bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mouse CD19-targeted CAR T cells with an optimized 4-1BB domain enhance protection against CD19+ leukemia and retain proliferation potential after long-term rest in vivo.
Authors: Li Gongbo , Beatty Nolan , Vishwasrao Paresh , Yu Bin , Park Paul , Zhang Yongliang , Davila Marco L 
Published: January 2023

Abstract
Chimeric antigen receptor (CAR) T cells have mediated exciting results for patients. The CAR is a novel antigen-receptor that includes an antigen-binding domain, derived from an antibody, linked to an intracellular T cell activation domain. This 1st generation CAR design worked well in vitro but mediated poor results in vivo. Modification of the CAR to include a co-stimulatory domain, classified as a 2nd generation CAR design, achieved optimal T cell persistence and tumor killing in mice, which served as the rationale for evaluation of CAR T cells in patients. The most common clinically evaluated CARs target CD19 and include CD28 or 4-1BB co-stimulatory domains. However, there has been no extensive comparison of 2nd generation CAR T cells in mice, which could serve as a model for how these domains impact CAR T cell function in patients. Therefore, we evaluated and compared the function of mouse CD19 targeted CARs modified with a CD28 or 4-1BB domain. In contrast to clinical results, mouse CD28 CAR T cells imparted better leukemia-killing and persistence than mouse 4-1BB CAR T cells. We identified a defect in CAR T cell signaling mediated by mouse 4-1BB and mutation of this domain restored CAR signaling and in vivo leukemia killing. Also, we determined that CAR T cells with the mutation-optimized 4-1BB co-stimulatory domain proliferated in response to antigen in the bone marrow (BM) after a &gt; 1-month period of rest, while CAR T cells with the CD28 co-stimulatory domain did not. We determined that this is likely due to the acquisition of an exhausted phenotype in the CD28 CAR T cells. Our results illustrate how co-stimulation modulates CAR T cell function in mice and suggest mechanisms for its impact on persistence and cancer-killing in patients.
Two-pronged Cell Therapy: Engineering NK cells to target CD22 and redirect bystander T cells to CD19 for the adoptive immunotherapy of B-cell malignancies
Authors: Szoor Arpad , Velasquez Mireya Paulina , Bonifant Challice Lee , Vaidya Abishek , Brunetti Lorenzo , Gundry Michael , Parihar Robin , Goodell Margaret , Gottschalk Stephen 
Published: January 2023

Abstract
CD19-CAR T cells have shown antitumor activity for B-cell derived malignancies, but they also have shown limitations regarding their toxicity and development of CD19- escape variants. Given their short lifespan and potent cytolytic function NK cells are attractive candidate effectors to express CARs. The goal of this project was to devise a strategy to genetically modify NKs that express a CAR to target CD22 and secrete a CD19-specific T-cell engager (CD19-ENG) to redirect T cells to B-cell leukemias. NKs were transduced with a retroviral vector encoding a CD22-CAR and/or CD19-ENG molecules. To mimic immune escape, CD19 or CD22 knockout (ko) leukemia cells (BV173) were generated. We performed coculture and cytotoxicity assays using CD22 and/or CD19+ targets with NKs +/− T cells. In the absence of T cells, CD22-CAR +/− CD19-ENG NK cells recognized tumor cells in an antigen-dependent manner as judged by cytokine production and tumor killing. Moreover, CD22 CAR +/− NK cells secreting engager molecules efficiently redirected T cells in the presence of CD19+ targets. In vivo, CD22-CAR/CD19-ENG NK cells recognized tumor cells in an antigen-dependent manner, redirected T cells to tumor cells, and induced significant regression of leukemia in xenograft models in comparison to mice treated with CD22-CAR NK cells or NK cells targeting irrelevant antigens. We have generated for the first time NK cells that kill B-cell malignancies through a CAR and simultaneously redirect bystander T cells to a 2nd B-cell antigen to enhance antitumor effects and prevent immune escape. Genetic modification of NK cells to enhance their antitumor activity and redirect bystander T cells may represent a promising addition to current cell therapies for B-cell malignancies.
S1P/PPARγ axis metabolically reprograms the T cells to modulate their anti-tumor potential
Authors: Chakraborty Paramita , Thyagarajan Krishnamurthy , Chatterjee Shilpak , Nganga Rose Nithenya , Selvam Shanmugam Panneer , Ogretmen Besim , Mehrotra Shikhar 
Published: January 2023

Abstract
Adoptive T cell transfer (ACT) holds promise in cancer therapy, and strategies to further improve its efficacy are being tested. It is reported that sphingosine 1-phosphate (S1P), generated by sphingosine kinase 1 (SphK1), regulates lymphocyte egress into circulation and also plays role in the differentiation of regulatory T cells and T helper-17 cells. Here, we examined the roles of SphK1/S1P signalling in regulating antitumor potential of T cells. We found that adoptive transfer of melanoma epitope gp100 reactive CD8 T cells (pMel) with SphK1 deficiency showed improved tumor control in murine melanoma model. Our data also indicated that Sphk1−/− T cells exhibited enhanced central memory phenotype, higher mitochondrial respiration and resistance to TGF-β mediated suppression. Importantly, Sphk1−/− T cells spontaneously secreted higher levels of IL17 compared to their wild type counterparts. Mechanistically, we demonstrate that SphK1/S1P signaling regulates IL17 generation through attenuation of PPARγ, which is down-regulated in Sphk1−/− T cells. In reciprocal studies, addition of S1P in culture media enhanced PPARγ expression, resulting in decreased IL17 production in Sphk1−/− T cells. Furthermore, down-regulation of PPARγ in Sphk1−/− T cell increased lipolysis to produce free fatty acids and metabolically reprogram T cells to depend on mitochondrial metabolism for meeting their energy demand. Thus, our data reveal that SphK1/S1P signaling regulates anti-tumor functions and persistence of T cells by controlling T cell energy metabolism through induction of lipolysis via PPARγ modulation. Overall, these data highlight the clinical potential of targeting SphK/S1P axis for improving efficacy of ACT for solid tumors.
Augmenting Ag presentation at the “CD4 memory checkpoint” can overcome age-associated “defects” in CD4 helper response and enhance vaccination.
Authors: Xia Jingya , Yi Kuang , Bautista Bianca L. , Vong Allen M. , Liang Jialing , Swain Susan L 
Published: January 2023

Abstract
Vaccines are the most efficient approach to prevent infection and transmission of viruses such as influenza, but they are inefficient in the elderly. Age-associated functional defects in naive CD4 T cells, lead to few Tfh and poor help for B cells and thus to reduced long-lived Ab and memory CD4 T cells. Therefore, a vaccination strategy that would enhance the response of aged CD4 T cells could promote B and T cell memory generation and more effective protection.
We found CD4 effectors generated by influenza A virus (IAV) made very few memory cells if they could not make autocrine IL-2 and that such IL-2 production depended on cognate Ag recognition at the effector stage to drive transition to memory, defining what we call the ‘memory checkpoint’. Full differentiation of CD4 effectors to Tfh/GC-Tfh effectors also requires such Ag recognition at this checkpoint.
Here we ask if providing adult or aged CD4 T cells with Ag/APC at the ‘memory checkpoint’ can augment the CD4 and Ab response to inactivated IAV (IIV) vaccine. We use a reductionist in vivo model with an irrelevant TcR transgenic host, where we can add the IAV-specific naïve CD4 T cells (young or aged from TcR transgenic donor) and vaccinate with inactivated IAV as the vaccine. We introduce Ag at the memory checkpoint. When with either young adult or aged CD4+ T cells, again recognize cognate Ag at the ‘memory checkpoint’, they yield more IAV-specific IgG memory B cells (MBCs), IgG-secreting long-lived plasma cells (LLPCs) and more tissue resident memory CD4 T cells. This suggests novel strategies that augment Ag presentation at this CD4 checkpoint may improve vaccine efficacy in both young and elderly.
Cereblon ablation increases T cell anti-tumor immunity
Authors: Hesterberg Rebecca , Beatty Matthew S , Epling-Burnette Pearlie K 
Published: January 2023

Abstract
Druggable targets that reinvigorate T cells may improve the efficacy of immunotherapy in melanoma or other cancers. T cells exist in a hostile tumor microenvironment associated with hypoxia, high lactic acid, and poor nutrient availability which limits their anti-tumor activity. We found that the genetic deficiency of Crbn, a novel E3 ubiquitin ligase substrate receptor, improves effector T cell function. Growth of subcutaneous B16 tumors was slower in Crbn−/− mice and the adoptive transfer of Crbn−/− T cells into mice with established B16 tumors lead to a significant growth reduction. While similar total tumor infiltrating lymphocytes (TIL) numbers and normal splenic populations were present, a significant increase in the proportions of CD44+ and PD1+ CD8+ TIL but not KLRG1+CD8+ TIL were present in Crbn−/− mice. These results suggest that CRBN represses antigen-dependent anti-tumor T cell immunity and represents a novel regulator of T cells in the tumor microenvironment. In melanoma, hypoxia is a mode of T cell suppression. Activated Crbn−/− T cells demonstrated higher extracellular acidification rate, oxygen consumption and glucose uptake compared to Crbn+/+ T cells. Under hypoxic conditions (2% O2), effector cell cytotoxicity is reduced at high densities of B16 melanoma compared to normal atmospheric oxygen (~21% O2) and proliferation is suppressed due to reduction of OxPhos. Using anti-CD3ɛ/anti-CD28 stimulation in hypoxic conditions, Crbn−/− T cells retained their proliferative capacity compared to Crbn+/+ T cells indicating that deletion of Crbn restores metabolic activity of T cells in a hostile tumor microenvironment and may serve as a novel drug target in immunotherapy.
5- Fluorouracil regulation of myeloid-derived suppressor cell differentiation in vitro and in vivo
Authors: Klement John David , Paschall Amy V , Savage Natasha M , Nayak-Kapoor Asha , Liu Kebin 
Published: January 2023

Abstract
The chemotherapeutic agent 5-fluorouracil (5-FU) is the standard therapy for patients with advanced colorectal cancer (CRC). 5-FU not only targets tumor cells for apoptosis but also induces apoptosis in myeloid cells, leading to myelosuppression, which has long been thought as a side effect of 5-FU therapy. Myeloid-derived suppressive cells (MDSCs) are a heterogeneous population of immature myeloid cells that exhibit potent suppressive activity to inhibit T and NK cell function. Recent studies have found that 5-FU suppresses MDSCs in mouse models. However, we observed that MDSCs still massively accumulate in human CRC patients after multiple rounds of 5-FU therapy. We hypothesize that a subset of the heterogeneous MDSCs is resistant to 5-FU and that 5-FU therapy selectively eliminates the sensitive MDSCs, enriching the 5-FU-resistant MDSCs. To test this hypothesis, we made use of both in vitro BM-derived MDSC (BM-MDSC) and in vivo mouse tumor models. We observed that 5-FU therapy significantly decreases CD11b+Gr1+ MDSC accumulation in an orthotopic colon cancer mouse model. BM-MDSCs can be induced by cytokines, including GM-CSF, G-CSF, M-CSF and IL-6, either alone or in combination. Cytological analysis indicates that various cytokines induce BM-MDSC to differentially acquire the morphological appearance of myeloid cell subsets, including macrophage- and granulocyte-like cells. Interestingly, all of these BM-derived MDSCs display resistance to 5-FU-induced apoptosis in vitro following exposure to 5-FU at doses as high as 10 μg/mL. Our data indicates that some MDSC subsets display resistance to 5-FU, providing a rationale to explain the enrichment of MDSCs in human CRC patients following successive treatments with 5-FU.
Induction of Nrf2/HO-1 pathway suppresses neuroinflammation in ischemic stroke
Authors: Yen Jui-Hung , Kuo Ping-Chang , Yu I-Chen , Scofield Barbara A , Brown Dennis , Curfman Eric T , Paraiso Hallel C , Chang Fen-Lei 
Published: January 2023

Abstract
Ischemic stroke induces reactive oxygen species, causing oxidative and inflammatory responses in ischemic brain. To date, recombinant tissue plasminogen activator is the only available therapy for the treatment of ischemic stroke. However, the treatment does not prevent oxidative stress and inflammation in the ischemic brain. Nrf2, a major regulator of oxidative responses, is a key transcription factor, which induces a wide set of phase II anti-oxidant enzymes, including HO-1, Gclc, and NQO1. In addition, Nrf2 has been shown to exert an anti-inflammatory effect through induction of HO-1. D3T, a sulfur-containing dithiolethione compound, is found in cruciferous vegetables and has been reported to induce anti-oxidant genes through activation of Nrf2. In this study, we evaluated the effect of D3T on the suppression of neuroinflammation in ischemic stroke. Our results show D3T attenuates brain infarct and ameliorates neurological deficits in stroke animals. In addition, D3T reduces CNS infiltrating inflammatory immune cells including neutrophils and monocytes in the ischemic brain. Moreover, D3T-induced suppression of inflammatory cytokine production was observed in wild-type but not in Nrf2-deficient microglia. Furthermore, the protective effect of D3T on the attenuation of ischemic brain infarct was abolished in Nrf2-deficient stroke animals and in the stroke animals administered with HO-1 inhibitor. Altogether, our results demonstrate that D3T-mediated suppression of inflammation in the ischemic brain is mediated through Nr2/HO-1 pathway, suggesting that targeting Nrf2/HO-1 pathway may be a promising therapeutic strategy for the amelioration of neuroinflammation in ischemic stroke.
Inflammatory signals from fat: Visfatin promotes pro-inflammatory activation and leukocyte interaction in endothelial cells
Authors: Singh Manindra , Benencia Fabian 
Published: January 2023

Abstract
Obesity is a global epidemic and is associated with a cluster of metabolic diseases that lead to premature mortality. Research in the past decade suggests that chronic, low-grade inflammation – a hallmark of obesity, is a major contributing factor for the pathophysiology of several cardiovascular and metabolic diseases, including atherosclerosis and type 2 diabetes. Evidence suggests that obesity leads to abnormal levels of adipokines, which is linked to insulin resistance and inflammation of the adipose tissue. Visfatin, an adipokine predominantly secreted by visceral adipose tissue and that circulates at increased levels in obesity has been associated with several inflammatory disorders and cancer. Whether visfatin functions as a pro-inflammatory or anti-inflammatory factor is still a subject of debate. We investigated the effect of visfatin on human endothelial cells at the level of gene expression, cytokine release, and interaction with leukocytes in vitro. We found that visfatin upregulates the gene expression of the cytokines – CCL2, IL6, GM-CSF, CXCL2, and CXCL8 (p&lt;0.05) at the transcript level. Additionally, visfatin significantly upregulated the secretion of CXC-cytokines – CXCL2 and CXCL8, in a dose and time-sensitive manner. Finally, we explored if visfatin treatment has any effect on endothelial-leukocyte interaction; we found that visfatin-treated endothelial cells exhibited pronounced adhesion to monocytes (p&lt;0.05). Visfatin also upregulated the expression of adhesion markers – ICAM1, VCAM1, and E-Selectin (p&lt;0.05). Overall, these results consolidate the notion that extracellular visfatin is a marker for vascular and adipose inflammation and a potential target for therapeutic intervention.
Disturbance of Renal Circadian Rhythm in Lupus Nephritis
Authors: Mishra Rakesh K , Bethunaickan Ramalingam , Zhang Weijia , Davidson Anne 
Published: January 2023

Abstract

Background
Circadian rhythm is a universal phenomenon that allows organisms to anticipate and respond to changes in their environment by regulating various biological processes in the number of tissues, including the kidney. We found a disturbance of expression of key clock transcriptional regulators in the kidneys of mice with SLE nephritis. The purpose of these experiments was to determine how this abnormality affected transcriptional regulation of renally expressed genes and immune cell homeostasis.

Methods
We studied NZB/W mice at the age of 12 weeks (without proteinuria) and 30 weeks (with proteinuria). Kidneys and blood were harvested at 4 hour intervals. Kidneys were perfused with PBS and used to generate RNA, protein lysates and single cells for flow cytometry analysis.

Results
Young NZB/W mice have normal circadian oscillation of peripheral WBCs. There was a marked increase of circulating renal macrophages in the nephritic mice but circadian rhythm of Ly6Chi vs Ly6Clo cells followed a normal pattern. Kidneys of young NZB/W mice displayed a normal circadian pattern of expression of the master transcriptional regulators of circadian rhythm including Bmal1, Clock, Per and Cry genes compared from the nephritic mice had attenuated rhythm of these genes with inversion of the normal pattern. We also examined levels of core clock proteins (BMAL1 & PER2) in the kidney of mice from young and nephritic group at different ZT times. As expected, there was significant reduction in BMAL-1 protein expression at various zeitgeber times in nephritic mice compared to young mice.

Conclusion
Circadian dysfunction in the setting of lupus nephritis may have important implications for the delivery and metabolism of therapeutic intervention.
TACI deficiency delays the onset of systemic lupus erythematosus symptoms and improves the survival of MRL/Lpr mouse
Authors: Liu Lunhua , Allman Windy R , Coleman Adam S , Takeda Kazuto , Akkoyunlu Mustafa 
Published: January 2023

Abstract
Systemic Lupus Erythematosus (SLE) is an autoimmune disease characterized by the production of autoantibodies and immune-complex deposition in organs such as skin and kidney. Ablation of BAFF with anti-BAFF antibodies alleviates SLE symptoms but the receptor responsible for the generation of pathogenic antibodies has been controversial. Since TACI is critical in the development of antibody secreting plasma cells, we evaluated the contribution of TACI in the formation of autoantibodies and the development of SLE in TACI deficient “lupus mouse”, generated by backcrossing the lupus prone Mrl/Lpr mouse with TACI−/− mouse. We found that TACI deficiency (Lpr-TACI−/−) increased the survival of lupus mouse as compared to the littermates, LPR-TACI +/+ and LPR-TACI +/− mice. The survival of Lpr-TACI−/− mice correlated with a delay in the development of anti-dsDNA and anti-SAM/RNP antibody titers. The prolonged survival of LPR-TACI −/− mice is likely a result of protection from kidney damage since proteinuria levels and IgG accumulation in the renal glomeruli were significantly less in LPR-TACI −/− mice as compared to the age-matched control mice with disease. Coinciding with the preserved kidney function, four month old LPR-TACI −/− mice kidney macrophages manifested M2-skewed phenotype, while macrophages harvested from kidneys of the age-matched control littermates were M1-skewed. Further supporting the protective role of M2-skewed macrophages, adoptively transferred LPR-TACI−/− macrophages improved the proteinuria and glomerulonephritis in control animals with kidney pathology. Collectively, LPR-TACI −/− mice experiments suggest that blocking TACI function may be a novel approach for the treatment of SLE disease.
Deciphering Interleukin 4-induced gene-1 as novel immune regulatory pathway in dendritic cell subsets
Authors: Fallarino Francesca , Gargaro Marco , Briseno Carlos , Murray Peter , Scalisi Giulia , Turco Antonella , Matino Davide , Puccetti Paolo , Murphy Theresa L , Murphy Kenneth M 
Published: January 2023

Abstract
Interleukin 4-induced gene-1 (IL4i1) is as L-phenylalanine oxidase initially described as an early IL-4-inducible gene in B cells [1]. Herein, we analyzed IL4I1 expression in different DC subsets and investigated the possible role of IL4I1 in T-cell regulation. By using a novel and highly specific antibody, reactive to mouse IL4I1, developed in our laboratory, we found that IL4i1 could be induced by IL-4 or CpG olognucleotides (CpGODN) only in classical DCs (cDC). IL4i1 induction, by IL-4 was prevented in cDCs isolated from AhR−/− mice. Moreover, IL-4–treated cDCs cultured with CD4+T cells favored the expansion of FoxP3+ CD4+ T cells (Treg) compared to untreated cDCs. This effect was abrogated in the presence of a small interfering RNA (siRNA) targeting IL4i1 but not by a control siRNA. Notably, IL-4-induced IL4i1 expression in cDCs required aryl hydrocarbon receptor (AhR) in these cells. IL4I1-mediated oxidative deamination of phenylalanine produces H2O2 and phenylpyruvate (PP). Recently we found that PP is a novel ligand of AhR. Phenylpyruvate administration in vivo significantly reduced disease severity in a murine model of multiple sclerosis, such effect was prevented in AhR−/− mice. Overall, these results point IL4i1 as a key enzyme in the regulation of immune responses.
Gestational hypothyroxinemia increases the immune response of the progeny that suffers experimental autoimmune encephalomyelitis.
Authors: Opazo Maria Cecilia , Bugueño Katherinne , Jara Evelyn , Haensgen Henny , Albornoz Eduardo , Gomez Felipe , Binzberger Rebecca , Toledo Paola , Blanco Noelia , Simon Felipe , Elorza Alvaro , Kalergis Alexis , Bueno Susan , Riedel Claudia 
Published: January 2023

Abstract
There are more evidences supporting the relevance of a healthy pregnancy over offspring imprinting. This is the case for the maternal endocrine status and the immune system. Hypothyroxinemia (Hpx) is a highly frequent thyroid hormone deficiency condition in pregnant women characterized by low levels of thyroxin (T4) with normal levels of triiodothyronine (T3) and thyroid stimulating hormone (TSH) in the blood. It is known that this condition is asymptomatic for the mother, however it impairs cognition in the offspring. In this work we have shown that the effects of gestational Hpx go beyond the central nervous system by also affecting the immune response. We observed that gestational Hpx increases the immune response in the offspring that suffers experimental autoimmune encephalomyelitis (EAE). EAE is an experimental model used to study multiple sclerosis (MS). We show here that mice C57BL/6 gestated in Hpx induced with EAE suffer earlier and more intense EAE symptoms compared to C57BL/6 mice gestated in euthyroidism. Even though, the population of T regulatory (Treg) cells in the spleen was similar in mice gestated in Hpx and mice gestated in euthyroidism, the Treg cells from mice gestated in Hpx have less capacity to suppress T effector cells and to secret IL-10. Mice gestated in Hpx have high basal levels of TNF-α and IL-17 and present increased brain blood barrier permeability. In this work we analyzed the population of Th-17 and capacity to secret IL-17 from intestine, meninges and spleen in mice gestated in Hpx that suffer EAE. These results are very novel and impact our knowledge of how the immune system of the offspring could be affected by the deficiency of maternal thyroid hormones during gestation.