bluuebunny/crossref_metadata_2025_split huggingface dataset:
Induction of multivalent and balanced T cell responses by modular nanoshell vaccine technology
Authors: Huang Pinghan , Tsai Hsiao-Han , Tat Vivian , Hsu Jason C , Aleksandra Drelich , Chen Hui-Wen , Hu Che-Ming Jack , Tseng Chien-Te 
Published: August 2023

Abstract
After the outbreak of COVID-19, cellular immunity has been shown to provide protection especially in breakthrough infection and B cell-deficient patients. Furthermore, durable cellular immunity targets conserved epitopes among SARS-CoV-2 variants or different coronavirus then inspiring an attractive strategy for vaccine development. However, how to simultaneously induce balanced T cell responses against multiple epitopes still needs to be elucidated. In this study, we sought to design a T cell-based vaccine that can adjust valency against different T cell epitopes. We selected seven SARS-CoV-2 specific short peptides as antigens, including I-Ab/H-2Kb/H-2Db restricted epitopes. Furthermore, we used PLGA nanoparticle as vaccine platform to achieve the precise loading of the indicated peptides. Mice were immunized with different combinations of peptides and splenocytes were used to access epitope specific-IFN gamma expressing CD8+ or CD4+ T cell responses. First, we confirmed the robust immunogenicity of each peptide. We then combined an equal amount of indicated epitope peptides and accessed the immunogenicity of each epitope in the combination. The results showed peptides compete with each other to induce T cell responses regardless of MHC allele and MHC class. In addition, dominant epitopes significantly suppressed the immunogenicity of the subdominant epitopes. Finally, by optimizing the antigen dose, nanoparticle vaccine induced more balanced T responses against all seven epitopes. Taking advantage of nanoparticle, our results reveal a viable strategy to induce multivalent T cell response. Furthermore, the result suggests that competition of T cell epitopes is one of the factors controlling cellular immunity.
Single-Chain B cell receptor mice as a novel tool for antibody generation and exploring B cell signaling
Authors: Borbet Timothy C , Zaldana Kimberly , Ilmain Juliana K , Chan Kun-Wei , Luo Christina C , Bryan Arielle M , Kirilov Milen , Kong Xiang-Peng , Bhabha Gira , Ekiert Damian C , Weng Wei , Sheiffele Paul , Koralov Sergei B 
Published: August 2023

Abstract
Conventional antibodies consist of a pair of heavy and light dimers, while heavy-chain only antibodies (HCAbs), like those found in camelid and shark species, lack a domain critical for light chain binding and develop without a light chain. HCAbs have unique physiochemical properties that confer advantages: they are smaller, have greater accessibility to their target antigens, higher hydrophilicity, higher tissue penetrance, and greater conformational stability under environmental stresses compared to conventional Abs. They are also highly amendable to biochemical engineering. We have generated a mouse model, that can produce HCAbs from the endogenous murine heavy chain locus. We have shown that these mice undergo efficient B cell development in the bone marrow and generate a diverse repertoire of B cells that populate all peripheral immune compartments. Single-cell sequencing of B cells revealed some biases in VH and JH usage, but highlighted the tremendous diversity of rearrangements that give rise to B cells bearing HCAbs. With these mice, we have been able to generate antigen specific HCAbs to the SARS-CoV2 spike protein that have a KD in the nM range, measured via bio-layer interferometry. Further, the IgG3dCH1 mice provide us with a novel model to understand how single domain B cell receptors (BCRs) function in comparison to conventional heavy/light chain BCRs. We have observed single-chain BCRs have distinct transcriptional profiles by single-cell sequencing and support higher plasma cell differentiation in vitro and in vivo. These data highlight the utility of our small animal platform for the generation of clinically relevant HCAbs, and as a model to study the biology of HCAb encoding B cells which remain poorly understood.
Bernard Levine Postdoctoral Fellowship, R43AI136141
Targeting CD70 using CAR NK cells to enhance NK cells cytolytic effect against osteosarcoma
Authors: Rav Emily Simon , Anjier Ariana , Acharya Sunil , Basar Rafet , Wang Yifei , Daw Najat , Rezvani Katy , Gopalakrishnan Vidya , Gorlick Richard , Kleinerman Eugenie S , Gordon Nancy 
Published: August 2023

Abstract

Background
Cure rates for osteosarcoma (OS) have not improved in &gt;30 years and no new effectivetherapies have been identified for metastatic disease. Natural killer (NK) cells have become an attractive tool for cancer immunotherapy. Efficay is improved by direct tumor targeting introducing a specific chimeric antigen receptor (CAR). Emerging knowledge suggests CD70 to be a viable candidate as expression is higher in lung metastases and it has been correlated with tumor escape from immune surveillance.

Objective
To assess whether using CD70 CAR-NK cells we can enhance trafficking, homing and proliferation of NK cells and therefore therapeutic efficacy against OS.

Methods
We analyzed the cBioportal database to verify CD70 expression in OS. Surface expression of CD70 on human OS cell lines and normal osteoblasts was determined by flow cytometry. CD70 CAR constructs were sequenced, verified and transduced into NK cells. CD70 CAR NK and control NK cells cytolytic activity against OS cells was evaluated by IncuCyte. In vitro cytokine release was measured upon CD70 CAR NK/NK cells exposure to OS cells (IsoPlexis).

Results
Database analysis confirmed OS CD70 gene amplification. Flow cytometry showed variable expression of CD70 on OS cells and no expression on human osteoblasts. There was an increase in CD70 CAR NK percent lysis against OS cells at the highest effector:target ratios compared to control NK cells. We found a predominant release in the chemoattractive/stimulatory cytokines upon OS cells exposure to CD70 CAR NK cells.

Conclusion
CD70 has immunotherapy potential against OS. CD70 CAR NK cells have increased cytolytic activity against OS cells in vitro. Cytolytic activity may be influenced by the release of specific cytokines.
CD4 depletion enhances the efficacy of Super2 IL-33 armored CAR T cell against solid tumors and leads memory formation of both endogenous and CAR T cells
Authors: Mohamed Asmaa , Ferry Shannon Lynn , Huang Yina H 
Published: August 2023

Abstract
Chimeric antigen receptor (CAR) T cells exhibit tremendous efficacy in patients with relapsed and recurrent liquid cancers. However, CAR T cells fail to control solid tumors because of their inability to infiltrate the suppressive tumor environment and their poor persistence in vivo. The development of 4th generation armored CAR T cells that employ direct tumor killing and release cytokines that can modulate the immune response to enhance tissue/tumor infiltration may improve CAR T cell persistence. Our lab has developed Super2 and IL-33 (S233) armored CAR T cells that significantly delayed B16F10 melanoma tumor growth and enhanced tumor infiltration of CARs and endogenous immune cells. Combination therapy of S233 CAR T Cells with anti-CTLA4 or anti-CD4 further enhanced this phenotype. S233 CAR T cells and anti-CD4 combination therapy promoted Melanoma-associated vitiligo (MAV), which correlates with development of tumor protective Tissue Resident Memory (TRM) cells. To assess whether CAR T cells differentiate into memory T cells, we treated tumor-bearing mice with S233 CAR T Cells with or without anti-CD4, resected their residual tumors and tracked memory T cell responses. We found that S233 CAR T cells persisted longer in the tumor draining lymph node and developed accelerated MAV when combined with anti-CD4. Both CAR T cells and endogenous T cells differentiated into central and effector memory but only endogenous T cells in the CD4 depleted group became TRM cells. While these findings provide insights on strategies to enhance long-term persistence of CAR T cells, additional studies are needed to induce CAR TRM cells and assess the durability of anti-tumor responses.
NIH grant T32-AI007363 NIH grant P30-CA023108
Persistent activation of TNFR1 on spinal cord interneurons contributes to immune dysfunction after spinal cord injury in rodents
Authors: Martynyuk Tetyana , Bracchi-Ricard Valerie , Ricard Jerome , Tom Veronica J , Bethea John R 
Published: August 2023

Abstract
Spinal cord injury (SCI) is a devastating condition with 250,000 to 500,000 new cases globally each year. Respiratory infections, e.g., pneumonia and influenza, are the leading cause of death after SCI. For instance, an individual with SCI is 37 times more likely to become infected and hospitalized from the flu. Yet, the field has a poor understanding of how immune dysfunction arises after trauma to the central nervous system (CNS) and how altered neuro-immune communication impacts an individual’s outcome to infection. In humans and rodents, SCI leads to maladaptive changes in the spinal-sympathetic reflex (SSR) circuit which is crucial to sympathetic function. Soluble Tumor Necrosis Factor (sTNF) is a pro-inflammatory cytokine that is elevated in the CNS after SCI and remains elevated for several months after injury. We recently demonstrated that blocking TNFR1 activation in the CNS, using XPro1595, decreased the maladaptive excitability of SpINs in the SSR circuit leading to improved immune response to infection after SCI. We hypothesize that hyper-excitability of glutamatergic SpINs after SCI due to persistent TNFR1 activation contributes to immune dysfunction. To test this, we utilized viral mediated targeted knock-down of TNFR1 on excitatory SpINs, and demonstrated that inhibiting TNFR1 on excitatory interneurons contributes to improved immune function, including an increase in virus-specific T cells and decrease in viral load remaining in the lung after infection.
Supported by grants from NIH (R01 NS111761)
Supported by grants from NIH (R01 NS111761)
IL-12 alters T-cell metabolism to augment anti-tumor immunotherapy
Authors: Mills Stephanie Ann , Dwyer Connor , Quinn Kristen , Oberholtzer Nathan , Chakraborty Paramita , Mehrotra Shikhar 
Published: August 2023

Abstract
To better limit tumor growth, adoptive T cell therapy (ACT), a cutting-edge new strategy for treating cancer, permits patients’ T cells to be modified. Long-lived central memory cells (Tcm), a specific subset of T cells, have been demonstrated through studies conducted in our group and others to be more effective at controlling tumor burden. Powerful inflammatory cytokine IL-12 has been extensively studied for its roles in the interaction of innate and adaptive immunity; however, therapeutic potential has not been optimized. We hypothesized that ex vivo administration of IL-12 may improve metabolic fitness and anti-tumor capabilities of T cells. Using a B16 murine melanoma model and T cells that are reactive to the melanoma epitope gp100, we found ex-vivo conditioning with IL-12 increased stem cell memory (Tscm) and central memory (Tcm) T cell populations. Additionally, we found unique metabolic signatures and proliferation patterns were dependent on IL-12 duration. We also observed enhanced engraftment and tumor control for up to 60 days after adoptively transferring these cells into animals with subcutaneous melanoma tumors. These metabolically conditioned T cells showed improved mitochondrial mass and function, elevated cytokine output, and perforin expression. Aside from switching from glycolysis to oxidative phosphorylation, these cells were also able to continue producing cytokines even when glucose or pyruvate levels dropped. Together, these findings imply that IL-12 modifies T cells’ reliance on glucose for energy while preserving their anti-tumor ability, potentially amplifying their ability to fight tumors in nutrient deprived microenvironments. IL-12 could therefore have a large impact on future ACT strategies.
R01 CA250458, R01 CA236379
Blockade of IL-18 signaling downregulates homing of effector CD4+ T cells to the gut mucosa and provides protection from experimental IBD
Authors: Leber Andrew J , Hontecillas Raquel , Juni Nuria Tubau , Fitch Sarah N , Bassaganya-Riera Josep 
Published: August 2023

Abstract
The immunological complexity of inflammatory bowel disease (IBD) results in persistent intestinal inflammation in millions of patients. As an epithelial cell-derived cytokine essential in the differentiation of CD4+ T helper cells, interleukin 18 (IL-18) is emerging as a therapeutic target for IBD. When naïve CD4+ T cells are stimulated with IL-18, the production of IFNγ and differentiation of Th1 cells increase. IL18R1−/− CD4+ T cells produce less IFNγ and inflammatory cytokines compared to WT. To examine the viability of targeting IL-18 in vivo, colitis was induced in WT, IL18−/−, and IL18R1−/− mice by DSS. In both knockouts, lower proportions of IFNγ- and IL17-producing CD4+ T cells were observed in the colon along with lesser overall severity relative to WT. To further investigate, we examined the interplay of antigen presenting cells and CD4+ T cells in vitro. IL18R1−/− Th1 cells had decreased α4β7 and GPR15 expression when co-cultured with WT BMDCs. Meanwhile, IL18R1−/− Tregs had decreased IFNγ and CCR9 expression when co-cultured with WT BMDCs that were stimulated with LPS and retinoic acid, suggesting that IL-18 may play a role in the differential gut homing of CD4+ T cell subsets. When CD4+ T cells are transferred to Rag2−/−, mice receiving IL18R1−/− T cells possess significantly lower proportions of α4β7+ Th1 cells while the proportion of Tregs in the colon are increased. Treatment of mice with DSS or Mdr1a−/− colitis with an oral IL-18 inhibitor (20 mg/kg) resulted in decreased Th1 cells and neutrophils, lower histological scores and improved disease severity. From converging data, we deduce that inhibition of IL-18 may alleviate exacerbated immune responses associated with IBD, yielding a favorable therapeutic.
Evaluating IL-27 production at the maternal-fetal interface and its role in innate antiviral immune defense during pregnancy
Authors: Barksdale Briah , Merlino Madeline S , Jurado Kellie Ann 
Published: August 2023

Abstract
During pregnancy, it is critical to protect the womb and developing fetus from encountered pathogens. A way to enact protection is through localized immune cells. Immune cells communicate via specialized effector molecules called cytokines. Interleukin-27 (IL-27) is a cytokine that can initiate a pro-inflammatory or anti-inflammatory response, dependent on other microenvironmental queues. Interestingly, IL-27 and its receptor (IL27RA) are known to be expressed in the placenta throughout pregnancy. This research aims to mechanistically describe the functional role of IL-27 at the maternal-fetal interface. To accomplish this aim, we will use human trophoblast organoids to determine the kinetics of IL-27 by trophoblast cells. Additionally, we will use a murine model of pregnancy to assess IL-27 production and localization at the maternal-fetal interface in individual concepti. Localized production will be visualized using immunohistochemistry of implantation site cross-sections from an IL-27 reporter (p28-GFP) mouse. Our second aim is to determine the impact of IL-27 signaling on viral replication in our model systems using Zika virus as a model pathogen. We will determine the contribution of IL-27 to antiviral activities by assessing viral burden through transcriptional and immunofluorescence readouts. We hypothesize that IL-27, through autocrine signaling, induces constitutive antiviral programming that directly restricts viral replication in trophoblasts due to its ability to induce STAT1 signaling. Overall, this project will elucidate the presence and functionality of IL-27 at the maternal-fetal interface within murine and human models.
Supported by grants from Pew Charitable Trusts Biomedical Scholars Program and Burroughs Wellcome fund
A novel heterodimer composed of IL-12p40 and CD5L promotes a unique cytokine profile from T cells
Authors: Hill Elizabeth M , Gerber Allison J , Fink Marc Y , Keegan Achsah Dorsey , Singh Nevil J 
Published: August 2023

Abstract
The IL-12 family of cytokines (IL-12, IL-23, IL-27, and IL-35) are heterodimers composed of α and β subunits. Both IL-12 and IL-23 utilize IL-12p40 (p40) as their β subunit yet exert distinct proinflammatory outcomes on the T cell response. Upon immune stimulation, the p40 monomer is also secreted from innate immune cells without its binding partners p35 and p19. In order to examine if p40 functions by interacting with additional partners, we generated a library of p40 binders using immunoprecipitation and mass spectrometry. Of the 19 potential binding partners identified biochemically, CD5L, a protein characterized for its role in regulating macrophages was the top hit. We generated a panel of dimer-specific monoclonal antibodies recognizing the p40-CD5L heterodimer. Using this reagent, we found that p40CD5L was present in the serum of both C57BL/6 and BALB/c mice at baseline and increased in the BAL of mice after allergic sensitization. Adoptive transfer and restimulation of transgenic T cells with recombinant p40CD5L resulted in significantly increased production of IL-4 and IL-10 – a unique cytokine profile that has not been identified with other p40-containing cytokines. From these results, we suggest that p40CD5L may function as a novel innate heterodimeric cytokine potentially promoting IL-4 and IL-10 responses.
Supported by grants from NIH (R21AI166330-02)
In Vitro Expansion of Antigen-Specific CD8+ T Cells from Human PBMCs Using Artificial T-cell Stimulating Microparticles
Authors: Sim Hajin , Livingston Natalie , Schneck Jonathan P 
Published: August 2023

Abstract
While current T cell-based immunotherapies have shown moderate success, clinical translation of these methods has been hindered by biological variability, limited efficacy, and high expense. Here, we create a novel T cell stimulating platform for antigen-specific CD8+ T cell activation and expansion, eliminating the extensive ex vivo expansion steps and thus making it accessible for many more cancer patients. The scaffold is formed using thiol-modified hyaluronic acid cross-linked with PEGDA, magnetic dextran nanoparticles, as well as DTT-reduced MHC-Ig dimer HLA-A*02 (A2) and anti-CD28. The gel is passed through a mesh device to form microparticles (MPs), which are then passively loaded with peptides (here, CMV and MART1) for ease of multiple expansions. They were plated with human PBMCs thawed overnight in culture media supplemented with human AB serum and a cocktail of cytokines (IL-1b, IL-2, IL-4, IL-6, and IFNγ). On days 7 and 14, cells were harvested and analyzed for proliferation of antigen-specific cells. After fourteen days of culturing PBMCs with A2 MPs, we could detect a significant expansion of antigen-specific T cells via flow cytometry. When stimulated with peptide-pulsed target cells for an intracellular cytokine assay, they expressed one or more of IFNγ, TNFα, and IL-2, demonstrating their polyfunctionality. We also observed a greater population of memory T cells, which indicates that these cells may persist longer in the body. We have developed a biomaterial-based T cell activation platform that provides critical signals, mimicking the lymph nodes. These MPs are effective in expanding antigen-specific T cells from human PBMCs, and these cells have robust killing of target splenocytes.
Design and Optimization of a Novel Proteomimetic Polymer Nanoplatform for the Co-delivery of Antigens with Adjuvants
Authors: Wang Max Mu , Choi MiRan , Zhang Bin , Gianneschi Nathan 
Published: August 2023

Abstract
We describe here a novel nanoplatform called the Protein-Like Polymer (PLP) with unique characteristics allowing for sustained delivery of tumor antigens in conjunction with STING agonists.

Methods
A library of PLP formulations were synthesized via ROMP and characterized. Cell uptake/functional assays were conducted with payload-specific T Cells. Ability of PLPs to co-deliver adjuvants was tested by electrostatically coupling 2′3′ cGAMP, forming stable nanostructures.

Results
Conjugating antigens to the polymer backbone using a cleavable disulfide linkage (reduces in APCs) resulted in increased endosomal localization and T cell proliferation/activation. Incorporating OEG side chains reduced enzymatic degradation while increasing immunogenicity and APC uptake. Increasing the density of side chains further improved vaccine efficacy and resistance to proteolysis. Antigen-PLP conjugates is effective only when paired with cells from their cognate system, demonstrating payload-specificity. Mice bearing established B16F10 tumors treated with gp100-PLPs resulted in increased survival time and reduced tumor burden with corresponding changes in immune cell profiles. Notably, STING-PLP complexes showed significantly smaller tumors at day 14 and allowed for subcutaneous administration of 2′3′ cGAMP. Studies looking at multiplexing pools of neoantigens onto one PLP and generation of immunological memory are ongoing.

Conclusion
The modularity of the PLP platform allows for complex nano-architectures including systems capable of delivering challenging compounds, ie small molecule STING agonists, subcutaneously through electrostatic coupling, highlighting its potential to revolutionize cancer vaccinology.

Supported by grants from the NIH (5F30CA257519-03)
Mechanisms underlying focused ultrasound-induced immunogenicity in melanoma
Authors: Pelka Sandra , Bandyopadhyay Sanmay , Pandey Sanjay , Guha Chandan , Macian Fernando 
Published: August 2023

Abstract
Advanced melanoma is an aggressive skin cancer with low survival rates. While the anti-tumor response relies on dendritic cells (DCs) to process tumor antigens, migrate to lymph nodes, and activate T-cells, within the tumor microenvironment (TME), DC activation is impeded. Subsequent immature DCs are a major mechanism underlying the ineffective T-cell response. We have shown that low-intensity focused ultrasound (LOFU), a novel therapy that delivers mechanical and thermal stresses to the tumor site, may enhance DC function in a B16 melanoma model. We hypothesize that LOFU enhances the dendritic cell-mediated anti-tumor response by inducing changes in tumor cell and dendritic cell autophagy and increasing pro-immunogenic factors in the TME. Autophagy is a lysosome-dependent process of cellular content degradation also implicated in the regulation of multivesicular body formation/degradation, and thus in the secretion of extracellular vesicles. Autophagy may subsequently modify the cargo of tumor-derived vesicles in a pro-immunogenic manner. We have found that LOFU treatment leads to increased LC3-II flux in melanoma cells and identified 82 proteins within melanoma-derived vesicles that are significantly enriched/reduced with LOFU, with enriched proteins partaking in processes such as antigen processing and intracellular trafficking. DCs pulsed with vesicles from LOFU-treated melanoma cells show enhanced activation of CD4+ and CD8+ T-cells, suggesting that LOFU may promote a DC-mediated anti-tumor response. Further understanding of DC involvement in the anti-tumor response will provide novel targets for the development of immunotherapies to improve survival and quality of life in advanced melanoma patients.
Supported by grant from the NIH (R01 CA226861)