bluuebunny/crossref_metadata_2025_split huggingface dataset:
Targeting γδ T cells for the immunotherapy of endometrial cancer
Authors: Cazzetta Valentina , Portuesi Rosalba , Fiamengo Barbara , Vitobello Domenico , Mikulak Joanna , Mavilio Domenico 
Published: August 2023

Abstract
Endometrial cancer (EC) is the most common gynecological neoplasm and the only gynecologic cancer with increasing incidence and mortality. EC is generally diagnosed at an early stage and is well managed with surgery and/or chemotherapy. However, EC patients with advanced/recurrent disease have a poor outcome and respond poorly to current treatments. Thus, there is the need to better investigate novel approaches. In this context, understanding the biology of the immune system is essential for recognition of new targets as well as optimization of existing immunotherapies.
Growing evidences highlight an important role of both adaptive and innate immunity in generating a suppressive environment. However, little is known about the specific role of unconventional gamma delta (γδ) T cells. Therefore, the aim of this project is to characterize the frequency, phenotype and function of γδ T cells in EC. We used flow cytometry to characterize γδ T cells in peripheral blood (PB) and tumor tissue of EC patients and in PB of healthy donors and analyzed two public datasets of single cell RNA-sequencing to deepen the transcriptional profile of γδ T cells. These analyses revealed that the main subsets of γδ T cells, namely Vδ1 and Vδ2 T cells, infiltrate the tumor of EC patients. Moreover, they are characterized by the upregulation of immune checkpoints, such as PD-1. However, upon stimulation, they maintain their anti-tumor potential producing cytokines, e.g. IFN-γ and TNF-α.
Overall, γδ T cells infiltrate EC and express immune checkpoints that can be targeted to implement the current immunotherapeutic strategies. Thus, further investigations are needed to exploit γδ T cells for EC therapy.
NCG-hIL15 humanized mice - an ideal model for human immune reconstitution of NK cells
Authors: Chen Santi Suryani , Liu Xing , Wu Meirong , Wang Huiyi , Yu Weiwei , Sun Hongyan , Ju Cunxiang , Wang Hongyu , Li Zhiying , Moore Mark , Zhao Jing , Xiang Gao 
Published: August 2023

Abstract
Immunodeficient NCG mice were unable to reconsituted human NK cells, but after knockin of human IL-15 (hIL-15), the NCG-hIL15 mice could assist the reconstitution of human NK cells. Compared with NCG, the level of hIL-15 was significantly increased in heterozygous and homozygous NCG-hIL15 mice. After transplanting human PBMC (huPBMC-NCG-hIL15) or NK cells into NCG-hIL15 (huNK-NCG-IL15), the level of peripheral blood hCD56+ NK cells in huNK-NCG-hIL15 mice were much higher than that in huPBMC-NCG-hIL15 mice. However, the reconstituted proportion of human CD3+ T cells in huNK-NCG-hIL15 mice was not comparable in huPBMC-NCG-hIL15 mice. The level of NK cell reconstitution is also highly dependent on the donor NK cells. The expression of perforin in the peripheral blood of huNK-NCG-hIL15 mice was significantly higher than that in huPBMC-NCG-hIL15 mice. In vitro functional analysis of NK cells from huPBNK-NCG-hIL15 and huPBMC-NCG-hIL15 showed Granzyme B expression in peripheral blood were comparable and human NK cells purified from splenocytes of huPBNK-NCG-hIL15 mice were cytotoxic upon coculture with Raji cells in the presence of Rituximab. Based on these in vitro data, we established huNK-NCG-hIL15 mice subcutaneously engrafted with Raji cells, and the in vivo efficacy of rituximab was evaluated. Efficacy study data showed that rituximab significantly inhibited the Raji tumor cells growth in huNK-NCG-hIL15 mice. Both huPBMC-NCG-hIL15 and huNK-NCG-hIL15 mouse model can rapidly reconstitute functional human NK cells compared to CD34+ HSC engrafted NCG-hIL15 mice. The development of huNK-NCG-hIL15 is an ideal mouse model to specifically evaluate the anti-tumor efficacy of drugs targeting human NK cells.
Microparticle-delivered stimulator of interferon genes (STING) agonist suppresses experimental autoimmune encephalomyelitis (EAE) via expansion of FoxP3+ CD4+ Tregs
Authors: Elzoheiry Manal Ali , Wang Yan , Kasimoglu Ezgi , Seyedsadr Maryamsadat , Batty Cole , Bachelder Eric , Kristy Ainslie , Markovic-Plese Silva 
Published: August 2023

Abstract

Background
The cGAS-cyclic GMP-AMP (cGAMP)-stimulator of intracellular STING receptors induces a powerful IFN-I response. Microparticles (MPs), synthesized biodegradable polymer, passively target the APCs, resulting in intracellular delivery of immuno-regulatory treatment with dose sparing and better tolerability. cGAMP MP treatment of EAE induced both IFN-I-dependent and -independent immunosuppressive responses through induction of IL-27 and IL-10.

Results
We show that cGAMP MP treatment reduced clinical disease scores of EAE after 3 doses. By flow cytometry, cGAMP MPs decreased absolute number of mononuclear cells in the CNS infiltrates, and the percentages of CD3− CD45high CD11+b Ly6C+ monocytes, CD11c+ and CD11c+ CD11b+ DCs, as well as their expression of CD80 and CD86, suggestive of tolerogenic phenotype. Moreover, we detected an increased percentage of FOXP3+CD4+ Tregs in CNS infiltrates, with increased expression of IL-10, IL-27R, and Granzyme B, and decreased percentage of IL-17A+CD4+ cells in cGAMP MP-treated mice. Furthermore, the supernatants of the in vitro co-cultured cGAMP MP-treated monocytes with CD3+CD4+, isolated from EAE spleenocytes, show increased IL-10 and IL-27 levels. Additionally, the in vitro D3+CD4+ cells show increased FOXP3+ expression with higher CTLA-4, and GITR in cGAMP treated group. Moreover, FOXP3cre+/+ IL-27RA fl/fl mice showed more aggressive disease compared to wild-type mice in attempt to test whether IL-27 induced effect of cGAMP MPs is mediated via Tregs.

Conclusion
cGAMP MPs suppress RREAE through induction of tolerogenic APCs and increase secretion of IL-10 and IL-27, which induce Treg expansion indicating its potential use as a tolerogenic therapy in patients with RRMS.
Suppression of IGRP-specific CD8 T cells following induction of tolerance to a Hybrid Insulin Peptide CD4 neoepitope
Authors: DiLisio James E , Reyes Kaitlin , Beard K Scott , Neef Tobias , Miller Stephen D , Baker Rocky L , Haskins Kathryn 
Published: August 2023

Abstract
An autoimmune-mediated death of insulin-producing beta cells, orchestrated by effector CD4 and CD8 T cells that recognize islet antigens, results in Type 1 Diabetes. A dominant CD4 islet autoantigen in the NOD mouse model of autoimmune diabetes is a neoepitope termed the 2.5 Hybrid Insulin Peptide (2.5HIP). By delivering the 2.5HIP as an antigen-specific immunotherapy on tolerogenic PLG-nanoparticles, islet grafts in diabetic NOD mice survived longer and cytokine production in autoreactive CD4 and CD8 T cells was suppressed, including the well-studied IGRP tetramer+ (tet+) CD8 T cells. Using both diabetic transplant recipients and prediabetic NOD mice treated with 2.5HIP nanoparticles, we examined mechanisms of peripheral tolerance induction to a dominant CD4 neoepitope and the impact on the function of IGRP CD8 T cells. Following induction of tolerance to the 2.5HIP, there was an increase in dysfunctional surface marker expression (PD1+ TIM3+) on effector 2.5HIP tet+ CD4 T cells in the spleen of treated mice. Along with a decrease in effector function, we observed an increase in the fraction of both Treg and Tr1 2.5HIP tet+ T cells expressing IL10 in the islets, spleen, and draining lymph nodes of tolerized mice. Concurrently, IGRP tet+ CD8 T cells accumulated in the draining lymph nodes and were less able to traffic and infiltrate islets. Of the IGRP tet+ cells that entered islets, there were fewer cytolytic CX3CR1+ effector cells in tolerized mice. The robust functional increase in both Treg and Tr1 2.5HIP tet+ T cells may provide a potential mechanism for the accumulation of IGRP tet+ CD8 T cells in the draining lymph node and explain their inefficient trafficking into, and function within, islets following tolerance induction.
NIH - T32 5T32DK120520-03, R01 2R01DK081166-11)
JDRF 2-SRA-2020-907-S-B
Artemotil mediates protection in experimental colitis through the induction of Type 1 regulatory T cells
Authors: Madan Upasna , Verma Bhawna , Binayke Akshay , Awasthi Amit 
Published: August 2023

Abstract
Artemotil (β-arteether) is a semi-synthetic derivative of artemisinin - a natural compound isolated from the plant Artemisia annua and used worldwide as a front-line anti-malarial drug. Besides its anti-malarial effect, several studies have reported its potential use as an anti-tumor and anti-inflammatory agent. Artemotil was found to suppress the generation and function of Th1 and Th17 while promoting the generation of Foxp3+ regulatory T cells (Tregs). The role of Artemotil in modulating the generation and functions of Type 1 regulatory T cells is not yet identified. Tr1 cells are one of the key cell types that are essential for inhibiting inflammatory response through IL-10. In this study, we report an additional role of Artemotil by selectively inducing the generation of Tr1 cells induced by IL-27 by upregulating the signature of Tr1 cells genes such as c-maf and Prdm1, IRF-1, and Batf. We found that co-administer of Artemotil with anti-CD3 antibody increases the induction of IL-10 and frequency of Tr1 cells while suppressing Th1 and Th17 cells in vivo associated with less severe T-cell-induced enteropathy and intestinal inflammation. We further show the anti-inflammatory role of Artemotil in the chronic T-cell-induced colitis model in which Artemotil treatment alleviates the signs of colitis associated with the increased frequencies of Tr1 cells. Taken together, these data suggest that Artemotil provides protection in T-cell-mediated colitis by increasing the expansion of Tr1 cells and inhibiting the generation of Th1 and Th17 cells.
This study was supported by grants from Science & Engineering Research Board (SERB), Department of Science and Technology (DST), Government of India (CRG/2018/002653)
Serum Albumin fused Flt3L expands a tolDC compartment and amplifies tolerogenic antigen immunotherapy for the prevention of pathogenic anti-drug antibody responses
Authors: Alpar Aaron T , Wallace Rachel P , Refvik Kirsten C , Hubbell Jeffrey 
Published: August 2023

Abstract
From antibodies to enzyme replacement therapies, biologics and protein drugs have become a major part of the medical market and revolutionized medicine. Repeated administration of many of these drugs, however, leads to humoral responses and potentially drug failure via neutralization, clearance, or immune related infusion reactions. Once pathogenic anti-drug antibodies are formed, patients must take broad immunosuppressive drugs, undergo continuous B cell depletion, or discontinue treatment entirely. Currently, no drugs are approved by the FDA to prevent the induction of anti-drug antibodies. Herein, we characterize the effect of our engineered cytokine, Flt3L-SA, in vivo and demonstrate how our molecule expands the dendritic cells while enhancing a tolerogenic, TGFβ expressing subpopulation. We then leverage these dendritic cells to attenuate the antibody response to the highly immunogenic drug Rasburicase and ultimately reduce antibodies by 100-fold when administered as part of a tolerogenic regime with low dose Rasburicase. We also show how this reduction in antibodies results in fewer and less severe infusion reactions upon final challenge, evidencing both a reduction in the antibody response as well as pathology. Future work will aim to determine the exact mechanism by which this tolerance is achieved as well as demonstrate efficacy in the context of a wide range of extremely immunogenic therapies, including enzyme replacement therapeutics for lysosomal storage disorders and AAV therapies.
Supported via the Chicago Immunoengineering Innovation Center and the Alper Family Foundation
Effects of BAMBI deficiency in the development of Systemic Lupus Erythematosus
Authors: Adrián Paula Pérez , Ruso Fulgencio , Cappitelli Vincenzo , Casado Victoria , Merino Jesús , Merino Ramón 
Published: August 2023

Abstract
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the production of autoantibodies and immune complexes deposition in various organs. Follicular CD4+ T cells play a crucial role in this autoimmune syndrome by activating autoreactive germinal center B cells. The transforming growth factor β (TGBβ) inhibitor BAMBI (Bone Morphogenetic Protein and Activin Membrane-Bound Inhibitor) is known to control the differentiation of CD4+ T lymphocytes into either tolerogenic Tregs or pathogenic Th17 cells. BAMBI absence in mutant mice or its pharmacological inhibition with the B101.37 monoclonal antibody promotes an increase in Treg differentiation and activity and at the same time, a reduction in Th17 cell differentiation, resulting in the inhibition of autoimmune disease development. Based on these findings, in the present study we have analysed the effects of BAMBI deficiency in the development of SLE. We have employed the chronic graft-versus-host disease lupus model induced after the transference of 40–60 × 106 lymphocytes from B6 or B6-BAMBI-KO mice into either Bm12 or Bm12-BAMBI-KO recipients. As reported previously, Bm12 mice receiving lymphocytes from B6 mice produce high levels of circulating IgG anti-DNA and anti-nucleosomal autoantibodies and exhibit linear glomerular IgG deposits in their kidneys. In contrast, both Bm12 mice injected with allogeneic B6-BAMBI-KO lymphocytes and Bm12-BAMBI-KO recipients receiving lymphocytes from B6 donors do not produce significant levels of circulating autoantibodies and do not reveal immune complexes deposits. Taken together, these results indicate that the absence of BAMBI in either donor or recipient lymphocytes protects mice against the development of SLE.
BAFF and APRIL signaling through B cell maturation antigen affects regulatory transitional B cells in multiple sclerosis
Authors: Kumar Gaurav , Massey Kaylea , Ko Rose M , Axtell Robert C 
Published: August 2023

Abstract
Multiple sclerosis (MS) is a debilitating neuro-autoimmune disease that can be stratified into relapsing remitting (RR) MS, secondary progressive (SP) MS, and primary progressive (PP) MS. B cells can have both inflammatory and regulatory functions in MS. Transitional B cells are considered to have regulatory functions in MS. BAFF and APRIL, two cytokines, signals through the receptors BAFFR, TACI and BCMA and are thought to drive the regulatory function of transitional B cells. Here, we assessed the frequencies and expression of BAFFR, TACI and BCMA on the surface of regulatory transitional B cells in the blood of RRMS, PPMS and healthy individuals along with soluble versions of these receptors in the plasma. We found that transitional B cells were decreased in both RRMS and PPMS patients. The surface expression of BCMA was significantly lower on transitional B cells from RRMS patients but not PPMS patients compared to healthy individuals with no change in the expression of TACI and BAFFR. In addition, we found elevated levels of soluble BCMA in blood of PPMS patients but not RRMS patients compared to healthy and no difference in soluble BAFFR and TACI. Our data suggests that reduced BCMA signaling is associated with reduced numbers of transitional B cells in RRMS and PPMS. In RRMS, expression of BCMA is reduced on regulatory transitional B cells. In PPMS patients, there are higher levels of soluble BCMA which could act as a blocker of APRIL signaling through BCMA on the surface of transitional B cells. Altogether, our data suggests that reduced BCMA signaling in transitional B cells is contributing to the MS disease and the dysregulation of the BCMA signaling in RRMS and PPMS may be through a different mechanism.
This study was supported by grants from the NMSS (RG-1602-07722), the NIH (R01AI137047 and R01EY027346) awarded to R.C. Axtell.
An albumin interleukin 33 fusion protein with enhanced lymph node retention suppresses murine models of multiple sclerosis
Authors: Budina Erica , Ishihara Jun , Solanki Aniruddhsingh , Nguyen Mindy , Hubbell Jeffrey A 
Published: August 2023

Abstract
Interleukin-33 (IL-33) is an immunoregulatory cytokine that suppresses pathogenic Th17 T cells. Recombinant IL-33 has been shown to prevent disease onset in experimental autoimmune encephalomyelitis (EAE), a murine model of multiple sclerosis. However, the systemic administration of IL-33 has not been clinically translated, in part because the cytokine’s short half-life makes it difficult to achieve therapeutically effective concentrations in the secondary lymphoid organs. To overcome this challenge, we have recombinantly fused mouse serum albumin (SA) to IL-33. ST2(IL-33R)-expressing mouse group 2 innate lymphoid cells treated with serum albumin fused IL-33 (SA IL-33) increased CD25 expression and IL-13 production in a dose-dependent manner. SA fusion prolonged IL-33 half-life and increased its concentration in the lymph nodes of naïve C57BL/6 mice. Prophylactic administration of SA IL-33 prevented the onset MOG-induced EAE, demonstrating equal efficacy to the clinically approved MS drug fingolimod. In mice with already-established EAE, administration of SA IL-33 reduced disease score and improved weight gain. At the cellular level, SA-IL-33 treatment reduced the frequency of CD45+ cells, including disease causing Rorgt+ CD4+ T cells, in the spinal cord. EAE mice treated with SA-IL-33 displayed an increase in ST2+ FoxP3+ CD25+ Tregs and Th2 T cells in both the spleen and spinal cord draining lymph nodes, suggesting that SA-IL-33 treatment suppresses EAE by expanding the protective type 2 response.
In vivo PK/PD assessment of invisibilized IgG cleaving protease for chronic treatment of autoimmune diseases
Authors: Sanmarco Liliana , Pellerin Alex , Green Tobias , Newton Andita , Borah Minasri , Anderson Jordan , Mojta Shanell , Hollfelder Melissa , Higginson-Scott Nathan , Otipoby Kevin , Mascanfroni Ivan 
Published: August 2023

Abstract
Pathogenic autoantibodies are key effectors of inflammation, promoting complement cascade activation, immune cell response, and the consequent tissue damage in autoimmune disease and transplant rejection.
Proteases that cleave IgG antibodies present a novel therapeutic opportunity to treat autoimmune diseases and antibody mediated transplant rejection. Using our IMPACT platform leveraging machine learning, we engineered IgG proteases with reduced immunogenicity for acute and chronic treatment of antibody-mediated diseases. In addition, the invisibilized protease was fused to a Fc to extend its half-life.
To evaluate these IgG proteases in vivo we developed a pharmacokinetics and pharmacodynamics (PK/PD) model. C57BL6 mice were dosed with intravenous immunoglobulin (IVIg) at different time points after protease treatment. Protease and IgG levels were quantified by MSD 15 minutes before and two hours after IVIg injection, respectively. IgG cleavage was detected at early time points which correlated with higher systemic drug levels. Additionally, the Fc-fused protease was detected at later time points with increased systemic enzymatic activity, as compared to the enzyme domain only. Finally, we evaluated the potential of the Fc-protease to cleave antigen bound IgG at the tissue level in a mouse anti–glomerular basement membrane (GBM) induced nephritis model. In summary, our in vivo studies established a PK/PD relationship between exposure and cleaved IgG while showing the benefit of fusing the protease domain to a Fc.
Tissue kallikrein suppresses interferon responses and reduces depressive-like behavior in lupus-prone mice
Authors: Sriram Uma , Nocito Cassandra , Lubinsky Cody , Hand Michelle , Seliga Alecia , Persidsky Yuri 
Published: August 2023

Abstract
Recent studies show that systemic immune responses, including Type I IFNs, affect neuroinflammation and neuropsychiatric responses. Major depression is the most common psychiatric presentation of NPSLE. Reports indicate that neuropsychiatric symptoms, especially depression and anxiety, may be common co-morbidities of childhood-onset of SLE and clinical studies have demonstrated that IFNα can cause depressive symptoms. The management of patients with neuropsychiatric lupus (NPSLE) continues to be a major therapeutic challenge. We have shown that the kallikrein-kinin system (KKS) [comprised of kallikreins (klks), bradykinins (BKs)] and angiotensin converting enzyme inhibitors (ACEi’s), suppressed Type I IFN responses in murine dendritic cells (DCs) from normal and lupus-prone mice and human peripheral blood mononuclear cells (PBMC). Other studies have shown that klk genes are decreased in lupus patients; giving exogenous klk1 ameliorated kidney pathology in mice. We administered (retro-orbitally) adenovirus carrying the kallikrein gene (klk1- a widely expressed tissue klk) in the MRL/lpr lupus-prone mice (at early disease onset) and analyzed immune responses and depressive-like behavior response. Klk1 improved depressive-like behavior, suppressed Interferon Responsive Genes, decreased splenic plasmacytoid DCs, and reduced plasma IFNa levels. Together, these findings suggest that kallikreins and other KKS molecules may be used to control IFNa production/responses and other inflammatory responses in SLE and NPSLE.
This work was supported by NIH Grant R21AR071679 and funds from the Department of Pathology and Laboratory Medicine.
Dynamic single cell lineage recording of endogenous viral specific CD8 T cells reveals multiple potential cell fate pathways
Authors: Abdullah Leena , Emiliani Francesco , Vaidya Chinmay , McKenna Aaron , Huang Yina H 
Published: August 2023

Abstract
In response to viral infection, antigen specific naïve CD8 T cells expand to give rise to a heterogenous pool of effector cells consisting of short-lived effector cells (SLECs) and memory precursor effector cells (MPECs). While these effector populations are phenotypically and functionally well characterized, we still don’t fully understand how they arise from antigen responsive naïve CD8 T cells. We have combined progressive lineage recording with single-cell RNA sequencing and T cell receptor sequencing to delineate the early differentiation of OVA-specific endogenous CD8 T cells in response to acute VSV infection. Transcriptional profiling of CD8 T cells captured at the peak of T cell response confirmed 8 distinct T cell states including a unique interferon responsive cluster. RNA Velocity trajectory analysis supported an asymmetric model of CD8 T cell differentiation where early effector cells gave rise to SLECs while MPECs differentiated into further memory precursors. Moreover, our CRISPR/Cas9-based lineage recorder uncovered T cell clones of various sizes and allowed us to follow up to 5 generations of differentiating CD8 T cells. We observed that expanded clones comprised of memory and effector CD8 T cells while medium size clones preferred memory or effector fate, suggesting that different clones follow different differentiation models. Thus, using RNA-seq based trajectory analyses and a dynamic lineage recorder we uncovered potential differentiation pathways taken by early viral specific CD8 T cells. Our single cell full length TCR-seq will further add to our current models and highlight TCR sequences with better memory potential in response to infection.
Supported by grants from NIH (R01 AI089805, R01 CA254042) and a training fellowship from the Burroughs Wellcome Fund.