bluuebunny/crossref_metadata_2025_split huggingface dataset:
Rational targeting of Cdc42 GTPase destabilizes Treg to promote anti-tumor T cell immunity
Authors: Nguyen Thi Thuy Phuong , Kalim Khalid , Zheng Yi , Guo Fukun 
Published: January 2023

Abstract
Targeting of regulatory T cells (Treg) is a long-sought approach to overcome tumor immune evasion. However, systemic depletion of Treg likely causes autoimmune diseases. Indeed, we have recently reported that Treg-specific homozygous deletion of Cdc42 GTPase caused a drastic reduction in Treg numbers and systemic inflammatory diseases. Here we show that heterozygous deletion of Cdc42 (Cdc42+/−) did not affect Treg homeostasis but destabilized them, leading to effector T cell reprogramming in these Treg and impaired function in suppressing effector T cells. Accordingly, Cdc42+/− mice had increased effector T cells. Intriguingly, Cdc42+/− mice showed a remarkable suppression of growth of mouse colon and pancreatic tumors, with no noticeable autoimmunity. Mechanistically, Cdc42+/− Treg exhibited a significantly increased expression of carbonic anhydrase 1 (CA1), reversal of which by CA1 shRNA or a CA inhibitor restored Treg stability and tumor growth. Pharmacological inactivation of Cdc42 by a chemical inhibitor, CASIN, destabilized wild type (WT) Treg in mice and triggered anti-tumor T cell immunity, with no detectable autoimmune responses. CASIN also destabilized WT Treg in vitro and adoptive transfer of CASIN-treated Treg with congenic effector T cells into Rag1−/− mice suppressed tumor growth, demonstrating that CASIN-induced Treg instability caused tumor suppression. Combined CASIN and Anti-PD-1 antibody led to an additive suppression of mouse tumors as well as human tumors in human immune cell-reconstituted NSG-SGM3 mice. Collectively, our data suggest that rational targeting of Cdc42-regulated Treg stability holds promise in counteracting tumor immune evasion.
Interferon gamma production by regulatory T cells is required for response to cancer immunotherapy
Authors: Gocher Angela Marie , Handu Sanah , Workman Creg J , Vignali Dario A. A. 
Published: January 2023

Abstract
We have shown that murine regulatory T cells (Tregs) produce the cytokine interferon gamma (IFNγ) during anti-PD1 immunotherapy and that IFNγ-driven Treg dysfunction is necessary for tumor clearance by anti-PD1. However, it is unknown how Treg production of IFNγ dictates response to cancer immunotherapy. It has been shown that IL12-induced production of IFNγ is required for response to anti-PD1 therapy. However, it has yet to be determined whether Tregs are a key responder to IL12. Thus, elucidating the interplay of IL12, IFNγ and Tregs in the tumor microenvironment (TME) is essential for understanding the mechanistic events required for response to immunotherapy. Our lab has generated two novel murine models that allow for Treg-restricted deletion of Ifng (IfngL/LFoxp3Cre-YFP) or Il12rb2 (Il12rb2L/LFoxp3Cre-YFP). These murine models were used to assess the contribution of Treg-derived IFNγ and response to IL12 on the growth of various syngeneic tumor models and response to checkpoint blockade, vaccination and tumor-specific antibodies. Surprisingly, mice with Ifng-deficient Tregs were completely resistant to anti-PD1 immunotherapy. Preliminary experiments suggest that IFNγ production by Tregs is an early requirement to produce an effector-like Treg state in order to mediate an effective anti-tumor response. Mice with Il12rb2-deficient Tregs had diminished therapeutic response to anti-PD1. These data suggest that IFNγ producing Tregs are required to shift the balance from an immunosuppressive TME to one that favors the reinvigoration of the anti-tumor response generated by cancer immunotherapies. Future studies will determine the mechanisms of resistance and whether IFNγ+ Tregs predict response to immunotherapy.
Influenza vaccines using liposomal formulations of toll-like receptor (TLR) 7/8 and 4 agonists as adjuvants
Authors: Cole Shelby L , Cybulski Van A , Whitacre Margaret , Lathrop Stephanie K , Bazin-Lee Helene , Burkhart David , Evans Jay T 
Published: January 2023

Abstract
Influenza virus infection is a global health concern and causes severe illness and thousands of deaths each year. While there are currently licensed vaccines available for both seasonal and pandemic influenza viruses there are several issues to consider; 1) current vaccines do not provide supra-seasonal or universal protection against influenza subtypes, 2) antigenic mismatch between the vaccine and circulating influenza strains impact vaccine efficacy, 3) and the efficacy of influenza seasonal vaccines could be improved in the elderly and immune compromised populations. Therefore, vaccine adjuvants capable of inducing more robust and broadly cross-protective immune responses to various subtypes of influenza virus is a critical unmet need. To address this challenge, we developed a synthetic dual TLR adjuvant combination system (TRAC-478) to improve influenza vaccine immunity. Preliminary data in human peripheral blood mononuclear cells demonstrate that combining our synthetic TLR7/8 and TLR4 agonists results in a synergistic innate immune responses. Influenza vaccine immunogenicity studies in both murine and pig models demonstrate the safety and efficacy of this new dual TLR adjuvant system. Using a co-encapsulated TRAC-478 liposome delivery system with a split-flu vaccine resulted in strong anti-influenza humoral, Th1 cell mediated immunity, and protection from a heterologous influenza challenge in mice. We hypothesize that using the TRAC-478 adjuvant system in combination with current influenza vaccines will induce a more robust and broadly cross-protective immune responses to circulating or pandemic influenza virus.
Influenza-specific ex vivo immune responses in ferrets commensurate with number of vaccinations despite decreased protection for repeat vaccines
Authors: Hofstetter Amelia R , Music Nedzad , Kamal Ram P. , York Ian A. 
Published: January 2023

Abstract
There is controversy in the literature whether consecutive year influenza vaccinations lead to decreased vaccine efficacy. Evidence for this effect is strongest in the context of repeated H3N2 vaccinations. As recently reported (NPJ Vaccines 4:28 (2019)), we recreated this phenomenon in the ferret model of influenza. Ferrets exposed to consecutive year vaccinations had significantly increased weight loss, viral shedding and fever after A/Hong Kong/4801/2014 (HK4801) virus challenge than ferrets only exposed to the 2017–2018 vaccine. One possible explanation for this phenomenon is that the repeated vaccination induces non-protective antibodies against influenza virus, which might include low avidity antibodies, non-neutralizing antibodies, and antibodies that otherwise do not elicit a protective immune response. Ferrets that received repeated vaccination had higher avidity antibodies to HK4801 HA at Day 2 and 14 post-infection than did the other two groups. Preliminary data do not support increased levels of HA stem-binding antibodies in repeat vaccinees’ serum. NA and NP are two additional viral proteins expressed on the cell surface, with greater or lesser evidence for their role in protective antibody responses, respectively. Serum reactivity to both proteins correlated with the number of vaccinations. Together, these results do not support a model of increased non-protective antibodies after repeat vaccination. Furthermore, the breadth of the T cell response against H3N2 viruses from different clades was similar between both vaccinated groups. Ongoing studies continue to explore the mechanism for the decreased protection observed in the repeat vaccination group compared with the single year vaccination group.
Understanding memory B cell responses to a chimeric-based universal influenza virus vaccine candidate
Authors: Lan Linda (Yu-Ling) , Li Lei , Huang Min , Henry Carole , Zheng Nai-Ying , Chen Yao-Qing , McNair Mahalia , Meade Philip , Guthmiller Jenna , Dugan Haley , Tepora Micah , Palm Anna-Karin , Shaw Dustin , Stamper Christopher , Khan Aly Azeem , Nachbagauer Raffael , García-Sastre Adolfo , Palese Peter , Krammer Florian , Wilson Patrick C 
Published: January 2023

Abstract
A universal influenza virus vaccine candidate trial was recently conducted that utilized novel influenza viruses expressing chimeric hemagglutinin (HA) proteins. The chimeric HAs present in the vaccines are composed of a typical stalk domain (from an H1 HA) paired with an exotic head from a strain to which humans are naive (avian H5 or H8). Given the conserved nature of the HA-stalk region, broadly neutralizing antibodies that target this region would confer protection against a wide range of influenza virus strains. The major objective of the trial is to determine whether the human immune response to influenza virus can be re-programmed to target universally conserved stalk epitopes. In this study, we collected antigen-specific memory B cells at day 28 post-vaccination at prime, boost and 1-year time points, and subsequently characterized both the transcriptome by single cell RNA-seq and antibody specificity by generating monoclonal antibodies (mAbs) from the sequenced immunoglobulin genes of the same B cells. Our research has suggested HA stalk-reactive B cells were selected after the boost time point. In addition, we have identified clonal expansions within an individual (referred to as private clones) and across multiple individuals (referred to as public clones) that have neutralizing capability and are able to bind divergent influenza strains. Furthermore, we have identified different memory B cell subsets induced by universal influenza virus vaccination at the single-cell level.
Simultaneous mRNA, protein, and immune repertoire profiling of antigen-specific T cells by single cell sequencing
Authors: Shi Xiaoshan , Siddique Imteaz , Nakamoto Margaret , Mortimer Stefanie 
Published: January 2023

Abstract
High-throughput single cell RNA-seq (scRNA-seq) has transformed our understanding of complex and heterogenous immune populations. New advances in scRNA-seq are expanding the molecules that can be profiled at the single cell level, such as oligo-conjugated antibody technologies that enable protein expression profiling alongside mRNA. Although the ability to sequence the immune repertoire can provide crucial insights into understanding the complexities of the adaptive immune system and advancing discoveries in immuno-oncology, the ability to extract this information from single cells requires new technology to profile regions of the mRNA that are missed by conventional 3′ scRNA-seq. In this study we utilized an ex vivo antigen stimulation system to measure antigen-specific T-cell activation and clonal amplification. Stimulated T cells from two donors were loaded onto the BD Rhapsody™ Single-Cell Analysis System to extract immune repertoire information in addition to gene and protein expression information from the same cells. In a single workflow, we profiled a panel of 400 mRNA targets, 20 BD® AbSeq protein markers associated with different status of T-cell activation, and the hypervariable region of T-cell receptors at the single cell resolution. The study is a proof of concept of combining mRNA, protein and immune repertoire analysis at the single cell level to deconvolute the heterogeneity and different activation of stimulated T cells.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
BD, the BD Logo, and Rhapsody are trademarks of Becton, Dickinson and Company or its affiliates. © 2019 BD. All rights reserved.
CAR T cells generated in situ using gene therapy in naturally activated lymph node T cells
Authors: Pease Larry R , Guttormson Matthew C , Parks Christopher A , Felts Sara J , Van Keulen Virginia P , Castro Sarah A , Evgin Laura , Vile Richard G , Barry Michael A 
Published: January 2023

Abstract
T cells expressing chimeric antigen receptors (CAR T cells) are manufactured outside the body using non-physiological activation and in vitro culture protocols. The procedure takes 3 weeks and costs approximately $450,000 to administer. We have devised a scheme using mouse models for manufacturing CAR T cells in just 3 days employing a natural, polyclonal anti-viral immune response inside a lymph node (LN) using off the shelf reagents that can be applied widely in populations. CAR T cells emerge from the LN within a week, invade solid tumor, and eliminate the targeted antigen-positive tumor cells. Polyclonal activation of T cells is achieved by modifying the antigen presenting properties of lymph node resident professional antigen presenting cells (APC) through viral expression of allogeneic major histocompatibility antigens. The ensuing polyclonal activation occurs in the context of virus activated APC within the normal aarchitecture of the LN and confers robust T cell activation signals directing the development of cytolytic T cells. During the course of T cell activation, the T cells are transduced in situ with retroviruses encoding selected CARs. This approach could be applied to any number of different CARs to target a variety of tumors. T cells activated in the lymph node are functionally distinct from those activated using standard CAR T manufacturing protocols. We anticipate that CAR T cells generated in situ to natural stimuli will exhibit more normal immune regulatory properties that will impact their therapeutic utility. Because our approach uses off the shelf reagents and is administered using standard clinical procedures, cost to patients should be lowered and access to therapy increased.
A single administration of Fc-fused recombinant human IL-7 induces expansions of T lymphocytes in healthy adults and cancer patients
Authors: Kim Sojeong , Heo Minkyu , Jeong Sohee , Woo Jungwon , Sung Young Chul , Shin Eui-Cheol , Park Su-Hyung 
Published: January 2023

Abstract
Interleukin-7 (IL-7) is a cytokine that plays important roles in T cell development and homeostasis. IL-7 is a potent agent for promoting T cell reconstruction in lymphopenia patients. In this study, we investigated the effects of GX-I7 (human IL-7 fused to hyFcTM for half-life extension and improved stability) in healthy volunteers (20~60 μg/kg, n=24) and advanced solid cancer patients who had no available effective treatments (60~1,200 μg/kg, n=33).
We observed significant increases in the number of circulating CD8+ and CD4+ T cells in healthy volunteers, the peak of which was seen on days 14-21. Importantly, naïve, TEM, and TCM subsets of CD8+ and CD4+ T cells increased, coupled with upregulation of Ki-67. TCR diversity of the naïve T cells tends to be increased in healthy adults by single administration of GX-I7. In addition, cytokine secretion assay using PBMCs revealed that Ag–specific (CMV-specific) T cell responses were maintained after GX-I7 administration in healthy volunteers. Moreover, in advanced solid cancer patients, GX-I7 administration significantly increased the number of all subsets (naïve, TEM, TCM and TEMRA) of both CD8+ and CD4+ T cells in a dose-dependent manner. More importantly, single GX-I7 administration increased the absolute lymphocyte count above the normal range in all patients with lymphopenic conditions (ALC &lt; 1,000/mm3).
In conclusion, following GX-I7 administration, dose-dependent increase of ALC and T cell subsets were observed. Our study suggests that GX-I7 as a T cell amplifier provides a unique opportunity for immuno-oncology combination strategies by reconstituting persistent T cell immunity, where anti-tumor effects are mediated by T cells.
Bispecific CAR T cells for multiple myeloma: natural ligand compared to tandem scFv design
Authors: Larson Rebecca , Castano Ana , Bouffard Amanda , Scarfo Irene , Kann Michael , Schmidts Andrea , Gallagher Kathleen , Maus Marcela Valderrama 
Published: January 2023

Abstract
One of the most promising candidates for Chimeric Antigen Receptor (CAR) T cell therapy beyond CAR19 is treatment of multiple myeloma with CAR T cells targeting B cell maturation antigen (BCMA). However, current reports of BCMA CAR in the clinic have a median progression free survival of 11.8 months, suggesting that targeting BCMA alone may not be sufficient. Evidence suggests patients treated with BCMA-targeted therapies may be thwarted by BCMA negative relapse. We have developed a novel second generation CAR T cell against another multiple myeloma target: transmembrane activator and CAML interactor (TACI). Mice were immunized against TACI and we designed a new scFv targeting TACI based on the resulting antibodies. Anti-TACI CAR T cells are cytotoxic in vitro and in vivo against multiple myeloma. To overcome single antigen loss, we designed bispecific tandem scFv CAR T cells targeting BCMA and TACI. These two antigens share a natural ligand, A Proliferation-Inducing Ligand (APRIL), which has also been used to design a dual-targeting CAR called TriPRIL. We show that these dual targeting CARs, based on tandem scFv or natural ligand design, have similar efficacy against wild type multiple myeloma models. However, this changes in the context of single antigen loss. We have characterized the proliferative (population doublings) and activation capability (CD69) of these CARs, as well as their memory (CCR7, CD45RA) and exhaustion phenotype (PD-1, TIM-3, LAG-3), with long term exposure to single antigen. Our studies show that sensitivity to antigen density differs between the tandem bispecifics and the natural ligand CAR. These data provide insight into how structural differences between dual-targeting CAR T cells affects their function.
The IL-15 receptor agonist N-803 combined with the anti-CD20 monoclonal antibody rituximab expands NK and CD8 T cells and alters single cell immune transcriptomes in a phase 1 clinical trial in lymphoma
Authors: Foltz-Stringfellow Jennifer Ann , Bartlett Nancy L. , McClain Ethan , Becker-Hapak Michelle , Hess Brian T. , Bachanova Veronika , Berrien-Elliott Melissa M. , Foster Mark , Schappe Timothy , Kahl Brad , Mehta-Shah Neha , Cashen Amanda F. , McDaniels Kristin , Moreno Chaz , Gao Feng , Griffith Obi , Griffith Malachi , Wagner Julia A. , Rock Amy D. , Soon-Shiong Patrick , Lee John , Petti Allegra A. , Fehniger Todd A 
Published: January 2023

Abstract
IL-15 is a cytokine that is crucial for the development, survival, and activation of NK cells, and pre-clinical studies demonstrated that IL-15 augments antibody-dependent cellular cytotoxicity. We hypothesized that IL-15 would augment anti-CD20 mAb (rituximab)-directed responses in patients with indolent non-Hodgkin’s lymphoma. To address this, we performed a first-in-human combination clinical trial of the IL-15 super agonist N-803 with rituximab, and investigated the impact of this novel immune combination on NK, CD8 T cells, and monocytes. The combination was safe and demonstrated clinical activity, including in patients refractory to rituximab. To understand the in vivo impact on the patients’ immune systems, we performed mass cytometry and cellular indexing of transcriptomes and epitopes by sequencing (CITE-seq), which measures single-cell RNA-seq and cell surface proteins simultaneously. N-803 induced expansion of NK (12.5-fold) and CD8 T cells (2-fold) in serial blood samples and upregulated CD38, chemokines, and MHC class II family member expression while maintaining CD16 expression on NK. On CD56dim NK, N-803 increased activating receptors, decreased CD57, and modulated transcription factor expression (AP-1, Eomes, CEBPB/D). CD56bright NK cells were primed, with increases in granzyme A, B, and K. Therapy also markedly altered the monocyte compartment, significantly upregulating type 1 interferon and interferon gamma pathways in CD14+ monocytes, and decreased the percent of CD16+ monocytes, suggestive of potential trafficking to the tumor. Collectively, our data demonstrates that N-803 plus rituximab induces complex immune activation, and supports N-803 combination with other mAbs and immunotherapies.
IFN-lambda 3 directly induces antiviral responses in human B and T cells leading to inhibition of HIV-1 infection of CD4+ T cells
Authors: Santer Deanna M. , Minty Gillian ES , Golec Dominic , Lu Julia , May Julia , Namdar Afshin , Shah Juhi , Elahi Shokrollah , Proud David , Joyce Michael , Tyrrell D. Lorne , Houghton Michael 
Published: January 2023

Abstract
Type III interferons (IFN-lambdas(λ)) are the most recently discovered interferon cytokine family that inhibit viruses by signaling through a unique IFN-λR1/IL-10RB heterodimeric receptor. Until now, IFN-λs were thought to primarily act on anatomical barrier epithelial cells, neutrophils and a subset of dendritic cells, although the majority of studies have been performed in mice. Here, we examine regulation of IFN-λR1 expression and downstream effects of IFN-λ3 stimulation of primary human blood immune cells and compare them to lung or liver epithelial cells. IFN-λ3 directly bound and upregulated IFN-stimulated gene (ISG) expression in freshly purified human B cells and CD8+ T cells, but not monocytes, neutrophils, natural killer cells and CD4+ T cells. Despite similar IFNLR1 transcript levels in B cells and lung epithelial cells, lung epithelial cells bound more IFN-λ3, which resulted in a 50-fold greater ISG induction when compared to B cells. The reduced response of B cells could be explained by the higher expression of the soluble splice variant of IFN-λR1, which inhibited ISG induction when added with IFN-λ3 to peripheral blood mononuclear cells. Human CD4+ T cells gained responsiveness to IFN-λ3 upon T-cell receptor stimulation since activation signals potently, and specifically upregulated membrane-bound IFN-λR1 expression. Importantly, IFN-λ3 treatment of activated CD4+ T cells caused a significant decrease in human immunodeficiency virus-1 (HIV-1) infection. Collectively, our data demonstrate that IFN-λ3 directly interacts with the human adaptive immune system, and thus can promote antiviral immunity at mucosal and peripheral sites, unlike what has been previously shown in published mouse models.
HIV-1 envelope glycan composition is a key determinant of efficient virus transmission via DC-SIGN and resistance to antibodies and inhibitory lectins
Authors: Hioe Catarina E , Jan Muzafar , Upadhyay Chitra 
Published: January 2023

Abstract
HIV-1 envelope (Env) undergoes extensive N-linked glycosylation that cloaks its surface with an assortment of oligomannose-, hybrid-, and complex-type glycans and enables virus interaction with carbohydrate-recognizing lectins. This study examined the importance of Env glycan heterogeneity for HIV-1 transmission via DC-SIGN, a mannose-binding lectin expressed on macrophages and immature dendritic cells. A diversity of glycan content was observed among HIV-1 strains and associated with varying degrees of trans-infection via DC-SIGN. When Env glycans were modified to display only oligomannose-type, DC-SIGN-mediated virus capture was enhanced. However, virus trans-infection was diminished due to increased degradation, which was alleviated by incorporation of hybrid-type glycans. HIV-1 strains also displayed varying sensitivity to trans-infection blockage by antiviral lectins GNA and GRFT, which correlated with sensitivity to neutralizing antibodies. Tier 1 viruses sensitive to neutralizing antibodies were more readily inhibited by GNA and GRFT than tier 2 and tier 3 viruses. Correspondingly, tier 1 Envs showed a greater reactivity with GNA and GRFT, which recognizes high-mannose-type N-glycans, suggesting enrichment of high-mannose type on tier 1 Envs. Of note, HIV-1 Env glycan content is influenced by amino-acid variations in the Env signal peptide (SP) and single-point mutations in the Env SP were sufficient to affect Env glycan content, leading to alterations in DC-SIGN-dependent trans-infection and virus sensitivity to antiviral lectins. Hence, SP variation and glycosylation that confer varied types of oligosaccharides to HIV-1 Env are critical determinants for virus fitness and phenotypic diversity.