bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inducible nitric oxide synthase (iNOS) mediated inhibition of T cell receptor (TCR) signaling is a mechanism for eosinophil dependent amelioration of alloimmunity in lung grafts
Authors: Onyema Oscar Okwudiri , Guo Yizhan , Mahgoub Bayan , Manafi Amir , Criswell Stacey , Lannigan Joanne , Jacobsen Elizabeth A , Gelman Andrew E , Kreisel Daniel , Krupnick Alexander S. 
Published: January 2023

Abstract
Eosinophils play a surprising role in downregulating lung graft-specific alloimmunity, even without immunosuppression. We demonstrate here, a mechanism used by eosinophils to downregulate alloimmune responses.
To study the mechanism of eosinophil dependent downregulation of alloimmune responses, we used a model of diphtheria toxin (DT) dependent ablation of eosinophils in iPHIL, a mouse model expressing DT receptor on the eosinophil-peroxidase gene locus. Fully MHC-mismatched Balb/c (H2Kd) lungs were engrafted to iPHIL (H2Kb) recipients without immunosuppression. We modeled alloreactivity in vitro, with mixed leukocyte cultures, comprising Th1-polarized eosinophils as modulators of T cells from C57BL/6J or Nur77-GFP mice stimulated by either allogeneic dendritic cells or anti-CD3/CD28 agonistic antibodies.
Ablation of eosinophils in non-immunosuppressed lung graft recipients leads to enhanced T cell activation and graft rejection. Similarly, in vitro, we observed a dose and contact dependent downregulation of T cell activation by eosinophils. While contact between eosinophils and T cells is mediated through the upregulation of PD-L1 on both wild-type and iNOS−/− eosinophils after TH-1 polarization, the inhibition of T cell activation is iNOS dependent and is directly linked to the iNOS dependent inhibition of the TCR signaling early gene, Nr4a1, monitored in Nur77-GFP mice.
Thus, following contact with T cells, iNOS-expressing eosinophils downregulate lung graft alloimmunity through a direct effect on TCR engagement early genes, leading to the inhibition of T cell activation. This provides an explanation for the surprising observation of lung-graft specific alloimmune downregulation by eosinophils.
Selenocompounds mediated upregulation of HLA class I expression enhanced mammaglobin-A peptide-specific cytotoxic T lymphocyte responses against breast cancer cells
Authors: Tiriveedhi Venkataswarup , Khandekar Durga , Amara Suneetha 
Published: January 2023

Abstract
Recent phase I DNA vaccine clinical trials with Mammaglobin-A (Mam-A), a human breast tumor-associated antigen (TAA), have shown to be safe and efficient. However, the success of cancer vaccines is limited by the diminished expression of HLA class I molecules on cancer cells. In our current communication, we studied the impact of various selenocompounds towards the expression of HLA class I molecules on breast cancer cells and their impact on the cytotoxicity of MamA2.1 (HLA-A2 immunodominant epitope of Mam-A) activated human CD8+T lymphocytes (hCTLs). We noted an enhanced HLA-A2 expression along with upregulation of components involved in antigen presentation machinery in all four breast cancer cell lines, namely AU565, UACC-812, MCF-7, and MDA-MB-231, following treatment with methyselenol producing methylseninic acid (MSA) and dimethylselenide (DMDSe). Furthermore, we have demonstrated enhanced cytotoxicity of MamA2.1 activated CTLs on HLA-A2+/Mam-A+ AU565 and UACC-812 cell lines following pre-treatment with MSA and DMDSe, while no significant toxicity was noted under similar conditions on HLA-A2+/Mam-A− MCF-7 and MDA-MB-231 breast cancer cell lines. Taken together, our data demonstrated that MSA and DMDSe potentiate effector cytotoxic responses following TAA vaccine specific activation of CD8+T lymphocytes, and thus suggesting their futuristic role as vaccine adjuvants in cancer immunotherapy.
Monoclonal antibodies for reducing opioid use disorders and overdose
Authors: Baehr Carly Anne , Pravetoni Marco 
Published: January 2023

Abstract
Vaccines and monoclonal antibodies (mAb) have been proposed as a therapeutic option for substance use disorders by selectively sequestering the target drug in serum and preventing drug distribution to the brain and drug-induced toxicity. Although there are multiple pharmacological interventions available to treat opioid use disorder, issues such as access, side effects, and compliance limit their clinical use. As an advantage over opioid receptor antagonists, opioid-specific antibodies do not interfere with endogenous opioid signaling. Whereas clinical efficacy of vaccines for substance use disorder is limited, as up to 70% of subjects do not achieve antibody levels sufficient for efficacy, passive immunization with opioid-specific mAb circumvents this limitation. We have developed a method to rapidly generate hybridomas expressing opioid-specific mAb by magnetic enrichment of opioid-specific B cells prior to fusion. Using this strategy, we identified murine hybridomas expressing mAb specific for oxycodone, morphine and fentanyl. Passive immunization with purified opioid-specific mAb was effective in reducing opioid distribution to the brain in mice. In this study, mAb consisting of different IgG subclasses were compared for efficacy against oxycodone. Sequencing of opioid-specific mAb showed limited variability in antigen-binding regions. Integration of functional, structural and sequencing characterization can aid development of next-generation mAbs for treatment of opioid use disorder, or to counteract other toxic chemicals.
Unbiased functional identification and therapeutic targeting of T cell neoantigens in a spontaneous murine squamous cell carcinoma
Authors: Dolina Joseph S , Lee Joey , Peters Bjoern , Schoenberger Stephen P 
Published: January 2023

Abstract
The comparative resistance of some cancers including head and neck squamous cell carcinoma (HNSCC) to checkpoint blockade is speculated to derive from the low frequency of expressed somatic mutations targeted by T cells as neoantigens (NeoAg). SCC VII, a spontaneously arising murine squamous carcinoma resembling human HNSCC in several key features, is likewise poorly immunogenic as irradiated tumor cells alone fail to induce protective immunity in syngeneic hosts. Justifying use of this model to identify NeoAgs, we confirm activated CD4+ and CD8+ T cells are detectable and essential for vaccine efficacy of SCC VII and polyI:C co-administration. Whole-exome sequencing tumor versus normal genome identified 39 nonsynonymous missense mutations that were synthesized into 81 representative 20-mers. NeoAg-specific CD4+ T cell IFN-γ responses were found against mutations of Pik3ca, Ctnnd1, and Otud5 while both CD4+ and CD8+ T cells produced IFN-γ when stimulated by a single Cltc mutation containing overlapping minimal epitopes during in vitro recall assays. NeoAgs eliciting CD4+ T cell responses alone were not protective in vivo. However, the more immunogenic Cltc NeoAg prophylactically protected hosts from tumor challenge and caused regression of established tumors as an immunotherapy. Anti-PD-1 combinatorial blockade resulted in synergistic tumor rejection by boosting Cltc-specific responses and increasing response diversification via epitope spreading. These data show that a functional NeoAg identification platform can be used to select immunotherapeutically relevant targets and filtration of neoepitopes that co-prime both CD4+ and CD8+ T cell responses is superior for practical intervention of poorly immunogenic tumors.
Use of small molecule inhibitors of metabolism to enhance T cell proliferation and phenotype for adoptive T cell therapies
Authors: Waller Alexandra Lauren , Funk Christopher Ronald , Ravindranathan Sruthi , Waller Edmund K 
Published: January 2023

Abstract
Adoptive T cell therapies including CAR T therapy are limited by poor in-vivo persistence and moderate anti-tumor efficacy. According to literature, metabolism plays a critical role in the phenotypic state and fate of T cells post-expansion. For instance, glycolytic T cells are more active and exhibit a terminal effector phenotype, while quiescent T cells (naïve and memory T cells) rely more on oxidative phosphorylation with better proliferative capacity. Therefore, this study aims to determine whether metabolic inhibitors used during ex vivo T cell expansion promote a more quiescent phenotype leading to T cells with increased in vivo persistence and efficacy. A screening was conducted using inhibitors of T cell metabolism to alter the balance between T cell activation and senescence, thereby enhancing T cells proliferative capacity. The two inhibitors screened were: idelalisib, an inhibitor of PI3K δ and ibrutinib, an inhibitor of Bruton’s tyrosine kinase.
The study involved culturing human T cells with the above-mentioned compounds for 15 days with anti- CD3/CD28 beads and 30 U/mL of IL-2. T cells cultured with idelalisib and ibrutinib had increased expansion (50- fold: idelalisib and 21-fold ibrutinib) when compared to control (cells with beads alone) with only 6-fold expansion. Phenotypic analysis performed using flow cytometry showed an increased percentage of CD27+ CD28+ CD8+ T cells in the idelalisib treated group and decreased number of senescent T cells that are double negative for CD27 and CD28.
Future experiments focus on other modulators of metabolism that may benefit T cell expansion. They will also focus on metabolic analysis of T cells using the Seahorse assay and in-depth analysis of the PI3K pathway using western blot.
Improving T cell functionality for adoptive T cell therapy in metastatic colorectal cancer
Authors: Ravindranathan Sruthi , Jajja Mohammad Raheel , Waller Alexandra , Bommireddy Ramireddy , Selvaraj Periasamy , Yang Lily , Waller Edmund K 
Published: January 2023

Abstract
Adoptive T cell therapy in solid tumors such as colorectal cancers are challenging due the low frequency of tumor infiltrating T cells and high molecular tumor heterogeneity. In this study, we are proposing to address this issue by developing a novel technology to expand patient tumor specific T cells ex-vivo, using patient’s matched tumor and PBMCs. Published data from our group shows T cells isolated from heavily treated lymphoma patients when treated with inhibitors of PI3K delta (idelalisib) and vasoactive intestinal peptide (VIPhyb) signaling results in decreased levels of senescent T cells and increased persistence in-vivo in NSG mice. Additionally, we have demonstrated that tumor membrane vesicles (TMVs) prepared from human tumor cells decorated with IL-12 and B7-1 (decorated TMVs), stimulate expansion of tumor antigen specific T cells with increased IFN gamma release. This study thus combines both strategies to obtain tumor antigen specific T cells with enhanced anti-tumor activity and in-vivo persistence.
Preliminary data was generated with PBMCs obtained from consented colon cancer patients expanded with decorated TMVs with/without VIPhyb and idelalisib. Results showed that on day 14 of T cell expansion, T cells expanded with decorated TMVs + VIPhyb + idelalisib resulted in significantly increased number of IFN gamma secreting CD8+ T cells (25% of T cells), when compared to T cells treated with decorated TMVs without VIPhyb and idelalisib (13% of T cells). On the other hand, in the absence of decorated TMVs, there were less IFN gamma secreting T cells, both in the CD4 and CD8 compartment. Further experiments are underway to test the persistence and efficacy of the expanded T cells in PDX mice with matched human colon cancer.
A novel agonist of CD137 immune checkpoint stimulator serves as a cancer immunoprevention agent with efficacy against various tumor types
Authors: Batra Lalit , Barsoumian Hampartsoum B. , Shrestha Pradeep , Hawthorne Jenci L. , Bowen William S. , Zhao Hong , Egilmez Nejat K. , Gomez-Gutierrez Jorge G. , Shirwan Haval , Yolcu Esma S. 
Published: January 2023

Abstract

Background
CD137 immune checkpoint stimulator plays an important role in T cell responses and has been targeted for cancer immunotherapy using agonistic Abs with shown toxicity. We herein report that a novel oligomeric form of the CD137 ligand, SA-4-1BBL, as a single agent has robust cancer immunoprevention efficacy against various tumors in mice without detectable toxicity, whereas an agonistic Ab to CD137 did not protect mice against tumor challenge.

Study Design
Mice (C57BL/6 or BALB/c) were pretreated s.c. with SA-4-1BBL protein or an agonistic Ab (3H3) to CD137 and subsequently challenged s.c. at various times (0 to 12 weeks) with different tumor cell lines. SA-4- 1BBL-treated mice without tumor challenge were euthanized 7 days later to harvest lymphoid tissues for immunophenotyping.

Results
Pretreatment with SA-4-1BBL protected mice against 4 different tumor types; 3LL lung, TC-1 cervical, B16-F10 melanoma, and A20 lymphoma. The conferred protection against tumors was dose-dependent and evolved within two weeks of SA-4-1BBL pretreatment and lasted for &gt; 8 weeks. In marked contrast, pretreatment with the agonistic Ab had no impact on the tumor growth. CD4+CD44+CD137+ T cells and NK cells producing IFN- γ were expanded by SA-4-1BBL. Depletion of CD4+ T and NK cells or blockade of IFN-γ totally abrogated the protective effect. Depletion of CD8+ T or B cells had no significant impact on tumor growth. Importantly, SA-4- 1BBL as a single agent also prevented post-surgical tumor recurrences.

Conclusion
This study demonstrates that SA-4-1BBL is a novel CD137 agonist with desired features for cancer immunoprevention and immunotherapy.
Teriflunomide prevents the activation-induced CD25 expression on canine CD4+ T cells
Authors: Jasiecka-Mikolajczyk Agnieszka , Socha Piotr , Jaroszewski Jerzy J. , Maślanka Tomasz 
Published: January 2023

Abstract
Teriflunomide (TER) is an immunomodulatory agent. First reports on the use of TER in dogs have already appeared. Immune mechanisms underlying the immunosuppressive and anti-inflammatory properties of TER are not completely understood. The study attempts to shed light on the involvement of CD4+ T cells in inducing these properties in dogs. This study investigates the effect of TER on CD25 and Foxp3 expression by canine CD4+T cells in the course of in vitro activation. CD25 is T cell activation marker, whereas Foxp3 is critical for the development and function of CD4+ regulatory T (Treg) cells. Peripheral blood mononuclear cells collected from clinically healthy dogs (n = 15) were exposed to TER at concentrations reflecting its plasma levels obtained at a typical dose and at a ten-fold lower concentration with PMA/ionomycin stimulation. The percentage of CD25-expressing cells within CD4+ T cell subset in the culture treated with TER 10−4 M did not differ significantly from the value of this parameter in untreated non-stimulated cells but was significantly lower than that in untreated stimulated cells. The exposure to TER 10−4 M led to a significant increase in the percentage of Foxp3-expressing cells within the CD4+ T cell subset. However, the exposure to the lower concentration of TER did not affect the evaluated parameters. The results indicate that TER at the concentration achieved at a therapeutic dose prevented the activation-induced CD25 expression and induced Foxp3 expression by canine CD4+ T cells. These results suggest that the generation of inducible Foxp3-expressing CD4+ T cells and inhibition of CD4+ T cell activation may constitute one of the mechanisms responsible for the immunosuppressive and anti-inflammatory properties of TER.
On The Evolutionary Origins Of Cd1d And The Type I, Semi-Invariant Natural Killer T Cells
Authors: Suryadevara Naveen Chandra , Kumar Amrendra , Chimski Paul , Oh Karin , Caster Courtney , Truman Richard R , Ren Liming , Criscitiello Mike , Joyce Sebastian 
Published: January 2023

Abstract
The evolutionary origins of CD1d molecules and the T cell receptors (TCR) that recognize them currently remain unknown. CD1d molecules control the functions of a specialized subset of lymphocytes called semi-invariant natural killer T (type I NKT) cells. Type I NKT cells recognize both the self and non-self agonist —α-galactosylceramide (αGC) and related glycolipids— when presented by CD1d molecules. NKT cells express an invariant TCR α-chain (iTCRα) resulting from the mouse Vα14 or human Vα24 to Jα18 rearrangement. iTCRα pairs with the mouse Vβ8 (Vβ7, Vβ6, Vβ2) or the orthologous human Vβ11 TCR β-chain to create a functional NKTCR. Our phylogenetic analyses revealed that whilst the mouse Cd1 homologs have Mesozoic origin in they they evolved as far back asin the anole lizards, the orthologues of Cd1d and that of Vα14/Vα24 and Jα18 gene segments emerged with the decent of mammals. Moreover, the known avian CD1 orthologs lack the α3 domain and both the reptilian and the avian CD1 lack a functional endo/lysosomal-recycling motif critical for NKT cell development and function. As well, monotreme genomes lack Cd1d, Vα14/Vα24 and Jα18 orthologs and the marsupial Cd1d, Vα14/Vα24 and Jα18 orthologs harbor indels that would render the encoded proteins non-functional. Consistent with these findings, one of the most divergent Cd1d gene isolated from the nine-banded armadillo Dasypus novemcinctus and ectopically expressed in a cell line presented αGC and activated mouse NKT cell hybridomas. Hence, we conclude that the CD1d-restricted αGC-presentation in the immune system evolved more recently as an eutherian innovation.
The melanomacrophage center is a potential evolutionary precursor to the mammalian germinal center
Authors: Steinel Natalie C 
Published: January 2023

Abstract
The melanomacrophage center (MMC), aggregates of highly pigmented melanomacrophages found within the lymphoid tissues of ectotherm vertebrates, is thought to be the evolutionary precursor to the mammalian germinal center (GC). Studies of cold-blooded vertebrates suggest that the MMC is the site of the humoral adaptive immune response, where B cells proliferate, generate high affinity antibody, and undergo affinity maturation. MMCs transiently increase in size/number following immunization or bacterial infection, supporting the hypothesis that the MMC response is likely linked to the adaptive immune response. However, while morphological studies and shared expression of a GC specific gene further support the idea that the MMC is a proto-GC, melanomacrophages perform many functions not associated with the humoral adaptive immune response. Therefore, to determine if the MMC is serving in a GC-like capacity or is a more generalist feature of ectotherm biology, we turned to the fish model system, three spine stickleback. Fish MMCs increase in size in response to immunization with NP-CGG in alum, but not control injection with adjuvant alone. Fish MMCs are also found in close proximity to lymphocytes and cells expressing the B/T cell activation marker GL7. These preliminary studies not only give further support to the notion of the MMC as a GC homolog, but also provide important tools to broaden the study of adaptive immunity in other fish, reptilian, and amphibian systems.
Sphingosine 1-phosphate lyase promotes the type I interferon-mediated innate immune response to influenza but is subjected to degradation by influenza A virus NS1
Authors: Wolf Jennifer J , Xia Chuan , Vijayan Madhuvanthi , Song Yuleum , Studstill Caleb J , Ngo Hanh , Johnson Marc C , Saba Julie D , Alexander Stephen , Hahm Bumsuk 
Published: January 2023

Abstract
Influenza viruses cause seasonal and pandemic influenza, remaining a threat to human health. The type I interferon (IFN)-mediated innate immune response is one of the central obstacles influenza A virus (IAV) must overcome to successfully replicate within the host. Here, we demonstrate that sphingosine 1-phosphate (S1P) lyase (SPL) enhances the type I IFN response, but this antiviral innate immunity is counteracted by IAV infection. Although SPL is an enzyme that metabolizes S1P, SPL was found to interact with IKKɛ and promote IKKɛ-mediated type I IFN responses. Thus, when SPL was knocked out of host cells, IAV replication increased and the elements of IKKɛ-induced type I IFN response decreased. However, IAV infection destabilized the SPL-mediated type I IFN response by inducing the degradation of SPL. Importantly, nonstructural protein 1 (NS1) of IAV triggered the depletion of SPL. SPL was ubiquitinated upon IAV infection or NS1 expression, whereas NS1-deficient IAV failed to elicit ubiquitination or downregulation of SPL. Transiently overexpressed SPL increased the levels of auto-phosphorylation of IKKɛ, resulting in enhanced expression of type I IFN and IFN-stimulated genes. However, this induction was markedly inhibited by IAV NS1. Collectively, this study reveals a pro-IFN function of SPL as well as a novel strategy employed by IAV to subvert the type I IFN response, providing new insights into the interplay between IAV and host innate immunity.
Influenza A virus (IAV) infection in humans leads to expansion of highly diverse CD8 T cell repertoires cross-reactive with Epstein Barr virus (EBV)
Authors: Gil Anna , Mishra Rabi , Song InYoung , Aslan Nuray , Ghersi Dario , Selin Liisa K. 
Published: January 2023

Abstract
The competence of T cell responses predominantly depends on how efficient T cell receptors (TCRs) are at recognizing antigenic epitopes. We show here that during acute severely symptomatic IAV infection there was an expansion of IAV-M1/EBV-BRLF1 and IAV-M1/EBV-BMLF1 double-tetramer+ cells directly ex vivo in 5 HLA-A2+ patients. We questioned whether this expansion specific to these two different cross-reactive responses would lead to alterations in the IAV-M158, EBV-BRLF1109 and -BMLF1280 TCR repertoires from before and during acute IAV infection. Using staining with VB mAb we found that T cell responses generated to these epitopes became surprisingly more polyclonal, with the sharing of Vb between M1, BMLF1 and BRLF1 populations which is not seen in healthy donor. Furthermore, by using next generation sequencing and single-cell analysis of TCRα and TCRβ repertoire of tetramer sorted IAV-M1 cells we showed dramatic changes in specific clonotype usage during acute IAV infection compared to before infection. In summary, these changes in TCR repertoire during acute symptomatic IAV infection suggest that during severe infection there is a preferential expansion of highly diverse cross-reactive responses between IAV and the persistent virus, EBV, which leads to permanent changes in TCR repertoires to both of these two viruses (NIH AI049320 and NIH AI109858).