bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract A123: CD14+ monocyte recruitment, transformation and support of tumor survival in a lung cancer model
Authors: Schenk Erin , Dietz Allan
Published: May 2016

Our previous work has shown that in multiple hematologic and solid malignancies, an increased presence of CD14+HLA-DRlo/neg cells in the peripheral blood are associated with systemic immune suppression and poor prognosis. We have also demonstrated that glioblastoma and non-Hodgkin lymphoma tumor cells can convert healthy donor CD14+ cells into CD14+HLA-DRlo/neg. Thus, a critical step toward the generation of these immune suppressive cells is the recruitment of CD14+ monocytes to the tumor. In this study we investigated the mechanism for monocyte recruitment by lung cancer cell lines, the effect these lines have on the monocytes, and the effect the monocytes have on the tumor. Monocyte recruitment by H23, a human lung adenocarcinoma cell line, was studied using freshly isolated CD14+ cells and measured in real time with live cell imaging. Monocytes from 10 of 10 unique healthy donors robustly migrated towards H23 supernatant in a dose dependent fashion. H23 produces CCL2 at levels of 9160 ± 1615 pg/mL measured by ELISA in addition to elaborating the monocyte recruitment factors CXCL1 and IL-8 seen by cytokine array. A 2 day co-culture of H23 with monocytes from 17 of 18 unique healthy volunteers resulted in downregulated monocyte cell surface expression of HLA-DR by 49.7% ± 22.1%, a phenotype known to mediate both local and systemic immune suppression. Most importantly, monocytes had a positive effect on H23 survival under stress conditions. H23 continuously exposed to cisplatin in culture had improved survival when co-cultured with monocytes for 2 days prior to cisplatin exposure. After 6 days of low serum growth conditions, H23 survival and proliferation was bolstered by the presence of monocytes in culture resulting in a 2 fold greater number of viable H23 cells compared to H23 cultured in low serum without monocytes. These experiments have been repeated and showed that 3 of 4 additional human lung cancer cell lines demonstrated improved survival in low serum growth conditions when co-cultured with monocytes. In conclusion, we have demonstrated the active recruitment of monocytes by lung cancer cell lines, the transformation of the monocytes to an immune suppressive phenotype and the monocyte cancer cross-talk that results in improved tumor survival when exposed to the likely in vivo conditions of a nutrient poor environment or chemotherapy. Taken together these results suggest that the recruitment of monocytes may confer additional survival benefit in the tumor microenvironment beyond local immune suppression and angiogenesis.

Citation Format: Erin Schenk, Allan Dietz. CD14+ monocyte recruitment, transformation and support of tumor survival in a lung cancer model. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A123.
Abstract A138: Identification of key immune regulators that control obesity-induced liver inflammation and diseases
Authors: Zhong Zhenyu , Umemura Atsushi , Liang Shuang , Karin Michael
Published: May 2016

Obesity has become a major risk factor for a number of human diseases, including cancer. Amongst different types of malignancies, obesity appears to have the highest association-rate with hepatocellular carcinoma (HCC), the dominant form of primary liver cancer. Obesity-induced sterile inflammation is the key switch mediating the transition from hepatic steatosis to steatohepatitis (non-alcoholic steatohepatitis, NASH) and HCC. However, the molecular mechanism underlying obesity-induced hepatic inflammation remains poorly understood. We show that fat accumulation increases liver damage and release of DAMP (damage-associated molecular patterns) from dying hepatocytes, to amplify inflammation and accelerate hepatic disease progression. These DAMP induce mitochondrial reactive oxygen species (mtROS) in macrophages, which direct inflammatory cytokine/chemokine production in a manner that requires activation of a calcium permeable channel, TRPM2 (transient receptor potential melastatin 2). Blockade of mtROS generation, removal of extracellular calcium or ablation of TRPM2 expression in macrophages abolished DAMP-induced inflammatory cytokine/chemokine production, suggesting that a “mtROS-TRPM2” pathway drives DAMP-induced inflammation. Moreover, we identified HMGB1 as the key DAMP released from lipid-damaged hepatocytes. By using HMGB1 neutralizing antibody, we found it suppressed HMGB1 expression in the liver of MUP-uPA mice (an animal model for human NASH), and markedly attenuated liver damage, accumulation of lipid droplets, inflammation, and fibrosis, which represent typical features of human NASH. These results also suggest that HMGB1 is a key component of a vicious autoregulatory cycle triggered by HFD consumption, that mains ER stress and liver inflammation, as well as elevated HMGB1 expression and release.

Citation Format: Zhenyu Zhong, Atsushi Umemura, Shuang Liang, Michael Karin. Identification of key immune regulators that control obesity-induced liver inflammation and diseases. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A138.
Abstract A156: Anti-p53 auto-antibody serum profiling using high-density peptide arrays
Authors: Lo Ken C. , Tan John C. , Sullivan Eric , Bannen Ryan , Richmond Todd , Grupp Florian , Weiser Stefan , Heindl Dieter , Stengele Klaus-Peter , Thomas Albert
Published: May 2016

Cancer is a result of a number of genetic alterations that disturb normal, controlled cell growth and differentiation. Mutational events leading to the activation of oncogenes or the inactivation of tumor-suppressor genes have been linked causally to the formation of tumors. p53 is one of the most important regulators of transcription, cellular cycle, DNA repair and apoptosis detected to date. Anti-p53 antibodies have been detected in the serum of cancer patients. This immune response is probably due to a self-immunization process linked to the strong immunogenicity of the p53 protein, and is associated predominantly with p53 missense mutation and p53 accumulation in the tumor. Auto-antibodies have also been proposed as potential diagnostic biomarkers for early stage diagnosis of cancers, since an increase in serum levels of certain auto-antibodies has been shown to precede the development of disease symptoms and correlate with cancer incidence for various cancers including breast and lung cancer.

Here we systematically evaluate reactivity of antibodies in p53 positive serum samples and identify reactive epitopes to normal and mutant peptides using a high-density (2.9 Million) peptide microarray. We assess the effect of linker length, peptide length, and flanking serines on antibody detection. We propose peptide array design parameters that can be applied to a whole proteome level to enable biomarker discovery and validation of novel auto-antibody epitopes associated with cancer.

Citation Format: Ken C. Lo, John C. Tan, Eric Sullivan, Ryan Bannen, Todd Richmond, Florian Grupp, Stefan Weiser, Dieter Heindl, Klaus-Peter Stengele, Albert Thomas. Anti-p53 auto-antibody serum profiling using high-density peptide arrays. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A156.
Abstract A166: Biogenesis and regulatory functions of super-enhancer RNAs in cancer cells of the immune system
Authors: Guo Yang Eric , Young Richard
Published: May 2016

Super-enhancers (SEs) are large (2 to >100 kilobases) DNA elements that control cell identity and disease. These unusual elements are occupied by high levels of key transcription factors, co-activators and chromatin regulators. SEs are hot spots for sequence variations associated with many human diseases, including cancer. Surprisingly, SEs are transcribed at high levels into RNAs (seRNAs). However, little is known about the biogenesis, structural identity and function of these seRNAs. Preliminary evidence suggests that seRNAs are components of SE complexes, composed of chromatin, RNAs and transcription factors, which play critical roles in controlling expression of key master cell-type-specific transcription factors and disease-causing genes. Our main objectives are to characterize the biogenesis of seRNAs, to determine the composition of RNA-protein complexes associated with seRNAs and to investigate the functions of seRNAs in gene regulation. We are focusing on the c-MYC and RUNX1 SEs, drivers of two cancers originating from B and T cells of the immune system: multiple myeloma (MM) and T-cell acute lymphoblastic leukemia (T-ALL), respectively. By uncovering the biogenesis and functions of seRNAs at the molecular level, our results may identify novel targets for new therapies for MM and T-ALL, as well as other cancers. New ways to modulate the immune response may also be revealed by our studies.

Citation Format: Yang Eric Guo, Richard Young. Biogenesis and regulatory functions of super-enhancer RNAs in cancer cells of the immune system. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A166.
Abstract A167: Functional analysis of DNA sensors in the interferon stimulatory DNA (ISD) pathway
Authors: Gray Elizabeth , Stetson Daniel
Published: May 2016

Detection of viral DNA and production of type I interferons (IFNs) are essential for antiviral immunity and immune responses against transformed cells; on the other hand, inappropriate immune responses to self DNA result in autoimmunity. Intracellular DNA sensing activates the STING-dependent IFN stimulatory DNA (ISD) pathway. Multiple DNA sensors have been proposed to activate the ISD pathway, including cyclic GMP-AMP synthase (cGAS) and a family of DNA binding receptors called the AIM2-like receptors (ALRs). Analysis of cGAS-deficient mice has revealed that cGAS is required for the IFN response to transfected DNA ligands, DNA viruses, and retroviruses. In contrast, while multiple studies have suggested that ALRs promote an optimal IFN response to various DNA pathogens, others have found that the IFN response is intact following ALR knockdown. Thus, while cGAS has clearly emerged as a key DNA sensor in the ISD pathway, the function of the ALRs remains unclear.

To define the relative contributions of the ALRs and cGAS to the ISD pathway, we generated mice lacking all thirteen ALR genes and cGAS-deficient mice. In addition, we have used CRISPR gene targeting to disrupt cGAS and all four human ALRs in human cells. Analysis of IFN induction following transfection of DNA ligands or DNA virus infection has revealed that cGAS is absolutely required for activation of the ISD pathway in both mouse and human cells, consistent with recent studies. In contrast, the IFN response to transfected DNA ligands and infection with a variety of DNA viruses is intact in ALR-deficient cells, suggesting that the ALRs may be dispensable for activation of the ISD pathway. We are currently exploring potential alternative functions of the ALR family of DNA binding receptors.

Citation Format: Elizabeth Gray, Daniel Stetson. Functional analysis of DNA sensors in the interferon stimulatory DNA (ISD) pathway. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A167.
Abstract A171: Long-term episomal gene transfer for safe engineering of T cells for adoptive cell therapy of cancer
Authors: Jin Chuan , Ramachandran Mohanraj , Fotaki Grammatiki , Nilsson Berith , Essand Magnus , Yu Di
Published: May 2016

Success in T-cell therapy of cancer is currently relying on long-term gene expression owing to retrovirus/lentivirus (RV/LV) integration. However, uncontrolled RV/LV integration in host cell genomes has the potential risk of causing insertional mutagenesis. Herein, we describe a novel episomal and long-term cell engineering method using non-integrating lentiviral (NILV) vector containing a scaffold matrix attachment region (S/MAR) element for either over-expression or down-regulation of genes. T-cells engineered with a NILV-S/MAR(CD19CAR) vector have similar levels of CAR expression as T-cells engineered with an integrating LV vector, even after numerous rounds of cell division. NILV-S/MAR-engineered CD19CAR-T-cells exhibited similar cytotoxic capacity upon CD19+ target cells recognition as LV-engineered T-cells. We propose that NILV-S/MAR vectors are superior to current options for enabling long-term gene expression without the risk of insertional mutagenesis.

Citation Format: Chuan Jin, Mohanraj Ramachandran, Grammatiki Fotaki, Berith Nilsson, Magnus Essand, Di Yu. Long-term episomal gene transfer for safe engineering of T cells for adoptive cell therapy of cancer. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A171.
Abstract A183: Molecular elucidation of the CBM complex in NF-kappaB activation by using single molecule imaging and structural studies
Authors: David Liron , Wu Hao
Published: May 2016

NF-kB family proteins are evolutionarily conserved master regulators of immune and inflammatory responses. NF-kB signaling has a crucial role in regulating the activation, proliferation and effectors functions of lymphocytes in adaptive immune responses. The CARMA1/Bcl10/MALT1 (CBM) complex is the central mediator of TCR and BCR induced NF-kB activation in lymphocytes. Mutations, chromosomal translocations and over-expressions of CBM component proteins have been shown to directly lead to non-Hodgkin's lymphomas. Elucidating the molecular basis of assembly and activation of the CBM complex could provide a platform for mechanistic understandings of the associated lymphomas and potential therapeutic target identification. Our research goals are to investigate the CBM mechanism in the NF-kB activation pathway by single molecule imaging and structural studies on the CARMA1/Bcl10, BCl10/MALT1 and CBM complex by combining electron microscopy and X-ray crystallography. We have succeeded in purifying the CARMA1/Bcl10/MALT1 complex and shown by negative staining EM that the CBM complexes form filamentous structures as well as BCl10 only. In vitro, polymerization assay indicated that CARMA1 nucleates Bcl10 filament assembly, which could promote downstream MALT1 activation. Based on the 3D map from negative stained electron microscopy and high-resolution individual structures of CARMA1/Bcl10 CARD domains, we reconstituted a structure model of CARMA1/Bcl10 filament. Single molecule imaging enables us to track after the filament formation of labeled BCl10 filaments and the nucleation of BCl10 by labeled CARMA1 and determine the filaments formation rates in vitro supported by kinetic studies. In addition, we could determine that polymerization BCl10 has polarity and the filament growth is only at one end. Moreover, Real time Super Resolution Microscopy (STORM) measurements showed a surprising funding that CARMA1(8-302) is segmented along the filaments and incorporated within BCl10 filaments. At the moment, we are pursuing the determination of cryo-EM structures of BCl10 and BCl10/ MALT1 complex. At the moment, we are processing BCl10 and BCl10/ MALT1 complex data sets that were collected with a K2 detector and by using Titan cryoEM microscope. BCl10 (1-205) structure was solved to a 6Å and the data is currently being further improved.

Citation Format: Liron David, Hao Wu. Molecular elucidation of the CBM complex in NF-kappaB activation by using single molecule imaging and structural studies. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A183.
Abstract B007: Targeting the ubiquitin signaling system for cancer immunotherapy
Authors: Kumar Suresh , Wang Liqing , Wu Jian , Wang Feng , Agarwal Saket , Kodrasov Matthew , Hancock Wayne , Mattern Michael
Published: May 2016

Studies have been performed to demonstrate that effectors of enzymes of the ubiquitin proteasome pathway associated with T cell activation are able to impair tumor survival by abrogating the immune evasion mechanism. Immune evasion is a collective term for the various strategies employed by tumors to suppress the host immune system's ability to recognize and destroy cancer cells. Significant advances in the understanding of the mechanisms of cancer immune evasion have led to the successful development of biologic immunotherapies that showed clinical efficacy against hematological as well as solid tumors. These include monoclonal antibodies, allogeneic hematopoietic stem cell transplantation, vaccination, cytokines, T cell checkpoint blockade and adoptive transfer of engineered T cells. Owing to the multifaceted nature of cancer immune evasion, however, several different approaches will likely be necessary for effective therapeutic management. We have adopted a dual targeting approach focused on the ubiquitin proteasome pathway to develop novel small molecule cancer immunotherapies. Using the UbiProTM discovery platform, we have identified inhibitors of a deubiquitylase (DUB) that is highly expressed in regulatory T cells (Tregs) and plays a critical role in promoting Treg functions. These inhibitors are shown to suppress Treg functions and promote T-effector cell function ex vivo and in vivo. In addition, we have identified inhibitors of an E3 ubiquitin ligase that negatively regulates T-effector cell function. These inhibitors are shown to activate T-cells and NK cell functions in vitro. A combination of such inhibitors should provide powerful anti-cancer immunotherapy.

Citation Format: Suresh Kumar, Liqing Wang, Jian Wu, Feng Wang, Saket Agarwal, Matthew Kodrasov, Wayne Hancock, Michael Mattern. Targeting the ubiquitin signaling system for cancer immunotherapy. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr B007.
Abstract B013: A soluble form of CD80 enhances anti-tumor immunity by inhibiting PDL1 immune suppression and does not suppress via CTLA-4
Authors: Horn Lucas , Clements Virginia , Ostrand-Rosenberg Suzanne
Published: May 2016

Tumor cells employ various methods of immune suppression to overcome anti-tumor immunity. One such method is that of the integral membrane protein programmed death ligand-1 (PDL1), which triggers apoptotic death or anergy upon binding programmed death-1 (PD1) on T cells. Treatment of PDL1+ tumor cells with a soluble form of the costimulatory molecule CD80 (CD80-Fc) prevented PDL1-mediated immune suppression by binding PDL1 and blocking interaction with PD1, thus restoring T cell activation. Additionally, CD80-Fc maintained activation of T cells in vitro more effectively than either PD1 or PDL1 antagonist monoclonal antibodies. Because CTLA-4 is also a receptor for CD80 and CD80-Fc has the potential to bind and deliver inhibitory signals into T cells via CTLA-4, it is important to assess CD80-Fc and CTLA-4 interactions. Healthy donor human PBMC stimulated with PHA for three days express CD28, PDL1, and CTLA-4 at the cell surface. To determine which of these receptors CD80-Fc binds to, we have blocked subsets of the receptors using very high concentrations of specific antibodies, and monitored CD80-Fc binding by flow cytometry. CD80-Fc binds to activated T cells blocked for CD28 and PDL1, PDL1 and CTLA-4, or CTLA-4 and CD28, and did not bind when all three receptors were blocked. These findings indicate that CD80-Fc binds to CTLA-4. However, inclusion of CTLA-4 antagonist antibodies L3D10 or Ipilimumab in co-cultures of human PBMC and PDL1+ C8161 human melanoma cells did not increase T cell production of IFNγ, suggesting that CD80-Fc is not suppressing through CTLA-4. Historically, CTLA-4 has been considered as a negative regulator of T cell function through its ability to deliver suppressive signals to T cells. Recent re-evaluations of CD80-CTLA-4 interactions suggest that CTLA-4 inhibits T cell function not by delivering intracellular signals, but by serving as a decoy receptor for CD80. Our results support this alternative functional mechanism and are consistent with the concept that CD80-Fc maintains T cell activation by saturating CTLA-4 receptors while simultaneously blocking PDL1 suppression and providing CD28 costimulation.

Citation Format: Lucas Horn, Virginia Clements, Suzanne Ostrand-Rosenberg. A soluble form of CD80 enhances anti-tumor immunity by inhibiting PDL1 immune suppression and does not suppress via CTLA-4. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr B013.
Abstract B042: Regulation of Lck activity by a cryptic phosphosite
Authors: Courtney Adam H. , Sjolin-Goodfellow Hanna E. , Ji Qinqin , Kadlecek Theresa A. , Salomon Arthur R. , Weiss Arthur
Published: May 2016

Src family kinases (SFKs) are initiators of receptor-mediated signaling events, enabling cells to perceive and interact with their environment. For immune cells SFKs are especially crucial because they couple antigen recognition to intracellular signaling pathways. In T cells, the SFK Lck phosphorylates the T cell antigen receptor (TCR) complex upon antigen encounter. Because Lck phosphorylates the TCR complex, its activity sets a critical threshold for T cell activation; yet how Lck activity is regulated in resting or stimulated cells is poorly understood. Through quantitative mass spectrometry (SILAC), we recently identified a cryptic Lck phosphosite (Y192) as sensitive to inhibition of the effector kinase Zap70. Because Zap70 is an Lck substrate, and dramatic reduction in phosphorylation of Y192 occurred upon Zap70 inhibition, we reasoned this site might function to regulate Lck in response to changes in Zap70 activity. To test this hypothesis we reconstituted Lck-deficient Jurkat T cells with Lck Y192 variants. Mutation of Y192 reveals that this site has a profound impact on Lck activity. Cells reconstituted with Y192E or Y192A show a marked reduction in Lck activity, characterized by hyperphosphorylation on Lck's inhibitory C-terminus (Y505) and a reduction in Lck autophosphorylation within the activation loop (Y394). Consistent with a defect in Lck activity, cells that express Lck Y192E/A variants, when assessed for hallmarks of T cell activation, reveal striking defects in TCR signaling. Interestingly, these defects are reminiscent of CD45-deficient T cells. Preliminary findings suggest that mutation of Y192 disrupts the ability of the phosphatase CD45 to dephosphorylate Lck's inhibitory C-terminus (Y505). Because of the profound effect that Y192 modification exerts on Lck activity, and therefore TCR signaling, we believe that phosphorylation of Y192 comprises a Zap70-dependent negative feedback mechanism that tunes Lck activity. Such a mechanism predicts that increases in Zap70 activity cause phosphorylation of Y192 within Lck's SH2 domain, favouring Lck inactivation by preventing CD45-mediated dephosphorylation of the inhibitory C-terminus.

Citation Format: Adam H. Courtney, Hanna E. Sjolin-Goodfellow, Qinqin Ji, Theresa A. Kadlecek, Arthur R. Salomon, Arthur Weiss. Regulation of Lck activity by a cryptic phosphosite. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr B042.
Abstract B058: Expansion and activation of CD103+ DC progenitors at the tumor site promote T cell accumulation in melanoma lesions and transform clinical response to BRAF and PD-L1 blockade
Authors: Salmon Hélène , Idoyaga Juliana , Rahman Adeeb , Palucka Anna Karolina , Bhardwaj Nina , Ginhoux Florent , Merad Miriam
Published: May 2016

A high number of melanoma lesions develop resistance to BRAF blockade or fail to respond to checkpoint inhibition therapy. Here we explored whether modulation of intratumoral APC can increase responses to BRAF or PD-L1 blockade in Braf-mutant and B16 melanoma. In both models, CD103+ DC were the only APC that transported intact antigens to the lymph nodes and primed tumor-specific CD8+ T cells. CD103+ DC expressed high PD-L1 levels, however PD-L1 or BRAF blockade led to partial response and minimal accumulation of intratumoral T cells. Strikingly, systemic Flt3L followed by intratumoral poly I:C injections expanded and activated CD103+ DC progenitors in the tumor mass, leading to dramatic expansion of intratumoral CD8+ T cells enhancing responses to BRAF and PD-L1 blockade and protecting mice from tumor rechallenge. Thus the paucity of activated CD103+ DC in tumors limit checkpoint blockade efficacy and Flt3L/poly I:C combination can significantly enhance clinical response to checkpoint and BRAF blockade.

Citation Format: Hélène Salmon, Juliana Idoyaga, Adeeb Rahman, Anna Karolina Palucka, Nina Bhardwaj, Florent Ginhoux, Miriam Merad. Expansion and activation of CD103+ DC progenitors at the tumor site promote T cell accumulation in melanoma lesions and transform clinical response to BRAF and PD-L1 blockade. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr B058.
Abstract B096: Oncolytic vaccination for HPV induced cancer
Authors: Atherton Matthew J. , Stephenson Kyle B. , Wan Yonghong , Stojdl David F. , Lichty Brian D.
Published: May 2016

Oncogenic human papilloma virus (HPV) is directly responsible for 5% of the world's cancer burden. HPV has been etiologically implicated in virtually all cases of cervical carcinoma and is increasingly responsible for head and neck cancers in the developed world with strains 16 and 18 considered high risk. Recently it was estimated that there were over half a million new cases of cervical cancer worldwide and in the same year this disease was projected to claim over 250,00 lives, making it the third most prevalent and fourth most lethal non- skin tumor in women. Current standard of care for advanced cervical carcinoma and head and neck cancers involve chemo-radiation with or without prior surgical resection resulting in significant morbidity without guaranteed cure. Screening programs and prophylactic vaccination are reducing the incidence of some HPV induced tumors, however lack of compliance and unvaccinated populations mean that significant illness and mortality still exists. We therefore sought to develop a novel pre-clinical immunotherapeutic for HPV induced cancers. An oncolytic vaccination strategy utilizing an attenuated Maraba MG1 rhabdovirus expressing inactive HPV antigens based on the E6 and E7 transforming proteins of strains 16 and 18 has been designed (MG1 E6E7). The TC1 cell line, which is a C57BL/6 syngeneic murine HPV cancer model, was acquired. Specific anti-E7 CD8+ T cell responses have been induced after oncolytic vaccination of mice generating sterilizing immunity against future engraftment with TC1 cells. Oncolytic vaccination of tumor free mice has generated between 24 and 50 million, combined blood and splenic CD8+ T cells per mouse, determined by interferon gamma production in response to a single E7 peptide quantified using intracellular staining and flow cytometry. Primed mice, boosted with MG1 E6E7 develop long lasting, marked and specific immunity against E7 and to a lesser extent E6. The prime: boost regimen has therapeutic efficacy against established subcutaneous TC1 tumors (mean volumes of 250 mm3) resulting in durable cures in this model. Depletion of CD8+ T cells impairs the activity of the vaccination protocol supporting the key role of cytotoxic lymphocytes in our treatment regimen. The data generated here support the pre-clinical activity of this oncolytic vaccination and pave the way for future clinical trials for the treatment of advanced metastatic HPV induced carcinomas.

Citation Format: Matthew J. Atherton, Kyle B. Stephenson, Yonghong Wan, David F. Stojdl, Brian D. Lichty. Oncolytic vaccination for HPV induced cancer. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr B096.
Abstract B124: Immunotherapy of acute myeloid leukemia using Vg9Vd2 T-cells
Authors: Parente-Pereira Ana C. , Whilding Lynsey M. , Krishnamurthy Pramila , Beatson Richard , Zabinski Tomasz , Barber Linda , Farzaneh Farzin , Mufti Ghulam , Maher John
Published: May 2016

Disease remission is successfully induced in the majority of patients with acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS), but is generally not sustained without stem cell transplantation. Adoptive immunotherapy offers an attractive option to consolidate AML remission. Here, we hypothesized that ex-vivo expanded Vg9Vd2 T-cells will mediate a therapeutically beneficial graft versus leukemia (GvL) effect in this disease.

Peripheral blood mononuclear cells from healthy donors and from patients with AML were activated in-vitro using zoledronic acid and a cytokine cocktail, containing IL-2. As a result, we observed an average 200-fold expansion of Vg9Vd2 T-cells from AML patients, over 15 days. Such expansion was only observed in patients without circulating AML blasts. Expanded patient-derived Vg9Vd2 T-cells exhibited a less differentiated phenotype than cells from healthy donors, indicated by higher expression of CD62L, CCR7 and CD27. A four hour Annexin V cytotoxicity assay using KG-1 and U937 leukemic cells showed that these Vg9Vd2 T-cells were cytotoxic and produced nanogram amounts of interferon-g; (IFN-g).

In a parallel approach, we investigated feasibility of developing a universal allogeneic Vg9Vd2 T-cell therapy. Feasibility of this approach has been demonstrated in a small trial in which haploidentical donor-derived Vg9Vd2 T-cells were safely infused and achieved an efficacy signal in patients with advanced haematological diseases. We have recently developed a novel method to expand Vg9Vd2 T-cells from healthy donors ex-vivo (“Method 2 (M2)” cells – patent protected), which improves their expansion over 2 weeks to 1700 fold. Cytotoxicity data shows that M2 cells destroy luciferase-expressing U937 and KG-1 target cells at 2-3 fold enhanced efficiency, accompanied by 2-3.5-fold increased IFN-g; release when compared to cells expanded using conventional approaches. Furthermore, M2 cells can achieve serial killing of AML cell lines through up to 4 cycles of re-stimulation. Treatment of SCID-Beige mice with an established U937 leukemic burden is also more effective using M2 expanded Vg9Vd2 T-cells, particularly when combined with zoledronic acid. Together these data highlight the promise of Vg9Vd2 T-cell immunotherapy of AML.

Citation Format: Ana C. Parente-Pereira, Lynsey M. Whilding, Pramila Krishnamurthy, Richard Beatson, Tomasz Zabinski, Linda Barber, Farzin Farzaneh, Ghulam Mufti, John Maher. Immunotherapy of acute myeloid leukemia using Vg9Vd2 T-cells. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr B124.
Abstract IA10: Combination checkpoint blockade
Authors: Wolchok Jedd D.
Published: May 2016

Given the activity noted with both CTLA-4 or PD-1 blockade, clinical trials are now investigating combination checkpoint blockade. The most mature data with a combination of ipilimumab + nivolumab in melanoma showed a response rate of 40% across dose cohorts with >50% in some cohorts in the context of manageable toxicity. Such responses are generally durable, even when treatment was stopped early for toxicity. Unlike in studies of PD-1 blockade monotherapy, there was no significant difference in clinical activity based on tumor expression of PD-L1. Phase 2 and 3 trials of this combination have similarly shown high rates of activity in melanoma with phase 1 programs in numerous other tumor types. Attention is being paid to the reasons underlying the efficacy of checkpoint blockade in certain malignancies. One hypothesis has been that cancers having a high mutational load may be more amenable to immune modulation by virtue of the larger number of potential neo-epitopes present, fostering baseline immune recognition that can then be potentiated by checkpoint blockade. We have found that melanoma patients having long term clinical activity with ipilimumab have a significantly greater median number of non-synonymous passenger mutations, compared with patients who do not respond or those who have only short-term regression. Strategies to enhance baseline immune reactivity are therefore necessary to investigate as means to improve the impact of checkpoint blockade on a broad spectrum of cancers.

Citation Format: Jedd D. Wolchok. Combination checkpoint blockade. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr IA10.
Abstract IA29: Towards synthetic immune cells for cancer immunotherapy
Authors: Figdor Carl G. , Mandal Subhra , Hammink Roel , Eggermont Loek , Weiden Jorieke , Voerman Dion , Tel Jurjen , Eksteen-Akeroyd Zaskia H. , Blank Kerstin , Rowan Alan E.
Published: May 2016

Most cellular therapies targeting cancer require culture of autologous cells. The dedicated facilities and personnel needed, the variability in quality of the product depending on the condition of the patient and state of disease, hamper widespread use. In an attempt to generate an of the shelf product to target cancer we exploited the ever expanding possibilities to build supramolecular structures offered by chemistry to mimic nature including the construction of artificial immune cells or functions thereof. Effective immunotherapy critically depends on efficient production of antigen-specific cytotoxic T-cells. Herein lies an opportunity for both chemists and immunologists to design and synthesize so-called artificial antigen presenting cells (aAPCs) that can generate in vivo T-cell expansion. We have designed a novel synthetic dendritic cell (sDC) that possesses essential features of natural DCs. Our sDC is based on a semi-flexible poly(isocyano peptide) polymer and, as proof of principle, carries both anti-CD3 antibodies for triggering the T cell receptor/CD3 complex as well as anti-CD28 antibodies as a co-stimulatory signal. Multiple copies of both antibodies facilitate multivalent binding similar to natural DCs. The high mobility of these polymer-bound antibodies, reminiscent of protein motility in a natural plasma membrane, enables receptor rearrangements to occur during T cell activation. We observed effective T cell activation at significantly lower antibody concentrations than freely soluble antibodies. We also demonstrate antigen specific T cell activation when MHC/peptide complexes were bound. These findings demonstrate the potential of multifunctional polymers for mimicking functions of natural DCs or other immune cells, paving the way for their exploitation in immunotherapeutic strategies.

Citation Format: Carl G. Figdor, Subhra Mandal, Roel Hammink, Loek Eggermont, Jorieke Weiden, Dion Voerman, Jurjen Tel, Zaskia H. Eksteen-Akeroyd, Kerstin Blank, Alan E. Rowan. Towards synthetic immune cells for cancer immunotherapy. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr IA29.
Infants Operated on for Necrotizing Enterocolitis: Towards Evidence-Based Pain Guidelines
Authors: Meesters Naomi J. , van Dijk Monique , Knibbe Catherijne A.J. , Keyzer-Dekker Claudia M.G. , Tibboel Dick , Simons Sinno H.P.
Published: May 2016

Background: Necrotizing enterocolitis (NEC) is known as an extremely painful childhood condition. Objectives: The objective of this study was to explore pain management around NEC-related surgery in our neonatal intensive care unit (NICU) from a chart review of prospectively collected data on 60 operated NEC patients admitted between 2008 and 2013 with a median (IQR) gestational age of 28.3 (25.5-31.6) weeks. Methods: Pain medication data and pain scores (i.e. COMFORTneo and Numerical Rating Scale pain and distress scores) from 72 h before until 72 h after surgery were collected. Results: Preoperatively, 95% of the patients received morphine versus 100% postoperatively, with a median dosage of 10.0 (IQR 9.7-14.5) and 16.9 (IQR 10.1-20.0) μg/kg/h, respectively. Postoperatively, 28 patients (46.7%) received additional fentanyl intermittently and 14 (23.3%) received midazolam, which was part of palliative treatment for 6 patients (42.9%). In patients receiving pain medication, median COMFORTneo scores were 10 (IQR 10-11) preoperatively and 11 (10-12) postoperatively. The pain scores were comparable with those of other patients admitted to the NICU in the same time period. Conclusions: Continuous morphine of 10 μg/kg/h preoperatively, with an increase to 15 μg/kg/h postoperatively, seems to constitute a good starting dose for further individualized pain management guided by pain scores.
Neuregulin 3 Knockout Mice Exhibit Behaviors Consistent with Psychotic Disorders
Authors: Hayes Lindsay N. , Shevelkin Alexey , Zeledon Mariela , Steel Gary , Chen Pei-Lung , Obie Cassandra , Pulver Ann , Avramopoulos Dimitrios , Valle David , Sawa Akira , Pletnikov Mikhail V.
Published: May 2016

Neuregulin 3 (NRG3) is a paralog of NRG1. Genetic studies in schizophrenia demonstrate that risk variants in NRG3 are associated with cognitive and psychotic symptom severity, and several intronic single nucleotide polymorphisms in NRG3 are associated with delusions in patients with schizophrenia. In order to gain insights into the biological function of the gene, we generated a novel Nrg3 knockout (KO) mouse model and tested for neurobehavioral phenotypes relevant to psychotic disorders. KO mice displayed novelty-induced hyperactivity, impaired prepulse inhibition of the acoustic startle response, and deficient fear conditioning. No gross cytoarchitectonic or layer abnormalities were noted in the brain of KO mice. Our findings suggest that deletion of the Nrg3 gene leads to alterations consistent with aspects of schizophrenia. We propose that KO mice will provide a valuable animal model to determine the role of the NRG3 in the molecular pathogenesis of schizophrenia and other psychotic disorders.
Management of Perforator Plunge in the Transverse Sinus
Authors: Beniwal Manish , Shukla Dhaval
Published: May 2016

Plunging is rare with the use of automatic-release perforators. We describe the management of a case of plunging in the transverse sinus in a child who underwent suboccipital craniotomy. The perforator got stuck in the bone and transverse sinus. The perforator-bone complex was removed in one piece with suturing of the transverse sinus.
Avaliação hematológica de cães naturalmente infectados por Leishmania (Leishmania) chagasi submetidos a tratamento com antimoniato de meglumina
Authors: Ikedagarcia Fabiana Augusta , Ciarlini Paulo César , Lopes Raimundo Souza , Marques Fábia Judice , Bomfim Suely Rgina Mogami , Lima Valéria Marçal Félix de , Perri Sílvia Helena Venturoli , Marcondes Mary
Published: May 2016

A presente pesquisa foi realizada com o objetivo de avaliar a resposta hematológica de cães com leishmaniose visceral submetidos a tratamento. Para tanto, sete animais naturalmente infectados por Leishmania sp. foram submetidos a um tratamento com 75 mg/kg de antimoniato de meglumina por via subcutânea, 12-12 h /3 semanas. Em todos os animais, uma contagem hematológica completa e punção biópsia aspirativa de medula óssea foi realizada para uma avaliação descritiva em até sete momentos: antes do tratamento, 30, 60, 90, 120, 150 e 180 dias após o início do tratamento. Antes do início do experimento foram observadas alterações hematológicas em quatro dos sete cães (57,1%), entre eles, anemia não regenerativa, linfopenia, linfocitose e monocitose. Durante o curso do experimento a ocorrência de leucocitose, como neutrofilia com desvio à esquerda e eosinofilia, foram observadas em alguns dos animais. Antes do início do tratamento (M1), a ocorrência de hipoplasia da série eritrocítica foi detectada pela citologia de medula óssea em dois animais (28,6%). Isto foi revertido por um aumento na quantidade de células eritróides progenitoras após a administração de antimoniato de meglumina. Desta forma, pode-se concluir que o tratamento promoveu a normalização das alterações hematológicas e recuperação da medula óssea
Propriedades tensiométricas comparadas entre fragmentos do centro tendíneo do diafragma, pericárdio fibroso e peritônio parietal de bovinos não conservados e conservados em glicerina
Authors: Guimarães Gregório Corrêa , Machado Márcia Rita Fernandes , Shimano Antônio Carlos , Terçariol César Augusto Sangaleti , Volpon José Batista , Daleck Carlos Roberto
Published: May 2016

Objetivou-se descrever o comportamento do centro tendíneo do diafragma, pericárdio fibroso e peritônio parietal de bovinos não conservados e conservados em glicerina a 98% durante 30, 60 e 90 dias submetidos a ensaios mecânicos de tração. Utilizou-se para tanto dez bovinos com idade entre 30 meses e 36 meses, mestiços, machos e fêmeas, coletando-se de cada animal fragmentos das membranas referidas. O centro tendíneo do diafragma e o peritônio parietal não sofreram alteração significante (p>;0,05) para os valores de tensão quando comparados os testes de resistência à tração do material não conservado com aqueles conservados em glicerina. Entretanto, todos os tecidos avaliados exibiram aumento significante (p£0,05) para os valores de alongamento quando conservados em glicerina 98% por até 90 dias. Verificou-se que o pericárdio fibroso é a membrana que suporta as maiores tensões, ou seja, maior força de tração por área de secção. Assim, concluiu-se que a glicerina é eficaz para conservação de membranas biológicas, além de alterar suas propriedades mecânicas.
Swan Song, or a Phoenix Rising
Authors: Ping YI
Published: May 2016

It is well-known that the basic ideas and principles of modern international law originated in Europe. In a period during the early twentieth century, however, a number of Chinese intellectuals examined and demonstrated that international law had already existed during the Spring and Autumn and the Warring States Periods in China. They committed themselves to exploring and carrying forward ancient Chinese international law as a way to maintain China’s rich cultural tradition in a global order governed by the West. This may be a swan song of the Chinese intellectuals who rose up against the oppression of imperialist powers and sought a more balanced order in the then-contemporary world. Were their efforts to be revived in another way, however, the swan song could be akin to a phoenix rising, initiating a time of renewal, bringing such ideas back into consideration.
A Theology of Transformation: Catholic Sisters and the Visual Practice of Church
Authors: Kryszak Jennifer E.
Published: May 2016

This article argues that visual practices, including image production and use, promote a theology of transformation. To discern the theological implications of these visual practices, this article employs ethnographic research and material analysis of images created and/or used by the Congregation of St. Joseph, a Roman Catholic women’s religious community in the United States. First, it examines the sisters’ prayer with or creation of images as a source of theological reflection. Second, it investigates the deployment of images in various ministries as a means of inviting others into the sisters’ vision of the church. Third, it assesses the commodification of images by the Congregation as a form of evangelisation that engages and challenges the global world. This article concludes that visual practices potentially inspire action for justice and compassion as well as reveal the challenge of manifesting a theology of transformation in a global and plural world.
Phellinotus, a new neotropical genus in the Hymenochaetaceae (Basidiomycota, Hymenochaetales)
Authors: DRECHSLER-SANTOS ELISANDRO RICARDO , ROBLEDO GERARDO LUCIO , LIMA-JÚNIOR NELSON C. , MALOSSO ELAINE , RECK MATEUS A. , GIBERTONI TATIANA B. , CAVALCANTI MARIA A. DE QUEIROZ , RAJCHENBERG MARIO
Published: May 2016

A new poroid genus with two conspicuous and common species growing on living Fabaceae trees is described from the Seasonally Dry Tropical Forests biome of Brazil. Both taxa in this forest pathogen genus resemble Phellinus rimosus macroscopically, but are distinguished by a dimitic hyphal system with skeletal hyphae present only in the trama of the tube layer while the context remains monomitic, and by the ellipsoid, thick-walled, adaxially flattened, yellow basidiospores that turn chestnut brown in KOH solution. Molecular and morphological studies of Brazilian specimens macroscopically similar to the Phellinus rimosus species complex were carried out to solve their phylogenetic relationships among the Hymenochaetaceae. Phellinotus gen. nov. with P. neoaridus sp. nov. as the genus type and P. piptadeniae comb. nov. are presented and described. Phylogenetically, Phellinotus is closely related to Arambarria, Inocutis, Fomitiporella and other taxonomically unresolved terminal clades, and unrelated to Fulvifomes and Phylloporia. Phellinotus and other genera of poroid Hymenochaetaceae that lack setae or setal hyphae and produce thick-walled, colored (pale yellow to rusty brown) basidiospores form a phylogenetic group here named the ‘phellinotus clade’. Our results indicate the need to include taxa from unexplored areas in order to get a thorough understanding of the phylogeny of the Hymenochaetaceae.
Oreocharis ninglangensis, a showy new species of Gesneriaceae from northwestern Yunnan in China
Authors: CHEN WEN-HONG , CHEN RUN-ZHENG , MÖLLER MICHAEL , WEN KAI , SHUI YU-MIN
Published: May 2016

A new species of Oreocharis (Gesneriaceae) from northwestern Yunnan in China is described and illustrated. The new species, Oreocharis ninglangensis, is most similar to O. delavayi, but it can be easily distinguished from it by conspicuous red stripes inside the corolla mouth and by the finely bullate leaf surface in the latter.