bluuebunny/crossref_metadata_2025_split huggingface dataset:
Use of an antigen specific T cell line to investigate unresponsiveness to PD-1 blockade.
Authors: Lodge P. Anne , Tjoa Benjamin A 
Published: January 2023

Abstract
Antibodies blocking the PD-1/PD-L1 interaction have shown great promise in cancer therapy. Inhibition of PD-1 signaling allows previously exhausted tumor specific T cells to resume proliferation and tumor elimination. Nivolumab is an antibody that binds to PD-1 and has been approved for treatment of melanoma, renal cell carcinoma and non-small cell lung cancer. In vitro studies show that nivolumab can enhance the secretion of IFNγ in a mixed lymphocyte reaction or recall antigen stimulation. To facilitate study of checkpoint inhibition, we attempted to use an antigen specific T cell line for an in vitro assay of PD-1 inhibition by nivolumab. We discovered that the myelin basic protein specific T cell line, BC3, is resistant to the effect of nivolumab in spite of expressing PD-1 and the presence of PD-L1 on the surface of the antigen presenting cells. Unresponsiveness to nivolumab is not donor specific but may reflect the nature of the response to specific antigens. In vitro responses to CMV and tetanus toxoid are enhanced by PD-1 blockade but reactivity to EBV nuclear antigen, as with the BC3 T cells, is not affected. Further study of these differences in sensitivity to nivolumab may allow better targeting of therapy to indications most likely to benefit.
hetIL-15 therapy results in tumor control by promoting tumor infiltration of CD8+ T cells with increased cytotoxic potential and decreased PD-1 expression.
Authors: Pavlakis George N , Ng Sin Man , Dimas Konstantinos , Valentin Antonio , Nagy Bethany , Alicea Candido , Felber Barbara K , Bergamaschi Cristina 
Published: January 2023

Abstract
Interleukin-15 (IL-15) is a common γ-chain cytokine that plays a significant role in the activation and proliferation of T and NK cells and holds great potential in fighting cancer. We have previously shown that bioactive IL-15 in vivo comprises a complex of the IL-15 chain with the soluble or cell-associated IL-15 receptor alpha chain that are together termed heterodimeric IL-15 (hetIL-15). We investigated the anti-tumor efficacy of hetIL-15 in several mouse cancer models. Repeated injections of hetIL-15 were effective in delaying tumor growth in the MC38 colon carcinoma and the B16 melanoma models. The beneficial effects of hetIL-15 therapy included a significantly reduced onset of lung metastasis in 4T1 breast cancer-bearing mice. We study the mechanisms of antitumor effects of hetIL-15 in several tumor models. hetIL-15 administration promoted a dramatic increase of tumor infiltration and persistence of CD8+ T cells, including tumor specific T cells, and resulted in an increased CD8+/Treg ratio. Tumor-resident CD8+ T cells showed features of effectors cells and were characterized by increased proliferation (Ki67+) and high cytotoxic potential (Granzyme B+). In the absence of hetIL-15, the smaller population of tumor-infiltrating T cells exhibited high levels of the exhaustion marker PD-1, potentially limiting their anti-cancer effectiveness. Provision of hetIL-15 resulted in a significant decrease in lymphocyte expression of PD-1, alleviating one potential mechanism for the exhaustion phenotype. Preclinical cancer studies support the use of hetIL-15 in tumor immunotherapy approaches to promote the development of anti-tumor responses by favoring effector over regulatory cells.
Identification of B7-H4 as a functional surface marker for monocytic myeloid-derived suppressor cells
Authors: Li Limin , Diao Wenli , Liu Ying , Li Shan , Liu Yuan , Zen Ke 
Published: January 2023

Abstract
Myeloid-derived suppressor cells (MDSC), a heterogeneous population of immature myeloid cells, have a strong ability to suppress T cell proliferation and function. However, so far there is no universal surface marker of MDSC for mice and humans. Here we report that B7-H4 is expressed in monocytic myeloid-derived suppressor cells (M-MDSCs, CD11b+Ly-6G−Ly-6Chigh) but not granulocytic myeloid-derived suppressor cells (G-MDSCs, CD11b+Ly-6GhighLy-6Cmid). Both M-MDSCs and G-MDSCs are isolated from tumor-bearing mice. Cell sorting of B7-H4-positive and B7-H4-negative cells from the M-MDSC fraction isolated from tumor-bearing mice shows that B7-H4-positive M-MDSCs have significantly stronger T cell suppressive function than B7-H4-negative M-MDSCs independent of the expression level of iNOS and Arginase 1. Consistent with this, M-MDSCs from B7-H4-deficiency mice display a reduced immunosuppressive function compared to M-MDSCs from wild-type mice. Furthermore, by inducing B7-H4-positive M-MDSCs from mouse bone marrow-derived MDSCs or from human peripheral blood mononuclear cells, glucocorticoid treatment significantly suppresses T cell proliferation and function. In summary, our findings identify B7-H4 as a novel functional surface marker for both mouse and human M-MDSCs.
Reelin promotes the adhesion and drug resistance of multiple myeloma cells via integrin β1 signaling and STAT3 activation
Authors: Ge Qing , Lin Liang , Lu Jin 
Published: January 2023

Abstract
Reelin is an extracellular matrix (ECM) protein that is essential for neuron migration and positioning. The expression of reelin in multiple myeloma (MM) cells and its association with cell adhesion and survival were investigated. Overexpression, siRNA knockdown, and the addition of recombinant protein of reelin were used to examine the function of reelin in MM cells. Clinically, high expression of reelin was negatively associated with progression-free survival and overall survival. Functionally, reelin promoted the adhesion of MM cells to fibronectin via activation of α5β1 integrin. The resulting phosphorylation of Focal Adhesion Kinase (FAK) led to the activation of Src/Syk/STAT3 and Akt, crucial signaling molecules involved in enhancing cell adhesion and protecting cells from drug-induced cell apoptosis. Reelin-induced phosphorylation of STAT3 and Akt could be suppressed by inhibitors against Src and Syk. The myeloma cell survival promoted by reelin could be inhibited by STAT3-specific siRNAs. Together, these findings indicate reelin’s important role in the activation of integrin β1 and STAT3/Akt pathways in multiple myeloma and highlight the therapeutic potential of targeting reelin/integrin/FAK axis.
Cancer cell-derived transforming growth factor (TGF)-β and myeloid cell-derived interleukin (IL)-1β synergize to promote type 2 inflammation and breast cancer progression
Authors: Wu Te-Chia , Xu Kangling , Banchereau Jacques , Palucka Anna Karolina 
Published: January 2023

Abstract
Human breast cancer displays type 2 inflammation that promotes cancer progression and can be linked with interleukin (IL)-13 signature. The underlying molecular mechanisms include cancer-derived thymic stromal lymphopoietin (TSLP). Here, we found that IL-13 correlates with high levels of IL-1β in primary breast cancers. IL-1β levels correlate with disease stage indicating association with cancer invasiveness. IL-1β induces TSLP promoter activation, enhanced transcription and TSLP protein production from breast cancer cell lines. Administration of IL-1 receptor (IL-1R) antagonist Anakinra prevents breast cancer progression in a humanized mouse model of breast cancer. Conversely, breast cancer cells induce IL-1β transcription and secretion in dendritic cells (DCs) and monocytes. This is mediated by cancer cell-derived transforming growth factor (TGF)-β. TGF-β activated kinase 1 (TAK1) is involved in caspase-1 activation and subsequent IL-1β production. Administration of TGF-β neutralizing antibody prevents breast cancer progression in vivo and this is associated with a blockade of IL-13 and IL-4 production in the tumor. Thus, TGF-β - IL-1β cross-talk drives type 2 inflammation and contributes to breast cancer progression.
BET bromodomain inhibition synergizes with immune checkpoint blockade to facilitate anti-tumor response in a murine model of non-small cell lung cancer harboring activating KRAS mutation
Authors: Adeegbe Dennis O , Freeman Gordon J , Wong Kwok-Kin 
Published: January 2023

Abstract
KRAS mutations accounts for about 30% of human non-small cell lung cancer (NSCLC) and represent one of key oncogenic drivers of lung cancer. While recent advances in targeted therapy have shown promise, effective therapy for lung cancer still remains a challenge. Given the demonstrated efficacy of BET bromodomain inhibitors in treatment of hematologic malignancies and the emerging clinical benefit of immunotherapeutic agents such as anti-PD1 antibody, we sought to determine whether a combinatorial approach involving these agents could synergize to promote anti-tumor response in genetically engineered mouse model of NSCLC harboring activating kras mutation. Upon administration of the BET bromodomain inhibitor JQ1, and/or α-PD-1 antibody, we evaluated by FACS, the dynamics of tumor-associated immune cells. Notably, treatment with JQ1 alone or in combination with α-PD1 resulted in increased CD8:Treg ratio in the tumors, likely due to the reduced proportions of Treg cells. Interestingly, while JQ1 treatment alone led to remarkable down-regulation of PD-1, α-PD1 treatment resulted in augmented T cell activation evidenced by increased expression of CD69 on tumor-CD8+ T cells. When tested ex-vivo, T cells isolated from tumors of double agent therapy showed the most enhanced ability to secrete IFN-γ relative to either agent alone. Lastly, in survival studies, the combination of both drugs was superior in reducing tumor burden and prolonged the survival of tumor-bearing mice demonstrating a synergistic effect between the two agents. Collectively, our findings highlight the therapeutic potential of combining BET bromodomain inhibition with immune checkpoint blockade for treatment of solid malignancies such as lung adenocarcinomas.
Cancer Immunosurveillance by Tissue-resident Innate Lymphoid Cells and Innate-like T Cells
Authors: Dadi Saida , Chhangawala Sagar , Whitlock Benjamin M , Franklin Ruth A , Luo Chong , Oh Soyoung A , Toure Ahmed , Pritykin Yuri , Huse Morgan , Leslie Christina S , Li Ming O 
Published: January 2023

Abstract
Malignancy can be suppressed by the immune system in a process termed immunosurveillance. However, to what extent immunosurveillance occurs in spontaneous cancers and the composition of participating cell types remain obscure. Here we show that cell transformation triggers a tissue-resident lymphocyte response in oncogene-induced murine cancer models. Non-circulating cytotoxic lymphocytes, derived from innate, TCRαβ and TCRγδ lineages, expand in early tumors. Characterized by high expression of NK1.1, CD49a and CD103, these cells share a gene expression signature distinct from those of conventional NK cells, T cells and invariant NKT cells. Generation of these lymphocytes is dependent on the cytokine IL-15, but not the transcription factor Nfil3 that is required for the differentiation of tumor-infiltrating NK cells, and IL-15, but not Nfil3, deficiency results in accelerated tumor growth. These findings reveal a tumor-elicited immunosurveillance mechanism that engages unconventional type 1-like innate lymphoid cells and type 1 innate-like T cells.
Polyguanine-conjugated antigens target scavenger receptors in dendritic cells and induce antitumor immunity against melanoma
Authors: Ciccotelli Jo Erika Narciso , Erdos Geza , Falo Louis D 
Published: January 2023

Abstract
We have developed a vaccine design involving the conjugation of polyguanine (Poly(dG)) to protein or peptide antigens for the specific targeting of scavenger receptors in dendritic cells with intrinsic adjuvant capabilities. We have shown that Poly(dG) conjugation to ovalbumin generates stable particle conjugates (Poly(dG)-OVA) that induce greater T cell (helper, cytotoxic, and memory) and antibody responses. In this study, we show that Poly(dG)-OVA treatment of DCs in vitro induces greater antigen internalization and expression of activation markers CD86, MHC II, and CCR7. In vivo, intradermal injection of Poly(dG)-OVA resulted in the formation of an antigen depot in the skin lasting for several days and an increase in recruited immune infiltrates. Increased association of Poly(dG)-OVA is observed with MHC II- and CD11c-expressing cells in the skin compared to OVA. This is accompanied by the expression of pro-inflammatory CCL2 and IL-6 in the skin. Poly(dG)-OVA was also able to internalize more in skin migrating DCs and was transported more efficiently to draining lymph nodes. Poly(dG)-OVA immunization of B16-OVA-bearing mice contributed to delayed tumor progression and improved survival. We also developed Poly(dG)-Mgp10025–33 and Poly(dG)-TRP2180–188 to study melanoma-specific responses. Immunization with the these conjugates led to significant peptide-specific in vivo cytotoxic T cell responses compared to unconjugated peptides. Mice with B16 tumors immunized with combined Poly(dG)-Mgp100 Poly(dG)-TRP2 had delayed tumor progression and improved survival. Our results show that Poly(dG) conjugation of antigens is an effective strategy for the development of antitumor vaccines with improved immune responses.
Mechanisms of activity of the combination TLR4 agonist and emulsion adjuvant GLA-SE
Authors: Orr Mark T , Desbien Anthony L , Cauwelaert Natasha D , Reed Steven G 
Published: January 2023

Abstract
GLA-SE is a safe and potent clinical stage vaccine adjuvant consisting of the synthetic TLR4 agonist glucopyranosyl lipid adjuvant (GLA) formulated in a squalene-in-water emulsion (SE). GLA-SE augments both TH1 and IgG2-skewed antibody responses to a variety of vaccine antigens. The adjuvanticity of GLA is critically dependent on its formulation in SE. Individually GLA promotes IgG2 responses with little detectable TH1 induction, whereas SE promotes IgG1 and TH2 responsiveness. The combination of GLA-SE mediates TH1 induction via MyD88 and TRIF signaling and activation of the inflammasome to produce IL-18. The TH1 induction activity also depends on an early burst of type I and II interferons, the latter of which is made by memory CD8 T cells, NK cells, and neutrophils.
In addition to TH1 responses GLA and SE synergize to promote the numbers of antigen-specific B cells and TFH cells quickly after immunization. This is reflected in a substantial increase in the IgG2-skewed antibody response. Promotion of B cell and TFH responses depends on IL-18 signaling and the presence of lymph node resident macrophages that take up the adjuvant. Disruption of either of these axes is sufficient to ablate the antigen-specific B cell and antibody responses. Surprisingly the generation of TFH cells did not depend on lymph node resident macrophages, whereas TH1 responses were dramatically reduced under these conditions. We identify GLA-SE as an adjuvant capable of inducing robust B cell and antibody response that operates through interaction with LN macrophages and dependent on IL-18 signaling but independent of the number of antigen-specific TFH cells.
Cutaneous vaccination with TLR2 agonists results in the suppression of Th1 responses
Authors: Johndrow Christopher T , Goldberg Michael F , Johnson Alison J , Porcelli Steven A 
Published: January 2023

Abstract
Novel effective vaccines are key to solving many global health problems. Past efforts to rationally design vaccines that elicit strong cell-mediated responses have largely failed. Vaccine adjuvants, particularly toll-like receptor (TLR) agonists, are a promising tool to shape immune responses by polarizing CD4 T cells to distinct fates. Use of these adjuvants to design novel vaccines requires the full characterization of their effects on CD4 T cell differentiation, particularly in regard to the route of vaccination. In a large scale in vitro screen of vaccine adjuvants, we demonstrated that Pam3CSK4, a synthetic mimic of bacterial lipoproteins with TLR1/2 agonist activity, strongly polarized CD4 T cells to the Th1 fate (IFN-γ+, T-bet+) supporting cell-mediated responses. We adapted a vaccination system to study CD4 T cell polarization in vivo using ovalbumin-specific transgenic CD4 T cells. This in vivo system showed a route-specific effect, with strong Th1 responses from intravenous immunization with ovalbumin plus Pam3CSK4, but a lack of Th1 responses with immunization through more clinically relevant cutaneous routes. Interestingly, Pam3CSK4 in combination with a known Th1 polarizing adjuvant, CpG ODN, showed little to no Th1 response suggesting a dominant suppressive mechanism. Furthermore, suppression of Th1 differentiation occurred in response to both TLR1/2 and TLR6/2 agonists. Using a variety of cell-ablative mouse models we were able to reverse suppression of Th1 responses and identify specific candidates of innate immune cells mediating suppression during vaccination with TLR2 agonists. Current work focuses on mechanisms of suppression by TLR2 agonists in the skin and relevance to improving vaccine design.
Performance of Keyhole Limpet Hemocyanin (KLH) as an Antigen Carrier for Protein Antigens Depends on KLH Property and Conjugation Route
Authors: Bi Shuguang , Bailey William , Brisson Catherine 
Published: January 2023

Abstract
KLH naturally exists either as high molecular weight (HMW) cylinders composed of 20 copies of subunits, or as separate subunits (Subunit). The molecular weight of KLH Subunit is about 400kDa. Owing mainly to its large size, and the abundant and complex carbohydrate structures on its peptide chains, KLH has been widely used for decades as an immune stimulant, antigen carrier, and vaccine component. But, how to characterize the properties of KLH as an antigen carrier through functional assays remains to be determined. At Stellar, we conjugated KLH of different formats and sources with ovalbumin (OVA) and fluorescence tag-conjugated OVA through different conjugation routes. Dendritic cells (DCs) derived from mouse bone marrow mononuclear cells and mouse DC line JAWSII were pulsed with these conjugates. By utilizing DC uptake assays and measuring the fluorescence intensity of DC lysate supernatant, the amount of KLH- OVA-fluorescence conjugates uptaken by DCs was quantitatively determined. With flow cytometry, both the DC maturation markers, and the amount of ovalbumin epitopes that are presented by MHC II molecules of DCs, were measured. To further compare the property of KLH-protein conjugates as potential vaccines, we vaccinated mice with OVA. OVA-specific memory T cells extracted from the vaccinated mice were stimulated with DCs that had been pulsed with KLH-OVA conjugates. T cell proliferation and cytokine release from these stimulated T cells were quantitatively measured and compared. To summarize, with the functional assays we designed, we found that the performance of KLH as a carrier for protein antigens depends on KLH property and conjugation route.
Development of a tri-agonist compound library used to determine optimal adjuvanticity of a Q fever vaccine
Authors: Tom Janine , Albin Tyler , Burkhardt Amanda M , Felgner Philip , Esser-Kahn Aaron 
Published: January 2023

Abstract
Vaccines are one of the most successful treatments for disease. However, the development of more effective vaccines is still necessary for diseases, such as malaria and HIV. Recently, researchers have demonstrated that administering a target antigen with multiple immune agonists, especially Toll-like receptor (TLR) agonists, can enhance specific immune responses. Thus, suggesting that vaccines will require synergistic activity of multiple agonists to effectively activate the immune system and eliminate disease. In addition, vaccine development would greatly benefit from understanding the effect distinct combinations of immune agonists have on immune activity. This information would allow us to more rationally design vaccines that elicit directed and prolonged immune responses. Previously, we probed immune system responses by conjugating three TLR agonists together. Testing our construct in vitro and in vivo, we were able to modulate innate immune and antibody responses, suggesting downstream changes in immune signaling and adaptive immune activation. Here, we synthesized a library of tri-agonist compounds and examined how different covalently linked TLR agonist combinations affected immune activation via NF-kB activity and cytokine production. We are currently testing our constructs on a Q fever vaccination model, where Q fever is a current bioterrorism threat, to examine changes in T and B cell populations, antibody responses, and efficacy of the tri-agonist adjuvants. From our studies, these tri-agonist scaffolds can inform us about immune system signaling and activation in order to study immune activation mechanisms. Therefore, allowing us to potentially design more effective vaccines.
Understanding the cellular impact of low dose rapamycin treatment.
Authors: Johnson Jenny Lynn , Thomas Paul Glyndwr , McGargill Maureen Ann 
Published: January 2023

Abstract
The need for improved influenza vaccines is highlighted every year through the infection of vaccinated patients and the emergence of new pandemic strains. Recent work from our lab showed that a low dose of rapamycin given during influenza vaccination significantly improved protection against subsequent infection with multiple strains. These data indicate that dampening aspects of the immune response paradoxically enhances immunity to multiple subtypes of influenza virus, which is the goal of a “universal” influenza vaccine. However, it is still unclear how the low dose of rapamycin enhances immunity to different influenza subtypes and what particular cellular population is important for this increased protection. In order to better understand how to generate the optimal immune response against multiple influenza subtypes, we are investigating how this low dose of rapamycin impacts B cell signaling. Interestingly, we found that a low dose of rapamycin blocks B cell class switching without having an impact on proliferation. Thus, we are investigating which components of the mTOR pathway are required for class switching, but not proliferation. Early work indicates that this dose of rapapmycin alters the speed and activation potential of several mTOR signaling components. Using this system, we hope to identify the signaling components and kinetics that generate the optimal immune response to multiple subtypes of influenza, which is important for generating a universal influenza vaccine.