bluuebunny/crossref_metadata_2025_split huggingface dataset:
Granzyme A-producing helper T cells are critical for lethal acute graft-versus-host disease
Authors: Olson Matthew Richard , Park Sungtae , Griesenauer Brad , Mehrpouya-Bahrami Pegah , Kazemian Majid , Imam Tanbeena , Srivastava Rajneesh , Hayes Tristan , Pardo Julian , Janga Sarath Chandra , Paczesny Sophie , Kaplan Mark H 
Published: January 2023

Abstract
Graft-versus-host disease (GVHD) is a deleterious allo-immune response that occurs in patients undergoing hematopoietic stem cell transplantation (HSCT) for treatment of blood-derived malignancies. CD4 T helper (Th) cells within the donor HSC pool recognize allogeneic HLA molecules and traffic to multiple organs, including the intestines where increased inflammation correlates with worsened clinical prognosis. Despite the known role of Th cells in GVHD, the means by which they promote disease remain unclear. In our studies, we have identified a novel class of Th cells that produce the serine protease granzyme A (GrA) within the intestines of mice succumbing to GVHD. GrA+ Th cells were distinct from other Th lineages and largely lacked co-expression of other Th lineage-associated cytokines or cytolytic mediators. Importantly, mice that received HSCT with GrA-deficient CD4 T cells exhibited significantly reduced morbidity and mortality as compared to controls, suggesting that Th-derived GrA is critical for GVHD progression. In vitro, differentiation of GrA+ Th cells was optimally induced with a combination of IL-6, IL-21 and IL-4 in a STAT3- and STAT6-dependent fashion. In vivo, STAT3- was also partially required for the differentiation of GrA+ Th cells during the GVHD response. In summary, GrA+ Th cells represent a novel STAT3-dependent Th cell type that is critical for allo-immune mediated disease.
Robust enhancement of tumor-dictated CD8 T cell trafficking and activation by QS21 adjuvanted vaccine
Authors: Sastry Jagannadha K , Nookala Sita M , Dorta-Estremera Stephanie M , Sierra Gloria , Yanamandra Ananta V. 
Published: January 2023

Abstract
Adjuvants enhance immune response to antigens and increase vaccine efficacy. We investigated the relative potency of adjuvants with diverse mechanism of action: a natural killer T cell activating ligand (alpha-galactosylcermide, αGalCer), a toll-like receptor 9 ligand (CpG B), a liposomal nanoparticle (R-DOTAP) and a plant saponin from Quillajira saponaria (QS21). We used OVA TCR transgenic model and also compared systemic vs mucosal routes of delivery for inducing antigen-specific CD8 T cell responses in naïve and syngeneic tumor-bearing mice. Wild type C57Bl/6 mice were vaccinated with OVA admixed with the adjuvants via subcutaneous (SC) or intranasal (IN) route 1d after adoptive transfer of the OT-I cells. Single cell suspensions of tissues 7d post-vaccination were analyzed using flow cytometry for OT-1 cells (CD45.1+Vα2+). Vaccination by either SC or IN route using QS21 adjuvant showed the most potent trafficking and activation of CD8+ T cells to both systemic and at mucosal sites, with the highest response observed in the lungs. Vaccination using the αGalCer by the IN route was relatively more effective than when delivered by the SC route, but the responses with αGalCer were lower compared to that with the QS21 adjuvant. However, vaccination of mice bearing B16F10-OVA resulted in preferential trafficking of OT-I cells to the tumors with much reduced numbers at the other tissues. Once again, these responses were robustly higher with QS21 vaccine delivered by either SC or IN route and the OT-1+ T cells from the tumors exhibited higher levels of functional and proliferating properties based on IFN-γ and Ki67 staining. Our results demonstrate that QS21 is a potent adjuvant for prophylactic as well as therapeutic vaccination applications.
Defining the signaling and downstream regulation of neutrophil-dendritic cell development for adjunctive therapy
Authors: Gui Michael , Fites J. Scott , Sykes David , Mansour Michael K , Klein Bruce S 
Published: January 2023

Abstract
Neutrophils are typically considered short-lived and terminally differentiated. However, recent studies have identified a poorly understood neutrophil subset that can differentiate into dendritic cells and become long-lasting “hybrids” of neutrophils and dendritic cells. These neutrophil-dendritic cells (PMN-DCs) maintain the functions of canonical neutrophils while gaining antigen-presenting properties of dendritic cells, enabling them to contribute to both innate and adaptive immunity. Although PMN-DCs have been characterized in various inflammatory conditions, the cues that drive their development are not fully understood. Granulocyte macrophage colony-stimulating factor (GM-CSF) has previously been implicated as an important signal for PMN-DC development in studies of murine and human neutrophils. In vivo neutralization suggests that GM-CSF is sufficient but not necessary to drive PMN-DC differentiation. To determine the requirement of GM-CSF we have deleted GM-CSF receptor using CRISPR editing of a murine neutrophil progenitor cell line. We have tested other implicated cytokines in vitro, including IL-3, IL-4, TNF-α and IFN-γ, however these signals alone do not induce differentiation of PMN-DCs but can augment or blunt PMN-DC differentiation. Previous work shows that PMN-DCs are potent effectors of antifungal immunity through both direct killing and antigen presentation; understanding of the developmental signals that drive PMN-DC differentiation is essential for any future therapies targeting PMN-DCs.
Monocytes outperform ex vivo generated dendritic cells as cellular vaccines to trigger cytotoxic T lymphocyte responses against cancer in pre-clinical models
Authors: Huang Min-Nung , Nicholson Lowell T. , Batich Kristen A. , Kopin David , Swartz Adam M. , Sampson John H , Gunn Michael Dee 
Published: January 2023

Abstract
Conventional dendritic cell (DC) vaccines using ex vivo generated DC have been widely tested in clinical trials in the treatment of human cancers. These DC vaccines have often displayed limited efficacy despite a complex manufacturing process. Although the efficacy of DC vaccines is generally thought to depend on their antigen-presenting cell function, several lines of evidence suggest that, instead of directly presenting antigen (Ag) to naïve T cells, administered DC trigger cytotoxic T lymphocyte (CTL) responses in vivo mainly by transferring Ag to lymphoid-resident conventional DC (cDC). We tested here whether monocytes, known to transfer Ag to cDC in vivo, can be directly loaded with tumor Ag to trigger robust anti-tumor CTL responses. We found that IV administered monocytes loaded with protein or peptide Ag triggered stronger Ag-specific CTL responses than conventional adjuvant-based and DC vaccines. In several murine cancer models, including intra-cranial glioblastoma, we showed that monocyte vaccination suppressed tumor growth better than conventional adjuvant-based, GVAX and DC vaccines. Such anti-tumor activities could be further enhanced with immune checkpoint inhibition. Ag-loaded monocytes did not directly activate naïve CD8+T cells. Instead, they formed stable physical contacts with and transferred Ag to splenic CD8+cDC. This Ag transfer required the formation of connexin 43-containing gap junctions between monocytes and CD8+cDC. Our findings demonstrated that gap junctions are an efficient Ag transfer pathway in vivo between monocytes and splenic CD8+cDC and suggested that IV administered Ag-loaded monocytes may serve as a simple and efficacious cellular vaccine platform for the treatment of human cancers.
Engineered Toxin Bodies delivering immunogenic MHC class I peptides to tumor cells summon polyfunctional and relevant CTL responses against cancers
Authors: Dekker Joseph D , Brieschke Brigitte , LeMar Sara , Cornelison Garrett L. , Iberg Aimee , Robinson Garrett L. , Chu Shaoyou , Willert Erin K , Ramos Hilario J. 
Published: January 2023

Abstract
Immunotherapeutic approaches such as CAR-T and T cell engaging bispecific antibodies have shown positive clinical outcomes and demonstrate the value of harnessing cytotoxic T lymphocyte (CTL) activity for tumor intervention. Here we describe Engineered Toxin Bodies (ETBs) with Antigen Seeding Technology (AST) as a unique tumor targeted therapeutic approach to recruit endogenous polyfunctional memory CTL responses against cancers.
Molecular Templates’ ETB platform combines antibody domains with a proprietary form of the Shiga like Toxin A (SLTA) subunit to specifically target cells and exploit SLTA’s intrinsic ability to self-internalize and destroy ribosomes. With the addition of a foreign class I antigenic peptide fused to an ETB (AST), antigens present in complex with endogenous MHC I to initiate a high avidity T cell response to target cells. Utilizing this approach, we demonstrate ETBs targeted to clinically relevant markers can seed antigenic peptides to hematological tumor, solid tumor, and primary cells.
Expanded CTLs from donors with preexisting Human Cytomegalovirus (CMV) reactivity phenotypically resembled memory expansion from natural infection. We next delivered class I CMV peptide to tumor cells by AST with ETBs targeted to PD-L1 and co-cultured tumor cells with expanded CTLs. This led to activation of CMV specific T cells, direct tumor-T cell engagement, and potent cell kill activity above that of parental ETBs. CTL phenotyping and cytokine profiling revealed polyfunctional T cell activation driven by AST enabled ETBs. Thus, ETBs delivering peptide via AST provide additive mechanisms of action: direct cell kill via ribosomal destruction, and antigen specific polyfunctional memory CTL activation.
Intradermal DNA Aβ42 immunization via needle-less Jet injection as potential immunotherapy for Alzheimer’s disease
Authors: Lambracht-Washington Doris , Fu Min , Rosenberg Roger N 
Published: January 2023

Abstract
The most common form of dementia found in the elderly is Alzheimer’s disease (AD) and AD progression has been strongly associated with accumulation of Amyloid beta 1–42 (Aβ42) and neurofibrillary tangles (tau) in brain. Amyloid pathology precedes tau pathology and an immune-mediated approach to reduce amyloid plaques without causing brain inflammation is highly desirable for possible clinical use. Genetic immunization, in which the immunizing agent is DNA encoding Aβ42, has great potential because the immune response to DNA delivered into the skin is generally non-inflammatory, and differs qualitatively from immune responses elicited by peptides, which are inflammatory with cellular production of IFNγ and IL-17. DNA immunization has been difficult in larger mammals and a potential barrier is the method for delivery of the DNA antigen. Over the last decade substantial progress has been made to increase the effectiveness of DNA delivery and immune responses, respectively. Recently, needless DNA Jet injection was approved by the FDA for a Zika vaccine. We are currently working on translating our established gene gun DNA immunization approach in mice into the clinically more feasible method of DNA delivery via jet injection. We have tested the Jet injector alone and in combination with electroporation in wild-type mice and New Zealand White rabbits as a large mammal. We found good antibody production and safe cellular immune responses (no inflammatory cytokines) in both species. Antibody levels and kinetics in mice and rabbits were very similar with about 30 μg of anti-Aβ antibodies after four immunizations. This study is designed to optimize DNA delivery for possible testing of the DNA Aβ42 vaccine for AD prevention in a future clinical trial.
Immunotherapy with tumor membrane vesicle-based cancer vaccine inhibits metastatic tumor growth in lung cancer model
Authors: Bommireddy Ramireddy , Huang Haley Lei , Munoz Luis E , Menon Ashwathi P , Ramalingam Suresh , Selvaraj Periasamy 
Published: January 2023

Abstract
Lung cancer is the leading cause of cancer related deaths accounting for approximately 30% of all cancer related mortalities affecting both men and women. Currently, there is no cure for the common non-small cell lung cancer (NSCLC) thus development of efficacious therapies is urgently needed. In this study, we investigate the efficacy of tumor membrane-based vaccine immunotherapy and immune checkpoint blockade antibodies lung cancer models LL/2 and CMT-167 models. LL/2 and CMT-167 tumor tissues were harvested from C57BL/6 mice and processed to generate tumor membrane vesicles (TMVs). TMVs were then modified with immunostimulatory molecules and used for immunization. Tumor growth was monitored every 3 days. Survival was assessed using a Kaplan-Meier survival curve and significance determined using a Log-rank test for comparison analysis. Metastasis of CMT-167 was assessed by clonogenic assay. TMV vaccine alone, or in combination, anti-PD-1 or anti-CTLA-4 antibodies neither decreased the tumor growth nor improved the survival of mice challenged with LL/2 tumor cells. The prophylactic vaccination of TMV vaccine prevents the CMT-167 tumor growth in mice and helps the mice develop immunological memory to the tumor cell challenge. In a therapeutic setting, the TMV vaccine inhibited tumor growth and induced primary tumor regression in 40% of mice and significantly reduced the metastatic tumor burden. Anti-tumor immunity induced by TMV vaccine requires CD4+ and NK cells albeit to a lesser extent but not CD8+ T cells for controlling the tumor growth. These models provide a valuable tool in analyzing the mechanism of interaction of immune cells with tumors that lead to either tumor regression or resistance to immunotherapies.
CD38hi B-regulatory (B-reg) cells maintain pathological immune tolerance in chronic lymphocytic leukemia (CLL)/B cell diseases: Potential therapeutic considerations
Authors: Manna Alak , Aulakh Sonikpreet , Sher Taimur , Ailawadhi Sikander , Chanan-Khan Asher , Paulus Aneel 
Published: January 2023

Abstract
Chronic Lymphocytic Leukemia (CLL) is considered a disease of antigen-naive CD5+CD23+CD19+κ/λ+ B-lymphocytes, in which the tumor microenvironment is highly immunosuppressive. Among several human B cell subsets, B10 [CD24hiCD27+CD38hi]/B regulatory cells (Bregs) are functionally discernable in patients with aggressive CLL as they support and regulate immunological tolerance via production of IL-10 and transforming growth factor β (TGF-β). In an ex vivo setting, we noted CD38hi CLL Bregs to promote transformation and expansion of CD4+CD25+FoxP3+Tregs; an effect that was abrogated in the presence of IL10/TGF-β neutralizing antibodies. In patients with CLL, Bregs also prohibit the expansion of cytotoxic T cells (CD8+T cells; Tc), natural killer (NK) cells and other pro-inflammatory lymphocytes (CD4+ helper T cells; Th1 and Th17). We noted that patients with CLL had a significantly higher % of Tregs (11.8±2.0) compared to healthy donors (1.9±0.3) and similar to Bregs, a substantial proportion of CLL Tregs had high CD38 expression (MFI=616.8±36.2). We demonstrated that an anti-CD38 therapeutic antibody was able to eliminate CD38+ Bregs and Tregs from CLL patients by immune effector mechanisms (ADCC, CDC, ADCP) as well as direct mitochondrial/FcγR-mediated apoptosis. Furthermore, in a PDX model of CLL, mice treated with anti-CD38 therapy had decreased % Bregs and Tregs but increased Th17 and CD8+ T-cells (vs. vehicle; p&lt;0.05). These observations carry implications beyond CLL for any Breg/Treg-dependent cancer or disease (i.e. Systemic Lupus Erythematosus, Rheumatoid arthritis, Sjögren’s syndrome), wherein CD38-targeted therapy could potentially rescue/improve an immune tolerant microenvironment by eradication of Bregs.
A novel preclinical immunocompetent CAR T cell mouse model for solid tumors
Authors: Sullivan Peter M , Arguedas-Jimenez Laura , Johnson Adam , Yokoyama Jason , Yuzefpolskiy Yevgeniy , Jensen Michael , Kalia Vandana , Sarkar Surojit 
Published: January 2023

Abstract
Chimeric antigen receptor (CAR) T cells therapies have made substantial progress in treating blood cancers in humans. However, CAR T therapies currently have several limitations, including the lack of a persistent anti-tumor memory population to limit relapse and poor efficacy against solid tumors. Many studies on CAR T therapies have used immunocompromised xenograft mouse models, limiting the translational impact of the results. Furthermore, mouse models have largely focused on cancers in the blood while neglecting solid tumors. To improve the relevance of preclinical studies of CAR T cell therapy and to understand the mechanisms underlying some of the current CAR T therapy limitations, we have created a novel immunocompetent mouse model. This model consists of three components: murine CD8 T cells expressing the human CD19 CAR, syngeneic murine tumor cell lines expressing a truncated human CD19 (CD19t) serving as the tumor antigen, and an immunocompetent mouse tolerized to human CD19t and the CD19 CAR. Our results demonstrate that the CD19 CAR T cells can efficiently target the CD19t-expressing tumor lines. Furthermore, tolerized recipient mice do not reject implanted syngeneic CD19t-expressing tumors or adoptively transferred CD19 CAR-expressing T cells. This model now provides a platform to study the in vivo biology of CAR T cells with respect to localization, exhaustion, and functional reinvigoration; to investigate best approaches to treating multiple solid tumors with distinct microenvironments; and to test the preclinical efficacy of novel CAR T strategies.
Covalent adaptor synNotch and chimeric antigen receptors (CARs) for programmable antigen targeting
Authors: Lomueller Jason Jakob , Butchy Adam , Tivon Yaniv , Kvorjak Michael , Miskov-Zivanov Natasa , Deiters Alexander , Finn Olivera J 
Published: January 2023

Abstract
Chimeric antigen receptors (CARs) are artificial T cell receptors that re-target patients’ T cells to specifically bind and kill tumor cells. Adoptive cell therapy with CAR T cells targeting CD19 has revolutionized treatment of refractory B cell acute lymphoblastic leukemia, and there is great interest in generating CAR T cells treating other cancers by targeting additional tumor antigens. Another promising class of engineered receptors are synthetic Notch (synNotch) receptors that can sense an antigen of interest on a neighboring cell and turn on expression of any transgene(s) of interest. To expand the targeting capabilities of these receptors, we have developed “universal” CAR and SynNotch receptors whose antigen-specificity can be re-directed by co-administered tumor-specific antibodies. Instead of directly targeting a tumor antigen, our universal receptors contain the SNAPtag self-labeling enzyme, which reacts with antibodies conjugated to benzylguanine (BG) to post-translationally assemble complete covalently associated antigen receptors. We demonstrate that the activation of SNAP CAR and SNAP-SynNotch receptors can be successfully re-targeted by several clinically relevant antibodies including: Rituximab, FMC63, Herceptin, and Cetuximab. SNAP-SynNotch cells demonstrate potent transgene activation, and SNAP-CAR T cells are capable of performing effector functions in a BG-antibody-directed antigen-specific manner. Additionally, the receptor response is titratable by BG-antibody dose. Finally, a continuous mathematical model was constructed to describe and optimize system activity. SNAP synNotch and SNAP CAR T cells provide a powerful new strategy to retarget engineered cells to multiple antigens.
Improving immunotherapy through immune checkpoint assays, CAR-T target display, and effector function optimization
Authors: Edgcomb Stephen , Zhu Jie , Tao Zhihua , Hsu Matthew , Kimbara Michelle , Oberle Suzan , Zhu Henry 
Published: January 2023

Abstract
Cancer and autoimmune disease lead to illness by damaging and evading a patients natural defenses. Immunotherapy methods are designed to reclaim and bolster the immune system to treat disease. This requires both highly specific binding of therapeutic agents to target molecules and optimizing techniques to harness the effector functions of the immune system. We describe methods for both these requirements. These include cell based systems for improving attachment to targets by assaying binding of therapeutic antibodies to immune checkpoint receptors and displaying molecular targets to CAR- T cells. They also include assays to improve immune effector functions such as Antibody-dependent cell-mediated cytotoxicity (ADCC) through cell based assays of IgG Fc binding to FCgRs.
Novel HIV DNA vaccine changes the immune hierarchy and elicits broad cytotoxic T cell responses targeting subdominant viral epitopes
Authors: Felber Barbara K. , Lu Zhongyan , Hu Xintao , Valentin Antonio , Rosati Margherita , Mullins James I. , Pavlakis George N. 
Published: January 2023

Abstract
HIV sequence diversity and the propensity of eliciting immunodominant responses targeting non-essential variable regions are hurdles in the development of an effective AIDS vaccine. We developed a DNA vaccine comprising conserved elements (CE) of Gag and Env. Macaques (RM) received the CE DNA prime/CE+full-length DNA co-delivery booster vaccine regimen via intramuscular injection followed by electroporation. Priming vaccination with CE DNA is critical to efficiently overcome the dominance imposed by Gag and Env variable regions. CE vaccine induced broad, potent and durable cytotoxic T cell responses targeting conserved segments. Gag CE-specific T cells showed robust anamnestic responses upon infection with SIVmac239 which led to the identification of CE-specific cytotoxic lymphocytes able to recognize several CE epitopes presented on SIV infected cells in vivo. The T cell responses induced by the HIV Env CE immunogen were also recalled upon SIV infection, leading to the identification of two cross-reactive epitopes between HIV and SIV Env. These data demonstrate that the CE DNA vaccines changed the normal immunodominance patterns, eliciting immune responses that included subdominant, highly conserved epitopes. These vaccine regimens augment cytotoxic T cell responses to highly conserved epitopes in the viral proteome and maximize response breadth. The HIV Gag CE vaccine regimen is evaluated for prevention and immunotherapy in clinical trials (HVTN119 and ACTG 5369).