bluuebunny/crossref_metadata_2025_split huggingface dataset:
Impact of curcumin and its metabolites on the immune response of type 1 diabetes-prone mice
Authors: Alard Pascale , Kosiewicz Michele M , Ma Jing 
Published: January 2023

Abstract
Recent studies have shown that diferuloylmethane (curcumin) ameliorates autoimmune diseases, including multiple sclerosis, rheumatoid arthritis, lupus and diabetes in animal and human models. We examined the effects of curcumin and its metabolites on the immune response in NOD mice. We assessed their ability to regulate pro-inflammatory cytokines and the differentiation of IFNγ-producing T cells. We also determined whether curcumin could affect Treg development and production of the anti-inflammatory cytokine, IL-10. NOD dendritic cells (DC) treated with curcumin in vitro produce decreased pro-inflammatory cytokines, including IL-12. Treatment of DC with ferulic acid and vanillin, two metabolites of curcumin, also downregulated production of IL-12. This effect was not due to an decrease in DC viability or increased production of IL-10. More importantly, NOD DC treated with curcumin and its metabolites induce fewer IFNγ-producing T cells, shown to be responsible for beta islet destruction. Mice treated with curcumin were protected from disease and this protection was associated with a decrease in IFNγ production in the spleen and pancreatic LN. Although CD103+ DC harvested from the mesenteric LN of curcumin-treated NOD mice were better at inducing in vitro Treg conversion, the levels of CD4+Foxp3+ and CD4+Foxp3+Neuropilin- cells in the pancreatic LN were actually impaired in those mice. Overall, our in vitro and in vivo data show that curcumin and its metabolites suppress the differentiation of IFNg-producing cells involved in type 1 diabetes development. Curcumin or its metabolites may have therapeutic potential for the prevention of type 1 diabetes.
Antigen targeted to the apoptotic pathway via dying erythrocytes leads to profound T cell exhaustion and tolerance
Authors: Watkins Elyse A , Roberts Jaeda , Dunaif Anya A , De Loera Robert , Bailey Luke , Hubbell Jeffrey 
Published: January 2023

Abstract
The current standard of care for autoimmune disease is administration of immunosuppressive agents that require a lifetime of treatment and leave patients susceptible to opportunistic infections. Thus, there is a great need for therapies that induce lasting and antigen-specific tolerance, which would require fewer treatments and allow the immune system to fully respond to pathogens. A promising target for the induction of antigen-specific tolerance is the apoptotic pathway. Antigen-presenting cells create a tolerogenic milieu upon recognition of apoptotic debris, and antigen presentation by APCs to T cells in the absence of inflammatory cues typically leads to anergy and deletion.
A large source of endogenous apoptotic cells is erythrocytes, or red blood cells. The purpose of this study is to discover antibody fragments (Fabs) that bind erythrocytes, and use these Fabs to deliver antigen to erythrocytes for processing via the apoptotic pathway and presentation to T cells for tolerance. Phage display on mouse erythrocytes yielded high affinity Fabs that lacked binding to peripheral blood mononuclear cells and induced no hematological changes. Erythrocyte-binding Fabs were recombinantly expressed with the model antigen ovalbumin and administered to mice. Targeting ovalbumin to erythrocytes led to robust abortive proliferation of antigen-specific T cells. Remaining cells appeared phenotypically exhausted, and had reduced capacity to produce inflammatory cytokines upon restimulation. Antigen presentation of erythrocyte-targeted antigen occurred in the spleen, because T cell exhaustion was completely reversed in splenectomized mice. Thus, targeting erythrocytes may induce lasting antigen-specific tolerance.
Novel compound JOAB-40 decreases inflammation in vitro and colitis severity in vivo by using the murine Dextran Sulfate Sodium (DSS) model
Authors: Szollosi Doreen E , Khajah Maitham , Hawai Sanaa , Ghoneim Ola , Bill Ashley , Daniels-Monk Mallory , Edafiogho Ivan O. 
Published: January 2023

Abstract
Crohn’s disease and ulcerative colitis are two disorders categorized as inflammatory bowel diseases (IBD). Patients with IBD may have inflammation extending the full thickness of the gut and non-gut organs mediated in part by Th-induced cytokines IL-1β, -2, -12, -13, -17, TNF-α, and IFN-Υ. To mimic this in vivo, DSS given in drinking water (3% w/v) can induce colitis in mice. Our previous work with novel anti-inflammatory enaminone E121 has shown that treating mice with E121 along with DSS-induced colitis (prophylactic approach) reduced colitis severity at gross and histological levels at the 30–60 mg/kg doses. Modifications made to E121 have resulted in new compounds (JOAB series) that include a piperazino ring which can not only decrease levels of pro-inflammatory cytokines but also inhibit chemokine signaling through CCR2. In vitro data suggest that JOAB-40 can reduce levels of TNF, IL-1, IL-6, and IL-12 in LPS-stimulated macrophages. Since JOAB-40 was successful in decreasing inflammation in a mouse model of hemarthrosis without toxicity, we assessed its efficacy in decreasing inflammation associated with colitis. Prophylactically, mice treated with JOAB-40 had a more profound reduction in severity of colitis at lower doses (1–10 mg/kg) than E121. At higher doses (30–60 mg/kg), both E121 and JOAB were equivalent in their potency in reducing colitis severity. When both agents were given after colitis induction (treatment approach), JOAB-40 reduced colitis severity at 10, 30, and 60 mg/kg doses (by 50% compared to vehicle), but E121 did not reduce colitis severity at histological level when used at similar dose range. These data suggest JOAB-40 is more effective in reducing colitis severity in mice compared to the parental E121 compound.
Contact-dependent suppression of FVIII-specific memory B cells by BAR-Tregs
Authors: Zhang Ai-Hong , De Paula Alves Sousa Alessandra , Venkatesha Shivaprasad H , Scott David W 
Published: January 2023

Abstract
Anti-drug antibody formation poses tremendous obstacles for optimal treatment of hemophilia A (HA). In this study, we sought to utilize chimeric receptor-modified natural regulatory T cells (Tregs) to target FVIII-specific memory B cells, which are responsible for persistent anti-FVIII neutralizing antibodies (inhibitors) in HA patients. Thus, CD4+CD25hiCD304+ natural Tregs were FACS sorted from naïve C57BL/6 mice and retrovirally transduced to express a chimeric B-cell antibody receptor (BAR) containing the immunodominant A2 domain of FVIII. Plasmablast-depleted (CD138neg) splenocytes from FVIII/IFA immunized FVIII-knockout HA mice served as the source for FVIII-specific memory B cells, which were then specifically stimulated in vitro with FVIII and enumerated in B-cell ELISPOT assays. Adding A2-BAR Tregs (1 per 150 splenocytes), but not conventional T cells, to the CD138− splenocytes significantly suppressed the formation of anti-FVIII producing cells, compared to the non-relevant OVA-BAR Tregs control group. Transwell experiments confirmed that the suppression was contact-dependent. Interestingly, even in the presence of antibody to FVIII (so-called inhibitors), similarly prepared CD4+CD25hiCD127low A2-BAR human natural Tregs completely suppressed polyclonal anti-FVIII Elispots formation, indicating potential by-stander suppression by A2-BAR Tregs on all FVIII-specific B cells in the local milieu. In conclusion, we demonstrated in vitro that FVIII domain-expressing BAR Tregs could efficiently target and suppress FVIII-specific memory B cells. Supported by a Bayer Hemophilia Grant [AHZ] and NIH HL126727 [DWS].
Mechanisms of “Peptide Alarm Therapy”: Can antiviral memory T cells directly kill cancer cells in an MHC Class I-dependent way?
Authors: Gavil Noah Veis , Joag Vineet , Rosato Pamela C. , Masopust David 
Published: January 2023

Abstract
“Peptide alarm therapy” is a novel cancer immunotherapy that repurposes antiviral memory CD8+ T cells within tumors to drive antitumor immunity. After a primary infection, antigen-specific CD8+ T cells establish a surveillance program that positions T cells throughout the entire body. In particular, resident-memory CD8+ T cells (Trm) permanently reside in all tissues after primary antigen exposure and survey their immediate surroundings for possible re-exposure. In addition to normal tissue, antiviral Trm populate solid tumors. In contrast to tumor-specific T cells, intratumoral antiviral Trm are not exhausted and demonstrate potent reactivation with cognate peptide. This re-activation of Trm in tumors leads to upregulation of effector molecules and chemokines, the local recruitment of circulating immune cells, and the increased migration of dendritic cells to the tumor-draining lymph node. The intratumoral injection of peptide, and the subsequent reactivation of antiviral Trm, induces tumor growth suppression and exhibits synergy with immune checkpoint blockade, specifically PD-L1. The mechanisms driving this antitumor immune response remain uncharacterized. In this study, we have tested whether viral peptide presentation by cancer cells is necessary for the efficacy of peptide alarm therapy. A targeted knock-out of the MHC Class I allele, H-2Kb, in the B16.F10 melanoma cell line has been generated and employed in combination with the OT-I Tg CD8+ T cells, which bind the SIINFEKL:H-2Kb peptide:MHCI complex. Our data suggest that peptide presentation by cancer cells is not necessary for the reactivation of intratumoral antiviral Trm, and does not reduce tumor growth suppression in the context of peptide alarm therapy.
Inhibition of TET (Ten-Eleven Translocation) enzymes enhances systemic anti-tumor immune responses in T cells
Authors: Seo Hyungseok , Yang Chao , Zhang Wade , Hogan Patrick , Rao Anjana 
Published: January 2023

Abstract
TET enzymes facilitate DNA demethylation by oxidizing 5-methylcytosine to 5-hydroxymethylcytosine and other oxidation products in DNA. Loss-of-function mutations in TET enzymes are associated with increased proliferation and increased stem cell function in many cellular lineages. Moreover, there are several hints that inhibition of TET enzymes in tumor-infiltrating T cells might enhance anti tumor immune responses in solid tumors. For instance, TET2 deficiency in mouse T cells increased the number of CD8+ central memory T cells, and TET deficiency human CD8 chimeric antigen receptor (CAR) T cells increased their antigen-specific proliferation and capacity to reject tumor cells in a patient with chronic lymphocytic leukemia. Furthermore, we previously showed that TET deficiency in regulatory T cells decreased the stability of Foxp3 expression; hence TET deficiency would be expected to counter the ability of regulatory T cells to suppress anti-tumor responses. In this study, we found that TET loss of function in CD8 TILs indeed slowed tumor growth and promoted anti tumor responses in a mouse CAR T cell model. The function of TET proteins and other Fe(II) and a-ketoglutarate dependent dioxygenases is inhibited by L-2-Hydroxyglutarate (L2HG), which is normally kept at low levels by the enzyme L-2HG dehydrogenase (L2HGDH). We found that when CAR T cells were rendered low in TET activity in any one of three ways: (i) deficiency of the TET enzymes, (ii) depletion of L2HGDH and (iii) pretreatment of L2HG, they showed decreased expression of inhibitory receptors as well as increased secretion of inflammatory cytokines. Based on these results, we suggest that TET inhibition in T cells might be a promising general approach in cancer immunotherapy.
Antigen-specificity/protein/mRNA single-cell sequencing profiling of bystander CD8+tumor-infiltrating lymphocytes (TILs) in renal cell carcinoma (RCC) reveals their transcriptomic similarities with tumor-specific T cells
Authors: Li Shamin , Simoni Yannick , Zhuang Summer , Newell Evan 
Published: January 2023

Abstract
Recently, using human colorectal and lung cancer, we showed that CD8+ T cells infiltrating tumor tissue (TILs) are not only specific for tumor antigens, but are also composed of CD8+TILs specific for cancer unrelated epitopes – called bystander – such as HCMV, EBV or flu epitopes. We also showed that the surface marker CD39 can be useful for discriminating bystander (CD39−) from tumor-specific (CD39+) CD8+TILs (Simoni et al, Nature, 2018). Here, our aim was to test these findings in human Renal cell Carcinoma (RCC) and to better understand the biology of these bystander CD8+TILs. Surprisingly, our primary CYTOF analyses showed heterogeneity within bystander CD8+TILs that possess various phenotypes including effector (CD45RO+), memory (CD45RO+ CCR7+), Trm (CD69+ CD103+/–), and senescent (CD57+ KLRG1+) cell features. Using targeted mRNA single-cell sequencing combined with BD AbSeq assay, we analyzed sorted CD8+TILs from RCC patients and identified bystander CD8+TILs using oligo-tagged tetramers. Gene signature analysis of these different subsets revealed transcriptomic similarities with the tumor-specific CD8+TILs. Our data provide a comprehensive perspective of cancer unrelated CD8+TILs in RCC and suggest functional roles for these cells in tumor immune responses, which could lead to new diagnostic and therapeutic strategies.
Amplification of neoantigen-specific antitumor responses using a long-lasting IL-2 fusion protein
Authors: Hernandez Rosmely , Toomer Kevin H , Põder Janika , LaPorte Kathryn M , Malek Thomas R 
Published: January 2023

Abstract
Low frequency of tumor-specific T cells represents one major reason for failed immunotherapy. IL-2 can boost tumor-reactive T cells but its benefit is limited due to toxicity and potential off-target effects. We developed a novel IL-2/CD25 fusion protein which exhibits more favorable pharmacology when compared to IL-2. At low doses, IL-2/CD25 targets the high affinity IL-2R and selectively boosts Tregs. Here we show that a single administration of high dose mouse IL-2/CD25 in conjunction with cognate peptide and LPS amplified tumor-specific Pmel-1 CD8+ and TRP-1 CD4+ Teff cells and Pmel-1 memory cells while only transiently increasing Tregs. This approach led to effective anti-tumor responses against B16.F10 melanoma. This scheme was modified to amplify endogenous polyclonal melanoma-reactive T cells, which is translationally more relevant. Development of tumor-specific T cells required priming and boosting with a mixture of B16.F10 neoantigens and poly (I:C) followed by application of IL-2/CD25 after the boost. This resulted in largely a CD4+ anti-neoantigen specific T cell response. This immunization scheme delayed B16.F10 growth and extend survival time not only when immunized mice were challenged with B16.F10 but also for mice with pre-established B16.F10. The immune response within the tumor microenvironment was accompanied by increased frequencies of granzyme B-expressing CD4+ and CD8+ T cells. Our data show that limited application of high-dose IL-2/CD25 in the context of neoantigen vaccines offers potential advantages over IL-2 for melanoma treatment by avoiding toxicities seen with IL-2 therapy and minimizing Treg-associated tolerance.
Immunometabolism regulation by PSGL-1 signaling in tumor-specific CD8 T cells
Authors: Hope Jennifer L , Otero Dennis C , Bae Eun-ah , Stairiker Christopher J , Palete Ashley B , Faso Hannah A , de Jong Petrus , Powis Garth , Bradley Linda M 
Published: January 2023

Abstract
We previously identified that the adhesion molecule P-selectin glycoprotein ligand-1 (PSGL-1) regulates T cell function and exhaustion in response to chronic viral infection and tumors. Subsequent studies have focused on investigating the role of PSGL-1 signaling in T cell responses, with an emphasis on understanding the mechanisms by which PSGL-1 regulates T cell exhaustion. Single-cell RNA-sequencing of tumor infiltrating PSGL-1−/− CD8+ T cells identified the upregulation and differential modulation of several genes associated with T cell metabolism and enhanced intratumoral responses, including Mtor, Hif1a, and Mki67 in Gzmb/Ifng double-positive cells from tumors and Tcf7 in both tumor draining and non-draining inguinal lymph nodes. These data suggest an important role for PSGL-1 signaling in the development and maintenance of effective anti-tumor T cell responses to melanoma. Using the Seahorse glycolysis stress test, we identified that both CD4+ and CD8+ PSGL-1−/− T cells demonstrate increased glycolysis after 72 hours of in vitro activation compared to wild-type T cells. In situ activation of PSGL-1−/− CD8+ T cells demonstrated that PSGL-1−/− CD8+ T cells have increased glycolysis and increased glycolytic capacity at both sub-optimal and optimal levels of α-CD3 stimulation, and 2-NBDG glucose uptake assays confirmed increased glucose uptake by PSGL-1−/− CD8+ T cells within two hours of stimulation. Importantly, this increased glycolytic phenotype does not come at the cost of CD8+ T cell stemness, as determined by TCF-1 staining. Taken together, these data show that PSGL-1 signaling has an intrinsic and immediate role in the development of T cell responses and their metabolic profile in response to melanoma tumors.
Metformin corrects immune dysfunction and modulates tumor gene expression in overweight individuals to yield context-specific improvement of lung cancer outcomes
Authors: Barbi Joseph J , Zollo Robert , Sass Stephanie , Srinivasan Aravind , Kannisto Eric , Patnaik Santosh , Yendamuri Sai 
Published: January 2023

Abstract
Non-small cell lung cancer (NSCLC) is the foremost cause of cancer-related mortality in the U.S. While early detection followed by surgery allows for the best possible NSCLC outcome, recurrence still occurs in 30% of these cases, and existing adjuvant therapies have largely failed to improve this statistic. Therefore, pressing need exists for more effective early stage NSCLC treatments. Metformin, a widely used diabetes drug with minimal side effects, is also known to show anti-cancer activity. A retrospective study of early and multistage NSCLC patients treated at Roswell Park revealed that the survival benefit of metformin is surprisingly restricted to overweight or obese patients with a body mass index (BMI)&gt;25kg/m2. In line with this, murine lung cancer models showed that metformin treatment markedly slowed tumor growth in obese mice while minimally affecting normal controls. Flow cytometric analysis of tumor-infiltrating cells from obese mice revealed indications of immune dysfunction including elevated frequencies of suppressor cells and high levels of the inhibitory checkpoint molecules PD-1 and CTLA-4. Strikingly, metformin reversed these obesity-driven changes while promoting an abundance of activated, memory T cells in tumor-associated tissues. Transcriptome analysis of tumor gene expression in both mice and patients found that metformin counteracts discrete metabolic pressures brought on by obesity capable of dampening anti-tumor immune responses including altered expression of solute transporters and biosynthetic pathways. Our findings suggest that metformin has the potential to improve NSCLC treatment of high-BMI patients by reshaping obesity’s impact on tumor gene expression and anti-tumor immunity.
Tumor RIPK1 signaling mediates intrinsic and extrinsic mechanisms of resistance to immune checkpoint blockade therapy
Authors: Cucolo Lisa , Qiu Jingya , Klapholz Max , Shi Junwei , Gilliland Gary , Jordan Martha , Minn Andy 
Published: January 2023

Abstract
Immune checkpoint blockade (ICB) therapy can result in impressive clinical responses of multiple cancer types, however resistance is common. Mechanisms of resistance include both tumor intrinsic evasion of immune mediated killing and tumor extrinsic features in the immune-microenvironment, such as poor infiltration of tumor reactive T cells and accumulation of immunosuppressive cells. By examining transcriptomic features associated with response or resistance to ICB, we found the expression of Receptor Interacting Protein Kinase 1 (RIPK1), a critical regulator of inflammation and cell death, is significantly higher in ICB resistant murine tumor cells. We hypothesized that RIPK1 expression in the tumor cells may influence both intrinsic and extrinsic mechanisms regulating response to ICB. To test this hypothesis, we genetically deleted RIPK1 in murine tumor cell lines and found improved overall survival upon treatment compared to tumors expressing RIPK1. Mechanistically, deletion of RIPK1 in the tumor cells results in favorable changes in the tumor microenvironment, including an increased frequency of effector T cells, a decreased frequency of immunosuppressive myeloid cells, and changes in inflammatory cytokine secretion that can impact these changes in immune infiltrate. Further, deletion of the apoptosis effector Caspase 8 reversed RIPK1 KO tumor sensitization to ICB, suggesting that tumor sensitization to cell death also contributes to improved response to ICB upon RIPK1 deletion. In conclusion, RIPK1 is an important regulator at the intersection of tumor intrinsic and extrinsic pathways governing response to ICB, and therefore is a potential target for combinatorial therapies to improve patient response to ICB.
Topoisomerase-I inhibitors induce the expression of clinically relevant immunomodulators in colon cancer cells
Authors: Samuel Temesgen , Shaddox Sage H , Bass Danielle , Bedi Deepa , Datta Pran 
Published: January 2023

Abstract
Immune checkpoint inhibitors have dramatically improved the treatment of certain cancers, but not of most colon cancer types. The reasons are unknown. Camptothecin (CPT), Topotecan (TOP) and Irinotecan (IRI) are chemotherapy drugs that non-specifically inhibit topoisomerase-I, causing side effects. Modulation of immune targets may synergize with topoisomerase inhibitors and reduce the drug dose and side effects. The aim of this study was to examine NF-κB pathway activation and the expression of immunomodulatory proteins after treatment. Colon cancer cells were grown in vitro and treated with either CPT, TOP or IRI. The expression of selected proteins identified through RNA analysis was examined by immunoblotting, ELISA or flow cytometry. We show that topoisomerase inhibitors activate NF-κB pathway with no correlation between peak activation and the concentrations that cause S-phase arrest. On the other hand, the drugs upregulated the immune checkpoint protein Programmed Death Ligand 1 (PD-L1) in colorectal cancer cells in a dose-dependent manner. Osteopontin (OPN) expression was also upregulated in cells grown and treated in medium with regular FBS, while those in charcoal-stripped FBS did not reveal a notable difference in OPN expression. Toll-like Receptor 9 (TLR-9) expression did not markedly vary among CPT treatments under both culture conditions. Topo-I protein expression was downregulated by the treatments, indicating drug-target engagement. Our results hint that PD-L1 and OPN may be potential molecular targets for combination therapy of colon cancer with topoisomerase-I inhibitors. The pros or cons of NF-κB activation by colon cancer cells after chemotherapy remains unknown, and is the subject of further studies.