bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD4 T cell phenotypes differentially modulate the CD8 T cell response in kidney cancer
Authors: Cardenas Maria A , Prokhnevska Nataliya , Jansen Caroline S , Master Viraj A , Kissick Haydn 
Published: January 2023

Abstract
CD8 T cell infiltration can independently predict survival and response to immunotherapy in kidney cancer patients. Given the importance of the CD8 T cell response in cancer, it is crucial to understand what signals promote their infiltration and support the maintenance of the anti-tumor response. The proportion of tumor infiltrating CD8 T cells, as measured by flow cytometry, was found to significantly correlate (R = 0.8, p&lt;0.0001) with tumor infiltrating CD4 T cells in 160 renal cell carcinoma patients, suggesting that CD4 T cells may promote the tumor-specific CD8 T cell response. We hypothesize that the phenotype of tumor infiltrating CD4 T cells affects CD8 T cell differentiation and may contribute to the efficiency of the anti-tumor response. We performed flow cytometry analysis on a subset of kidney cancer patients to further characterize the CD4 populations in the tumor tissue. T-regulatory cells (Treg), defined by their master transcription factor FOXP3, accounted for 15.5 ± 7.2 % of the tumor infiltrating CD4 T cells. Importantly, the Treg population negatively correlated with CD8 differentiation and expression of effector-like molecules. Additionally, we found a Th1-like population expressing high levels of EOMES and GZMK that have previously been associated with cytolytic capacities. In conclusion, we found various subsets of CD4 T cells in the tumor that may differentially modulate CD8 T cell function and influence the anti-tumor response.
Unsuccessful checkpoint blockade and vaccination therapy despite expansion of tumor neoantigen-specific CD8 T cells in the Lewis Lung Carcinoma (LLC) model
Authors: Li Shamin , Simoni Yannick , Zhuang Summer , Ma Shaokang , Chee Jonathan , Redwood Alec , Robinson Bruce , Newell Evan 
Published: January 2023

Abstract
The most promising cancer immunotherapies to date target immune checkpoint receptors, such as PD-1 and CTLA-4. However, T cell checkpoint blockade has no inherent tumor specificity and shows uneven response, with up to 70–80% of patients resistant to these therapies. Here, we use a spontaneous mouse model resistant to immunotherapy to explore how immune checkpoint blockade (ICB) affects tumor-specific T cells and understand why these cells fail to eradicate tumor cells after treatment. Using combinatorial tetramer staining by CyTOF, we identified a new tumor-specific neoantigen in Lewis Lung Carcinoma (LLC). These tumor-specific CD8+ tumor-infiltrating lymphocytes (TILs) strongly exhibit an activation phenotype and can also be detected in different peripheral compartments at lower frequency. Surprisingly, despite an absence of tumor regression, we observed a remarkable expansion of tumor-specific CD8+ TILs after ICB with anti-PD1, anti-CTLA4, or neoantigen peptide vaccination. This expansion is associated with phenotypic changes, including enrichment of exhaustion markers and CD39 expression indicating chronic TCR activation and confirming recognition of tumor cells. Current single-cell RNA sequencing and TCR profiling analysis will correlate transcriptomics and clonality of these tumor-specific T cells after cancer therapies with anti-tumor response and elucidate their functionalities within the tumor microenvironment. Overall this work provides a new model for studying neoantigens in tumors and revisits the role of neoantigen-specific T cells in the context of ICB, which will have important implications in designing future immunotherapeutic strategies.
Lymphoma mutations in HVEM (TNFRSF14) disrupt bi-directional activation of BTLA and CD160 singaling within the tumor microenvironment
Authors: Sedy John R , Lin Wai , Huang Sarah , Balmert M. Olivia , Nguyen Jennifer , Moshayedi Aref , Ware Carl 
Published: January 2023

Abstract
Herpesvirus entry mediator (HVEM; TNFRSF14) is frequently mutated in human cancer with a high degree of mutations in follicular and diffuse large B cell lymphoma. In lymphocytes, HVEM receptors are activated by its ligands LIGHT (TNFSF14) and Lymphotoxin-α, and engage in bi-directional signaling with the receptors B and T lymphocyte attenuator (BTLA) and CD160. We hypothesized that mutations in HVEM disrupted binding to either of its ligands resulting in altered cell-intrinsic HVEM signaling in lymphoma, or in altered activation of BTLA or CD160 in tumor infiltrating lymphocytes. Here, we show that point mutations identified in human lymphoma were localized to the extracellular domain and specifically target ligand binding, resulting in preferential loss of HVEM interactions with CD160 and BTLA compared to LIGHT (TNFSF14). Disrupted binding correlated with loss of receptor functionality resulting in abrogated cell-intrinsic HVEM activation of NF-κB, as well as reduced CD160 activation of ERK and AKT phosphorylation by HVEM ligation. We next identified HVEM-activated CD160 and BTLA signaling pathways through AP/MS and phospho-proteomic analysis in NK and T cells. Activation of CD160 versus BTLA resulted in contrasting phosphorylation status in a number of phospho-proteins, illustrating opposing functions of these receptors. In a mouse model of lymphoma we tested the impact of BTLA versus CD160 in regulating antigen-specific anti-tumor responses, demonstrating regulatory effects of these receptors in vivo. Together these data illustrate how selective pressures within the tumor microenvironment may drive cancer mutations that alter HVEM network signaling in vivo, and indicate potential targets for onco-immunotherapy.
Upregulated 4-1BB (CD137) expression on skin T cells from a mouse model of precancerous skin
Authors: Leggatt Graham R , Salim Nazhifah , Mattarollo Stephen 
Published: January 2023

Abstract
Non-melanoma skin cancer (NMSC), including both squamous cell carcinoma (SCC) and basal cell carcinoma (BCC), is a commonly diagnosed cancer in Australia. The role of the immune system in the development of NMSC is still poorly understood. Using a mouse model of precancerous, hyperplastic skin, we have investigated the expression and targeting of the T cell co-stimulatory molecule, 4-1BB (CD137). Our data shows that 4-1BB is expressed on both CD8 and non-CD8 T cells isolated from precancerous mouse skin with limited expression on T cells in normal skin or skin-draining lymph nodes. In contrast, the ligand for 4-1BB, 4-1BBL, was poorly expressed in skin and draining lymph nodes from precancerous or normal skin. Intradermal injection of antibodies against 4-1BB was more effective at labelling target cells within the hyperplastic skin than intravenous injection. Agonist antibodies against 4-1BB showed some enhancement of skin T cell function in vitro. The data suggests that 4-1BB expression is upregulated on T cells within precancerous mouse skin and may represent a promising target for immunotherapy against precancerous skin lesions.
Inhibition of PI3Kδ improves tumor specific T cell immunity and metabolic fitness
Authors: Rangel Rivera Guillermo O , Dwyer Connor , Arthontoulis Dimitrios , Knochelmann Hannah , Smith Aubrey S , Reyes Amalia Rivera , Wyatt Megan , Paulos Chrystal M 
Published: January 2023

Abstract
Durable responses have been observed with adoptive T cell therapy (ACT) in some patients. However, current protocols used to expand T cells often exhibit suboptimal tumor control. Failure in these therapies has been attributed to premature differentiation and impaired metabolism of the infused T cells. Previous work done in our lab showed that reduced PI3Kδ signaling improved ACT. Because PI3Kγ and PI3Kδ have critical regulatory roles in T cell differentiation and function, we tested whether inhibiting PI3Kγ could recapitulate or synergize PI3Kδ blockade. To test this, we primed melanoma specific CD8+ pmel-1 T cells in the presence of PI3Kγ (IPI-459), PI3Kδ (CAL101 or TGR-1202) or PI3Kγ/δ (IPI-145) inhibitors and infused them into B16F10 tumor bearing mice. Sole inhibition of PI3Kδ or PI3Kγ in vitro promoted greater tumor immunity and survival compared to dual inhibition. To understand how PI3Kδ or PI3Kγ blockade improved T cell therapy, we assessed their phenotype. CAL101 treatment produced more CD62Lhi CD44lo T cells compared to IPI-459, while TGR-1202 enriched mostly CD62LhiCD44hi T cells. Because decreased T cell differentiation is associated with mitochondrial metabolism, we focused on CAL101 treated T cells to study their metabolism. We found that CAL101 decreased glucose uptake and increased mitochondrial respiration in vitro, indicating augmented mitochondrial function. These findings indicate that blocking PI3Kδ is sufficient to mediate lasting tumor immunity of adoptively transferred T cells by preventing premature differentiation and improving mitochondrial fitness. Our data suggest that addition of CAL101 to ACT expansion protocols could greatly improve T cell therapies for solid tumors.
Illuminating the onco-GPCRome: targeting the prostaglandin receptor-Gαs signaling axis as a novel immune checkpoint in cancer
Authors: Wu Victoria H , Wenzel Alexander , Yung Bryan , Wang Zhiyong , Mesirov Jill P. , Gutkind J. Silvio 
Published: January 2023

Abstract
Recent advances in checkpoint blockade immunotherapy (CBI) have revolutionized cancer treatment, but the limited response rates in most cancers suggest that new approaches and targets are clearly needed to fully elucidate the underlying biology of dysfunctional CD8 T cells and achieve durable responses. G-protein coupled receptors (GPCRs) are the most intensively studied drug targets as they play key roles in many physiological processes, and they have remained longstanding favorable pharmacological targets. Using novel computational approaches integrating multiple single cell RNAseq databases, we first deconvoluted tumor-infiltrating T cell heterogeneity to generate a transcriptomic GPCR signature based on coupling specificity and receptor family. Preliminary data shows an upregulation of prostaglandin receptors, PTGER2 and PTGER4, on exhausted T cells, suggesting that these GPCRs, coupled to the Gαs G protein α subunit, may contribute to diminishing anti-tumor cytotoxicity. Expression and stimulation of EP2 and EP4 on highly activated CD8 T cells and subsequent Gαs signaling decreases cytokine secretion and CXCR3-mediated migration. Additionally, blockade of EP2 and EP4 significantly decreases tumor growth in multiple immune competent, syngeneic tumor models. Altogether, we hypothesize that activation of Gαs downstream signaling cascades may be dampening the anti-tumor cytotoxicity of CD8 T cells, thereby limiting the effectiveness of PD-1 and CTLA-4 blockade. EP2 and EP4 and Gαs signaling may represent promising candidates as immune checkpoints that can be targeted in combination with CBI as part of novel multimodality precision immunotherapy approaches to reactivate the immune system to destroy tumors.
Novel lipid A mimetics, BECC438 and BECC470, act as potent adjuvants in bacterial and viral subunit vaccines
Authors: Harberts Erin , Varisco David , Haupt Robert , Jain Akshay , Middaugh Charles R. , Ernst Robert K. 
Published: January 2023

Abstract
Infectious diseases continue to be the leading cause of morbidity and mortality worldwide, an urgent need exists to develop effective vaccines against a variety of pathogens. Effective vaccines contain target pathogen antigens and adjuvant(s) which effect the magnitude, duration and mechanism of action of the immune response. Bacterial enzyme combinatorial chemistry (BECC) was developed to synthesize custom bacterial lipid A that are partial agonists for innate immune receptor Toll-like receptor 4 (TLR4). BECC is a robust mechanism for design of lipid A mimetics that drive specific activating immunity while avoiding excessive adverse host reactions. Initial screenings identify two lead adjuvant candidates, BECC438 and BECC470, that are formulated with bacterial and viral antigens, including Yersinia pestis (rF1-V), Staphlococcus alpha toxin, and HPV (VLPs). Mice are immunized on two-week intervals with several squalene or aqueous based BECC formulations. Antigen-specific serum antibody titers measured by ELISA for total IgG, IgG1, and IgG2c(2a) for BECC formulations are significantly more balanced than those currently approved for clinical use. Pathogen challenge models also show increased levels of efficacy/survival. Both BECC compounds allow for durable antibody production, antigen sparing, dose sparing, adjuvant sparing, production of T follicular helper cells, and provide protection against a heterologous challenge in viral infection models. Additionally, BECC adjuvanted vaccines with the model antigen ovalbumin show balanced and durable antibody responses in multiple murine backgrounds C57BL6, BALBc, and CD-1. BECC molecules are effective components of next-generation infectious disease vaccines.
Vaccine-elicited T cells rely on a distinct transcriptional program from infection-elicited T cells
Authors: Thompson Scott Beckett , Ivanova Daria Leonidovna , Klarquist Jared , Scott-Browne James , Kedl Ross M. 
Published: January 2023

Abstract
Protection from pathogens relies on both humoral (antibody-mediated) and cellular (T cell-mediated) responses. While infections robustly elicit both of these types of immunity, currently approved vaccine adjuvants largely fail to induce significant T cell responses. However, recent work by our lab and others suggests that the mechanisms governing vaccine-elicited T cells (Tvac) may be substantially different than those governing infection-elicited T cells (Tinf). We have recently demonstrated that optimal subunit vaccine-elicited T cell responses rely on different cytokine signals (IL-27 and 15) and metabolic function (oxidative phosphorylation vs. aerobic glycolysis) leading to phenotypically and functionally different outcomes (memory vs. effector). Our goal was to investigate the transcriptional programming that promotes Tvac formation. Using a combined ATACseq, bulk RNA-seq and single-cell RNA-seq approach, we compared Tvac and Tinf at various time points post-immunization or infectious challenge. Our data indicate that the chromatin organization and gene expression of Tvac markedly diverges from Tinf even at early time points. Furthermore, single-cell analysis indicates that Tvac represent a unique subset of activated/memory T cells, distinct from Tinf. Remarkably, while displaying a gene signature similar to highly proliferative cells, Tvac also share transcriptional properties with recently identified stem cell memory T cells, such as oxidative phosphorylation and transcription factor TCF1 gene signatures. Taken together, our data have identified a novel transcriptional program that supports a robust vaccine-elicited T cell response with implications for future therapeutic vaccine design.
The quality of heat shock protein-mediated responses is determined by the antigen presenting cell
Authors: Sedlacek Abigail L , Kinner Lauren , Binder Robert 
Published: January 2023

Abstract
Immunization with Heat shock proteins (HSPs) elicits a spectrum of immune responses including non-antigen specific immunoregulatory responses and proinflammatory antigen specific responses. While some of the aspects of these mechanisms have been determined, many questions remain. HSPs interact with their receptor, CD91, which is expressed on the surface of antigen presenting cells (APCs). In response to HSPs, APCs upregulate expression of costimulatory molecules, release cytokines, and cross-present HSP-chaperoned peptides. This culminates in activation of T and NK cells, which are both required for an optimal Th1 response. We have uncovered unique signaling cascades in macrophages (MΦ), conventional dendritic cells (cDCs), and plasmacytoid dendritic cells (pDCs) which include NF-kB and p38 MAPK and result in different downstream immune responses. In MΦ, p38 MAPK mediates phosphorylation of STAT1 at both phosphorylation sites, Y701 and S737. Phosphorylation of STAT1 leads to the secretion of CXCL10 and downstream activation of NK cells. In contrast, cDCs fail to phosphorylate STAT1 at Y701, produce reduced amounts of CXCL10 and do not activate NK cells, thus yielding sub-optimal T cell responses. Alternatively, activation of pDCs by HSPs induces surface expression of NRP1. NRP1 expression allows for engagement and activation of regulatory T cells and downstream suppression of immune responses. Collectively, our data demonstrates that targeting HSPs to particular APCs is critical in elucidating the desired immune response and needs to be considered in future clinical applications.
Using lymph node slices to study vaccine adjuvants ex vivo
Authors: Ball Alexander , Catterton Megan A , Pompano Rebecca R 
Published: January 2023

Abstract
Despite the success of vaccines in preventing infectious diseases, there is a growing need for the development of new vaccine adjuvants. We recently demonstrated that live lymph node (LN) slices serve as a unique platform to study adaptive immunity ex vivo. These slices enable visualization of changes in the spatial organization and activation state of the tissue during an immune response. Here we hypothesized that different adjuvants induce unique cytokine and spatial profiles within tissue, and we examined the influence of three adjuvants (alum, CFA, & poly I:C) to test this. We transferred ovalbumin (OVA)-specific CD4+ T cells (OTII) into C57BL/6 mice before vaccinating the mice with OVA protein and adjuvant. LNs were collected on days 1, 4, and 7, sliced on a vibratome, and cultured overnight with either the intact OVA protein or PBS. The supernatant was analyzed for cytokine secretion by ELISA (IFNγ, TNFα, and IL-4) from cultured slices. Slices were immunostained for activation markers, CD69 and CD22, and analyzed with fluorescent imaging. Cytokine secretion was greatest in slices from vaccinated mice that had been co-cultured with the OVA protein in comparison to slices that were either cultured or immunized with PBS. The different adjuvants induced unique cytokine profiles in slices both in terms of timing and magnitude as TNFα, IFNγ, and IL-4 peaked on different days from one another within the same vaccination scheme as well as between adjuvants. The spatiotemporal distribution of T cell and B cell activation markers also differed between adjuvants. This study demonstrates that LN slices can analyze the unique influences of different adjuvants and highlights slices as a potential screen for novel vaccine adjuvants.
A microneedle vaccination with glycoprotein E of Varicella Zoster Virus elicits antibody production and polyfuctional T cells in mice
Authors: Cha Hye-Ran , Shim Doo-Hee , Lee Jaemyun 
Published: January 2023

Abstract
A microneedle has several benefits in that it is painless, easy to use and preventable from potential cross-contamination by using conventional needle. Besides these advantages, vaccination with microneedle may provide comparable or even higher immune response than vaccination through conventional intramuscular route. Here we have been developing herpes zoster vaccine using microneedle coated with recombinant glycoprotein E (gE) of VZV, which is essential in virus replication and cell-to-cell spread. Microneedles were coated by gE and three different adjuvants: double-mutant heat-labile toxin (dmLT), the combination of TLR1/2 and TLR3 ligands (Lpampo), or mutant cholera toxin A subunit (CTA). C57BL/6 mice were immunized with the coated microneedles two times at 2-week intervals. The overall increase in gE specific antibody titers was observed among adjuvanted groups compare to the non-adjuvanted group. Moreover, the humoral response was IgG1-dominated. Hence, a decreasing tendency of IgG1/IgG2c ratio was observed in gE adjuvanted with dmLT or Lpampo groups whereas relatively high ratio was detected in CTA group. This indicates that Th1 immune response complemented Th2 response in former groups. By conducting ELISPOT, we measured the amount of IFN-gamma secreting T cells as cell-mediated immune response is significant in immunogenicity of HZ vaccination. Moreover, polyfunctional CD4 T cells were increased after injection with gE +Lpampo coated microneedle. In consistent with humoral response, dmLT or Lpampo adjuvanted group exhibited enhanced cell-mediated immune response. Taken together, delivery of HZ subunit vaccine with microneedle may provide a potent immune response against reactivation of VZV.
Utilising nanoparticles to enhance the vaccine induced T cell immune response
Authors: Wilson Kirsty L , Powles Liam , Tsirikis Peter , Xiang Sue D , Selomulya Cordelia , Henderson Edward , Corrie Simon , Plebanski Magdalena 
Published: January 2023

Abstract
Vaccines are a powerful and cost-effective tool to limit the spread of disease, however, there are still some complex diseases for which we don’t yet have a successful and long-lasting vaccine, for example malaria and cancers. One reason for this is the difficult choice of target antigen, though there is also a need for potent adjuvants/delivery systems capable of inducing both antibody and T cell responses. Current licensed adjuvants, for example Alum, mainly elicit strong antibody responses, therefore, novel vaccine delivery systems need to be developed. One such system is by utilising nanoparticles to increase the vaccine induced immune response. Inorganic carboxylated polystyrene nanoparticles in the viral size range (40–50nm) are non-inflammatory, stable in solution, and target dendritic cells. Importantly, these nanoparticles elicit potent antibody, CD4+ T cell and CD8+ T cell responses to a range of antigens in murine studies, including to malaria and cancer peptides. Using fluorescent nanoparticles allows us to assess the interaction of these nanoparticles with specific cell subsets, such as antigen presenting cells, by flow cytometry. Furthermore, we can analyse the biodistribution of the nanoparticles using small animal imaging via the IVIS. Ideally, for maximum translation capacity into human vaccines these nanoformulations would be biodegradable. In addition to our model polystyrene nanovaccines we have been investigating biodegradable nanoparticles made from other materials, i.e. iron oxide. Comparable to polystyrene nanoparticles, biodegradable nanovaccines are also endocytosed by dendritic cells and are capable of inducing antibodies and cytokine secretion from both CD4+ T cells and CD8+ T cells.