bluuebunny/crossref_metadata_2025_split huggingface dataset:
Overcoming barriers to solid tumor immunotherapy using natural killer cell therapies designed to mimic intraepithelial group 1 innate lymphoid cells
Authors: Horowitz Nina Beryl , Hickey John , Nolan Garry P , Sunwoo John B 
Published: January 2023

Abstract
Our work aims to design cell-based immunotherapies for maximum solid tumor efficacy by leveraging insights from tissue-resident cells and single-cell RNA sequencing of innate immune cell subsets present within patient tumors. We discovered that by co-culturing peripheral blood natural killer cells (pbNKs) with irradiated epithelial tumor cells, we induced high expression of surface integrins, increased cytotoxicity and IFNg production, decreased sensitivity to TGFb, and significantly enhanced solid tumoroid infiltration. The cells were profiled using CyTOF and we determined that they closely resembled intraepithelial group 1 innate lymphoid cells, so we refer to them as ieILC1-like cells. We quantified the cytotoxicity of ieILC1-like cells against a variety of target cell lines and determined that they are broadly cytotoxic and capable of antibody-dependent cellular cytotoxicity, which was assessed using cetuximab. Tumor-infiltrating capacity was modeled using 3D tumoroids grown from single-cell suspensions of epithelial tumor cell lines in basement membrane extracts; ieILC1-like NK cells or pbNK cells were fluorescently labeled, added to tumoroids, and imaged using confocal microscopy. We stimulated the cells in the presence or absence of overnight TGFb exposure to determine the extent of TGFb-mediated immunosuppression. ieILC1-like cells were more resistant to TGFb and produced significantly more IFNg after stimulation than their pbNK counterparts. Preliminary in vivo work indicates that ieILC1-like cells perform comparably to K562-expanded NK cells. Thus, ieILC1-like NK cells represent a novel class of cell therapy that are more capable of infiltrating solid tumors and resisting their immunosuppressive cues.
Supported by NIH R35DE030054.
Evaluating potential toxicities of cancer immunotherapies using autoimmune animal models.
Authors: Okpasuo Onyekachi Juliet , Cattin-Roy Alexis N , Karasseva Natalia , Mahadevappa Chandrashekara Puthanapura , Radhakrishnan Vijay , Zaghouani Habib , Pages Diana Gil 
Published: January 2023

Abstract
During the past few decades, cancer immunotherapy has recorded breakthroughs involving therapeutic agents that mediate immune response against several tumor types. However, an important consideration with immunotherapy treatment in the clinic has been the emergence of immune related adverse events (irAEs), that could lead to frequent hospitalization, premature therapy cessation and fatalities, thus limiting the efficacy of these therapies. Additionally, despite the clinical benefits of combining therapies, it has been observed that severity of toxicity increases with combination of therapy. Although, these irAEs are commonly seen in patients receiving immunotherapy, such events have been rarely observed in preclinical mouse models due to their nature: inbred, resistant, and short study duration. Here, we report the use of EAE autoimmune mouse model to evaluate potential toxicities of a novel cancer immunotherapy: anti-CD3ɛ mono-Fab, that is effective as a single therapy and could synergize with checkpoint inhibitors for superior anti-metastatic tumor response. EAE model groups treated with combination therapies showed more disease severity recapitulating the higher grade of irAEs seen in patients receiving combination therapies. The data suggest that autoimmune mouse models could serve as an effective tool to study the severity of toxicity associated with immunotherapeutic agents, and to determine optimized combinatorial regimens for best therapeutic performance.
Supported by NCI U01CA244314
Overcoming AML T cell therapy barriers with engineered proteins
Authors: Ruan Aitong , Chiu Edison , Oda Shannon K 
Published: January 2023

Abstract
Leukemia is the most common pediatric cancer and the second leading cause of cancer deaths in children. Adoptive cell therapy (ACT) has been validated in pediatric B cell acute lymphoblastic leukemia, where engineered cancer-targeting T cells can effectively eradicate tumor and tumor recurrence, generally more safely than drug and radiation therapy. However, ACT is not approved for patients with pediatric AML, other leukemias or solid tumors. Of note, pediatric AML patients have a worse 5-year survival rate than ALL.
AML cells can severely diminish ACT efficacy by expressing ligands that activate T cell inhibitory receptors and by downregulating costimulatory ligand expression. To address these issues, we developed fusion proteins that combine the ectodomain of an inhibitory T cell receptor with an intracellular costimulatory signaling domain. We have shown that this effectively “replaces a brake with an accelerator” in CD8 T cells, and have now developed FPs with different costimulatory domains to confer different attributes to T cells. Despite improved outcomes with single FP T cell therapy, we hypothesized that combining differently costimulated T cells would provide a synergistically diverse T cell response and improve outcomes. We tested 1:1 pairwise combinations of differently costimulated T cells that are also transduced with a tumor-specific TCR in serial killing over-time (SKO) assays and in vivo persistence studies. In survival studies, dual T cell therapy resulted in enhanced survival. We propose that a synergistically diverse T cell therapy provides enhanced antitumor efficacy over a monogenic cell product.
Adjuvant AS01 induces monocyte activation that favors T cell expansion and diversification in primary human cells
Authors: Smith Carson L , Canaday David H 
Published: January 2023

Abstract
Vaccines are less effective in older adults, and efforts to improve vaccine efficacy generally show limited clinical improvement. However, the vaccine Shingrix, developed by GlaxoSmithKline, is FDA approved and recommended for adults over the age of 50 for the prevention of shingles. Shingrix contains the novel adjuvant AS01 and shows remarkably high (&gt;90%) effectiveness at both preventing shingles and reducing the development of postherpetic neuralgia, even in adults over 80. We seek to examine AS01-induced activation of myeloid cells and determine their impacts on T cell memory differentiation and function in an effort to understand how this vaccine’s high clinical efficacy is produced.
We use peripheral blood samples to examine AS01-induced innate immune cell activation, including interactions with T cells once activated. We find that AS01 induces costimulatory marker upregulation and cytokine production in myeloid dendritic cells and monocytes in a synergistic manner, with the most upregulation seen in monocytes. Naïve and memory CD4 T cells incubated with AS01-treated monocytes have increased frequencies of TH17 (CCR6+ CXCR3−) cells and decreased frequencies of TH1 cells compared to controls. Naïve CD4 T cells proliferate more when incubated with AS01-treated monocytes than with controls. AS01-treated monocytes additionally induce 2x less IFN-γ and IL-5 in response to mitogen (SEB) from CD4 T cells, suggesting a modulation of response.
We propose that AS01 influences monocytes to diversify the existing memory T cell repertoire and skews naïve T cells towards more cytotoxic phenotypes such as TH17, contributing to greater clinical efficacy in preventing shingles in vivo.
Supported by VA Merit grant (CX002060)
Improving the metabolic efficiency of human T cells through AMPK agonist treatment
Authors: Braverman Erica , McQuaid Margaret , Byersdorfer Craig A 
Published: January 2023

Abstract

BACKGROUND
Adoptive immunotherapies are limited by the increased differentiation and metabolic exhaustion of the T cell product. The cellular energy sensor AMP-activated protein kinase (AMPK) controls many metabolic pathways. We hypothesized that increasing AMPK signaling during T cell expansion would create more metabolically efficient T cells.

RESULTS
Human T cells were stimulated in vitro, expanded with or without AMPK agonist A769662, and restimulated under varying glucose concentrations. Agonist treatment increased T cell expansion by manual cell counts (1.5× increase in cell number, p&lt;0.01), and produced a higher percentage of CD62L+CD27+ cells by flow cytometry (10% increase, p&lt;0.01). Twenty-four hours after re-stimulation, agonist-treated cells exhibited decreased basal oxygen consumption and extracellular acidification rates (45% and 20% decrease, p&lt;0.001) but increased spare respiratory capacity (SRC) (97% increase, p&lt;0.001). At 72 hours, control cells decreased proliferation in a glucose dose-dependent manner, as measured by BrDU uptake, whereas treated cells maintained a higher level of proliferation despite limited glucose availability.

DISCUSSION
Increasing AMPK signaling during T cell expansion improves doubling time while maintaining higher CD27 positivity, suggesting a greater yield of less differentiated cells. Upon re-stimulation, treated cells initiate less metabolic activity but maintain greater proliferation and SRC under glucose-limiting conditions. These results suggest that greater metabolic efficiency in agonist-treated cells allows for continued function despite nutrient restriction, a trait which could confer significant advantages to adoptive immunotherapies.
Supported by grants from NIH (5K12 HD052892) and the St. Baldrick’s Foundation (St. Baldrick’s Fellowship Grant)
Phenotypic and transcriptomic profiles of tumor-conditioned macrophages in dogs, and the translational value of canine cancer immunology research.
Authors: Kerboeuf Mikael , Haaland Anita Haug , Moe Lars , Argyle David , Boysen Preben 
Published: January 2023

Abstract
Tumor-associated macrophages (TAMs) are heterogeneous and abundantly present in the tumor stroma. They derive from tissue-resident macrophages and recruited monocytes and play a central role in several aspects of cancer. In murine models, TAMs promote invasion, intravasation, circulatory survival and extravasation of tumor cells. TAMs also promote tumor progression and angiogenesis, remodel the tumor microenvironment, and modulate the adaptive immune system. Evidence suggests TAMs are vital for establishing the premetastatic niche. Historically, family-owned dogs have proven to be reliable cancer models. In contrast to laboratory animals, dogs develop cancer naturally and commonly, co-inhabit our environment, are genetically outbred and immunologically experienced. We hypothesized that canine tumor conditioned monocyte-derived macrophages (TCMs) share many characteristics with M2 macrophages and have a tumor-promoting phenotype as in humans. We collected blood from healthy dogs, isolated peripheral blood monocytes and differentiated them in vitro using tumor conditioned media from three canine cancer lines and M-CSF. Three TCM populations were generated and compared with M1 and M2 macrophages. We assessed the macrophages using multicolor flow cytometry and RNA-sequencing. All TCM showed an increased expression of two M2 markers (CD209a and FcɛRI), while 2/3 TCM populations had increased expression of two additional M2 markers (CD206 and CD11d). Transcriptomic analysis is ongoing. Results so far support the hypothesis that canine TCMs share many similarities with human TCMs, further warranting the use of dogs as cancer immunology models.
Supported by grant from Agria Animal Insurances (N-2020-0061) and The Research fund for cancer in dogs (Oslo, Norway)
Discovery of an organized nasopharynx-associated lymphoid tissue in the nasal cavity of rainbow trout and its role in secondary adaptive immune responses to nasal vaccines
Authors: Salinas Irene , Garcia Benjamin , Dong Fen , Casadei Elisa 
Published: January 2023

Abstract
The olfactory organ of all vertebrates is constantly exposed to pathogens. The nasal mucosa is therefore equipped with a local immune system that protects the host from infection. In teleost fish, we previously described the presence of a diffuse nasopharynx-associated lymphoid tissue (NALT) found in the olfactory lamellae. This diffuse NALT (d-NALT) consists of myeloid and lymphoid cells but lacks an organized structure. Rainbow trout d-NALT mounts quick innate immune responses as well as adaptive immune responses to nasal vaccines. Here we report the discovery of a novel lymphoid structure located in the inner epithelial lining of the nasal cavity, opposite to the tips of the olfactory lamellae, in rainbow trout. This lymphoid structure represents the first description of an organized NALT (O-NALT) in teleost fish. Rainbow trout O-NALT is rich in B and T lymphocytes at the steady state with a proportion of approximately 8:3:1 IgM+ B cells, CD8+ T cells and IgT+ cells, respectively. Using laser capture microdissection we show that compared to d-NALT, O-NALT is highly enriched in expression of CD4, CD8 and IgM but not IgT. Strikingly, O-NALT expresses 8-fold higher levels of activation-induced cytidine deaminase aidc mRNA compared to d-NALT. Following primary and secondary intranasal vaccination with a live attenuated viral vaccine against infectious hematopoietic necrosis (IHN), trout O-NALT was expanded with increased numbers of B and T cell lymphocytes. Our results indicate that teleost fish posses organized lymphoid structures in the nasal cavity that participate in the maturation of the adaptive immune response and that need to be considered as important players in the response to mucosal vaccines.
This work was supported by USDA NIFA award # 2020-67015-31457
Altered transcriptome responses in the lungs of preweaned calves supplemented with a yeast fermentate during a viral-bacterial coinfection
Authors: McDonald Paiton O , Maina Teresia Wangari , Samuel Beulah Esther Rani , McGill Jodi L 
Published: January 2023

Abstract
Bovine respiratory disease is a complex syndrome which contributes to severe and often fatal pneumonia in beef and dairy cattle. Saccharomyces cerevisiae fermentation products (SCFP) are feed ingredients that have been previously shown to alter the immune response and outcome of a respiratory viral infection in calves. The objective of this study was to determine the effect of SCFP supplementation on the lung transcriptome in calves responding to a co-infection with bovine respiratory syncytial virus (BRSV) and Pasteurella multocida (PM). Twenty-eight, 1–2 day old calves were assigned to two groups: 1) control diet; or 2) SCFP treated diet. Calves were infected with BRSV on day 21, followed 6 days later by intratracheal inoculation with PM. Calves were euthanized on day 10 post-viral infection. There were no differences in viral or bacterial burden between treatment groups, but SCFP treated calves tended to have less severe lung pathology. Transcriptome analysis of lung tissue and bronchoalveolar lavage samples revealed over-represented genes related to membrane integrity, pathogen binding, and complement activation which presumably contributes to the host’s resiliency and response to the coinfection. Results from this study suggest that supplementing preweaned calves with SCFP may modulate the innate and adaptive immune response in mucosal sites. These findings will contribute to a better understanding of the underlying host response to bovine respiratory disease.
Supported by funding from Diamond V Mills, Inc., Cedar Rapids, IA
Mapping the spatial biology of COVID-19 immunopathology in placenta tissues
Authors: Ma Ning , Braubach Oliver , Rebutini Patricia Zadorosnei , Machado-Souza Cleber , Stonoga Emanuele Therezinha Schueda , de Noronha Lucia , Souza-Fonseca-Guimaraes Fernando , Kulasinghe Arutha 
Published: January 2023

Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that emerged in late 2019 has spread globally, causing a pandemic of respiratory illness. The virus primarily affects the lungs, where it induces respiratory distress syndrome ranging from mild to acute. However, a growing body of evidence shows COVID-19 pathology in other organs that also carry the ACE-2 receptor, including the placenta. Most newborns delivered from COVID-19 positive mothers test negative following delivery, suggesting that there are protective mechanisms within the placenta. In this study, we compared a unique collection of placental tissue samples from COVID-19 positive and gestational age-matched non-COVID-19 samples. Using AKOYA’s ultrahigh-plex Phenocycler assay, we mapped dozens of protein biomarkers in situ with single-cell and subcellular spatial resolution to produce unbiased and comprehensive spatial biomarker maps of potential placental pathobiology associated with COVID-19 infections. Our antibody panel was aimed at in-depth identification of immune cell lineages, activation states, immune checkpoints, and tissue structures; infected cells were identified via the deployment of two antibodies directed at COVID-19 spike and capsid proteins, as well as an antibody directed against the ACE-2 receptor. All imaging data were obtained at single-cell resolution across whole tissue samples, and cellular phenotypes were subsequently identified via unsupervised clustering methods. These data are the first of their kind and will provide additional insight into the pathobiology of COVID-19.
TLR2 senses spike protein of SARS-CoV-2 to trigger inflammation
Authors: Zaki Hasan , Khan Shahanshah 
Published: January 2023

Abstract
The pathogenesis of COVID-19 involves cell death caused by SARS-CoV-2 infection and hyperinflammatory response. Therefore, understanding the mechanism of inflammatory response during SARS-CoV-2 is the key to develop strategies to reduce COVID-19 complications. We investigated what viral ligand is responsible for activation of innate immune cells by stimulating human and mouse macrophages with recombinant spike (S), membrane (M), envelope (E), and neucleocapsid (N) proteins. Interestingly, we observed that S protein, its subunits and S-trimer, but not M, N, and E proteins, induced inflammatory cytokines and chemokines. Similar to immune cells, human and mouse lung epithelial cells also responded to S protein to produce inflammatory molecules. Since SARS-CoV-2 dose not infect immune cells, we interrogated whether virally infected epithelial cells stimulate macrophages. We observed that S protein-expressed lung epithelial cells induced inflammatory responses in macrophages when cocultured. Notably, S protein induced inflammation independent of ACE2 receptor. Digging dipper, we wanted to explore what pattern recognition receptor senses S protein to trigger inflammation. Our investigation revealed that TLR2, but not TLR4, is the immune sensor for S protein which in association with either TLR1 or TLR6 activates the NF-kB pathway via MyD88. Consistently, S-induced inflammatory response was completely absent in Tlr2−/− mice. Blocking TLR2 chemically or though gene manipulation also abrogated S-induced inflammation in vitro. Together these data demonstrate the mechanism of cytokine storm during SARS-CoV-2 infection, and suggest that TLR2 could be a potential therapeutic target for COVID-19.
Supported by R01 DK125352
Children with Multi-System Inflammatory Syndrome (MIS-C) develop functionally competent T cell memory against SARS-CoV-2 following recovery
Authors: Rybkina Ksenia , Bradley Marissa C , Bell Joseph N , Meng Wenzhao , DiLorenzo Michael P , Anderson Brett R , Pethe Kalpana , Luning-Prak Eline , Farber Donna L , Connors Thomas J 
Published: January 2023

Abstract
While pediatric SARS-CoV-2 infection is largely spared from severe COVID-19, a small fraction of children develop a life-threatening condition known as MIS-C, which is associated with an aberrant peripheral blood immunophenotype; namely within adaptive immune cell populations. Little is known about how MIS-C impacts memory generation that would be imperative for future SARS-CoV-2 responses. Here, we used high dimensional flow cytometry and TCR sequencing to assess the peripheral blood T cell profile during acute MIS-C (n=17), following recovery (MIS-C-R) up to 9 months post-MIS-C (n=9), and relative to age-matched convalescent, mild pediatric COVID-19 (non-MIS-C; n=5) and uninfected controls (n=3). We also applied antigen-specific analyses to determine the magnitude and functional capacity of SARS-CoV-2-specific T cells in MIS-C-R (n=10) and non-MIS-C children (n=16). Flow cytometry showed unique T cell composition in MIS-C which resolved in recovery: lacking persistence of exhausted CD8+ T cells observed during MIS-C, and reestablishment of the naïve T cell pool. TCR sequencing showed decreased clonal diversity and distinct TCR Vβ usage in acute MIS-C compared to MIS-C-R. Results from antigen-specific analyses show that while T cell memory is similar phenotypically, MIS-C-R children have greater frequencies of SARS-CoV-2-specific CD4+ T cells than non-MIS-C up to 9 months since diagnosis. Meanwhile, SARS-CoV-2-specific CD8+ T cells in MIS-C children were more cytokine-producing, while more cytotoxic in non-MIS-C children. Together, these results reveal a restoration in global T cell phenotype in MIS-C-R children, as well as the robust production of functionally competent SARS-CoV-2 specific T cell memory.
This work is supported by grants from NIH (K23 AI141686, 3U01 AI100119-10S1)
Investigating the role of inflammasome activation by dengue virus non-structural protein 1 during dengue infection.
Authors: Wong Marcus P , Juan Evan Y W , Blanc Sophie F , Biering Scott B , Wang Phoebe , Beatty Robert , Harris Eva 
Published: January 2023

Abstract
Dengue virus (DENV), consisting of serotypes 1–4, is the most medically important flavivirus, causing an estimated 50–100 million dengue cases globally every year. Most symptomatic DENV infections present as an acute febrile illness; however, severe complications include vascular leakage leading to hemorrhage and hypovolemic shock. DENV non-structural protein 1 (NS1) is secreted from infected cells and has been implicated as a major driver of dengue pathogenesis, activating immune cells and acting directly on endothelial barriers, causing them to become pathologically hyperpermeable. While recent work has delved into the mechanisms behind the endothelial cell-intrinsic pathway of DENV NS1-induced vascular leak, much less is known about how DENV NS1 interacts with immune cells and what role this activation plays in DENV infection. We have discovered that DENV NS1 can trigger activation of the inflammasome, a family of cytosolic innate immune sensors that react to danger-associated molecular patterns. DENV NS1 induces the release of the pro-inflammatory cytokine IL-1β in human and murine monocytes in a caspase-1 dependent manner that is independent of NLRP3. Additionally, we found that caspase-1-deficient, but not NLRP3-deficient, mice are more susceptible to infection in a murine model of DENV infection. These results indicate that the inflammasome acts as a sensor of the DENV viral toxin NS1 and plays a protective role during DENV infections
This work was supported by NIAID/NIH grant R01 AI24493 (E.H.)