bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mitofission impairs the anti-tumor activity of NK cells in tumor microenvironment
Authors: Zheng Xiaohu , Sun Rui , Tian Zhigang , Wei Haiming 
Published: January 2023

Abstract
The survival and functional performance of NK cells is a metabolically demanding process. However, there is largely unknown how inappropriate metabolic reprogramming underlies disease microenvironments leads to NK dysfunction. Here, through analysis of human liver cancer samples (n=102) and benign liver samples (n=22), we found tumor infiltrative NK (TINK) cells present small and fragmented mitochondria in the cytoplasm, while normal liver NK and peripheral NK cells have big and tubular mitochondria. We also observed hypoxic tumor microenvironment is the main factor to induce NK cell excessive mitochondrial fission into fragmentation and that dependent on increasing Drp1 activity. Consequently, the fragmentation causes the metabolic disturbance and losing of TINK cells and that predicts poor survival in liver cancer patients. Revising mitochondrial morphology by targeting Drp1 improves mitochondrial metabolism and anti-tumor capacity of NK cells. Thus, disease-related mitochondrial restructuring drives NK cell exhaust and that is manipulated to the benefit of temper or enhance immunity.
Interactions of natural killer cells and pancreatic cancer cells in simulated microgravity
Authors: Gregg Randal K , Falwell Harrison , Bludorn Grant , Laken Jordan A , Boos William F , Arnold Jennifer P , Bradley Jillian H 
Published: January 2023

Abstract
One of the major concerns of long-duration spaceflight is the exposure of astronauts to space radiation and the potential for cancer development. Natural killer (NK) cells continuously survey the body for such abnormalities and upon detection are activated to produce pro-inflammatory cytokines as well as cytotoxins which induce apoptosis of the cancer cells. This study employed the rotary cell culture system to generate a simulated microgravity (SMG) environment in order to assess its effect upon the growth of human cancer cells and their susceptibility to lysis by NK cells. We found that SMG diminishes the numbers of both pancreatic cancer cells and NK92MI, a human NK cell line, over a 7 d culture period. Additionally, SMG NK92MI cells had a ~50% decrease in cytotoxicity against PANC-1 pancreatic cancer cells. Surprisingly, a long-term SMG culture of up to 14 d resulted in complete restoration of NK92MI cell cytotoxic function. Interestingly, SMG cultured NK92MI cells produced more IFN-gamma than Static NK cells throughout the time periods assessed. In comparison, 7 d culture of pancreatic cancer cells in SMG led to a ~50% reduction in the ability of either Static or SMG NK92MI cells to induce apoptosis in the SMG cultured cancer cells. Therefore, over a two-week period in SMG culture, NK92MI cells were able to adapt to the environment and recover cytotoxic function, however, pancreatic cancer cells cultured in SMG acquired resistance to lysis by the NK cells. Hence, cancer cells developing in astronauts in microgravity might have a greater potential to resist NK cell control than in patients here on Earth which upon elucidation may aid in the identification of relevant new tumor targets for therapeutics.
Interferon associated adaptive immune resistance represented by immune checkpoint expression and immune cell localization in high-grade serous ovarian cancer
Authors: Li Deyang , Vidotto Thiago , Shakfa Noor , Asante Afrakoma-Afriyie , Peterson Nicole , de Ladurantaye Manon , Mes-Masson Anne-Marie , Francis Julie-Ann , Koti Madhuri 
Published: January 2023

Abstract
High-grade serous carcinoma (HGSC) is a deadly malignancy leading to ~70% of the 140,000 ovarian cancer deaths globally each year. Resistance to platinum chemotherapy followed by recurrence and an incurable disease occurs in most HGSC patients. Contemporary immune checkpoint blockade therapies have shown minimal efficacy in this cancer. Our previous investigations established that tumour interferon (IFN) activation status and CD8+ T cell density are predictors of chemotherapy response in HGSC. Furthermore, we also showed that IFN induced chemokine CXCL10 is a key determinant of increased survival via immune cell recruitment in the tumour immune microenvironment (TIME). Given that signal transducer and activator of transcription 1 (STAT1) is central to the feed forward loops of cellular IFN responses, we investigated STAT1 associated transcriptomic alterations and spatial profiles of immune cells in 204 pre-treatment HGSC tumours.
RNA-sequencing based whole transcriptomic profiling revealed that higher STAT1 expression significantly correlated with higher immunomodulatory gene expression, including immune checkpoints and activators, in both chemotherapy sensitive and resistant tumours. Findings were independently validated in a cohort of 379 HGSC tumour RNA-Seq profiles from The Cancer Genome Atlas Network ovarian cancer dataset. Multiplex immunofluorescence based spatial profiling of CD8+ T cells, FoxP3+ T regulatory T cells, CD68+M1, and CD163+ M2 macrophages and expression of PD-L1, PD-1, IDO1 immune checkpoints was performed. Findings from our study provide evidence for IFN mediated adaptive immune resistance in the HGSC TIME and will potentially inform the design of rational chemo-immunotherapy approaches.
Virally delivered cytokine therapy drives immune equilibrium in melanoma
Authors: Walsh Michael , Lenehan Patrick , Ali Lestat , Dougan Michael , Knipe David , Dougan Stephanie K 
Published: January 2023

Abstract
Although a modified, replicating herpes simplex virus (HSV) encoding GM-CSF was FDA-approved for the treatment of melanoma (T-VEC), which was revolutionary for the field of oncolytic virotherapy, there are several areas for improvement. First, the live virus cannot be safely administered to immunocompromised patients. Second, GM-CSF induces immune-suppressive myeloid cells and is likely not the optimal insert. To this end, our lab has used a non-replicating HSV virus, termed d106S, to serve as a safe viral vector for the treatment of cancers. We have repurposed the non-lytic HSV-1 d106S vaccine vector for local delivery of IL-12. Our replication-defective d106S virus releases a large burst of IL-12 locally within the tumor environment, synergizing with a type I IFN response induced by the virus. We have shown that d106S-IL12 induces regression and long-term stable immune equilibrium in murine B16 melanoma. These results are promising, demonstrating that the d106S vector can deliver immunotherapeutic cargo and induce shrinkage of established tumors. Intriguingly, most mice do not fully clear their tumors but establish an equilibrium phase and withdrawal of therapy eventually leads to tumor outgrowth. We have profiled the immune response induced by d106S-IL12 at separate time points, identifying several points of possible intervention, including blockade of innate inflammatory cytokines IL-1β, TNFα and IL-6. Indeed, blockade of each of these inflammatory cytokines provides a boost to the response initiated by d106S-IL12, which we have further profiled with single-cell RNA-sequencing.
A flagellin-adjuvanted cancer vaccine combined with photodynamic therapy enhanced anti-tumor immune response in B16-F10 melanoma cancer model
Authors: Hwang Hye suk , Lee Shee Eun , Rhee Joon Haeng 
Published: January 2023

Abstract
Immune checkpoint inhibitors have now become a standard therapy for malignant melanoma. However, immune checkpoint inhibitor monotherapy has shown effective in only a limited number of patients. Photodynamic (PDT) and immune combined therapy is a promising modality for the treatment of metastatic melanoma. Herein, we report the combination therapy with PDT and flagellin adjuvanted- Tyrp1/Trp2/gp100 peptides based vaccination (Fla-vax) in mouse models of B16-F10 melanoma cancer. We used the bilateral tumor model to determine whether the Fla-vax could induce systemic anti-tumor effects. The combination therapy with PDT and Fla-vax sufficiently induced systemic anti-tumor immune responses for local and abscopal tumor control,, with a significant increase in the systemic IFNγ/TNFα secretion from cytotoxic T lymphocytes (CTLs) and tumor-infiltrated effector memory CD8+ T cells. It was also found that the combined therapy greatly induced the number of migratory CD103+ DCs which is secreting CXCL10. The therapeutic benefit of PDT and Fla-vax proved to be CD8+ T-cell dependent, and the antitumor activity was additive with that provided by programmed death-1 (PD-1)-targeted immunotherapy. These results suggest the combination of active immunotherapy with tumor ablation by PDT as a feasible novel treatment strategy for advanced melanoma cancer.
Innate immune cell modulation using cargo-less nanoparticles to ameliorate severe inflammation
Authors: Lasola Jackline Joy Martin , Kamdem Henry , McDaniel Michael , Pearson Ryan M. 
Published: January 2023

Abstract
1.6 million people are diagnosed with sepsis each year in the US with a mortality rate of 25-30%. Attempts to improve patient outcomes include strategies to abrogate the overexaggerated immune response seen in sepsis but have thus far failed. It has been hypothesized that these have collectively been unsuccessful as they target a single molecular pathway and do not address redundancies in immune activation pathways. We have shown that nanomaterials are inherently potent immunomodulators that can broadly reduce the release of inflammatory cytokines. As such, we aimed to understand the physical and biological mechanisms that underlie the macrophage response to nanomaterials in the setting of LPS stimulation. PLA-based particle formulations were prepared with similar sizes (~500 nm) and different charges depending on surfactant (−50 to −20 mV). Particles prepared with an anionic surfactant (PEMA) are more effective than those with a neutral surfactant (PVA) at inhibiting inflammatory cytokine secretion in a manner independent of NF-κB activation. Additional studies show that PLA-PVA particles colocalize with TLR agonist at the cell surface, while agonist localization was abolished in cells treated with PLA-PEMA particles. LPS-induced endotoxemia studies show that i.p. administration of PLA-PEMA particles prior to challenge with LPS significantly improved survival of mice compared to those challenged with LPS alone. This work contributes to the fundamental understanding of the physicochemical properties of biomaterials and their modulation of immune cell responsiveness towards activating stimuli and suggests that nanomaterials are novel immune modulators in cases of acute and potentially life-threatening inflammation.
In-silico based screening system to develop allosteric modulators of Bruton’s tyrosine kinase
Authors: Berthelette Alyssa , Kolli Nilima , Amatya Neha , Andreotti Amy , Berg Leslie , Kang Joonsoo , Reboldi Andrea 
Published: January 2023

Abstract
Ibrutinib is a covalent inhibitor that binds to the Bruton’s tyrosine kinase (BTK) ATP-dependent active site and it is currently used for treating a variety of B cell tumors. A high rate of drug resistance develops in Ibrutinib-treated patients through mutations at the BTK kinase domain, resulting in more aggressive diseases. Thus, there is significant interest in identifying modulators that inhibit BTK in an active site-independent manner, specifically via allosteric modulation. We used an in-silico system to identify small molecules that bind outside of the BTK active site. Candidate compounds were then tested for direct binding to full length BTK protein using Saturated Transfer Difference Nuclear Magnetic Resonance (STD-NMR) and were subsequently used in primary mouse B cell-based assays to determine their functional relevance. Ex vivo B cells were triggered via the BCR in the presence of compounds and signaling events downstream of the BCR, including Nur77 upregulation and calcium flux, were measured by flow cytometry. A series of candidate allosteric BTK inhibitors were then confirmed to retain suppressive activity in an ABC-DLBC lymphoma line that carries the Ibrutinib resistant mutation. Additionally, primary human CLL, but not healthy B cells, were susceptible to candidate inhibitor-mediated death. These results indicate that a screening approach integrating in silico docking, NMR measurements, and cellular assays with primary B cells can be used to identify BTK inhibitors that act outside of the kinase domain. Allosteric BTK inhibitors will broaden the range of treatment options for B cell cancers, possibly overcoming or preventing the Ibrutinib-induced resistance when used as a combination therapy.
Targeting of ITK-SYK fusion oncogene through in-silico generated and biologically validated ITK-PH domain modulators
Authors: Ferraj Enxhi , Kolli Nilima , Berthelette Alyssa , Frascoli Michela , Andreotti Amy , Berg Leslie J. , Reboldi Andrea , Kang Joonsoo 
Published: January 2023

Abstract
A subset of human Peripheral T cell Lymphomas, Not Otherwise Specified (PTCL-NOS), harbors a chromosomal translocation of interleukin-2 (IL-2) inducible T cell kinase (ITK) and spleen tyrosine kinase (SYK), giving rise to the composite ITK-SYK protein. ITK, a TEC family member kinase, plays an essential role in TCR and CD28 signaling. SYK signaling pathway is more broadly linked to activation of diverse lymphoid and myeloid cell types. ITK-SYK fusion protein consists of the PH domain of ITK and the SYK kinase domain allowing localization to the signalosome complex at the membrane via the PH domain and constitutive activity in T cells. Given the ubiquitous activity of SYK, it is predicted that targeting the ITK-PH domain of the fusion protein with small molecules can be a precise and less toxic approach to suppress ITK-SYK+PTCL-NOS in vivo. To this end, we performed a computational screen of small molecule libraries to identify candidate ITK-PH domain interacting compounds. A thermal shift assay was used to verify the specificity of binding of candidates to the target protein domain. Cellular assays tested small molecule activity in human and mouse T cells constitutively expressing the ITK-SYK fusion protein. We showed that the first-pass small molecule candidates specific to ITK-PH domain inhibit cytokine production driven by ITK-SYK fusion protein in human and mouse T cells. Our results demonstrate the efficiency of the virtual screening to identify drug candidates with biological activity in lymphocytes. Development of drugs that target ITK-PH domain is a rationale strategy towards first-in-class therapeutics for this subset of PTCL-NOS with very poor prognoses.
Host-directed therapy in foals reduces the severity of clinical pneumonia against Rhodococcus equi intrabronchial challenge
Authors: Bordin Angela Ilha , Giguere Steeve , Bray Jocelyne , Berghaus Londa , Hook Magnus , Scott Brenton , Johnson Rena , Cohen Noah 
Published: January 2023

Abstract
Pneumonia caused by Rhodococcus equi, an intracellular bacterium that infects macrophages, is an important cause of disease and death in immunocompromised hosts, especially foals. No vaccine is available, and control is based on antibiotic treatment of subclinical cases, increasing the pressure for emergence of antibiotic resistance and indicating novel control approaches are needed. TLR agonists have immunomodulatory function. Our first objective was to evaluate the ex vivo effects of treatment of neonatal foal alveolar macrophages (AMs) with a combination of TLR2 and TLR9 agonists (PUL-042). Using bronchoalveolar lavage (BAL), we collected AMs from 48 foals and determined the killing of intracellular R. equi using the ratio of CFUs at 48 hours (T48) to post-phagocytosis (T0) (i.e, T48:T0). Concentration of pro- and anti-inflammatory cytokines were measured in AM culture supernatant, BAL fluid, and serum. Compared to control, PUL-042 decreased TNF-a in BAL fluid but increased IL-6, IFN-g, and TNF-a in AMs after ex vivo infection with R. equi, suggesting that nebulization induced an immune response upon infection that is favorable for killing intracellular pathogens. We also evaluated the in vivo the efficacy of PUL-042 in reducing the severity of clinical pneumonia in foals after intrabronchial challenge with R. equi. Infected foals nebulized with PUL-042 had a shorter duration of clinical signs of pneumonia and of presence of lung abscesses when compared to foals that did not receive nebulization, consistent with our ex vivo findings. Our results show potential for enhancing neonatal immune responses against respiratory pathogens and reducing severity of clinical pneumonia following intrabronchial challenge with R. equi.
Rapid characterization of MHC class I diversity in desert bighorn sheep reveals population-specific allele expression
Authors: Dolan Brian P , Beechler Brianna R. , Dugovich Brian S. , Epps Clinton W. , Jolles Anna E. 
Published: January 2023

Abstract
Diversity of MHC class I alleles within animal populations is necessary to prevent pathogen escape from adaptive immune responses. However, demonstrating this phenomenon is difficult especially in wild animal species where the effect of pathogen escape is more profound than in laboratory animal models. Desert bighorn sheep (Ovis canadensis nelsoni) are a charismatic megafauna native to the southwest of North America which are threatened by a number of infectious diseases and, more recently, paranasal tumors. We sought to develop a relatively inexpensive and rapid way to identify the diversity of MHC class I genes and alleles within individual bighorn sheep. Both DNA and RNA were isolated from leukocytes collected from 154 adult bighorn sheep from across different mountain ranges in Southern California. Previously identified primers from domestic sheep were used to amplify exons 2 and 3 from OMHC I genes while appending adapters for PacBio circular consensus sequencing. PCR amplicons from individual animals were then subjected to a second round of PCR to append index sequences to allow assigning of individual sequences to unique animals. We successfully identified over 40 unique MHC class I sequences expressed by bighorn sheep. Between 2 and 5 loci were amplified in each individual animal. Analysis of gDNA revealed several pseudogenes and other DNA sequences which were not transcribed. Finally, we demonstrate that particular DNA sequences are found within particular populations, suggesting that as bighorn populations fragment, there is a loss of MHC class I diversity.
Assessment of immunocontraceptive vaccine for population control: evaluation of a recombinant fusion GnRH vaccine in mice
Authors: Chang Ai-Mei , Hou Ding-Liang , Chen Chen-Chih , Ke Guan-Ming , Lee Jai-Wei 
Published: January 2023

Abstract
Gonadotropin-releasing hormone (GnRH) plays a pivotal role in regulating the reproductive endocrine system. Accumulated lines of evidence indicated that GnRH immunocontraception vaccine is a potential tool for control and management of animal population. However, the poor immunogenicity of GnRH peptide has limited its efficacy and field application. In the present study, a modified antigen, containing 8 GnRH repeats and immunostimulatory fusion protein, was formulated an oil-based MONTANIDE™ ISA 206 VG adjuvant to enhance the immunogenicity of the GnRH vaccine. The vaccine was vaccinated in 4-week-old male ICR mice (n = 10) and boosted twice, 4 and 8 weeks, respectively after the primary immunization. Mice injected with saline served as the unimmunized control (n = 10). Results demonstrated that the serum GnRH-specific antibody in vaccinated mice was significantly (p &lt; 0.05) increased 5 weeks, peaked at 12 weeks, and remained high until 18 weeks after the primary immunization. In comparison with the control, vaccinated mice had significantly (p &lt; 0.05) reduced serum testosterone concentration (0.856 ± 0.411 v.s. 160.77 ± 1. 865 ng/ml) and hampered development of testicles (weight and spermatogenesis) at 24 weeks after the primary immunization. Finally, a mating test indicated that the conception rate of female mice housed with vaccinated and unvaccinated males was 29% and 75%, respectively. The recombinant GnRH vaccine may be a potential approach for controlling animal population through immunocontraception.
Resilience of High Immune Response (HIR) Genetics in the Context of Climate Change: Effects of Heat Stress on Cattle with Diverse Immune Response Genotypes
Authors: Mallard Bonnie A , Husseini Nasrin , Cartwright Shannon , Livernois Ali , Hodgins Doug , Altvater-Hughes Tess , Beard Shannon , Karrow Niel , Canovas Angela , Schmied Julie 
Published: January 2023

Abstract
As the temperature-humidity index (THI) increases, animals are less able to regulate body temperature leading to decreases in production and immune function. The High Immune Response (HIR™) technology classifies animals based on estimated breeding values (EBVs) of their IR capacity as high (H), average (A), or low (L). H-responders with their unique IR genotype have less disease than A or L, but have not been evaluated for adaptation to heat stress. Therefore, rectal temperatures of beef & dairy cows with known IR classification were recorded during THI&lt; 74 (normal) and THI&gt;74 (above normal). Results show that 50% of beef & 65% of dairy cows had body temperature above 39.2 at THI&gt;74. However, cows that ranked H for antibody response were significantly better at regulating temperature and respiration at THI&gt;74, than A or L. In vitro experiments showed variability in PBMC function when cells were heat stressed (42□, 4hrs). PBMC were stimulated with ConA to measure proliferation or LPS to assess nitric oxide (NO) production. HSP70 was measured in control and heat treated cells. Dairy results indicated that H responders had significantly greater cell proliferation across all treatments and greater NO and HSP production with significance depending on the treatment. Beef results indicate that HSP70 from unstimulated PBMC increased after 1 or 2 heat treatments, and the magnitude of the change varied by IR class. Conversely, PBMCs challenged with LPS produced more NO before than after heat treatments. These results indicate that cattle with diverse immune response genotypes vary in their ability to withstand heat stress, and that it should be possible to identify resilient individuals with both greater immunity and adaptation to climate warming.