bluuebunny/crossref_metadata_2025_split huggingface dataset:
A novel form of 4-1BB ligand has better efficacy than agonistic 4-1BB antibody as well as TLR agonists in a peptide based cancer vaccine (41.5)
Authors: Sharma Rajesh K. , Elpek Kutlu G. , Yolcu Esma S. , Schabowsky Rich-Henry , Zhao Hong , Bandura-Morgan Laura L. , Shirwan Haval 
Published: January 2023

Abstract
Toll like receptor (TLR) agonists, MPL and CpG-ODN, have been the subject of intense preclinical and clinical investigations as most promising vaccine adjuvants. Given the paramount importance of costimulation in the modulation of immune responses, we hypothesized that agonistic costimulatory ligands may serve effective alternatives to TLR agonists for vaccine development. Unlike TLR agonists, costimulatory ligands primarily function as cell membrane-bound proteins and lack activity in soluble form. We, therefore, used core streptavidin as a chaperon to generate costimulatory ligands that function in soluble forms. We compared the activity of a novel form of 4-1BBL composed of the extracellular domain of murine 4-1BBL fused to the C-terminus of core streptavidin (SA-4-1BBL) with an agonistic Abs to 4-1BB and TLR agonists as components of peptide based vaccines in a cervical cancer mouse model. Vaccination with SA-4-1BBL resulted in better efficacy than agonistic Ab as well as MPL/LPS/CpG in the eradication of established tumors. Therapeutic efficacy of SA-4-1BBL was associated with enhanced tumor specific primary and memory responses and reversal of T cell anergy. Importantly, therapeutic efficacy was achieved in the absence of sever toxicity associated with the agonistic Ab. Taken together, these data demonstrate the feasibility of using SA-4-1BBL as an effective alternative to TLR agonists or agonistic Abs to 4-1BB as immunomodulatory component of therapeutic cancer vaccines.
Funded in parts by NIH R43 CA 109866 (HS), R43 AI071618 (HS, ESY)
R41 CA121665 (HS), KLCRP (HS), KSTC-145-402-22 (HS).
Amplifying TLR2 signals in Tumor-specific T cells Lowers the Activation Threshold to a Weakly Immunogenic Tumor Antigen and Enhances Antitumor Activity (41.50)
Authors: Davila Eduardo , Zheng Liquin , Geng Degui , Asprodites Nicole 
Published: January 2023

Abstract
Although toll-like receptor (TLR) agonists are primarily believed to potentiate T cell responses by activating antigen-presenting cells, we recently demonstrated that TLR2 engagement on CD8 T cells enhanced proliferation and increased the production of several effector molecules. Here, we examined the antitumor activity of TLR2-stimulated pmel CD8 T cells, which recognize the weakly immunogenic melanoma tumor antigen gp100. TLR2 engagement on pmel T cells but not TLR2-/-pmel or MyD88-/-pmel cells reduced the activation threshold up to 100-fold as measured by IFN-γ, granzyme B, perforin and IL-2 production and cytolytic activity. Adoptive T cell transfer of pmel T cells in wild type or MyD88 knockout mice and injection with TLR2 ligand improved the antitumor activity against an established melanoma tumor as compared with mice treated with TLR2-/-pmel or MyD88-/-pmel T cells and TLR2 ligand. Furthermore, TLR2-ligated T cells persisted longer than did control T cells. Overexpressing TLR2 in pmel T cells improved effector responses above that of T cells expressing wild type levels of TLR2. These data highlight an important and physiologically relevant role for activating TLR2-MyD88 signals within T cells and could enhance immunotherapeutic approaches to treat cancer patients by augmenting T cell responses to weakly immunogenic TAs. Supported by a NIH (1P20 RR021970) and the Louisiana Cancer Research Consortium.
Continuous Intra-Lymphatic Dendritic Cell Infusion in Patients with Advanced Melanoma (41.51)
Authors: Kalinski Pawel , Edington Howard , Brown Charles , Whiteside Theresa , Butterfield Lisa , Bartlett David , Kirkwood John 
Published: January 2023

Abstract
Continuous intralymphatic delivery of dendritic cells (DCs), using implantable lymphatic cannulas, allows to efficiently deliver "non-exhausted" DCs to draining lymph nodes without the disruption of nodal structures, and to recreate the physiologic kinetics of DC accumulation observed during physiologic immune responses.
We have completed initial feasibility and safety evaluation of semi-continuous intralymphatic infusions of antigen-loaded DCs (a total of 300,000 DCs over 4 days) in six patients with stage IIIb-IV melanoma in clinical trial UPCI 03-118. Four of the initial six patients received two four-day-long courses of the intralymphatic DC infusions, completing the protocol. Two patients did not complete the treatment for reasons unrelated to the protocol.
Already at this ultra-low dose of DCs, we have observed evidence of clinical efficacy of DC infusions in 3 of 4 patients with stage IV melanoma who completed both courses of treatment. We observed one near-complete response (currently &gt;20 months), and two disease stabilizations (10 months; and &gt; 7 months), with only one patient progressing. We have recently initiated the evaluation of "high-dose" intralymphatic infusions (total of 3 million DCs over 4 days).
Our data demonstrate the feasibility of prolonged intralymphatic delivery of biotherapeutic agents and provide preliminary indication that DCs can be clinically effective in patients with advanced cancer at the doses 100-fold lower than previously considered to be necessary.
Dissecting the integration of Natural Killer cell receptor signaling (41.52)
Authors: Xu Bin , Kaiser Brett , Strong Roland K. 
Published: January 2023

Abstract
Natural Killer cell is a major component of the innate immune system. It plays important roles in rejection of cancerous and viral-infected cells. NK cells express numerous surface receptors that can be activating or inhibitory. To investigate how activating and inhibitory NK receptors integrate signals in the decision to activate or to be tolerant, we studied the activating/inhibitory effects of NKG2D-MIC and NKG2A/CD94-peptide-HLA-E pathways, both alone or in combination using human NKL leukemia cell line and in vitro purified protein ligands. Our results demonstrate NKG2D receptors have antibody-induced cell activation effect. MIC ligand induces NKG2D receptor down-regulation on cell surface but alone is not sufficient to induce receptor activation. Surprisingly, NKG2A/CD94 ligand HLA-E/peptide potentiates NKG2D receptor activation. Possible molecular signaling mechanisms will be discussed. Supported by NIH grant AI48675.
Photodynamic Therapy Elicited Adaptive Immunity is Critically Dependent on IL-17 Bioactivity (41.56)
Authors: Brackett Craig , Maier Patricia , Ramsey Kimberley , Gollnick Sandra 
Published: January 2023

Abstract
Photodynamic therapy (PDT) is a FDA-approved local cancer treatment of early stages of cancer and palliation of advanced disease. A hallmark of PDT is the induction of acute local inflammation characterized by increased infiltration of innate immune cells into the treated tumor bed, which is dominated by CD11b+Ly-6GhiF4/80- leukocytes that morphologically resemble neutrophils. We have previously shown that enhanced anti-tumor CD8+ T cell activation following PDT is dependent upon the presence of CD11b+Ly-6GhiF4/80- leukocytes in the tumor and tumor draining lymph nodes (TDLNs). We now show that PDT promotes the differential expression of IL-23 and IL-17 within the treated tumor bed. Blocking the action of IL-17 using IL-17RA-/- mice does not diminish bone marrow release of CD11b+Ly-6GhiF4/80- leukocytes or their migration to the tumor post PDT but does abrogate accumulation of these cells in the TDLN. Furthermore, blocking IL-17 inhibits post-PDT activation of tumor specific CD8+ T cells. These findings suggest that IL-17 differentially regulates CD11b+Ly-6GhiF4/80- leukocyte migration and indicates that CD11b+Ly-6GhiF4/80- leukocyte accumulation in the TDLN is critical for induction of T cell activation. Neutrophils are capable of informing and shaping the immune response by influencing DC, monocyte, and lymphocyte activation. Understanding this behavior of neutrophils holds promise for the rationale design of PDT regimens to optimally enhance anti-tumor immunity in a clinical setting.
Tumor IL-10 forms an immediate non-redundant blockade to CpG-induced activation of tumor-infiltrating DC and macrophages (41.61)
Authors: Stewart Charles A , Scheetz Loretta , Trinchieri Giorgio 
Published: January 2023

Abstract
Toll like receptors (TLR) are a major class of pathogen recognition receptors that trigger potent dendritic cell DC activation and present an important immunotherapeutic target. Combination therapy of established tumors by local injection of CpG oligonucleotides (TLR9 agonists) and blockade of IL-10 signaling results in tumor rejection in mice. Using the MCA38 colon carcinoma and B16 melanoma models, CpG treatment alone drastically reduces tumor growth for the duration of treatment, but does not lead to tumor rejection. Local tumor injection of MCA38 tumors with CpG oligonucleotides and intra-peritoneal anti-IL10R antibody led to rapid production of CCL2, TNF- ƒÑ and IL-6 that were detectable in serum after 2 hours. 24 hours after CpG treatment, IL-10, IL-12, and IFN-ƒx were also detectable in serum. Cytokine production at both 2 hour and 24 hour time-points was partially dependent upon blockade of IL-10 signaling. To examine activation of tumor-infiltrating leukocytes, ex vivo cultures of tumor-isolated cells were set up at time-points following CpG injection. Significant production of TNF- ƒÑ, IL-6 and IL-10 was observed only at 2 hours with blockade of IL-10 signaling. The transient period of cytokine production was matched at the cellular level: Prior to treatment MCA38 tumors comprise of ~40% tumor-infiltrating leukocytes (TIL) which are mostly CD11b+CD11c+ cells characteristic of the macrophage or DC lineage. 24 hours following treatment, TILs were predominantly CD11b+Ly6G+ cells. Our results show that CpG treatment of tumors induces rapid activation of tumor-infiltrating cells that is highly potentiated at early time-points by the blockade of IL-10 signaling.
Novel phytol-based adjuvants in vaccine formulations: Evaluation of safety and efficacy in induction and persistence of specific immune responses (41.64)
Authors: Aachoui Youssef , Ghosh Swapan K 
Published: January 2023

Abstract
Adjuvants function as "immunologists' dirty tricks" for enhancement of immune response. However, no two adjuvants work in a similar fashion. Adjuvant efficacy depends on how they influence microenvironment, antigen presentation and retention of immunogens. We are developing water-in-oil emulsion-type adjuvants from phytol (J Immune Based Ther Vaccines 2006, 5:4; ibid 2007; 5: 3). Phytol is a diterpene present in chlorophylls, alpha-tocopherols etc. Phytol derivatives we developed are hydrogenated/nitrated and glycosylated products (US patent pending). Our previous study showed that one phytol derivative PHIS-01(hydrogenated) is an excellent non-toxic adjuvant. In this study, we investigated the safety and efficacy of two other phytol compounds, PHIS-02 and PHIS-03 relative to PHIS-01 and other adjuvants. Using phthalate- keyhole limpet hemocyanin (KLH) conjugate as the immunogen emulsified with these adjuvants, we assessed titer, specificity, and isotypic profiles of anti-phthalate antibodies induced. In addition, we studied their effects on cellular immunity such as T cell proliferation and cytokine production. Preliminary results show that (1) phytol adjuvants significantly augment antibody response of isotypes IgG1 and IgG2a, (2) promote effective T cell proliferation, and (3) exhibit low toxicity with no adverse autoimmune anti-DNA response. We also observed that these adjuvants exert apoptotic/necrotic effects on mouse lymphoma cells 2C3 and A20 in a concentration-dependent manner.
Rapid isolation of pure, functional and bead-free T cells directly from whole blood using Dynabeads® FlowCompTM technology (42.11)
Authors: Moeller Anne-Sophie W , Kierulf Bente , Reed Berit , Neurauter Axl A , Schjetne Karoline W 
Published: January 2023

Abstract
BACKGROUND: Fast and gentle isolation of pure cells directly from whole blood is beneficial for both basic research, drug discovery, patient monitoring (e.g. analysis of circulating tumor cells, prenatal diagnostic, T cell numeration in HIV, chimerism analysis in transplantation) and would render the time-consuming gradient centrifugation step superfluous.
AIM: Develop a fast and gentle protocol for isolation of functional and bead-free human CD3+, CD4+ or CD8+ T cells directly from whole blood.
METHODS: Peripheral blood was drawn from healthy donors. Antibody (specific for human CD3, CD4 or CD8) conjugated to a modified biotin was added to label the cells. In the next step, Dynabeads coated with modified streptavidin were added and target cells were captured when placed on the magnet. After resuspension with FlowCompTM Release Buffer the bead-free cells were analyzed using flow cytometry and tested in functional assays.
RESULTS: Highly pure and viable human T cells were isolated in less than 50 minutes directly from whole blood using Dynabeads® FlowCompTM. Isolated T cells readily proliferate in response to stimulation with Dynabeads® CD3/CD28 T Cell ExpanderTM as measured by incorporation of EdU (EdU Click-iT). We demonstrate that isolated T cells were activated both in a polyclonal and antigen-specific manner, and that isolated cytotoxic CD8+ T cells kill target cells.
ImmPort FLOCK: Automated cell population identification in high dimensional flow cytometry data (42.17)
Authors: Scheuermann Richard H. , Qian Yu , Wei Chungwen , Sanz Inaki 
Published: January 2023

Abstract
Historically, investigators have analyzed flow cytometry (FCM) data using manual gating strategies and two-dimensional dot plot visualization to identify and compare specific cell populations between samples. However, this approach is subjective and does not scale well as the number of parameters detectable with different fluorochrome reagents increases. We have developed a software system for automated population discovery in multidimensional FCM data - FLOw Clustering without K (FLOCK), which specifically takes into account the unique features of FCM data, including non-spherical parameter distributions and sparse cell populations. FLOCK includes the following steps: data cleansing, data shrinking, data normalization, informative dimension selection, dense hyper-region identification, histogram partitioning, centroid calculation, clustering, visualization, population statistics calculation, cell ontology mapping. As a result, FLOCK automatically and objectively determines the number of cell populations detectable in a sample and assigns population membership to each cell event, from the multi-parameter flow cytometry data, in the absence of manual gating. Our study of normal human peripheral blood has identified 17 unique B lymphocyte populations based on the expression of IgD, IgG, CD24, CD27, CD38 & B220. Supported by NIH N01AI40076.
PGC nanocarrier-formulated native GLP-1 prevents diabetes in NOD mice (42.3)
Authors: Reichstetter Sandra , Reed Lynn , Lai Man-Shun , Castillo Gerardo M , Bolotin Elijah M 
Published: January 2023

Abstract
Formulation of peptides in a nanocarrier made up of protected-graft-copolymer (PGC) can greatly enhance pharmacokinetics and biodistribution of the peptides in vivo. Glucagon-like peptide 1 (GLP-1) is β-cell protective and decreases the severity symptoms of diabetes in Type 2 and streptozotocin models. We hypothesized that PGC-formulated GLP-1 can prevent Type 1 diabetes in NOD mice. 4-5 week old female NOD mice [n=25] were treated for eight weeks with 3mg/kg PGC-formulated GLP-1 administered s.c. either once or three times per week. After the treatment period, the mice were monitored for signs of diabetes until they reached 36 weeks of age. A group of NOD mice treated with anti-CD3 antibodies (0.5mg/kg i.p. once at treatment week 0 and once at treatment week 1) was used as a positive control for diabetes prevention. At 36 weeks of age, 48% of the mice treated with saline had developed diabetes. Anti-CD3 treatment prevented 50% of the diabetes cases in the mice with only 24% of the mice in this group having developed diabetes at 36 weeks of age. Once a week GLP-1 treatment (3mg/kg s.c.) was as effective in preventing diabetes as anti-CD3 treatment with only 28% of mice being diabetic at 36 weeks of age. Surprisingly, there was no difference between the three times per week GLP-1 treatment group (3mg/kg s.c.) and the saline control: 54.1% of the mice treated three times per week became diabetic, which was not statistically significantly different form the 48% diabetes incidence in the saline group.
T regulatory and prostate cancer cell-specific drug targeting using novel XPclad© nanoparticles (42.7)
Authors: Singh Rajesh , Singh Shailesh , Sharma Praveen K , Lillard, Jr. James W 
Published: January 2023

Abstract
In our study, we determined the utility of cisplatin-loaded biodegradable XPclad nanoparticles coated with polycaprolactone-polyethylene glycol copolymers conjugated to folate. We hypothesized that these 28 ± 5 nm sized particles would enhance the therapeutic efficacy of cisplatin through targeted delivery to both prostate tumors and CD4+CD25HiFoxP3+ T cells (Treg) cells. PCa cell lines (PC3 and LNCaP), but not normal prostatic epithelial cells (PrEC and RWPE-1), expressed high levels of folate receptor α (FRα/FR1). Similarly, Treg, but not CD4+CD25LoFoxP3- T cells (T helper), cells expressed FRδ/FR4. Treatment using 20 μM of cisplatin solution resulted in &gt; 90% cell death of PCa cells, PrEC, and RWPE-1 cells as well as purified Treg and T helper cells after 24 hr of culture. However, XPclad nanoparticles, containing cisplatin (equivalent to a 200 nM dose) induced the same level of apoptosis in PCa and Treg cells as cisplatin solution, but did not affect PrEC or RWPE-1 cells or T helper cell death. We also show that PC3 tumor-bearing mice that received either cisplatin solution (8 mg/kg/week) or cisplatin-encapsulated folate-coated XPclad nanoparticles (equivalent cisplatin dose = 0.08 mg/kg/week) had significant tumor regression. While mice receiving cisplatin solution had significant higher kidney and liver toxicities, XPclad nanoparticle-treated mice did not. In conclusion, our studies show that novel XPclad nanoparticles enhance tumor therapy efficacy with lower toxicities and suggest that FR-targeted nanotherapy cause the destruction of prostate tumor cells and Treg cells.
TNFR-TNFL superfamily member interactions induced by combined TLR/CD40 stimulation act as Signal 3 for enhanced CD8+ T cell immunity (44.10)
Authors: Sanchez Phillip J , Kedl Ross M 
Published: January 2023

Abstract
Previously, we have shown that antigen-specific CD8+ T cell expansion following TLR agonist/anti-CD40 immunization is dependent on the enhanced expression of CD70 on dendritic cells. The dependence on CD27-CD70 interactions was confirmed by using a blocking anti-CD70 antibody. We now show that in CD27-/- mice, the antigen-specific CD8+ T cell response to TLR agonist/anti-CD40 immunization is intact. However, administration of a blocking anti-OX40L antibody during immunization attenuates the CD8+ T cell expansion. Thus, similar to the wild-type C57BL/6 mice, the enhanced primary expansion of antigen-specific CD8+ T cells in response to TLR agonist/anti-CD40 vaccination is dependent on TNF receptor (TNFR) - TNF ligand (TNFL) superfamily interactions. Furthermore, TLR agonist/anti-CD40 vaccination is not affected by depletion of IL-12 using a neutralizing anti-IL-12p40 antibody in either wild-type or CD27-/- mice. Together, these data suggest that TNFR-TFNL interactions, facilitated by combined TLR agonist/anti-CD40 vaccination, can act as Signal 3 for CD8+ T cell immunity in the absence of IL-12.
This research is supported by a grant from the NIH.
Immunotherapeutics effects of IL-7 during a chronic viral infection (44.13)
Authors: Nanjappa Som Gowda , Kim Eui Ho , Sullivan Jeremy , Morre Michel , Suresh M 
Published: January 2023

Abstract
Viral persistence in chronic infections is associated with ineffective T cell responses. Here, we evaluated the therapeutic use of IL-7 to enhance T cell responses during a chronic infection with lymphocytic choriomeningitis virus (LCMV). Mice infected with Clone 13 strain of LCMV were treated with IL-7 at various intervals spanning the first 30 days after infection. Our studies show that the ability of IL-7 to enhance T cell responses to a chronic viral infection is constrained by high viral load. Early in the infection (days 0-15 PI), when the viral load is very high, IL-7 administration induced minimal enhancement of T cell responses to LCMV. However, with reducing viral load (after day 15 PI), IL-7 augmented virus-specific T cell responses, albeit temporarily. An extended duration of IL-7 treatment after the initial phase of T cell expansion (days 8-30 PI), markedly enhanced the number of virus-specific CD8 T cells. Moreover, extended IL-7 therapy altered the cell surface phenotype, and augmented the cytokine-producing ability of LCMV-specific T cells. Surprisingly, despite the augmentation of virus-specific T cell responses, IL-7 therapy had a modest effect on circulating viral load in chronically infected mice. In summary, these findings suggested that IL-7 administration could be used as an adjunct to anti-viral immunotherapy to bolster T cell responses to chronic viral infections. This work was supported by PHS grant AI68841 to M. Suresh.
MCMV induces salivary gland dysfunction in autoimmune-prone NZM2328 mice (44.17)
Authors: Carroll Virginia , Lundgren Alyssa , Brown Michael G 
Published: January 2023

Abstract
Loss of secretions from the salivary gland (SG) is a disease feature of Sjogren's Syndrome (SS) as well as certain viral infections, including HCV and HIV. We have found that murine cytomegalovirus (MCMV) infection of autoimmune-prone NZM2328 mice serves as a model for virus-induced exocrinopathy, displaying many SS-like disease features. Interestingly, MCMV can induce severe SG dysfunction within 2-7 days after intraperitoneal infection. MCMV replication within the gland was not directly responsible for dysfunction since NZM2328 mice secreted normally 7-14 days after low dose infection, despite high virus within the gland. Since the low dose regimen limited viral burden elsewhere in the animal, such as the spleen, SG dysfunction seemed to correspond with systemic viral load. Surprisingly, lymphocytes were not required for MCMV-induced dysfunction at day 7, including NK, T and B cells. At day 2, morphological alterations were noted in acinar epithelium and flow cytometric analysis revealed rapid changes in glandular monocytic populations. Ongoing experiments are examining a potential causal link between systemic immune responses and/or infiltrating inflammatory monocytes with morphological and functional changes within the salivary gland. Supported by NIH R01AI50072 and a Sjogren's Syndrome Foundation Student Fellowship