Furthermore, HCQ treatment reversed the pristane-induced reduction in the Firmicutes/Bacteroidetes (F/B) ratio. These findings underscore the roles of HCQ in facilitating efferocytosis.
Tumor-associated KLRG1+ Tregs drive a dominant immunosuppressive program in Non-small Cell Lung Cancer
Authors: Oluwatola Oluwaseyi , Oseni Saheed , Bag Arup , Adeegbe Dennis , Pilon-Thomas Shari 
Published: November 2024

Abstract
Killer cell lectin-like receptor G1 (KLRG1) is a co-inhibitory receptor expressed on T and NK cells. While KLRG1’s role in effector T and NK cells has been extensively documented, its impact on regulatory T cells (Tregs) remains poorly understood. This study aims to characterize and compare the immunosuppressive potential of KLRG1+ Tregs in contrast to KLRG1- Tregs and their therapeutic targeting in non-small cell lung cancer (NSCLC).
CTV-labelled effector T cells were cultured with TAMs and tumor digests in the presence or absence of each Treg subpopulation for 5 days in a Treg suppression assay. For in vivo studies, T cells with either KLRG1+ or KLRG1- Tregs were adoptively transferred into lung tumor-bearing mice. Intracellular staining for GZB, IFN-γ and TNF-α was performed on the processed tumor cells. Lastly, cohorts of tumor-bearing KP mice were treated with a KLRG1-blocking antibody for 6 weeks.
Our study revealed that KLRG1+ Tregs exhibit robust suppressive functions both in vitro and in vivo, contingent upon intact KLRG1 signaling. Blockade of KLRG1 in vitro and in vivo profoundly arrested tumor growth, augmented effector T cell activity, and significantly prolonged the survival of tumor-bearing mice.
In conclusion, KLRG1+ Tregs drives immunosuppressive activity in the tumor microenvironment; this provides novel insights into the functional dynamics of KLRG1+ Tregs and underscores their therapeutic potential for boosting anti-tumor T cell responses in NSCLC.
CAR T cells against TRP1 significantly reduce melanoma growth in vitro and in vivo
Authors: Boucher Justin , Austin Anna , Kostenko Elena , Guevara-Patino Jose 
Published: November 2024

Abstract
CAR T cells have shown great efficacy for patients with liquid malignancies. However, CAR T cell treatments have had limited efficacy in patients with solid lesions, such as melanoma. One reason is the lack of suitable targets specific to the tumor antigen, which also do not cause toxicity. TRP1 is abundantly expressed in melanoma cells as well as in healthy melanocytes. While targeting healthy melanocytes may cause vitiligo, that condition is not life threatening. Previous work shows monoclonal antibodies against TRP1 reject melanoma tumors and lung metastases. Therefore, we designed a CAR with an scFv against human TRP1 along with 41BB and CD3z costimulation. When we examined CAR T cell cytotoxicity in vitro, we found a significant reduction in melanoma target cells after 48hrs co-culture with CAR T cells at 10:1 and 5:1 E:T ratios compared to untransduced (UT) cells. CAR T cell cytokine production after melanoma co-culture also showed significantly higher IFNg compared to UT cells. To examine CAR T cell efficacy in vivo, we challenged NSG mice intradermally with melanoma tumor cells. Four days after the tumor challenge, mice were given CAR T or UT cells by tail vein injection. We found that mice given anti-TRP1 CAR T cells had significantly reduced tumor area by day 10 and had significantly increased survival compared to mice given UT cells. This work shows that TRP1 can be effectively targeted by CAR T cells and increase the survival of mice with melanoma.