bluuebunny/crossref_metadata_2025_split huggingface dataset:
Combination CpG and MPL agonists heighten vaccine-induced protection against Influenza A virus challenge
Authors: Lampe Anna T , Hain Jacey L , Brown Deborah M 
Published: January 2023

Abstract
Current strategies for Influenza A Virus (IAV) vaccination result in antibodies against strain specific viral coat proteins. These antibodies do not provide protection against heterologous IAV strains and leave the population vulnerable to pandemics of highly pathogenic IAV. Vaccines may be improved by targeting T cells, which unlike B cells, are able to recognize inner, conserved IAV antigens, such as nucleoprotein (NP), increasing the potential for protection against different IAV stains. During live IAV infection multiple pattern recognition receptors (PRRs) on innate immune cells are activated; therefore vaccines triggering multiple PRRs may elicit better protection. As dendritic cells are the most potent antigen presenting cells for T cells, bone marrow derived dendritic cells (BMDCs) were activated by combined agonists that trigger separate, yet complementary PRR signaling pathways. Here we show that combined PRR agonists MPL and CpG, which activate TLR4 and TLR9 respectively, increased in vitro cytokine and chemokine expression by BMDCs. Further, intramuscular vaccination of mice with IAV pdm09 H1N1 hemagglutinin (HA) protein plus combined CpG and MPL, as adjuvants, induced the formation of pdm09 HA specific IgG2c antibodies. Combined CpG and MPL recipient mice experienced less weight loss and lower mortality after lethal challenge with pdm09 H1N1 compared to mice receiving lone agonists, suggesting that combined agonists improve protection. T cell activation in response to combined CpG and MPL is currently being investigated, as well as responses generated via mucosal, intranasal vaccination. Thus combined agonists that promote B and T cell mediated immunity could be incorporated into next generation IAV vaccines.
Biodegradable nanoparticle delivery of inactivated swine influenza virus vaccine provides heterologous cell-mediated immune response in pigs
Authors: Dhakal Santosh , Hiremath Jagadish , Bondra Kathryn , Lakshmanappa Yashavanth Shaan , Shyu Duan-Liang , Ouyang Kang , Kang Kyung-il , Binjawadagi Basavaraj , Goodman Jonathan , Tabynov Kairat , Krakowka Steven , Narasimhan Balaji , Lee Chang Won , Renukaradhya Gourapura J 
Published: January 2023

Abstract
Swine influenza virus (SwIV) causes considerable economic loss to pig industry, and some SwIV are zoonotic. This study was conducted to evaluate the cross-protective efficacy of PLGA (poly lactic-co-glycolic acid) nanoparticles (NPs) encapsulated SwIV vaccine in pigs. Killed SwIV H1N2 (δ lineage) antigens (KAg) were encapsulated in PLGA NPs of 200–300 nm (PLGA-KAg NPs), and influenza antibody-free pigs were prime-boost vaccinated intranasally as mist and challenged using a heterologous, virulent and zoonotic SwIV H1N1 (γ lineage). PLGA-KAg NPs induced maturation of pig macrophages and dendritic cells in vitro. In vaccinated pigs, PLGA-KAg NPs induced antigen specific lymphocyte proliferation and enhanced the frequency of T-helper/memory cells and cytotoxic T cells in peripheral blood mononuclear cells (PBMCs). In virus challenged pigs, the PLGA-KAg NPs vaccine rescued virus induced clinical fever, reduced the gross lung pathology, reduced the virus load in the lung sections with complete clearance of the virus from the lungs of most of the pigs; but the nasal virus shedding was not reduced. Immunologically, at post-challenge day 6 in a recall response in PBMCs of PLGA KAg NPs vaccinated pigs, a significant increase in IFN-γ secreting T cells against both vaccine and challenge viruses were detected. However, humoral immune response in those pigs was not augmented. In conclusion, intranasal delivery of PLGA NPs based SwIV induced cross-protective response through specific cell-mediated response. Future studies are aimed at boosting the mucosal antibody response.
Protein kinase D1 is essential for Saccharopolyspora rectivirgula-induced pulmonary inflammation and alveolitis.
Authors: Yoon Taewon , Kim Young-In , Fitzpatrick Elizabeth , Yi Ae-Kyung 
Published: January 2023

Abstract
Saccharopolyspora rectivirgula (SR) causes farmers’ lung disease, a form of hypersensitivity pneumonitis (HP). The initial proinflammatory responses to SR is dependent on Toll-like receptors (TLR) 2 and 9, and their signaling adaptor MyD88. We found previously that protein kinase D1 (PKD1), a serine/threonine kinase, can be activated through a MyD88-dependent TLR signaling pathway and plays a pivotal role in TLR/MyD88-dependent production of proinflammatory mediators. SR activates PKD1 via a MyD88-dependent manner in murine cells and SR-induced alveolitis is substantially suppressed by pretreatment with a PKC/PKD inhibitor Gö6976. However, it is currently unknown whether PKD1 plays a regulatory role in SR-mediated proinflammatory responses and alveolitis in vivo. In the present study, to further understand contribution of PKD1 in the SR-induced pulmonary inflammation, tamoxifen-inducible PKD1-deficient mice were generated. Wild-type mice and PKD1-deficient mice were challenged intranasally with SR. We found that the levels of proinflammatory mediators (including TNFα, IL-6, IL-12, and IFNγ), alveolitis, and neutrophil influx in the bronchoalveolar lavage fluid and interstitial lung tissue after single intranasal expose to SR were significantly lower in PKD1-deficient mice compared to those in wild-type mice. Our results demonstrate that PKD1 is essential for SR-mediated proinflammatory responses and neutrophil influx in the lung. Our findings also suggest that PKD1 might be one of the critical factors that contributes to the development of HP, and could be a therapeutic target for certain forms of HP, such as farmers’ lung disease.
Differential nutrient uptake by lung resident and spleen effector memory T cells following influenza virus infection
Authors: Cartwright Emily Kathleen , Kohlmeier Jacob E 
Published: January 2023

Abstract
Pulmonary virus infection generates a distinct population of CD8 T cells that reside within the lungs following antigen clearance. Resident memory T cells (TRM) are critical mediators of host defense, however, their ability to provide protection wanes over time. The protective immunity provided directly corresponds to the number of antigen-specific CD8+ TRM within the lungs at the time of secondary infection. Mechanisms underlying the differential loss of CD8 lung TRM remain poorly understood. Recent work has highlighted the importance of cellular metabolism in relation to cellular function and survival, but little is known about the metabolic phenotype of lung TRM following influenza infection. We investigated whether metabolic differences could explain the differential loss of lung TRM compared to circulating TEM. Using fluorescently labeled glucose and palmitate, we compared the nutrient uptake capacity of nucleoprotein (NP)-specific CD8 T cells in bronchealveolar lavage (BAL), mediastinal lymph node (mLN), spleen and lung TRM in vivo following influenza infection. We find lung TRM have differential capacity for nutrient uptake compared to NP-specific cells in other tissues. In addition, we utilized the Seahorse XF analyzer to measure oxygen consumption rate (OCR) and determine the spare respiratory capacity (SRC) of lung TRM compared to circulating TCM and TEM. We find that lung TRM tend to have lower OCR at baseline, as well as decreased SRC. Taken together, these data show that lung TRM have a different metabolic profile than circulating memory T cells. This work highlights the need to understand the differential metabolic requirements of TRM, which could facilitate future vaccine design strategies aimed at these pathways.
Glucagon-like peptide-1 receptor signaling attenuates RSV-induced type 2 responses and immunopathology
Authors: Bloodworth Melissa Harintho , Zhang Jian , Hotard Anne L. , Moore Martin L. , Hartert Tina V. , Niswender Kevin D. , Peebles R. Stokes 
Published: January 2023

Abstract
Glucagon-like peptide-1 receptor (GLP-1R) agonists are a well-accepted and safe treatment for Type II diabetes. Although GLP-1R agonists mainly act to potentiate insulin and suppress glucagon secretion, recent evidence suggests that GLP-1R signaling also has anti-inflammatory effects. Severe RSV-associated illness is partially caused by type 2-associated immunopathology. We hypothesized that GLP-1R signaling inhibits type 2-mediated immunopathology during infection with RSV 12/12-6, a strain of RSV that was isolated from a hospitalized infant with severe lower respiratory tract infection and bronchiolitis. We show here that GLP-1R agonist treatment decreased airway inflammation, airway reactivity, and airway mucus production in RSV 12/12-6-infected mice. GLP-1R agonist treatment decreased total lung IL-13 and IL-33 levels, with concurrent decreases in lung IL-13-producing group 2 innate lymphoid cell (ILC2), CD4+ Th2 cell, and basophil numbers as well as IL-33-producing epithelial cells. The GLP-1R agonist prevented airway inflammation, and did not impact viral load, anti-viral interferon and antibody production during secondary RSV infection. Relative to the respective mock-infected groups, RSV-infected GLP-1R agonist treated mice did not have increased weight loss compared to vehicle treated mice. A phenome-wide association study (PheWAS) identified a link between GLP-1R signaling and acute bronchitis and bronchiolitis in humans. These studies demonstrate that GLP-1R signaling protects against type 2-mediated immunopathology during RSV infection. GLP-1R agonists are the first known FDA-approved agents to inhibit IL-33 and may represent a novel treatment strategy for RSV bronchiolitis.
Induction of arginase-1 in MDSC requires exposure to CD3/CD28 activated T cells
Authors: Abdelaal Ahmed Mansour , Bian Zhen , Culpepper Courtney , Liu Yuan 
Published: January 2023

Abstract
Tumor-induced myeloid derived suppressive cells (MDSC) have been reported to inhibit T cell responses. In our study MDSC inhibit T cell proliferation through arginase-1 dependent manner. However, surprisingly we observe that freshly isolated MDSC from the bone marrow of tumor bearing mice do not constitutively express arginase-1 until after exposure to T cell proliferation system. The aim of this study is to determine the mechanism by which arginase-1 is induced in MDSC following exposure to a proliferative T cell environment. We show that treatment of MDSC with culture supernatant isolated from T cells activated with CD3/CD28 antibodies successfully induced arginase-1 expression and this process is independent of IL-10 and IFNγ. This suggest that arginase-1 induction in MDSC can occur independently of cell-cell contact. Interestingly IL-2, Con A or PMA activated T-cell supernatant as well as supernatant from multiple cancer cell lines failed to induce arginase-1 in MDSC. We also show that M-MDSC express higher levels of arginase-1 than G-MDSC after co-culture with CD3/CD28 activated T cells as well as its supernatant. In addition, other bone marrow cells have shown potential to express arginase-1 following exposure to the same conditions. For example, we observe that healthy Ly6C+ monocytes but not mature granulocytes successfully express arginase-1. These data demonstrate that T cells activated through stimulation of TCR elicit the immunosuppressive function of MDSC through induction of arginase-1.
Immune tolerance to recombinant immunotoxin LMB-100 using synthetic vaccine particles encapsulating rapamycin
Authors: Mazor Ronit , King Emily , Kishimoto Takashi Kei , Pastan Ira 
Published: January 2023

Abstract
Protein and cell based therapeutics have great potential to treat many human diseases. However, they often elicit an immune response that blocks their efficacy. Synthetic Vaccine Particles containing Rapamycin (SVP-R) is a novel approach to mitigate immunogenicity and induce specific immune tolerance.
We evaluated the efficacy of SVP-R to reduce anti-drug antibody (ADA) formation to a recombinant immunotoxin, LMB-100, a genetically engineered protein designed for cancer therapy. Due to the bacterial origin of the toxin, LMB-100 is highly foreign to the human immune system. Almost all patients treated with LMB-100 made ADAs after two cycles (6 doses) of treatment that neutralized the drug and greatly lowered efficacy.
Using an immune-competent mouse model we found that combining SVP-R with LMB-100 dramatically decreased ADA titers by 98%. The decrease persisted for more than six weeks despite additional challenges with LMB-100, indicating development of immunological tolerance to LMB-100. Transfer of splenocytes from tolerized mice to naïve mice, and challenge with LMB-100 resulted in a 66% decrease in titer, indicating that the immune tolerance is transferable.
SVP-R are being evaluated in humans to prevent ADA to Pegsiticase, an enzyme for refractory gout treatment. Our data indicates that combining SVP-R with LMB-100 should be useful in treating cancer by allowing more treatment cycles and better efficacy. This approach can be used to increase the efficacy of other immunogenic agents such as CAR-T cells, antibody drug conjugates and viral gene therapy vectors.
Phosphorylation of Y248 in the ITSM of PD-1 is indicative of PD-1-mediated inhibitory function
Authors: Bardhan Kankana , Weaver Jessica D , strauss laura , Patsoukis Nikolaos , Boussiotis Vassiliki A 
Published: January 2023

Abstract
PD-1 is an inhibitory receptor expressed on activated T-cells. SHP-2 tyrosine phosphatase interacts with PD-1 and is critical for PD-1-mediated inhibition. The cytoplasmic tail of PD-1 contains an immunoreceptor tyrosine based inhibitory motif (ITIM) and an immunoreceptor tyrosine-based switch motif (ITSM). It has been reported that SHP-2 may interact with either ITIM or ITSM of PD-1. We sought to identify the precise motif of PD-1 that is mandatory for SHP-2 interaction and PD-1 inhibitory function. Using GST-SHP-2 fusion protein we determined that PD-1 interacted with SHP-2 after PD-1 ligation with simultaneous TCR/CD3-mediated activation. This interaction required phosphorylation of the ITSM and was abrogated when the ITSM tyrosine Y248 was mutated to phenylalanine. In contrast, when the ITIM tyrosine Y223 was mutated, interaction of PD-1 with SHP-2 remained unaffected. Based on these findings, we hypothesized that phosphorylation of Y248 might be indicative of PD-1-mediated inhibitory signaling. We generated an antibody specific for Y248 and examined expression and function of PD-1pY248+ T cells in peripheral blood from healthy humans. PD-1pY248+ cells were detected within the CD4 but mostly CD8T cell populations, mainly in central memory and effector memory subsets and in much lower extent in terminally differentiated effectors. Although PD-1high did not correlate with altered ability of CD8 T cells to produce effector cytokines, PD-1pY248+ expression correlated with impaired production of IFN-g and TNFa in response to TCR/CD3+CD28-mediated stimulation. Thus, PD-1pY248 might serve as a biomarker indicative of PD-1 mediated inhibitory signaling in patients with chronic infections and cancer.
Multimeric Soluble 4-1BBL as a T Cell Stimulator for Adoptive Immunotherapy.
Authors: Kornbluth Richard S , Hamilton Victoria Snarsky , Stone Geoffrey W 
Published: January 2023

Abstract
Members of the TNF SuperFamily of ligands (TNFSFs) have significant potential as immuno-oncology agents. The TNFSFs are trimeric membrane proteins that can be cleaved into soluble single trimers. While the soluble single trimers can be easily prepared and studied, they have little or no activity in vivo. This deficiency is caused by the need to cluster their cognate receptors in the plane of the membrane in order to induce a supramolecular signaling complex on the cytoplasmic side of the plasma membrane. For the TNFSF ligands, this requires that they be used as many-trimer multimers that mimic the natural expression of many trimers on the surface of stimulating cells. To meet this need, we prepared fusion proteins comprised of the extracellular domains of TNFSF ligands joined to a natural protein that provides a multimerization scaffold. When surfactant protein D (SP-D) is used as a scaffold, the result is a 4-trimer TNFSF ligand product (UltraLigand™). Our published studies have described such multimeric forms of CD40L, OX40L, GITRL, CD27L/CD70, BAFF, RANKL, and TRAIL and shown that they are highly active in vitro and in vivo. As an extension of this work, 4-trimer forms of murine and human 4-1BBL (CD137L, TNFSF9) were constructed and expressed in CHO cells. As a co-stimulatory molecule, SP-D-4-1BBL (Ultra4-1BBL) activated both CD4+ and CD8+ T cells and promoted the growth of NK cells in vitro. Given the interest in 4-1BB (CD137) as a marker of therapeutically effective tumor-infiltrating lymphocytes (TILs), SP-D-4-1BBL should be a useful should be a useful growth factor for TIL manufacturing and T cell culturing in general.
Inhibitory cytokines mark distinct subpopulations of intratumoral regulatory T cells
Authors: Yano Hiroshi , Sawant Deepali V , Liao Mengting , Bruno Tullia , Chikina Maria , Workman Creg J , Vignali Dario AA 
Published: January 2023

Abstract
Regulatory T cells (Tregs) play a crucial role in the maintenance of self-tolerance and the resolution of inflammation; however, they also negatively regulate anti-tumor immunity by contributing to the immunosuppressive tumor microenvironment. Our previous studies have shown that the inhibitory cytokine interleukin-35 (IL35), a member of the IL12 cytokine family, is preferentially expressed by activated Tregs and the IL35 expression is required for the maximum suppressive activity of Tregs.
To investigate the role of Treg-derived IL35 in the tumor microenvironment, we generated a Treg-specific IL35 reporter knockin mouse as well as a Treg-specific IL35 deletion mouse. We demonstrated that IL-35+ Tregs are preferentially enriched in the B16 melanoma tumor microenvironment and actively involved in tumorigenesis by promoting the expression of multiple inhibitory receptors in CD8+ tumor-infiltrating T lymphocytes (TILs). However, the impact of IL35 relative to IL10 and the phenotype of IL35+ Tregs relative to other Treg subpopulations in the tumor microenvironment have not been fully elucidated.
We have assessed the expression of IL35 and IL10 in intratumoral Tregs and the phenotype and function of cytokine producing subpopulations using a triple Foxp3/Ebi3/Il10 reporter knockin mouse. We have also assessed IL35/IL10 expression in Tregs infiltrating several human tumors. Interestingly, there are distinct IL35-/IL10-expressing Treg subpopulations in mouse and human tumors. We continue to characterize the phenotype and function of intratumoral Treg subpopulations in mouse and human tumors.
Cancer testis autoantigens as germ cell autoantigens continuously egress from normal testes and maintain Treg-dependent tolerance
Authors: Tung Kenneth S K , Harakal Jessica , Qiao Hui , Rival Claudia , Paul Liesbeth , Hardy Daniel M. , Cheng C. Yan , Goldberg Erwin 
Published: January 2023

Abstract
We investigated tolerance for: 1) testis antigens (mitotic germ cells and somatic cells) located external to the blood-testis barrier (BTB) and 2) testis antigens (sperm and meiotic germ cells) located internal to the BTB. Transiently Treg-depleted B6AF1-DEREG mice produced autoantibodies to mitotic germ cells known to express NY-ESO-1 and MAGE-A (target cancer testis antigens (CTA) in cancer vaccine trials) and to Leydig cells. Unexpectedly, Treg-depleted mice also produced autoantibodies to sperm antigens. These plasma membrane and cytoplasmic sperm antigens continuously egressed from normal seminiferous tubules via the residual bodies. They were recognized by infused circulating antibodies to form local immune complexes outside the BTB and were sensed by antigen-specific T cells in regional lymph nodes. In contrast, sequestered sperm antigens, excluded from residual bodies, were not tolerogenic. Therefore, we provided direct evidence against the “complete testis antigen sequestration” dogma and established Tregs as a critical physiological tolerance mechanism for non-sequestered testis autoantigens. These findings are germane to both autoimmunity and tumor immunity. We found that vasectomized mice responded to sequestered sperm antigens, whereas Treg-depleted mice with autoimmune orchitis responded to non-sequestered sperm antigens. We also detected both tolerogenic and non-tolerogenic sperm antigens as CTA in many types of human cancer. We posit that the existence of two sets of CTA as normal germ cells antigens with distinct tolerogenic properties will impact the outcome of anti-tumor immunity in male patients and will influence the selection of optimal immunogens in cancer vaccine development.
Nonredundant role for GM-CSF in autoimmune pathology of the neureretina in the absence of IFN-γ and IL-17A
Authors: Bing So Jin , Silver Phyllis , Jittayasothorn Yingyos , Horai Reiko , Caspi Rachel R 
Published: January 2023

Abstract
Autoimmune uveitis is a complex group of sight-threatening CNS diseases caused by activated retina-specific Th1 or Th17 cells that acquired the ability to cross the blood-retinal barrier, but their respective contribution to disease is not fully understood. In this study, we used mice deficient in IL-17A, IFN-γ (GKO), or both (DKO) to dissect the role of Th1 and Th17 lineage-specific cytokines in pathogenesis of uveitis. Experimental autoimmune uveitis (EAU) was induced by active immunization with the retinal autoantigen IRBP in CFA. IL-17A−/− mice were protected from EAU, whereas GKO mice had exacerbated disease. Surprisingly, DKO mice were fully susceptible to EAU with scores similar to WT controls. The frequency of macrophages and production of proinflammatory cytokines (GM-CSF, IL-6, and TNF-α) was increased in the spleen and eyes of DKO mice. To test whether other T cell lineage-related cytokines contribute to pathology in the absence of IL-17A and IFN-γ, EAU-challenged DKO mice were treated with blocking Abs to IL-17F, IL-22, TNF-α, or GM-CSF. Blockade of IL-17F, IL-22, or TNF-α in DKO mice did not affect the severity of EAU. By contrast, treatment of DKO mice with an anti-GM-CSF Ab during the expression stage of disease significantly suppressed disease severity in DKO, but not in WT mice. These results suggest that IL-17F, IL-22 or TNF-α individually are dispensable, but GM-CSF appears to play a major and nonredundant role as a pathogenic cytokine in EAU when IFN-γ and IL-17A are both absent. Our results highlight that ‘classical’ Th1 and Th17 cells are not the only pathogenic effectors, and that other inflammatory cytokine(s), such as GM-CSF, can also drive uveitis.