bluuebunny/crossref_metadata_2025_split huggingface dataset:
Natural killer: dendritic cell activation using anti-CD137 mAb improves cetuximab mediated innate and adaptive immune responses in head and neck cancer (TUM7P.948)
Authors: Srivastava Raghvendra , Jie Hyun-bae , Li Jing , Ferris Robert 
Published: January 2023

Abstract
EGFR-specific mAb cetuximab mediated activation of NK by FcγRIIIa cross-linking is important for anti-tumor immunity.Following cetuximab activation,NK express costimulatory receptor CD137, which can be ligated to improve activity of cetuximab-activated NK,which promote DC maturation and CD8+ T-cell priming, leading to tumor antigen spreading and Th1 cytokine release through NK-DC cross-talk. We show that the clinically available CD137 agonist mAb augments cetuximab-mediated ADCC in FcγRIIIa-V/F donors,but not in poor binding FcγRIIIa-F/F.We show that cetuximab-activated NK induce CD137 expression on DC, which can be further stimulated by CD137 mAb, leading to enhanced DC maturation and cross-presentation capacity.In a prospective phase II trial of neoadjuvant cetuximab in stage III/IV HNC patients, intratumoral activated NK cells also upregulated CD137, which was influenced by FcγRIIIa-V/F polymorphism. Moreover, cetuximab treatment induces CD137 expression on intratumoral activated CTL.We show that FcγRIIIaV/F polymorphism on NK cells greatly influences their induction of CD137 expression, as does the EGFR receptor density on HNC.Thus, careful selection of HNC patients in combination immunotherapy using cetuximab plus CD137 mAb may be necessary, using EGFR level of expression and FcgammaRIIIa genotype, to optimize NK:DC priming of tumor antigen specific CTL.Results suggest a beneficial effect of combination of cetuximab and CD137 mAb in currently designed clinical trials.
Organotypic brain slice culture reveals fractalkine signaling alters the kinetics of microglia and medulloblastoma migration in the CNS tumor microenvironment (TUM7P.953)
Authors: Dorand Dixon , Barkauskas Deborah , Nthale Joseph , Myers Jay , Petrositue Agne , Huang Alex 
Published: January 2023

Abstract
Medulloblastoma (MB) is the most common pediatric Central Nervous System (CNS) malignancy and is capable of metastasis to distant sites. Little is known about the interactions between MB and the highly selective CNS immune environment; therefore, we investigated the role of microglia, the endogenous immune cells of the CNS, in MB microenvironment. Microglia reside in the brain parenchyma and help maintain the homeostatic immunosuppressive environment partially due to their expression of CX3CR1 for which Fractalkine (FKN) is the only known ligand. FKN is constitutively expressed by neurons and astrocytes leading to tonic inhibition of microglial activation. Our Enzyme Linked Immunosorbent Assay (ELISA) results demonstrate that MB can produce as much as 2.5 ng/mL/1x106 tumor cells/24hrs of FKN. Organotypic brain slice culture coupled with 2 photon laser scanning microscopy reveals that genetic deletion of CX3CR1 results in an increase basal microglia speed from 0.071 μm/min in CX3CR1+/- mice to 0.123 μm/min in CX3CR1-/- mice. In the presence of MB, microglia speed increases to 0.154 μm/min in CX3CR1+/- mice to 0.203 μm/min in CX3CR1-/- mice. This data supports the role of FKN signaling as an inhibitor of microglial activation. Interestingly, MB tumor cells move faster in brain slices of CX3CR1-/- mice (0.301 μm/min) compared to CX3CR1+/- mice (0.144 μm/min), further suggesting that FKN signaling may play a role in MB invasion as well as microglia movement.
New complement roles revealed by immune gene profiling in endometriosis and endometriosis-associated ovarian cancer (TUM7P.957)
Authors: Vlad Anda , Suryawanshi Swati , Huang Xin , Budiu Raluca , Elishaev Esther , Kim SungHwan , Tseng George , Edwards Robert 
Published: January 2023

Abstract
Endometriosis is a largely benign, chronic inflammatory disease that may predispose to some of the ovarian tumors histotypes, collectively called endometriosis-associated ovarian cancers (EAOC). Although it is widely accepted that chronic inflammation drives cancer, the immune deregulation in chronic precursors of ovarian cancer has not been systematically studied. We performed here the first comprehensive gene profile of the tissue immune microenvironment in normal endometrium, benign endometriosis and EAOC (n=133), using Nanostring and the nCounter® GX Human Immunology Kit comprising probes for 511 immune genes. We found that the complement pathway was most prominently upregulated, suggesting its involvement in the transition from chronic endometriosis to atypical (premalignant) endometriosis and to EAOC. Complement protein expression was confirmed via immunohistochemistry, which identified upregulation of complement in epithelial cells. To explore the roles of complement in early EAOC carcinogenesis, we derived several novel ovarian cell lines with conditional mutations that can be turned on and off, using the Cre-loxP system. Using this in vitro model we demonstrate that genetic instability triggers upregulation of complement genes in epithelial cells, but without an advantage on cell death. These findings reveal an early link between epithelium and innate immune environment and further support a paradigm shift on complement roles in supporting cancer growth.
Anchoring of TLR agonists in the plasma membrane of B cells for enhanced cellular vaccines. (VAC10P.962)
Authors: Szeto Gregory , Van Egeren Debra , Alejandro Brian , Liu Haipeng , Irvine Darrell 
Published: January 2023

Abstract
B cells are a unique lymphocyte subset that can integrate adaptive and innate receptor signaling. B cells can serve as cellular vaccines if appropriately activated (e.g., by TLR ligands) and loaded with antigen. However, activation can also lead to changes in homing receptor expression that abrogate their ability to traffic efficiently to lymphoid organs. To overcome this issue, we present a strategy to provide sustained TLR stimulation to B cells using brief (15 min pulse) exposure to lipid-tailed CpG oligonucleotides (lipoCpGs), which results in stable insertion of the TLR ligand in the plasma membranes of primary murine B cells. LipoCpG internalization was studied by flow cytometry and confocal microscopy, revealing that nearly 100% of lipoCpG was surface-localized for at least 2 h. LipoCpG-pulsed B cells exhibited sustained activation for 3 days in vitro as determined by expression of MHC I, MHC II, CD86 and CD69; expression was equivalent to levels on B cells continuously incubated with soluble CpG. Pulsed B cells did not significantly activate unpulsed B cells in a mixed coculture, indicating that inserted lipoCpG acts in an autocrine fashion. Finally, LipoCpG-pulsed B cells incubated with ovalbumin drove proliferation of OT-I CD8+ T cells in vitro. These data suggest that inserted lipoCpG provides strong activation signaling, potentially enabling rapid cell priming and prolonged in situ activation, facilitating development of B cell-based immunotherapies.
Targeting antigen-loss variant tumors through CD134 plus CD137 dual costimulation-mediated bystander programming (VAC10P.964)
Authors: Mittal Payal , Wang Xi , St. Rose Marie-Clare , Vella Anthony , Adler Adam 
Published: January 2023

Abstract
Tumor-specific cytotoxic CD8 T cell-based therapies can be efficacious in cancer patients. Nevertheless, disease progression often resumes due to outgrowth of antigen-loss variants (ALVs) that lack expression of the targeted MHC class I-restricted epitopes. Co-administration of agonists to the TNF/TNFR costimulatory family members CD134 (OX40) and CD137 (4-1BB) can program antigen-stimulated CD4 T cells to differentiate into cytotoxic Th1 cells, which can directly target MHC II+ tumors and provide classical helper function that facilitates effector differentiation in CD8 T cells responding to a linked epitope. Interestingly, these specific cytotoxic CD4 Th1 cells also imprint antigen-non-responding T cells with a similar ability to express IFN-γ and cytotoxic molecules. Because these bystander-programmed effector T cells are polyclonal, we currently hypothesized that they contain specificities capable of targeting ALVs. In both lymphoma and melanoma models, dual costimulation therapy was markedly enhanced when specific CD4 T cells were engaged to differentiate into cytotoxic Th1 effectors. This augmentation occurred not only when the tumor expressed the MHC class II-restricted epitope recognized by the specific cytotoxic CD4 Th1 cells, but importantly also with tumors modeling ALVs that lack the targeted epitope. Taken together, dual-costimulated tumor-non-specific CD4 T cells provide an alternate form of help that facilitates targeting of ALVs.
Development of an in vitro system for screening adjuvant combinations for induction of specific patterns of CD4 T cell differentiation (VAC10P.967)
Authors: Goldberg Michael , Kadolsky Ulrich , Yates Andrew , Porcelli Steven 
Published: January 2023

Abstract
Adjuvants are a critical component of many vaccines, but their mechanisms of action remain poorly understood. While great progress has been made in defining several pathways that are activated by various adjuvants, their impact on adaptive immunity remains largely unpredictable. Here, we developed an experimental system to screen multiple defined compounds with known adjuvant activity on the strength and quality of antigen-specific murine CD4 T cell priming in vitro. By screening combinations of suboptimal concentrations of 30 adjuvants, we identified combinations that showed remarkable synergy in priming naive CD4 T cells toward TH1, TH2, TH17 or TfH phenotypes. By applying a variety of computational algorithms to data from this in vitro approach, we identified synergy between agonists of different pattern recognition receptor families for induction of specific functional programs in CD4 T cells during priming. These were ranked by strength of polarization and with respect to robustness under different culture conditions. Immunizations in mice showed strong correlations between in vitro ranking and in vivo outcomes of antigen specific CD4 T cell responses. Our approach provides a method for analyzing large numbers of adjuvant combinations to predict their influence on the outcome of CD4 T cell priming in vivo. This should assist in the development of new adjuvants that enable more predictable and functionally polarized CD4 T cell responses following vaccination.
Neutralization of murine myeloid suppressor cells enhances the efficacy of GD2-specific CAR T cells directed against human sarcoma in a xenograft model (VAC11P.1010)
Authors: Highfill Steven , Smith Jillian , Long Adrienne , Cui Yongzhi , Orentas Rimas , Mackall Crystal 
Published: January 2023

Abstract
Genetically engineered T-cells expressing chimeric antigen receptors directly target cell surface antigens on tumor cells. CARs have shown remarkable activity in clinical trials for hematologic malignancies but remain unproven for the treatment of solid tumors. The disialoganglioside GD2 is considered a tumor-associated antigen for neuroblastoma and we have found that GD2 is expressed on pediatric sarcomas. Analysis of primary human tumor tissue samples taken from 18/18 patients with osteosarcoma and alveolar rhabdomyosarcoma (2/15) demonstrate robust expression of GD2 on the cell surface. Based on this finding, we developed assays to test the ability of GD2-CAR T-cells to target human sarcomas. We find that GD2-CAR T-cells specifically lyse GD2-expressing solid tumors in vitro even at low E:T, but fail to induce a response in vivo. In studying these xenografted tumors we discovered an expansion of murine CD11b+Gr1+ MDSC that inhibit human T-cell responses in vitro. This lead us to adopt a combinatorial therapeutic strategy in which we inhibited the suppressive potential of MDSC by administration of all-trans retinoic acid followed by the infusion of GD2-specific CAR T-cells. This resulted in significant improvements in overall survival and tumor growth. The results presented here suggest that ATRA coadministration with CAR T-cell therapy for solid tumors may yield a beneficial additive effect through the modulation of myeloid derived suppressor cells.
Defining the role of the novel CD4+CD26high T cell subset in adoptive cancer immunotherapy (VAC11P.1001)
Authors: Bailey Stefanie , Nelson Michelle , Schwartz Kristina , Bowers Jacob , Paulos Chrystal 
Published: January 2023

Abstract
Th17 cells mediate exquisite tumor eradication upon transfer into mice bearing large, established melanoma. Unfortunately, the polarizing cytokines (TGF-β, IL-6, etc.) required to generate Th17 cells are not yet FDA approved for clinical use. Therefore, we sought to determine if these IL-17-producing CD4+ cells could be enriched from peripheral blood. Herein, we found that functional Th17 cells could be isolated from blood, without the need for polarization, by using the surface marker CD26. Excitingly, very few (5e4 cells) of these CD26-rich cells, termed CD26high T cells, displayed striking antitumor activity and survival upon transfer into melanoma-bearing mice compared to CD26negative or bulk CD4+ T cells. Of translational significance, we found that human CD26high cells had heightened cytotoxicity and caused greater cancer cell death in vitro. Interestingly, both murine and human CD26high T cells boast a unique phenotype compared to their bulk CD4+ or CD26negative counterparts, including: enhanced expression of the cytokines IL-17, IFN-γ, and IL-22, transcription factors RORγ and Tbet, and cytokine receptors CD25, CD122, and CD127. Collectively, these findings describe a novel T cell subset that has promise in cell-based cancer therapies due to its unique polyfunctionality and engraftment. Moreover, enrichment of CD26high T cells from peripheral blood bypasses the need for polarization, thereby enhancing the translational potential for Th17 cells in immunotherapy.
Human anti-MUC1 antibodies elicited by a prophylactic cancer vaccine for mAb and CAR-modified T cell immunotherapies (VAC11P.1004)
Authors: Lohmueller Jason , Finn Olivera 
Published: January 2023

Abstract
MUC1 is a tumor-associated protein that is abnormally expressed on all human adenocarcinomas. In our recent clinical trial, individuals at-risk for colonic adenoma received a prophylactic MUC1 cancer vaccine. Many individuals responded by producing high-affinity anti-MUC1 antibodies. We isolated a panel of these antibodies and seek to develop them as antibody and CAR T cell cancer immunotherapies. Our collaborators used a novel method combining proteomics and next-gen DNA sequencing to identify and recombinantly produce 12 high-affinity antibodies from the sera of vaccinated individuals. By ELISA we determined that the antibodies bind with high-affinity to diverse epitopes within the MUC1 vaccine peptide. We also found that several of the antibodies can stain MUC1+ pancreas, colon, and breast cancer cell lines but not a cell line expressing normal MUC1. We then determined that the antibodies could stain MUC1+ human pancreas and colon tumor samples but not normal tissues. Having established tumor-specificity we are now assessing the antibodies in a series of functional assays. Finally, we have constructed CAR lentiviral constructs using the variable regions of the antibodies as targeting domains and are testing their ability to mediate MUC1-specific target cell lysis. As the antibodies are human-derived, indication of efficacy in pre-clinical testing has the potential to directly enable clinical trials and hopefully approval of these reagents for therapy of cancer patients.
Reduced ectonucleotidase expression on TGF-β independent Th17 improves tumor control (VAC11P.1006)
Authors: Chatterjee Shilpak , Kesarwani Pravin , Thyagarajan Krishnamurthy , Soloshchenko Myroslawa , Webster Christine , Paulos Chrystal , Mehrotra Shikhar 
Published: January 2023

Abstract
Amongst various effector CD4+ T cells, T helper (Th)-17 subsets hold promise in adoptive T cell transfer therapy for cancer. However, ex vivo programming of Th17 cells in presence of TGF-β increases cell surface expression of ectonucleotidases CD39 and CD73, that in turn increases susceptibility to immunosuppression and reduced effector phenotype. Our data shows that ATP mediated suppression of IFN-γ production by Th17 cells can be overcome either by genetic ablation of CD73 or generating TGF-β independent Th17 in presence of IL-1β. An in vivo comparison of anti-tumor potential between TGF-β dependent and independent Th17 showed that Th17 cells generated in presence of IL-1β are more efficacious than TGF-β dependent Th17 cells in controlling B16 melanoma. IL-1β dependent Th17 cells are also highly polyfunctional and express high level of pathogenic genes, such as T-bet and granzyme B that results in increased cytotoxic potential. It is likely that effector property of IL-1β dependent Th17 is due to their high glycolytic capacity, since generating IL-1β dependent Th17 cells in pyruvate containing media impaired glycolysis and its anti-tumor potential. Thus, our data suggests that owing to TGF-β induced ectonucleotidase expression, ex vivo culture conditions for generating Th17 cells need to be reconsidered in favour of TGF-β independence for exploiting their full potential in adoptive T cell therapy.
Inhibition of glycogen synthase kinase 3 beta promotes long term cytokine-free survival of chimeric antigen receptor-expressing T cells (VAC11P.1008)
Authors: Sengupta Sadhak , Sampath Prakash 
Published: January 2023

Abstract
Application of chimeric antigen receptors (CAR), where an antibody or ligand-binding domain is fused with the zeta signaling chain of the T cell receptor (TCR), is a novel and alternative strategy to bypass the failure of cytotoxic immune response induction by most tumor cells. The resulting CAR+ T cells are redirected by the neo-specificity to attack tumors expressing the surface antigen or receptors recog¬nized by the gene-modified TCRs. Although CARs have shown impressive preclinical efficacy, translational success is restricted due to the limited viability of the T cells in absence of IL2 consequent to preconditioning chemotherapy. Although multiple doses of T cell transfer and systemic administration of IL2 are effective for short time periods, unregulated overdose of T cells and IL2 result in immune toxicity. In this study we have identified that inhibition of glycogen synthase kinase 3 beta (GSK3β) maintained increased survival of the CAR+ T cells in the absence of IL2, upon activation with target-specific tumor cells. Increased CAR+ T cell proliferation was observed upon GSK3β-inhibition in comparison to non-treated CAR+ T cells. GSK3β-inhibited CAR+ T cells also showed increased CD127 (IL7R) expression compared to the bystanders as well as non-treated CAR+ T cells indicating a possible development of memory population. This is a fundamental progress towards the development of a cancer immunotherapy strategy that will avoid the complications arising from IL2-toxicity.
Autologous tumor vaccine modified with recombinant new castle disease virus expressing IL-7 promotes anti-tumor immune response (VAC12P.1022)
Authors: Liu Haiyan , Zhao Lixiang , Mei Yu , Sun Qing , Guo Linghua , Liu Xiufan 
Published: January 2023

Abstract
Autologous tumor cell modified with non-lytic Newcastle Disease Virus (ATV/NDV) is a promising vaccine for cancer immunotherapy. IL-7 plays a critical role in lymphocyte development and homeostasis. In order to improve the efficacy of ATV/NDV, we inserted IL-7 gene into the genome of non-lytic NDV strain LX using reverse genetic system. The insertion of the IL-7 gene neither affected the main features of NDV replication nor its tumor selectivity. The gene product was biologically active and stable. Then we tested the anti-tumor effects of the autologous tumor vaccine modified with LX/(IL-7) in the melanoma model. We showed that B16-F10 cells modified with LX/IL-7 induced a strong anti-tumor activity against melanoma both in prophylaxis and therapeutic models. The IFN-γ production and the cytotoxicity of tumor-specific CD8+ T cells were significantly enhanced after immunization with B16-F10 cells modified with LX/(IL-7) in both models. Although the tumor infiltrating CD4+ T cells and CD8+ T cells were both increased and their IFN-γ productions were also up-regulated, the anti-tumor activity of the tumor vaccine modified with LX/(IL-7) was dependent on CD8+ T cells. Our results demonstrated that the autologous tumor vaccine modified with NDV/(IL-7) could promote the anti-tumor immune responses mediated by CD8+ T cells and significantly improve the efficacy of the ATV/NDV.
Adenoviral-vectored tuberculosis vaccine AERAS-402 enhances T cell cytokine response in TB+ patients. (VAC12P.1026)
Authors: Graves Andrew , van Zyl-Smit Richard , Esmail Ali , Bateman Mary , Dawson Rodney , Goldin Jonathan , van Rikxoort Eva , Douoguih Macaya , Pau Maria , Sadoff Jerald , Bennett Sean , Snowden Margaret , Gearhart Jacqueline , Ishmukhamedov Sadritdin , Schrager Lewis , McClain J. , Bateman Eric , Hokey David 
Published: January 2023

Abstract
Development of a vaccine to prevent pulmonary tuberculosis remains a public health priority. AERAS-402 is a live, replication-deficient adenovirus 35-vectored TB vaccine expressing the mycobacterial antigens 85A, 85B, and TB10.4. While this vaccine has been shown to be safe in TB uninfected patients, little is known about the performance of this vaccine in TB-experienced populations. The safety and immunogenicity of AERAS-402 was evaluated in a Phase 2a randomized, placebo-controlled, double-blinded dose-escalation study in HIV-negative South African adults with either active or recent pulmonary TB infection. Peripheral blood mononuclear cells isolated from trial patients were assessed via intracellular cytokine staining (ICS) examining T cells for interferon-γ (IFN-γ), TNF, and IL-2 production. While modest CD4+ T cell responses were observed, the ICS data indicate the induction of a robust, dominant CD8+ T cell response not observed in placebo patients, including instances of total cytokine responsive cells approaching 6% of CD8+ T cells in some subjects. Polyfunctional analysis revealed that vaccine-specific T cells were primarily secreting IFN-γ either alone or in combination with TNF following stimulation. These data suggest that AERAS-402 is able to boost T cell responses in patients with a history of TB infection.
Gp96-Ig DNA transfer into muscle by electroporation induces antigen specific cellular immune responses (VAC12P.1027)
Authors: Strbo Natasa , Gonzalez Louis , Podack Eckhard 
Published: January 2023

Abstract
The secreted gp96 vaccine approach is based on the use of cellular vaccines (293 cells) that continuously secrete gp96-Ig associated with antigenic peptides that are cross presented to CD8 T cells in vivo. Cellular gp96 vaccines are highly active in generation CTL for cancer therapy and SIV prophylaxis. Here, we investigate the use of gp96-Ig cDNA as vaccine platform. An optimized electroporation approach was used to deliver the B45 plasmid encoding secreted gp96-Ig and HIV-gag-gfp or ovalbumin genes. The B45 plasmid is a bovine papilloma virus derived vector from which the potentially transforming genes E5, E6, and E7 have been removed as well as the L1 and L2 viral capsid genes. The plasmid (50ug) was injected into the muscle (musculus tibialis anterior) of C57/Bl6 mice followed by electric field pulse that induced temporary and reversible pores in the muscle cell membrane. 48h after i.m. injection and EP-delivery, we found gfp (HIV-gag-gfp) expression in the muscle cells. To determine gp96-Ig secretion and cross presentation to CD8 cells we used cDNA for ovalbumin as antigen in conjunction with gp96-Ig and expansion of OT-I TCR-transgenic cells as readout. Vaccination by electroporation increased the frequency of OT-I in the spleen and in the mucosal compartment (lamina propria, small intestine).This simple, efficient and reproducible method of delivery may have great potential for further development of DNA gp96-Ig vaccination.