bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhaled IL-10 Suppresses Lung Tumorigenesis via Abrogation of the Inflammatory Macrophage - T17-cell Axis
Authors: Li Qingsheng , Egilmez Nejat K. 
Published: January 2023

Abstract
Lung tumorigenesis has been linked to chronic type 17 inflammation in both murine models and patients. Interleukin-10 is a pleiotropic regulatory cytokine with profound inhibitory effects on type 17 responses. Intratracheal administration of a sustained-release IL-10 formulation resulted in suppression of tumor development in the LSL-K-ras spontaneous lung cancer model. Analysis of lung lymphocyte populations demonstrated that the antitumor activity of IL-10 was associated with a rapid 3 to 5-fold decline in IL-17-producing T-cell populations. Type 17 T-cells did not express detectable levels of IL-10RA suggesting that IL-10 was not directly acting on T-cells. In contrast, select myeloid cell populations including macrophages and a small subset of dendritic cells were found to express IL-10RA in the lung. In tumor-bearing control mice macrophages displayed a pro-inflammatory phenotype that reverted to a naive-like state following IL-10 treatment. Importantly, depletion of phagocytes resulted in a 3 to 5-fold decline in type 17 T-cell numbers with a concurrent 7-fold reduction in tumor burden. These findings demonstrate that in the LSL-K-ras model lung tumorigenesis is driven by the inflammatory macrophage - type 17 T-cell axis and that IL-10 abrogates this process via its direct activity on macrophages. Inhaled IL-10 formulations may be of utility in prophylaxis of lung cancer in high-risk patients.
Population dynamics of tumor-specific CD8 T cell differentiation from plastic to fixed dysfunctional states
Authors: Philip Mary , Pilkinton Mark , Ramesh Ram , McDonnell Wyatt J. , Gangula Rama D. , Camara Steven , Chopra Abha , Schietinger Andrea , Mallal Simon Alexander 
Published: January 2023

Abstract
Tumor-specific CD8 T cells (TST) encountering cognate antigen in tumors differentiate to a dysfunctional state characterized by expression of multiple inhibitory receptors and failure to make effector cytokines. We demonstrated that TST differentiate through two discrete chromatin states; TST were initially in a plastic chromatin state and could be functionally rescued but then transitioned to a fixed chromatin state resistant to therapeutic reprogramming. T cells harboring these discrete chromatin states could be identified with distinct, novel membrane protein expression profiles, permitting prospective identification of reprogrammable tumor-specific T cells from bulk tumor-infiltrating T cell populations. Importantly, human tumor-infiltrating PD1-high CD8 T cells were in a similar chromatin accessibility state as murine fixed dysfunctional T cells. Based on our studies, two models could describe TST differentiation: (i) TST progress en-masse from the plastic to fixed state, potentially through a transitional state or (ii) individual TST within a population exist in either the plastic or fixed dysfunctional state; initially, most TST are in the plastic state, however over time plastic T cells fail to proliferate or die while the fixed dysfunctional TST preferentially proliferate and/or survive. To test these models, we carried out single-cell RNA-Sequencing on TST at various time points during tumor progression. Determining the population dynamics of TST differentiation through dysfunctional states in tumors could inform our strategies for TST rescue, for example, by selecting rare plastic TST from bulk populations versus novel strategies to reprogram the epigenome of fixed dysfunctional TST.
Blockade of CD25 reduces regulatory T cell expansion and some manifestations of IL-2 mediated toxicity but does not improve anti-tumor efficacy in animal models of IL-2 immunotherapy
Authors: Harding Fiona A. , Alvarez-Jares Hamsell , Belmar Nicole , Choi Donghee , Forsyth Charles , Fox Melvin , Sheridan James , Stickler Marcia , Tang Anthony , Hollenbaugh Diane 
Published: January 2023

Abstract
High dose IL-2 (HD IL-2) is an approved treatment for melanoma and RCC that can result in long term disease remission, but is associated with severe adverse side effects. Adverse effects may be mediated by non-specific activation of cells expressing the high affinity CD25-containing IL-2 receptor. HD IL-2 also results in the expansion of regulatory T cells (Treg) that may negatively impact tumor immunotherapy. Blocking IL-2 binding to CD25 was explored as a mechanism to reduce expansion of Tregs and adverse effects. We created a high affinity anti-mouse CD25 mAb that effectively blocks the interaction of IL-2 with CD25 and carries the N297A Fc region modification to reduce FcgR binding. Saturation of CD25 ameliorates some but not all biomarkers of IL-2 toxicity. These include behavioral signs and serum ALT elevation. However, lung and spleen weight increase induced by IL-2 was not affected by CD25 blockade. CD25 blockade reduced IL-2 mediated expansion of Tregs. A saturating dose of CD25 antibody prior to 5-6 days of IL-2 therapy resulted in a complete loss of IL-2-mediated tumor efficacy in the CT26 model. CD25 blockade and IL-2 expanded CD8+ and NK cells while reducing blood and TIL Tregs, but did not impact primary tumor growth in the 4T1 or B16F10 tumor models. The loss of anti-tumor activity in the CT26 model correlated with reduced CD8+ CD25+ expansion in the blood and the TIL populations. Expansion of CD25 negative CD8+ and NK cells with concomitant Treg cell suppression in three different syngeneic tumor models did not result in improved anti-tumor efficacy. This result has implications for IL-2 and IL-15 engineering, and suggests that IL-2 mediated toxicity cannot be completely controlled by re-directing IL-2 to the CD122/CD132 receptor.
Prediction of hosts who are more likely to respond to PD-1 inhibitors/blocking antibodies
Authors: Daftarian Pirouz M. , George Marybeth , Tleugabulova Mayra Cruz , Magcase Dennie , Ouerkaxi Wushouer , Delcommenne Marc C. 
Published: January 2023

Abstract
A major challenge in the development of immune checkpoint blockades (ICB) is the variability in outcome of such therapeutics. This leads to a large number of non-responder patients receiving costly therapies that are also associated with some toxicities. Surrogates of efficacy are needed both to select the right patient population for the clinical trials and in later stages, to identify the right patients for the right ICB, ideally with assays that are rapid, high throughput and noninvasive. T cell clonal expansions caused by ICB are mainly local. As a result, the expanded T cells are mostly found in the tumor deposit and hard to detect in the peripheral blood. We optimized an antigen recall assay that assesses the ability of PD-1 blocking antibody to further enhance antigen specific CD8 T cell expansion and thereby discriminates responders from non-responders. A simple version of this assay has been used for functional screening of ICBs. By improving each component of the assay and its readout, we have increased the “signal to noise” window allowing us to rapidly detect PD-1 blockades’ effects on the host. We use a cocktail of optimized recall peptides, a characterized conditioned media and 96-well plates to support rapid (7 days) expansion of memory T cells. Finally, as a readout system, cocktail of MHC class I tetramers generated by peptide exchange was used to detect the overall expanded clonotypes. Currently, we have optimized the assay with commercially available anti-PD-1 and anti-PD-L1 antibodies. This assay system has the potential to assist in predicting those who are more likely to respond to ICB therapies. We have initiated blinded studies using clinical samples for clinical validation.
Changes of immune profile following nanosecond electric pulse treatment for pancreatic cancer
Authors: Guo Siqi , Burcus Niculina I. , Hornef James , Jing Yu , Jiang Chunqi , Heller Richard , Beebe Stephen J. 
Published: January 2023

Abstract
Pancreatic cancer is a notorious malignancy with a very poor prognosis. Without significant advance in the treatment and early diagnosis, pancreatic cancer will continue to be a serious global health issue. Nanosecond electric pulses (nsEPs) or nanosecond pulsed electric fields (nsPEFs) have been demonstrated to result in completely local ablation and protection from the second tumor challenge in several cancer models. Here, we assessed the potential advantages of nsEPs for pancreatic cancer in a syngeneic mouse Pan02 pancreatic cancer model and characterized its immune profile. We found that a single nsEP treatment could achieve complete tumor regression without any mortality. Animals with partial regression prolonged survival in those treated with nsEPs more than those treated with irreversible electroporation (IRE). Low rate of abscopal effect (10%) and vaccine-like protective effect (1/15) was observed in animals treated with nsEPs but not with surgery or IRE. Significant reduction of both T regulatory cells and myeloid derived suppressor cells (MDSCs) was seen in the blood of tumor animals after nsEP treatment. Heterogeneous T cell responses following nsEP treatment were found. Very few T cells were infiltrated in the tumor but large amount of MDSCs existed in tumor. The number of dendritic cells in the tumor was increased and activated with upregulation of multiple activation markers following nsEP treatment. Our findings could help design a more effective combinational nsEP therapeutic strategy for pancreatic cancer.
Tissue specific CD91+ APCs determine effectiveness of immunosurveillance mediated by HSPs
Authors: Sedlacek Abigail L. , Kinner Lauren B. , Wang Yifei , Mizes Alicia P. , Binder Robert J. 
Published: January 2023

Abstract
Gp96, an immunogenic Heat Shock Protein, is capable of eliciting potent immune responses against its chaperoned antigens and is being investigated as a potential cancer immunotherapy. Gp96 induces immune responses by activating antigen presenting cells (APCs) through CD91 which elicits downstream T and NK cell activation. T cell activation following gp96 is well understood, including the necessary APCs. Conversely, NK cell activation is less understood, even though NK cells are required for full T cell activation. We have determined that macrophages (MΦ) are capable of inducing IFNγ production by NK cells but, inversely, dendritic cells (DC) are not, despite similar expression of CD91. Thus, we investigated MΦ and DC production of soluble mediators of NK cell activation. IP10 was identified as a chemokine which is more greatly induced in MΦ than DC in response to gp96. Blocking the IP10 receptor, CXCR3, prevents IFNγ production by NK cells when cultured with gp96 activated MΦ. IP10 is STAT1 induced, thus we examined STAT1 phosphorylation (STAT1p) in MΦ and DC in response to gp96. STAT1 is phosphorylated in MΦ, but gp96 fails to induce STAT1p of tyrosine in DC. In STAT1 signaling, p38 is proximally upstream of STAT1p and interestingly, both MΦ and DC phosphorylate p38 to similar levels, suggesting that the difference between MΦ and DC is downstream of p38. There are several phosphatases downstream of CD91 which could be inhibiting STAT1p. To further support this, we inhibited tyrosine phosphatases using pervanadate, resulting in DC STAT1p in response to gp96. Collectively, these data indicate that gp96 activation of MΦ may result in a more robust immune response by activating NK cells which is a critical consideration for clinical applications.
The prevalent Boxer MHC class Ia allele Dog Leukocyte Antigen (DLA)-88*034:01 presents 9-mer peptides with a defined binding motif
Authors: Hess Paul R. , Nemec Paige S. , Kapatos Alexander , Holmes Jennifer C. 
Published: January 2023

Abstract
Adoptive cell transfer (ACT) for chemoresistant malignancies can induce dramatic remissions and prolong life, but factors that determine T-cell efficacy and off-target/-tumor events associated with therapy remain incompletely understood. Modeling ACT in spontaneous canine cancer models could provide critical insights and propel development; however, relevant peptides from tumor-specific antigens (TSAs) have yet to be reported in dogs. Discovery of TSA epitopes restricted by a high-prevalence MHC allele can be facilitated by in silico analysis using a defined binding motif and a large peptide dataset to train prediction software. We sought to characterize the binding preferences of the DLA-88*034:01 allele, carried by 82% of Boxers, an inbred, popular breed with increased risks for glioma and lymphoma, important ACT targets. We hypothesized that DLA-88*034:01 presents diverse self-peptides with conserved residue preferences at ≥1 position(s) that can be inferred by pattern analysis. To test this prediction, DLA-88*034:01-FLAG was immunoprecipitated from large-scale cultures of a canine cell clone, PN62, and alkaline-eluted peptides were sequenced by liquid chromatography-tandem mass spectrometry. DLA-88*034:01 prefers peptides that are 9 amino acids (AAs) in length. Analysis of 640 9-mers showed a preference for acidic AAs at position(P)1, non-bulky hydrophobic AAs at P2, and basic AAs at P4. At P6 and P9, H and F were highly favored, respectively. Bulky hydrophobic and charged AAs were unfavorable at P2, as were W, M and Y at P8. This dataset will allow more accurate prediction of TSA-derived peptides recognized by CTL in a majority of Boxers with glioma or lymphoma, and should foster development of an ACT model in this breed.
PD-1/PD-L1 Monoclonal Antibody Development for Canine Cancer Therapy
Authors: Choi Jin Wook , Withers Sita S. , Sciammas Roger , Rebhun Robert B. , McSorley Stephen J. 
Published: January 2023

Abstract
PD-1 is an inhibitory receptor that binds to PD-L1 on the surface of tumor and antigen presenting cells in the tumor microenvironment, causing suppression of T-cell activity and tumor progression. Human clinical trials with anti PD-1 monoclonal antibodies, nivolumab and pembrolizumab, showed an objective response rate (ORR) of 30% to 40% with melanoma and 87% with relapsed or refractory Hodgkin’s lymphoma. Here, we have explored the development of canine PD-1/PD-L1 reagents for potential veterinary applications. An S2 insect cell expression system was utilized to produce recombinant canine PD-1Ig and PD-L1Ig and antigen-specific B cells were enriched using PD-1 and PD-L1 tetramers. Enriched B cells were fused and resulting hybridomas were screened for specific reactivity to PD-1 and PD-L1. Then, antibodies capable of blocking PD-1Ig and PD-L1Ig were selected. A total of 5 monoclonal antibodies were generated from the two rounds of immunizations and are being studied for in vitro activity in T cell assays. These canine reagents have multiple diagnostic and therapeutic applications in the context of canine oncology.
Porcine macrophage-like cells permit viral replication, produce inflammatory mediators, and undergo apoptosis following infection with Rift Valley fever virus MP-12
Authors: Noronha Leela E. , Smolensky Dmitriy , Cox Victoria , Schirtzinger Erin E. , Chitko-McKown Carol G. , Fawver Zachary , Wilson William C. 
Published: January 2023

Abstract
Rift Valley fever virus (RVFV) is a mosquito-borne virus that can cause severe disease in a variety of domestic animal species as well as in humans. Outbreaks occur primarily in Sub-Saharan Africa with ruminant livestock most affected and outcomes that include abortion storms and high mortality among young animals. Clinical disease is not known to occur in domestic pigs (Sus scrofa); however, a limited number of studies have detected circulating antibodies to RVFV among wild and domestic members of the pig family (Suidae). Experimentally, RVFV has been shown to replicate in some porcine epithelial and neuronal cells in vitro, and induce a viremia in a fraction of pigs inoculated in vivo. No studies to date have investigated the behavior of RVFV in porcine cells of immune origin, whereas in humans and mice, macrophages and dendritic cells have been implicated as having roles in RVFV pathogenicity. Here, we use a porcine monocyte-derived macrophage-like cell line, CΔ2+, to investigate porcine innate immune cell permissiveness to MP-12, an attenuated form of RVFV. MP-12 caused observable cytopathic effects in CΔ2+ cells and reduced cell viability in dose- and time-dependent manners. At 24 hours, the reduced viability was coincident with apoptosis, with necrosis also occurring at a higher multiplicity of infection. At early time points, MP-12 replicated at levels comparable to cells derived from sheep kidney—a target organ of a highly susceptible host species. Infected CΔ2+ cells also upregulated transcription of inflammatory mediators. This work provides the first evidence that porcine immune cells are permissive to RVFV, and indicates that additional studies are warranted to better understand the dynamics of RVFV in suids.
Leukocyte immune-type receptor expressing cells increase in response to Edwardsiella ictaluri infection in Channel Catfish, Ictalurus punctatus
Authors: Blackmon Laura Elizabeth , Quiniou Sylvie , Wilson Melanie , Bengten Eva 
Published: January 2023

Abstract
Unique to teleosts, leukocyte immune-type receptors (LITRs) represent a large polygenic, polymorphic family of immunoregulatory receptors. Monoclonal antibodies (mAb), CC34 and CC41, were developed against a catfish clonal CTL line and a catfish NK cell line, respectively. Both recognize members of the LITR family. One subset of LITRs, defined by reactivity with mAb CC41, is a distinct marker of a cytotoxic cell population, which expands during the antiviral response to Channel Catfish Virus (CCV). It is hypothesized that these mAb CC41-reactive LITRs function as NK-like receptors and recognize stress induced ligands on target cells. Edwardsiella ictaluri, an intracellular pathogen, primarily establishes infection in pronephros (head kidney) monocytes and may elicit a strong cytotoxic response. To explore this, small fish (&lt;1kg) were infected with E. ictaluri via bath immersion (1×107cfu/ml). Following infection, whole blood was collected and analyzed daily by flow cytometry for mAb CC34 and CC41 staining. In parallel, an in vitro model was developed to monitor the number of CC34 and CC41-reactive cells in response to E. ictaluri infection. Briefly, head kidney monocytes and peripheral blood lymphocytes (PBL) were isolated from catfish of either susceptible or resistant background. Head kidney cells were infected with E. ictaluri (MOI 10) and then mixed with PBL from the same individual catfish and analyzed daily by flow cytometry. There was an increase in staining with both mAbs CC34 and CC41 by day 3 after infection both in vivo and in vitro. This study highlights the utilization of these mAb reagents to monitor cytotoxic cell response to E. ictaluri infection.
Priming of CD8 T cells in established chronic viral infection directs a distinct differentiation and functional program for long-term immunity
Authors: Snell Laura M. , Brooks David G. 
Published: January 2023

Abstract
CD8 T cell exhaustion impedes control of chronic virus infection, yet how CD8 T cell fate decisions and function are instilled and progressively controlled throughout infection is unclear. We demonstrate that in contrast to T cells primed at the onset of persistent viral infection, CD8 T cells primed after the chronic infection is established preferentially generate into memory-like cells that resist contraction, are less exhausted and have heightened responsiveness to immunotherapy. Mechanistically, viral persistence down-regulates APC activation machinery, decreasing T cell intrinsic TCR and CD28 signaling upon priming. The lower signal intensity provides the required threshold for activation and proliferation, but alters transcriptional programming and cellular differentiation to enable long-term retention of antiviral functions despite high levels of virus replication. In the absence of strong antigenic signaling, IL-21 acquires the ability to induce functional effectors, while preserving the memory-like reservoir. Analogous mechanisms also govern differentiation of anti-tumor CD8 T cells in the tumor microenvironment during chronic viral infection, undermining tumor control. Thus, dynamic changes in signal intensity and IL-21 usage during chronic infection function as fate-assigning rheostats to resist exhaustion and program long-term immunity.
MHC class II antigen-processing chaperone H2-O shapes CD4 T cell receptor repertoire
Authors: Song Nianbin , Welsh Robin A. , Boronina Tatiana , Cole Robert , Foss Catherine , Sadegh-Nasseri Scheherazade 
Published: January 2023

Abstract
HLA-DO (DO) is a MHC class II-encoded, nonpolymorphic heterodimeric protein in humans (known as H2-O in mice). It is expressed only in thymic medulla, B cells and some subsets of dendritic cells. Although DO is found to form a stable complex with HLA—DM, its physical function remains unknown. To solve this mystery, our lab previously proposed a model suggesting that the DO facilitates the binding of DM-resistant peptides and inhibits the binding of DM-sensitive peptides to the HLA-DR1 molecules in vitro. Based on this model, we expected to see lower density of pMHC expression on mTEC cells in thymus of DO-KO mice, leading to faulty negative selection and different CD4 TCR repertoires as compared to DO-WT mice. By developing a “Prime-Restim” strategy based on the principle of mixed lymphocyte reaction, we observed the proliferation of primed DO-WT CD4 T cells in responding to repeated in vitro stimulations by antigen presenting cells from DO-KO mice and vice versa. This strategy led to enrichment of some clonal populations in DO-KO mice reactive to self-peptides presented by B cells from DO-WT mice. To identify the self-antigens these T cell clones recognize, we developed CD4 T cell hybridomas from them and screened their specificities with peptides eluted from B cells of DO-KO and DO-WT mice. We also identified a peptide from an autoimmunity-associated protein that eluted only from DO-KO B cells but not seen from DO-WT B cells. These results demonstrate that in the absence of DO, the peptide repertoire of MHC II in the DO-KO mice is different from the peptide repertoire of MHC II in the DO-WT mice. These findings support the idea that the expression of DO in thymic medulla might lead to better thymic deletion of self-reactive CD4 T cells.