bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunogenicity and the CXCL12-coat control T cell accumulation in murine pancreatic cancer
Authors: Yan Ran , Wang Zhikai , Gao Ya , Janowitz Tobias , Fearon Douglas T 
Published: January 2023

Abstract
Improving the efficacy of immunotherapies in pancreatic cancer can increase patient survival rate. Microsatellite-stable pancreatic adenocarcinoma (PDA) exclude T cells and resist immunotherapy. This may occur by cancer cells coating themselves with CXCL12, a chemokine produced by cancer associated fibroblasts (CAFs). However, PDA with microsatellite instability (MSI-H) is infiltrated with T cells and responsive to PD-1 checkpoint inhibition. We found that human MSI-H pancreatic cancer cells display the CXCL12-coat, suggesting additional factors control pancreatic cancer immune accessibility. We hypothesized that expression of neoantigens by MSI-H PDA is an independent variable that overcomes T cell exclusion mediated by the CXCL12-coat. To test this, we established a mouse hepatic PDA metastases model in which PDA cells express doxycycline-induced ovalbumin (OVA), and adoptively transferred OVA-specific OT-I CD8+ T cells report intra-tumoral accumulation of T cells. Inducing OVA expression in established metastases increases intra-tumoral accumulation and activation of OT-I cells, despite the presence of the CXCL12-coat. We are currently investigating whether high- and low-affinity antigen-expressing cancers are subject to T cell mediated elimination in the presence and absence of the CXCL12-coat, respectively. The CXCL12-coat on PDA cells and the expression of neoantigens can independently control immune response. As the formation of the CXCL12-coat is facilitated by the Transglutaminase-2 (TGM2) and can be inhibited by TGM2 inhibitors (Z Wang et al., bioRxiv, 2019), these findings reveal potential combination therapies to overcome pancreatic cancer immunotherapy resistance.
Improving Specific Targeting of Tumors Through Bispecific SNIPER Antibodies
Authors: Erbe Amy K , Hernandez Reinier , Gerhardt Daniel , Hammer Bonnie , Felder Mildred , Bercher Mark , Dennin Jennifer , Massey Christopher , VandenHeuvel Sabrina , Feils Arika S , Heck Mackenzie , Hank Jacquelyn A , Glasser Bryan , Green Roland , Sondel Paul M 
Published: January 2023

Abstract
GD2 is expressed on neuroblastomas, as well as melanomas, small cell lung cancers and sarcomas. Anti-GD2 mAb can be used to treat these cancers and is part of the standard care for neuroblastoma. Expressed minimally on normal tissues, GD2 is expressed on some nerve cells. Thus, anti-GD2 treatment can cause neuropathic pain. To increase tumor specificity, we developed a bispecific SNIPER antibody, INV721, to simultaneously target 2 tumor antigens. One arm of INV721 is specific to GD2 and the other arm is specific to B7H3. B7H3 is overexpressed on multiple tumor types, including those listed above, with minimal expression on most normal cells (some expression on liver cells), and absent on nerve cells. The Fab arms targeting GD2 and B7H3 are each low-moderate affinity, such that INV721 will only bind with high affinity when both arms of INV721 bind to their antigens on the same cell. This high-affinity binding requires expression of both GD2 and B7H3, resulting in high-specificity of the SNIPER to tumor cells. To test the in vivo binding affinity of our bispecific antibody, INV721 was radiolabeled with 89Zr. Mice bearing GD2/B7H3-expressing tumors were intravenously injected with 89Zr-labeled INV721 and its longitudinal in vivo biodistribution was monitored via positron emission tomography imaging. 89Zr-INV721- showed elevated and persistent accumulation in the tumor with minimal uptake in normal tissues. 89Zr-radiolabeled isotype control antibody displayed significantly lower tumor uptake demonstrating the specificity of INV721. The goal of this SNIPER-antibody is to enhance the tumor-specific delivery of therapeutic mAbs, which may decrease toxicity and improve efficacy for cancers expressing both GD2 and B7H3.
Synthetic long peptide vaccines possessing a universal helper epitope can unmask the therapeutic effects of MHC I-restricted neoepitopes
Authors: Swartz Adam , Riccione Katherine , Congdon Kendra , Sanchez-Perez Luis A , Nair Smita K , Sampson John H 
Published: January 2023

Abstract
Synthetic long peptide (SLP) vaccines targeting neoantigens arising from patient-specific missense mutations may offer a means to address the antigenic heterogeneity of glioblastoma (GBM) tumors by simultaneously targeting a multitude of tumor-specific antigens expressed throughout these heterogenous tumors. This approach is challenged by uncertainties pertaining to vaccine design, including peptide composition required for immunogenicity and efficacy. To this end, we investigated the mechanism of an efficacious SLP vaccine targeting the neoantigen Odc1, expressed within the mouse astrocytoma SMA560. This led to the identification of three generalizable principles governing the effectiveness of neoantigen-targeting SLPs: (1) SLPs containing an MHC I-restricted neoepitope may activate neoantigen-reactive CD8+ T cells, which drive direct antitumor effects; (2) to induce robust neoantigen-reactive CD8+ T-cell responses, CD40L-mediated T cell "help" is required; and, (3) CD40L interactions are conferred by an SLP only when a “helper” epitope is physically conjoined to an MHC I-restricted neoepitope. To leverage these findings for clinical translation, we developed a rationally-designed vaccine comprised of an MHC I-restricted neoepitope linked to a universal “helper” epitope. This design was capable of maintaining the effects of the Odc1 vaccine and, remarkably, unmasked the therapeutic effects of, otherwise, poorly immunogenic MHC I-restricted neoepitopes. Together, these findings are significant because they elucidate mechanisms required for efficacious SLP vaccines and demonstrate a clinically-tractable approach with the potential to expand the therapeutic breadth of neoantigen vaccines.
An in vivo humanized mouse model for determining donor-dependent cytokine release syndrome in response to checkpoint and bispecific antibody therapy
Authors: Draheim Kyle M , Yang Jiwon , Jiao Jing , Cheng Mingshan , Keck James G 
Published: January 2023

Abstract
Although immuno-oncology therapies such as checkpoint inhibitors, bispecific antibodies and CAR-T cells have been successfully used as cancer treatments, they can have severe adverse effects such as cytokine release syndrome (CRS). The animal models and in vitro human PBMC assays presently in use do not reliably predict CRS in patients. Currently, the only widely accepted predictors of CRS are cancer burden and therapeutic dose. Despite this, most pre-clinical assays used to evaluate CRS do not incorporate cancer cells. A predictive assay that can screen for patient/cancer/therapy combinations at risk for developing CRS would improve the safety of immuno-oncology drug development.
We have developed a novel, rapid, sensitive and reproducible in vivo PBMC humanized mouse model that is able to differentiate human PBMC donors based on individual cytokine release response. Using this model, we evaluated cytokine release in response to immune checkpoint inhibitors and bispecific T-cell engaging antibodies. Additionally, this mouse model uses luciferase labelled cell lines to allow the measurement of tumor burden at the time of treatment as well as assess short term tumor response. We have ongoing efforts to further develop this assay in order to evaluate efficacy and CRS of at the same time, and progress will be discussed. This mouse model has immediate utility in current and future drug development.
Vaccine efficacy against primary and metastatic cancer with in vitro-generated CD103+ conventional dendritic cells
Authors: Zhou Yifan , Slone Natalie , Chrisikos Taylor T , Kyrysyuk Oleksandr , Babcock Rachel Lauren , Medik Yusra B. , Li Haiyan S , Kleinerman Eugenie S , Watowich Stephanie S 
Published: January 2023

Abstract
Type 1 conventional dendritic cells (cDC1s) possess efficient antigen presentation and cross-presentation activity, yet the efficacy of immunotherapy utilizingthis population is understudied. We used in vitro-generated CD103+cDC1s in vaccination strategies to test their ability to control melanoma and osteosarcoma tumors. In vitro-generated CD103+cDC1s produced cDC1-associated factors such as IL-12p70 and CXCL10, and demonstrated antigen cross-presentation activity upon stimulation with the Toll-like receptor 3 (TLR3) agonist polyinosinic:polycytidylic acid (poly I:C). In vitro-generated CD103+cDC1s also migrated to tumor-draining lymph nodes (TdLNs) following poly I:C treatment and intratumoral (i.t.) delivery. Vaccination with poly I:C-activated and tumor antigen-loaded CD103+cDC1s enhanced tumor infiltration of tumor antigen-specific and IFN-g+CD8+ T cells, and suppressed melanoma and osteosarcoma growth. CD103+cDC1s showed superior anti-tumor efficacy compared to vaccination with monocyte-derived DCs (MoDCs), and led to complete regression of 100% of osteosarcoma tumors in combination with CTLA-4 antibody-mediated checkpoint blockade. In addition, CD103+cDC1s effectively protected mice from pulmonary melanoma and osteosarcoma metastases. Our data indicate an in vitro-generated CD103+cDC1 vaccine elicits systemic and long-lasting tumor-specific T cell-mediated cytotoxicity, which restrains primary and metastatic tumor growth. The CD103+cDC1 vaccine was superior to MoDCs and enhanced response to immune checkpoint blockade. These results indicate the potential for new immunotherapies based on use of cDC1s alone or in combination with checkpoint blockade.
Oncolytic peptide LTX-315 enhances anti-melanoma immunity by inducing MyD88-dependent maturation of dendritic cells
Authors: Yang De , Li Xiao-Qing , Alam Md Masud , Rekdal Øystein , Oppenheim Joost J 
Published: January 2023

Abstract
LTX-315 is a synthetic cationic oncolytic peptide with potent anticancer cytotoxicity. LTX-315 induces both immunogenic tumor cell death and the generation of tumor-specific immune response. Given the central role of dendritic cell (DC) maturation in the induction of antigen-specific immune responses, we investigated the effect of LTX-315 treatment on the maturation of tumor-infiltrating DCs (TiDCs) and the generation of anti-melanoma immunity. The results reveal that LTX-315 induces the maturation of DCs through three distinct mechanisms: 1) release of DAMPs/alarmins; 2) complexes of LTX-315 with nucleic acids that act immunostimulatory; and 3) direct induction of the maturation of both conventional and plasmacytoid DCs by triggering TLR7. Importantly, LTX-315-induced maturation of DCs in vitro and in vivo was dependent on the presence of the signal transducer MyD88 as well as distinct TLRs. Our data also substantiate the notion that reagents like LTX-315 with both tumor cell cytotoxic and immunostimulaing capacities are more effective immunotherapeutics.
Reprogramming leukemia cells into antigen presenting cells as a novel cancer vaccination immunotherapy
Authors: Linde Miles H , Dove Christopher G , Gurev Sarah F , Phan Paul , Zhao Feifei , Gars Eric J , Marshall Payton L , Miller Lindsay P , Majeti Ravindra 
Published: January 2023

Abstract
B-cell acute lymphoblastic leukemia (B-ALL) is an aggressive hematopoietic neoplasm characterized by recurrent genetic lesions resulting in malignant transformation. Despite an arrest in B cell maturation, human B-ALL blasts retain the capacity for reprogramming to the myeloid lineage. To test the therapeutic implications of reprogramming, we generated murine models of B-ALL capable of myeloid lineage reprogramming via ectopic expression of CEBPα and PU.1. Once reprogrammed, B-ALL cells acquired an APC phenotype and stimulated antigen-specific T cells. In vivo B-ALL reprogramming in immunodeficient mice led to a modest survival benefit, however, reprogramming in immunocompetent mice led to tumor eradication and protection from subsequent re-challenge, demonstrating successful generation of durable, systemic, and curative immunity. This therapeutic benefit is dependent on the presence of both CD4+ and CD8+ T cells, and characterized by an increased frequency of bone marrow-infiltrating T cells and oligoclonal T cell expansion. Moreover, local myeloid reprogramming of a primary tumor drove systemic immune activation and eradication of distant, non-reprogrammed lesions. Our data suggests that B-ALL cells reprogrammed to the myeloid lineage are potent APCs capable of presenting tumor-associated antigens, and in vivo reprogramming elicits robust, durable, tumor-eradicating, and systemic T cell-mediated immunity. Recent efforts have further identified sarcoma and carcinoma models amenable to myeloid-lineage reprogramming. Thus, reprogramming of malignant cells into APCs represents a novel immunotherapeutic strategy with potential clinical utility in the treatment of a broad array of human malignancies.
Evaluation of a live attenuated influenza A vaccine expressing an IgA-inducing peptide in swine
Authors: Brand Meghan J Wymore , Gauger Phillip , Ferreri Lucas , Cáceres C. Joaquín , Perez Daniel , Vincent Amy , Rajão Daniela 
Published: January 2023

Abstract
Influenza A (IAV) is a significant disease for both human and swine health, as well as a zoonotic concern. The pig is an ideal model for human disease and evaluation of novel IAV vaccines. The objective of this study was to evaluate a bivalent (H1N1 and H3N2) live attenuated influenza A vaccine (LAIV) containing the IgA-inducing peptide gene (IGIP) in swine. Pigs were vaccinated with IGIP-LAIV or LAIV, challenged with heterologous H1N1 or H3N2, and necropsied at 5 or 14 days post infection (DPI). Peripheral blood mononuclear cells (PBMCs) and cells isolated from bronchoalveolar lavage fluid (BALF) were evaluated by flow cytometry and assessed for IgA-secreting cells (IgA-SCs) by ELISPOT. Pigs vaccinated with IGIP-LAIV and LAIV had decreased lesion scores compared to non-vaccinated pigs after challenge with either virus, and there was a trend for decreased lesion scores in the IGIP-LAIV compared to LAIV group. At 0 DPI, PBMC CD21+B cells were increased in IGIP-LAIV pigs compared to LAIV and naïve pigs. There was no difference in circulating total IgA-secreting cells between any groups post-challenge, though at 14 days post boost both the IGIP-LAIV and LAIV groups had increased virus-specific IgA-SCs to both vaccine viruses. In BALF at 5 DPI, the IGIP-LAIV and LAIV had increased total IgA-SCs, as well as increased virus-specific IgA-secreting cells to both vaccine viruses regardless of challenge virus. In conclusion, IGIP-LAIV and LAIV protected from heterologous challenge and induced virus-specific IgA-SCs in blood and BALF. IGIP-LAIV increased circulating B cells and had a trend for improved protection compared to LAIV, though no difference was observed in IgA-SCs between vaccines. Thus, the IGIP-LAIV represents an alternative LAIV vaccine.
The transcriptional landscape of porcine peripheral blood immune cells
Authors: Herrera-Uribe Juber , Byrne Kristen A , Liu Haibo , Becker Sage , Loving Crystal L , Tuggle Christopher K 
Published: January 2023

Abstract
Blood consists of different immune cells with essential functions given by distinct molecular signatures including transcriptomic profiles. The porcine immune system shares many similarities with that in human; however, the porcine immune cell transcriptome has not been as comprehensively studied. To identify distinct transcriptomic profiles of major porcine peripheral blood immune cells, we employed magnetic separation and fluorescence activated cell sorting methods to isolate 9 different cell types from blood PBMCs from two healthy pigs, representing monocytes, neutrophils, NK cells and specific populations of T and B cells. We obtained deep individual cell type transcriptomes using RNA-seq, detecting up to 10,974 genes. Principal component analysis revealed appropriate clustering of replicate samples and the separation of T, B, NK and myeloid cell groups. Pairwise comparisons revealed significant expression of a large number of genes specific for monocytes, neutrophils and NK cells only, while enrichment analysis identified highly differentially expressed genes for all cell types. Gene Ontology term analysis showed high enrichment of biological processes related to the nature of each cell type (i.e., B cell antigen presentation enriched in B cell transcriptome). The gene enrichment and specific genes of porcine immune cells is being further validated in silico to human datasets using gene set enrichment analysis (GSEA), and in vitro using qRT-PCR analysis. Taken together, the gene expression profiles reported here is the first comprehensive transcriptomic study of circulating porcine immune cell types and provides a valuable resource to elucidate molecular markers for porcine immune cell identity and function.
Mast cell histamine mediates intestinal immune response to early life stress in piglets via histamine 1 receptor (H1R)
Authors: Thelen Kyan , Wilson Neco , Berrios-Vazquez Glorian , Moeser Adam J 
Published: January 2023

Abstract
Mast cells are important immune modulators of stress-related gastrointestinal (GI) disorders. Our previous studies investigating early life stress in piglets showed that histamine, a major mast cell granule mediator is released within minutes of early weaning stress and is followed by increased intestinal mucosal expression of histamine receptor subtypes. The precise contribution of histamine to stress-induced GI immune responses is unknown. The objective of this study was to test the hypothesis that stress-induced intestinal immune response is mediated through the histamine 1 receptor (H1R). Fifteen-day-old female Yorkshire piglets were administered either saline vehicle or the H1R antagonist, desloratadine (2 mg/kg; intramuscular), 30 minutes prior to early weaning stress. At weaning, piglets were weaned from their dams and housed in nursery pens with ad libitum access to water. At 24 hours post-weaning, mid-jejunum and mesenteric lymph nodes (MLN) were collected for subsequent qPCR gene expression as well as IHC localization and protein expression analyses. Unweaned control piglets remained with the sow and were collected immediately at weaning. Gene expression for H1R was not influenced by weaning; however, immunohistochemical analysis of H1R revealed increased expression and localization in lamina propria cells and myofibroblasts in jejunal villi and expression in the longitudinal and circular muscle layers in early weaned piglets. In the MLN, weaning increased the expression of TNFα, TGFβ, and β-Integrin. This response was reduced in the group treated with the H1R antagonist. Together, these data demonstrate that histamine via H1R plays an important role in early weaning stress-induced intestinal immune responses.
The Immune response of Susceptible or Resistant Bat Species to White Nose Fungus
Authors: Lupfer Christopher R , Anderson Briana N. , Naffa Keslie S. , Tomasi Tomas E. 
Published: January 2023

Abstract
North American bats are being decimated by White Nose Syndrome (WNS), a disease caused by the psychrophilic fungus Pseudogymnoascus destructans (P.d.). Bats are an important part of the ecosystem and have global economic impact through consuming insect pests or pollination of crops. Thus, we need to understand why some bat species are susceptible and some resistant to WNS to develop strategies to protect bats from this disease. However, WNS has decreased populations of some temperate bat species to levels where conducting research on live bats must be curtailed. We have generated transcriptomic profiles derived from primary wing fibroblast cell cultures of resistant and susceptible bat species that were exposed to P.d. Our data indicate that fibroblasts from susceptible species of bats have an enhanced immune response whereas the immune response in resistant species is muted. Importantly, these results were confirmed in samples collected from live bats prior to population declines. We have now developed qRT-PCR assays to test the activity of several immune genes that were upregulated in the cell-cultures (transcriptomics analysis). Generally, we found that wing tissue from tricolored bats (WNS-susceptible) late in hibernation showed increases in gene expression compared to pre-inoculation with Pd, but wing tissue from big brown bats (WNS-resistant) did not change (or decreased). These results, though preliminary, suggest that wing fibroblast cell-cultures may indeed be a good surrogate for using living bats in some kinds of WNS research. Our data also suggest that the cause for high mortality in WNS susceptible bat species is immune activation instead of immune tolerance.
Characterization of IL7R α in Xenopus tropicalis: implications for the evolution of cytokine signaling pathways
Authors: Appasamy Pierette M , Lambert Lisa A. , Sammons John C. , Daskalova Kristina G. 
Published: January 2023

Abstract
Signaling through the IL7 receptor is essential for development of B and T cells in humans and mice, as evidenced by the SCID-like phenotype seen in the absence of IL7/IL7 receptor signaling. The IL7R signaling system is active in lower vertebrates but has not previously been examined in Xenopus frogs. The IL7R is a heterodimer comprised of the IL7R α chain and the “common” γ chain that is shared by other IL2 family cytokines that utilize the JAK-STAT pathway. We have recently cloned the IL7R α cDNA from Xenopus tropicalis. Comparison of the predicted amino acid sequence of the Xenopus IL7R α indicates closest homology with the zebrafish IL7R α, and significant homology with other characterized IL7R α including those from humans, mice, chickens, alligators, primitive caecilians, green sea turtles, and anole lizards. Analysis of the predicted amino acid sequence indicates an intact WSXWS motif (common to all IL7R family members). The Box 1 motif, normally required for JAK interaction, has an asparagine replacing aspartic acid (similar variations are found in the zebrafish and anole lizard IL7R α sequences). There are at least five potential N glycosylation sites. There is a 16 amino acid insert unique to the Xenopus sequence that is not found in the reported IL7R α sequence of any other animal that we have examined. Our attempts to identify an IL-7 gene in the Xenopus genome have been unfruitful. Synteny analysis indicate there is no gene with homology to IL-7 at the expected position in the Xenopus genome. Interestingly, we have identified a transposon in place of the IL-7 gene at the expected position. The distribution of IL7R α in various organs of X. tropicalis is now being examined, and as well predictions of the 3D structure of the Xenopus IL7R α protein.