bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract B011: Human IL-2 mutant exhibits a higher antitumor efficacy and lower toxicity than wildtype IL-2 in different preclinical contexts
Authors: Carmenate Tania , Montalvo Magela , Rojas Gertrudis , Fuentes Dasha , Leon Kalet 
Published: November 2016

Abstract
High dose IL-2 is registered for the treatment of melanoma and renal cell carcinoma, however this therapy has limited efficacy and severe toxicity. It has been shown that in patients HD IL-2 drives preferential expansion of regulatory T cells, which dampen the antitumor immunity. This results explain at least partially the limited efficacy of this therapy. In this work, we characterize a human IL-2 mutant which differs from wildtype IL-2 by four mutations at the interface with the a-subunit of IL-2R. In vitro, IL-2 mutant induces proliferation of CD8+CD44hi and NK1.1 cells as efficiently as wtIL-2 does, but it shows a reduced capacity to induce proliferation of CD4+Foxp3+ regulatory T cells. In vivo, IL-2 mutant clearly induces preferential proliferation of CD8+CD44hi witha quite reduced proliferation of CD4+Foxp3+ regulatory T cells. The IL-2 mutant shows higher antimetastatic effect than wtIL-2 in several transplantable tumor models. i.e. the experimental metastasis model of MB16F0; the experimental and spontaneous metastasis models for the mouse pulmonary carcinoma 3LL-D1222 and the metastasis mammary carcinoma 4T1. Moreover initial data also support a higher potency of this IL2 mutant, when used in combination with other immune-therapies. Relevantly, the IL-2 mutant also exhibits lower lung and liver toxicity than does wtIL-2 when used at high doses in mice. The IL-2 mutant described could be a good candidate for improving cancer therapy based on IL-2.
Citation Format: Tania Carmenate, Magela Montalvo, Gertrudis Rojas, Dasha Fuentes, Kalet Leon. Human IL-2 mutant exhibits a higher antitumor efficacy and lower toxicity than wildtype IL-2 in different preclinical contexts [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B011.
Abstract B049: Immunotherapy of EBV associated gastric cancer using EBV specific T cells
Authors: Moon Jae Seung , Kim Jung Ho , Lee Sang Kyou 
Published: November 2016

Abstract
Epstein-Barr virus (EBV)-associated gastric cancer (EBVaGC) is mediated by EBV-infected malignant cells in gastric intestinal tract. About 10% of gastric carcinoma are EBVaGC worldwide, and over 85,000 patients are developed EBVaGC annually. EBVaGC shows several different aspects with EBV-negative gastric cancer such as clinicopathology, CpG island methylation status and expression of EBV-related genes. Therefore, immunotherapy to treat EBVaGC would be a good approach because expression pattern of EBVaGC is evidently different with that of EBV-negative cells in stomach. Adoptive T cell transfer has been effective for treatment of EBV-positive nasopharyngeal carcinoma (NPC) and lymphomas, which express similar EBV antigens to those of EBVaGC cells. Thus, we wanted to use system of NPC clinical trials to EBVaGC. Targeted EBV antigens expressed both in NPC and EBVaGC were latent membrane proteins (LMPs)-1,2 and EBV-determined nuclear antigen (EBNA) 1. To generate EBV-specific cytotoxic T lymphocytes (EBV-CTLs), we firstly isolated peripheral blood mononuclear cells (PBMCs) from healthy human donor blood by using Ficoll and sorted dendritic cells (DCs) to deliver LMP2A and EBNA1 expressing-vectors by nucleofection into DCs. Nucleofected DCs were cocultured with PBMCs in order to cross-present EBV antigens toward CD4+, CD8+ T cells in DC maturation condition. Generated EBV-CTLs were incubated with IL-2, 7, 15-supplemented CTL media for inducing memory-like cells and expanded for 14 days to apply to adoptive transfer of EBVaGC model. Cytotoxicity of EBV-CTLs were specific for EBV-expressing gastric cancer cell line (SNU 719) in vitro, not EBV-negative gastric cancer cell line (MKN45). To confirm efficacy of EBV-CTLs in mouse model, we developed EBVaGC mouse model using SNU719 and injected 1 × 107 cells/mouse once a week for three times. EBV-CTLs had an effect on the reduction of tumor size of EBVaGC and effectively migrated into tumor site. Also, administered EBV-CTLs stably existed in blood and tumor tissue even over 3 days analyzed by FACS and immunohistochemistry. In conclusion, EBV-CTLs represented improved anti-tumor activity to treat EBVaGC and specific lysis to EBV-expressing cells. Also, any side effects or autoimmune diseases were not detected. Therefore, EBV-CTLs would be likely to play important roles in targeting many EBV-associated cancers and be applied to clinical trials.
Note: This abstract was not presented at the conference.
Citation Format: Jae Seung Moon, Jung Ho Kim, Sang Kyou Lee. Immunotherapy of EBV associated gastric cancer using EBV specific T cells [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B049.
Abstract B064: Induction of protective antitumor immunity through attenuation of endoplasmic reticulum aminopeptidase 1 function
Authors: Sugiyarto Gessa , Bailey Ian , Elliott Tim , James Edward 
Published: November 2016

Abstract
Background: The ability to generate potent anti-tumor CTL responses is fundamental to the rejection of many tumors. However many potent anti-tumor responses are suppressed by regulatory T cells (Treg) preventing this. Recent clinical trial studies, using checkpoint inhibitors have shown great promise in inducing protective anti-tumor responses albeit in a minority of patients. In the mouse CT26 tumor model, removal of Treg suppression uncovers potent cryptic anti-GSW11 CTL, which are immunodominant and protective. The generation of GSW11 peptide in the ER is susceptible to destruction by endoplasmic reticulum aminopeptidase 1 (Erap1) significantly reducing the amount presented at the cell surface to T cells. Here we investigate the ability to overcome Treg suppression by modulating the function of Erap1.
Methods: GSW11-specific T cells were characterised from CT26 challenged Treg replete and depleted mice by flow cytometry. Attenuation of Erap1 function was achieved using either the inhibitor Leucine-thiol or siRNA and its effect on GSW11 generation determined by T cell activation assays and reverse phase HPLC.
Results: We identified that Treg suppressed GSW11-specific T cells exhibit a clonally exhausted phenotype in CT26 challenged mice. Examination of GSW11 peptide generation showed that attenuation of Erap1 function by inhibition or siRNA significantly increased the generation of GSW11 (∼75-fold increase). Strikingly, Treg replete mice challenged with Erap1 siRNA CT26 were able overcome Treg suppression and induce GSW11-specific T cell responses resulting in tumor protection. In addition, in vivo intra-tumoral administration of Erap1 inhibitor in established CT26 tumors resulted in the attenuation or regression of tumor growth in the majority of mice.
Conclusions: These results reveal the therapeutic potential of modulating Erap1 function to alter tumor-specific antigen presentation to overcome clonal exhaustion and generate potent anti-tumor T cell immunity.
Citation Format: Gessa Sugiyarto, Ian Bailey, Tim Elliott, Edward James. Induction of protective antitumor immunity through attenuation of endoplasmic reticulum aminopeptidase 1 function [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B064.
Abstract B069: The best is not enough—when it is alone: Current clinical doses of humanized anti-Her2 antibodies should be applied in combination to delay development of trastuzumab resistance by additively enhancing ADCC
Authors: Tóth Gábor , Szöőr Árpád , Simon László , Yarden Yosef , Szöllősi János , Vereb György 
Published: November 2016

Abstract
Although the recently concluded CLEOPATRA trial showed clinical benefits of combining trastuzumab and pertuzumab for treating HER2-positive metastatic breast cancer, trastuzumab monotherapy is still the mainstay in adjuvant settings. Since trastuzumab resistance occurs in over half of these cancers, we examined the mechanisms by which treatment of intrinsically trastuzumab-resistant and -sensitive tumors can benefit from the combination of these antibodies. F(ab')2 of both trastuzumab and pertuzumab were generated and validated in order to separately analyze antibody-dependent cell-mediated cytotoxicity (ADCC)-based and direct biological effects of the antibodies.Compared to monotherapy, combination of the two antibodies at clinically permitted doses enhanced the recruitment of natural killer cells responsible for ADCC, and significantly delayed the outgrowth of xenografts from intrinsically trastuzumab-resistant JIMT-1 cells. Antibody dose-response curves of in vitro ADCC showed that antibody-mediated killing can be saturated, and the two antibodies exert an additive effect at sub-saturation doses. Thus, the additive effect in vivo indicates that therapeutic tissue levels likely do not saturate ADCC. Additionally, isobole studies with the in vitro trastuzumab-sensitive BT-474 cells showed that the direct biological effect of combined treatment is additive, and surpasses the maximum effect of either monotherapy. Our results suggest the combined therapy is expected to give results that are superior to monotherapy, whatever the type of HER2-positive tumor may be.The combination of both antibodies at maximum clinically approved doses should thus be administered to patients to recruit maximum ADCC and cause maximum direct biological growth inhibition.
Citation Format: Gábor Tóth, Árpád Szöőr, László Simon, Yosef Yarden, János Szöllősi, György Vereb. The best is not enough—when it is alone: Current clinical doses of humanized anti-Her2 antibodies should be applied in combination to delay development of trastuzumab resistance by additively enhancing ADCC [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B069.
Abstract B073: Protein nanoparticles for controllable T cell activation
Authors: Lajoie Marc J. , Butterfield Gabriel , Gray Elizabeth , DaPron Kate , Stetson Daniel , Baker David , King Neil 
Published: November 2016

Abstract
The immunosuppressive nature of cancer prevents a sustained response to T-cell-inducing vaccines. We hypothesize that co-delivering antigens and adjuvants directly to sub-compartments of antigen presenting cells could elicit specific CTL and Th1 activation, providing more control over the immune response. To accomplish this goal, we are developing protein nanoparticles capable of delivering payloads consisting of RNA, protein, and small molecules to target cells. One class of protein nanoparticles is produced in E. coli and is capable of packaging its own RNA to enable evolution of improved functions such as increased RNA packaging, improved serum stability, and pH-mediated disassembly for endosomal payload delivery. Another class of protein nanoparticles can bud from HEK293F cells containing small molecule adjuvants and mRNA-encoded antigens. By pseudotyping these enveloped protein nanoparticles with viral fusion proteins, we can deliver nucleic acids to the cytosol of target cells where they are translated to produce the desired protein antigen. These non-viral delivery devices could improve our ability to control immune responses against cancer.
Citation Format: Marc J. Lajoie, Gabriel Butterfield, Elizabeth Gray, Kate DaPron, Daniel Stetson, David Baker, Neil King. Protein nanoparticles for controllable T cell activation [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B073.
Abstract B083: Novel mRNA manufacturing techniques using modification of nucleosides and purification improves the efficacy of mRNA chimeric antigen receptor T cells
Authors: Foster Jessica , Choudhari Namrata , Barrett David , Resnick Adam 
Published: November 2016

Abstract
Chimeric Antigen Receptor (CAR) T-cells made with mRNA offer a transient and safe alternative to viral CARs, mitigating the concern for persistent unwanted side effects from constitutively active T-cells. Previous studies have shown that mRNA CARs are transiently effective, but lack CAR persistence and have struggled to show success across tumor types. We hypothesized the efficacy of mRNA CARs could be improved by utilizing recent advancements in RNA technology including the use of modified nucleosides and novel purification methods of the RNA to prevent double stranding that induces toxicity. Using the established CD19 CAR model in B-cell leukemia, we created mRNA CARs using previously described methods and compared them to mRNA CARs created using the modifications of pseudo-uridine or N-methyl pseudo-uridine instead of uridine, and purification to remove any aberrant double stranded mRNA. Comparing the modified and purified mRNA CAR T cells to those created using prior methods of making mRNA, the novel mRNA CAR T cells showed a two-fold increase in expression of the CAR on their surface initially, as well as a two-fold improvement in cytotoxic killing of leukemia cells that persisted for up to five days. The modified and purified mRNA CAR T cells also showed reduced expression of negative checkpoint regulators compared to original RNA CAR T cells. In vivo studies using a leukemia mouse model with a single dose of CAR T cells also showed marked improvement using modified and purified mRNA CAR T cells, with purified RNA showing the most robust 2-log enhanced suppression of leukemic burden. Our results provide a novel technique that can be easily incorporated into RNA production for use in clinical trials, and poise RNA CARs for increased efficacy as new CAR targets emerge and are being tested.
Citation Format: Jessica Foster, Namrata Choudhari, David Barrett, Adam Resnick. Novel mRNA manufacturing techniques using modification of nucleosides and purification improves the efficacy of mRNA chimeric antigen receptor T cells [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B083.
Abstract B090: Evaluation of a novel impedance-based assay for label free and dynamic assessment of CAR-T constructs efficacy against B-cell leukemia and lymphoma
Authors: Xue Chelsea , Cerignoli Fabio , Senyukov Vladimir , Pierog Piotr , Miller Thomas , Shaw Amy , Xi Biao , Muir Lincoln , Abassi Yama , Kassim Sadik 
Published: November 2016

Abstract
Immunotherapy is emerging as a promising approach in cancer treatment, allowing specific targeting of cancer cells based on surface marker expression. In vitro characterization of the effectiveness of the treatment is a necessary step before moving into clinical trials, but currently technologies rely on endpoint assays that are unable to capture the complexity and dynamics of the in vivo immune response. Here we present the adaptation of an impedance-based technology to measure immune-mediated cell killing of B lymphoid and lymphoma tumor cells. Impedance technology is able to detect alterations in cell adhesion, morphology and cell number of adherent cells by measuring changes in conductance of microelectrodes embedded in 96 and 384-wells cell culture plates. The variation is reported through an impedance-related parameter termed Cell Index (CI). The platform enables label-free and continuous monitoring of cell viability, leaving the same cells still available for secondary end point assays. To adapt such adhesion-based technology to monitor non-adherent B cells, we developed a strategy where the bottom of the wells is coated with an anti-CD40 antibody. The coating allows specific adhesion and retention of B cells and measurement of changes in impedance that are proportional to cell number. Using primary T lymphocytes expressing Chimeric Antigen Receptors (CAR) against CD19 or unrelated targets, we were able to demonstrate specific killing of tumor B cell lines and primary cells also at very low effector:target ratios. The results were also confirmed by flow cytometry. Taken together our results demonstrate the high sensitivity of this high throughput approach when compared to end point assays like 51Cr release or flow cytometry, improving the workflow for therapy validation and optimization in the context of personalized medicine.
Citation Format: Chelsea Xue, Fabio Cerignoli, Vladimir Senyukov, Piotr Pierog, Thomas Miller, Amy Shaw, Biao Xi, Lincoln Muir, Yama Abassi, Sadik Kassim. Evaluation of a novel impedance-based assay for label free and dynamic assessment of CAR-T constructs efficacy against B-cell leukemia and lymphoma [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B090.
Abstract B097: Analyze and recover functionally defined single immune cell clones through Opto-Electro-Positioning technology
Authors: Wang Xiaohua , Bronevetsky Yelena , Stadler Guido , Beaumont Kristin , Bao Xiaoyan Robert , Shimizu Saki , Smith Duane , Chapman Kevin 
Published: November 2016

Abstract
Cancer patients mount protective innate and adaptive immune responses against tumor cells. Since such responses are able to control or even cure eliminate cancer if harnessed properly, identifying clonal adaptive immune cells targeting tumor cells with designated function is extremely valuable but remains challenging. We provide evidence here that the combination of the Opto-Electro-Positioning (OEP) technology and the nanofluidic culture enables high throughput screening and harvesting functionally defined immune cells through single cell based analysis. Immune cells can be maneuvered deterministically through OEP into defined nanofluidic culture space which supports efficient antigen specific T/B cell clonal expansion and functional profiling ex vivo. The viable T/B cell colonies with designated functions can subsequently be harvested after analysis for downstream applications including rapid expansion, antigen receptor sequencing, and epigenetic analysis etc. Integration of these two basic technologies with computer controlled automation leads to our Beacon platform that allows researchers to screen tens of thousands of live single immune cells simultaneously. The system is also exceptionally efficient in investigating immune cell population from the tumor biopsies that are typically of low cellularity. In summary, the OEP and nanofluidic device based Beacon solution provides researchers an “all-purpose” single cell based discovery platform for both basic and clinical oncology investigations.
Citation Format: Xiaohua Wang, Yelena Bronevetsky, Guido Stadler, Kristin Beaumont, Xiaoyan Robert Bao, Saki Shimizu, Duane Smith, Kevin Chapman. Analyze and recover functionally defined single immune cell clones through Opto-Electro-Positioning technology [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B097.
Abstract B101: Mechanism of T cell activation by phase separation
Authors: Su Xiaolei , Ditlev Jonathon , Hui Enfu , Rosen Michael , Vale Ronald 
Published: November 2016

Abstract
Activation of many cell surface receptors triggers the reorganization of downstream signaling molecules into micron- or submicron-sized clusters. However, the functional consequences of such clustering have been unclear. We biochemically reconstituted a 12-component signaling pathway on model membranes, beginning with T cell receptor (TCR) activation and ending with actin assembly. When TCR phosphorylation was triggered, downstream signaling proteins spontaneously separated into liquid-like clusters that promoted signaling outputs both in vitro and in human Jurkat T cells. Reconstituted clusters were enriched in kinases but excluded phosphatases and enhanced actin filament assembly by recruiting and organizing actin regulators. These results demonstrate that protein phase separation can create a distinct physical and biochemical compartment that facilitates signaling.
Note: This abstract was not presented at the conference.
Citation Format: Xiaolei Su, Jonathon Ditlev, Enfu Hui, Michael Rosen, Ronald Vale. Mechanism of T cell activation by phase separation [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B101.
Abstract B102: A microengineered human lymph node-on-a-chip to study cancer immunotherapy
Authors: Goyal Girija , Ingber Donald 
Published: November 2016

Abstract
Preclinical studies in cancer immunotherapy, such as those involving checkpoint blockade are usually conducted and validated in mice; however, rodent immune responses do not faithfully mimic those observed in humans. The overall goal of this project is to leverage recent advances in human organ-on-chip technology to create a human lymph-node-on-chip (hLN-on-chip) that exhibits complex human immune functionality. For maximum applicability, we have started with human peripheral blood mononuclear cells (PBMC) such that these devices may be widely used as non-invasive surrogates for patient testing. We have optimized matrix, cellular composition, chip design and microfluidic perfusion to induce B cell clusters reminiscent of follicles found in the lymph node. T and B cell function is maintained in the hLN-on-chip as evidenced by IL-2 and immunoglobulin M production in response to heat killed formalin fixed Staph aureus Cowan I (SAC), commonly used to induce a T cell dependent antibody response. Current studies are focused on defining the alterations in sensitivity and activity of the T cells. For instance, lymph node resident T and B cells are known to be polarized. Thus, we have tested if coinhibitory receptors such as CTLA-4, known to be important in anti-tumor immunity are also polarized under tissue like conditions. We also present preliminary data on optimizing the perfusion of the lymphocyte parenchyma with microengineered channels lined with lymphatic and vascular endothelial cells to model antigen delivery and immune cell trafficking. These devices will enable near real-time assessment of secreted factors, cell trafficking and high-resolution imaging of dynamic cellular interactions that occur in the human lymph node, and hence, they should prove invaluable in studying human cancer response to anti-cancer immunotherapies in the future.
Citation Format: Girija Goyal, Donald Ingber. A microengineered human lymph node-on-a-chip to study cancer immunotherapy [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B102.