bluuebunny/crossref_metadata_2025_split huggingface dataset:
Delivery of tumor-targeted immunotoxin by tumor-specific T cells (155.19)
Authors: Swanson Anna , Wu Danli , Kuiatse Isere , Kim Hong-Sung , Heslop Helen , Brenner Malcolm , Yotnda Patricia 
Published: January 2023

Abstract
Immunotherapy of cancer using monoclonal antibodies and adoptive transfer of lymphocytes is showing considerable promise. Based on experience with other cancer therapeutics immunotherapeutic modalities, targeting discrete molecular components of the tumor, will prove superior to a single effector system. “Multitargeting” approaches will not only prevent tumor immune escape, they will also allow safer and potent anticancer therapies. We have engineered antigen specific T cells so that they can target tumor cells directly, and then release a potent toxin linked to a monoclonal antibody molecule thereby targeting an independent tumor antigen to the tumor reactive T cell. These engineered T cells were tested in vitro and in vivo in a xenograft model of B-lymphoma. We found that tumor antigen specific T cells retain their anti-tumor activity and tumor homing abilities even when transduced with a gene encoding a chimeric monoclonal antibody-toxin. The release of the targeted immunotoxin molecule occurs only after the T cell has engaged its tumor target. The combination of these two distinct effector systems (T cells and immunotoxin) targeting two distinct tumor antigens have superior anti-tumor activity than a single system targeting a single antigen, with no greater toxicity. By combining two highly specific and partially effective strategies for tumor destruction, we achieved selective and more effective tumor killing.
Bivalent HPV16/18 L1 VLP vaccine induces cross-neutralizing antibodies that may mediate cross-protection (155.27)
Authors: Pinto Ligia , Kemp Troy , Hildesheim Allan , Safaeian Mahboobeh , Pan Yuanji , Porras Carolina , Schiller John , Lowy Douglas , Rolando Herrero 
Published: January 2023

Abstract
Human papillomavirus (HPV) L1 VLP-based vaccines are protective against HPV vaccine-related types; however, correlates of protection have not been defined. We observed that vaccination with the HPV-16/18 L1 VLP vaccine induced cross-neutralizing antibodies for HPV types for which evidence of vaccine efficacy has been demonstrated (HPV31/45) but not for other types for which no cross-protection was observed (HPV52/58). In addition, HPV31/45 cross-neutralizing titers showed a significant increase with number of doses (HPV31, p&lt;0.001; HPV45, p&lt;0.001) and correlated with HPV16/18 neutralizing titers, respectively. Cross-neutralizing antibody titers were much lower than those observed for the homologous vaccines types but persisted over 36 months of follow-up. Increases in antibody avidity occurred with increased doses of vaccine but levels of avidity did not correlate well with antibody titers. Our findings support the hypothesis that cross-neutralizing antibodies are effectors of the cross-protection observed for the HPV16/18 vaccine. Further studies are needed to better understand the potential role of avidity in protection against infection.
Development of three murine models of Staphylococcus aureus infection to aid in the characterization of a broadly protective vaccine and elucidation of immune responses to different routes of infection (155.29)
Authors: Prabhakara Ranjani , Mocca Christopher , Brady Rebecca , Plaut Roger , Burns Drusilla , Stibitz Scott , Merkel Tod 
Published: January 2023

Abstract
Staphylococcus aureus is a common etiological agent of nosocomial and community acquired infections. Pan-antibiotic resistant strains of S. aureus have emerged at an alarming rate, making vaccination an important defense against S. aureus. S. aureus causes a variety of infections that differ in route of infection and disease progression, but previous experimental vaccines have focused on only one type of infection. Hence, multiple animal models are an important tool in studying broadly protective S. aureus vaccines. We sought to develop several murine models that mimic both planktonic and biofilm-mediated S. aureus disease. We focused on three infections that are of the greatest concern: biofilm-mediated implant infection, skin and soft tissue infection (SSTI), and sepsis. We have reproducibly generated a local, chronic, biofilm-mediated implant infection in 100% of C57BL/6 mice, and have determined the minimum dose that results in a chronic infection rate of 100%. We have also made progress towards determining the LD50, maximum non-lethal dose, and minimum lethal dose of S. aureus in a BALB/c sepsis model, as well as reproducibly produced SSTI in BALB/c mice. We plan to use these models to elucidate the memory responses associated with each infection type, and to characterize putative vaccines. If protective immune responses are elicited after vaccination in each of these models, those antigens will be combined into a multivalent vaccine to be tested in all three models.
Yet another phenotypic strategy for defining human IL-17 + CD4 T cells (155.33)
Authors: Li Yongqing , Yee Cassian 
Published: January 2023

Abstract
IL-17-secreting T cells represent a distinct lineage of CD4+ T effector cells (Th17) essential in the pathogenesis of numerous inflammatory disorders, autoimmune diseases and tumor immunity. A means to define the population of human Th17 cells has been confounded by the heterogeneity and plasticity of helper and regulatory T cell subsets. Although combinations of chemokine and surface receptors have been associated with the Th17 phenotype, these markers are not unique to Th17 T cells. In this study, we adapt a murine-based strategy to enrich for a population of human IL-17-producting CD4 T cells from PBMC. Isolation of IL-17+ and IL-17-negative T cell clones provided uniform lineage-specific populations that enabled us to identify differentially expressed genes. Consistent with previously published data, enhanced expression of IL-23R, RORC and CCR4 were well-represented in IL-17+ CD4 T cell clones confirming their lineage specificity. A closer analysis of these clones reveals a functional phenotype that excludes Rhodamine and differentially expresses CCR6 and CD161. We extrapolate these findings to the unselected PBMC population and using a combination of surface and functional markers, demonstrate that a uniquely defined, non-destructive strategy can be used to enrich for IL-17+, Treg-resistant T cells; furthermore, antigen-specific IL-17+ effectors can be expanded using this approach to achieve numbers of cells sufficient for adoptive cellular therapy of cancer.
A modified form of a tumor-associated phosphorylated peptide derived from BCAR3 generates CD8 T cells that are highly crossreactive to the natural phosphopeptide and capable of recognizing tumor cells (156.13)
Authors: Obeng Rebecca , Zarling Angela , Engelhard Victor 
Published: January 2023

Abstract
Using metal ion affinity chromatography and mass spectrometry, we identified a phosphopeptide (BCAR3pT130) derived from BCAR3 among numerous phosphopeptides presented by HLA-A2 on human melanoma cells. BCAR3 activates the PI3K/Akt pathway and mediates migration and estrogen resistance in breast cancer cells, processes that are associated with malignancy. In evaluating the immunotherapeutic use of BCAR3pT130, we found that BCAR3pT130 had a moderate HLA-A2 binding affinity but was nonimmunogenic in HLA-A2 transgenic mice. Substitution of the P9 anchor residue Leu with Val had no impact on HLA-A2 binding affinity but the modified BCAR3pT130 proved to be significantly immunogenic. This suggests that the modification altered the conformation of the phosphopeptide and that the immune response against the natural phosphopeptide is restricted by tolerance. CD8 T cells generated using the modified BCAR3pT130 produced IFNγ, TNFα, and were cytotoxic based on CD107a expression. The T cells were equally reactive with the natural and modified phosphopeptides. This suggests that the natural BCAR3pT130 phosphopeptide is a weak agonist and that the sequence modification creates a stronger agonist. These T cells also recognized endogenously processed BCAR3pT130 on HLA-A2 human melanoma and breast cancer cells. Thus, the association of BCAR3 with malignant processes makes BCAR3pT130 an appealing target for cancer immunotherapy that can be evaluated using the modified form of the phosphoepitope.
IGKV3 proteins as candidate “off-the-shelf” vaccines for kappa-light chain-restricted B cell non-Hodgkin’s lymphomas (156.15)
Authors: Dolcetti Riccardo , Martorelli Debora , Guidoboni Massimo , De Re Valli , Muraro Elena , Turrini Riccardo , Merlo Anna , Pasini Elisa , Caggiari Laura , Romagnoli Luca , Spina Michele , Mortarini Roberta , Gasparotto Daniela , Mazzucato Mario , Carbone Antonino , Rosato Antonio , Anichini Andrea 
Published: January 2023

Abstract
An increasing set of B-cell non-Hodgkin’s lymphomas (B-NHLs) show a biased usage of IGKV3-20 and IGKV3-15 immunoglobulin genes, but it is not known whether the corresponding proteins are immunogenic in the majority of patients. We herein demonstrate that IGK+ lymphoma patients show spontaneous T cell responses to these proteins and that IGKV3-specific cytotoxic T lymphocytes (CTLs) can be easily induced ex vivo. IGKV3-20-specific CTLs cross-react against different IGKV3 proteins, an effect mediated by the presence of 21 shared, sometimes promiscuous, T cell epitopes, presented by the most common HLA class I allele products, thus assuring a broad HLA coverage of IGKV3-based vaccines. Many natural variants of these epitopes are carried by IGK light chains expressed by a broad spectrum of B-NHLs and we show that IGKV3-20-specific CTLs cross-react also against several of these variant epitopes. Both humoral and CTL specific responses were induced by KLH-conjugated IGKV3-20 protein in HLA-A2-transgenic mice and co-injection of IGKV3-20-specific CTLs with IGKV3-20+ lymphoma cells into SCID mice totally prevented tumor growth, thus confirming the ability of these effectors to mediate efficient cytotoxic responses also in vivo. These results provide the rationale to exploit IGKV3 proteins as “off-the-shelf” vaccines for a large fraction of lymphoma patients.
Bi-specific TCR anti-CD3 fusion for the immunotherapy of malignant melanoma (156.16)
Authors: Jakobsen Bent , Ashfield Rebecca , Bossi Giovanna , Adams Katy , Harper Jane , Buisson Sandrine , Paston Samantha , Baker Debbie , Williams Daniel , McGrath Yvonne , Hassan Namir 
Published: January 2023

Abstract
The human immune system can theoretically identify malignant cells by ‘scanning’ peptide antigens presented on the cell surface by Class I HLA molecules, and indeed many cancer patients generate CD8 cyto-toxic T cell responses to tumour-specific antigens. However, the majority of patients fail to clear tumours, since T cell avidity for self-antigens tends to be weak, and cancer cells employ escape mechanisms for avoiding destruction by T cells. We have engineered novel, bi-functional protein therapeutics termed ImmTACs which re-direct the immune system to target and destroy tumour cells with a high degree of potency and specificity. IMCgp100 comprises a soluble, high affinity T cell Receptor specific for the HLA-A2 presented melanoma-associated epitope gp100 280-288, fused to an anti-CD3 scFv domain. The anti-CD3 moiety activates a polyclonal T cell response when targeted to melanoma cells presenting as few as twenty gp100 epitopes, predominantly through activation of effector memory CD8 lymphocytes, leading to killing of the tumour cells. Extensive in vitro testing of IMCgp100 with a panel of tumour cells and normal human tissue cells demonstrates pico-Molar potency against cancer cells and specificity for HLA-A2 gp100 positive cells. The agent is able to inhibit tumour growth in mouse xenograft models. A phase I dose-escalation trial in late stage melanoma patients was initiated in October 2010 and interim results will be presented at this meeting.
MHC II-restricted peptides derived from invariant chain negative breast cancer cell-based vaccines activate healthy donor T-cells (156.17)
Authors: Chornoguz Olesya , Grmai Lydia , Gapeev Alexei , O’Neill Michael , Ostrand-Rosenberg Suzanne 
Published: January 2023

Abstract
We have generated cell-based MHC II vaccines to activate patients’ CD4+ T-cell to facilitate tumor immunity. This was based on the hypothesis that invariant chain (Ii) negative, MHC II and CD80 positive tumor cells present novel MHC II peptides and circumvent patients’ tolerance to their cancer. To confirm that absence of Ii facilitates presentation of novel immunogenic tumor peptides we sequenced MHC II peptides from MCF10/DR7/CD80, MCF10/CIITA/CD80/Ii siRNA (Ii-) and MCF10/DR7/CD80/Ii, MCF10/CIITA/CD80 (Ii+) vaccines using LC-MS/MS. 116 Peptides were identified for MCF10/DR7/CD80, 228 peptides for MCF10/DR7/CD80/Ii. 52 Peptides were in both cell lines. 108 Peptides were identified for MCF10/CIITA/CD80, 28 peptides for MCF10/CIITA/CD80/Ii siRNA, 6 peptides in both cell lines. This confirms that Ii+ and Ii- MHC II vaccines present distinct peptide repertoires and agrees with in vitro studies showing that human MHC II Ii- and Ii+ vaccines activate distinct T-cell subsets. 7 peptides with the highest MHC II binding affinity were tested for their ability to activate tumor-specific CD4+ T-cells. 5 of the peptides activated CD4+ T-cells to the same extent as breast cancer peptide Her2 p776, (measured by IFNγ production). This is the first study to compare the human MHC II peptide repertoire in the absence or presence of Ii and to identify novel immunogenic MHC II-restricted peptides that are potential therapeutic reagents for cancer patients.
Requirements for common γ chain family cytokines for adoptively transferred γδ T cell function and homeostasis in vivo (156.19)
Authors: Kakimi Kazuhiro , Izumi Takamichi , Kondo Makoto , Fujieda Nao , Kondo Atsushi , Morishita Takeharu , Maekawa Ryuji , Matsushita Hirokazu 
Published: January 2023

Abstract
We have conducted clinical trials of adoptive γδ T cell therapy for the treatment of esophageal, colorectal, pancreatic and non-small-cell lung cancers, and cholangiocarcinoma. Patients received ≥1x109 γδ T cells every 2 weeks. These cells had been expanded ex vivo for 14 days by culture with zoledronate (5 μM) and IL-2 (1000 IU/ml). Infused γδ T cells gradually accumulated in patients’ peripheral blood and accounted for 10% of PBMC as late as 3 months after the final injection, even without IL-2 administration in vivo. These cells maintained the ability to release cytotoxic granules (detected by CD107 assay) and produce IFN-γ. To determine the factors contributing to γδ T cell survival in vivo, we investigated their expression of receptors for common γ chain (γc) family cytokines at the time of infusion. IL-2Rβ and γc, but not IL-7Rα, IL-15Rα or IL-21R, were present on these cells, and although the IL-2Rα was upregulated at the initiation of culture, it was only very weakly expressed at the time of transfer. When γδ T cells were maintained for longer periods in culture, IL-15 but not low dose IL-2 and IL-7 supported their survival. These results suggest that tissue-specific γc-dependent signals in response to IL-15 might affect the survival and function of infused γδ T cells, and hence materially influence the success of immunotherapy.
Activating and tolerizing signaling “fingerprint” in T cells responding to DC/tumor MUC1 vaccine (156.23)
Authors: Reichenbach Dawn , Finn Olivera 
Published: January 2023

Abstract
Early biomarkers of efficacy of vaccines are lacking. We hypothesize that intracellular signaling in T cells following administration of the vaccine yields a “fingerprint” predictive of an effective or a tolerogenic immune response and could be used as a biomarker. Our lab has explored immune responses to human tumor MUC1 peptide vaccine in wildtype (WT) versus human MUC1 transgenic mice (MUC1.Tg) and found that they were significantly lower in MUC1Tg. To “fingerprint” signaling differences between the effective immune response in WT versus the lower (tolerogenic) response in MUC1.Tg mice, we vaccinated both with DC loaded with MUC1 peptide, harvested spleens (n=3) at 1,6,12, and 24 hours post vaccination and analyzed phosphorylation downstream of TCR and costimulatory molecules, as well as activation of cell survival, proliferation, and apoptosis signaling cascades. We saw early increases in pSTAT1, pSTAT4, and p38MAPK with a trend towards higher phosphorylation in WT mice that did not reach statistical significance. There was also a trend towards higher phosphorylation in WT mice in pLCK and pAKT downstream of the TCR and costimulatory molecules. Interestingly, at certain timepoints we observed increases in signaling in MUC1.Tg mice, which may serve as compensatory mechanisms that we will further investigate. We are challenging these two vaccinated groups of mice with MUC1+ tumor in an attempt to correlate early T cell activation with later protection from tumor growth.
Increased lethal cytokine-mediated toxicity following immunotherapy in aged versus young mice (156.24)
Authors: Bouchlaka Myriam , Alderson Kory , Sckisel Gail , Chen Mingyi , Murphy William 
Published: January 2023

Abstract
We and others have previously demonstrated that agonistic αCD40 and IL2 immunotherapy (IT) treatment in young mice results in synergistic anti-tumor effects in multiple tumor models. However, efficacy of this and other ITs can be limited by their induction of systemic toxicity. The majority of mouse tumor studies use younger mice (8-12 weeks) which do not accurately represent the age of a typical cancer patient (&gt;50 yrs or 12 months in a mouse). Therefore, we sought to determine the potential toxicity of IT in aged versus young mice. Within 2-3 days after IT treatment, marked toxicity was observed in aged (17-22 months) and middle aged mice (9-12 months) resulting in 100% mortality as opposed to no mortality in young mice (2-4 months). Aged mice had significantly higher serum levels of TNFα, IFNγ, IL6 and IL17a after IT compared to young treated mice, with a proportionate increase with age. Histological evaluation of aged treated mice revealed disseminated lymphocytic infiltrates to the gut, lung and liver; multiple organ damage with hepatic piecemeal necrosis; extensive GI mucosal damage and peribronchiolitis in the lung compared to young treated mice. Finally, IT-treated aged IFNγ-receptor and TNFα-KO mice showed significant decreases in mortality compared to treated WT-aged mice. This data suggests that IT with αCD40/IL2 induces a cytokine storm culminating in multiple organ damage and rapid lethality that correlates with increased serum cytokines and worsens with age.
Cytokine-based cancer immunotherapies induce differential effects on CD4+ and CD8+ T cell phenotype and function (156.9)
Authors: Sckisel Gail , Hsiao Hui-Hua , Bouchlaka Myriam , Wilkins Danice , Tietze Julia , Blazar Bruce , Murphy William 
Published: January 2023

Abstract
We have recently observed that cytokine-based immunotherapy (IT) regimens induced an Ag-nonspecific expansion of the memory T cell pool which was responsible for anti-tumor efficacy. We therefore sought to determine the effects of systemic IT on subsequent Ag-specific responses. During IT, there was a significant expansion of the memory CD8 T cell population with the cells in active cell cycle. Splenocytes from treated mice proliferated less in response to TCR engagement, yet they proliferated to a greater extent when stimulated with IL2 alone. Flow cytometric analysis of stimulated splenocytes revealed that CD8 T cells from IT treated mice appropriately upregulated activation markers (ie, CD25, CD44, NKG2D) and expanded normally. Conversely, CD4 T cells from the same mice exhibited markedly less proliferation, activation marker upregulation, and IFNγ production in response to the same stimuli. In a vaccine model where mice were immunized against allogeneic splenocytes while undergoing IT, CD8 T cells were capable of responding to antigen earlier and more robustly upon secondary in vitro restimulation than immunized, untreated mice. Together, these data show that immune stimulatory therapies allow CD8 T cells to function as Ag-specific or nonspecific effectors while functionally exhausting CD4 T cells. We also show that neither CD4 exhaustion after IT, nor the nonspecific expansion of memory CD8 T cells impairs the ability of CD8 T cells to generate new Ag-specific responses.
Activation of the inflammsome inhibits the development of cell-mediated immunity (157.10)
Authors: Sauer John-Demian , Pereyre Sabine , Portnoy Daniel 
Published: January 2023

Abstract
The inflammasome is a multiprotein complex that leads to the activation of Caspase-1, resulting in secretion of IL-1β and IL-18 as well as the induction of a programmed cell death termed pyroptosis. To understand the role of the inflammasome in the development of CD8+ T-cell mediated adaptive immune responses, we utilized Listeria monocytogenes as a model system. Wild type L. monocytogenes infection resulted in negligible activation of the inflammsome, therefore we engineered a strain of L. monocytogenes to robustly activate the inflammasome. A strain of L. monocytogenes (LpFlaA-L. monocytogenes) engineered to ectopically express Legionella pneumophila flagellin, a well described agonist of the Nlrc4 inflammsome, resulted in strong activation of the inflammasome in vitro. Activation of the inflammasome resulted in attenuation of L. monocytogenes virulence during acute infection. In addition, mice immunized with LpFlaA-L. monocytogenes failed to provide protection against a subsequent lethal challenge. Protection from challenge was rescued in Caspase-1 mice immunized with LpFlaA-L. monocytogenes. In addition to lack of protection, there was a decrease in the development of both antigen specific CD4+- and CD8+-IFNγ producing T-cells following immunization with LpFlaA-L. monocytogenes compared to wild type immunization. Taken together our data suggests that activation of the inflammasome is detrimental to the development of cell-mediated adaptive immune responses.
RIG-LIKE receptor signaling modulates the innate immune response and astrocyte activation after injury to the spinal cord (157.6)
Authors: de Rivero Vaccari Juan Pablo , Minkiewicz Julia , Wang Xiaoliang , German Ramon , Marcillo Alex , Dietrich W. , Keane Robert 
Published: January 2023

Abstract
Injury to the central nervous system (CNS) induces a glial response, in which astrocytes become activated and produce inflammatory mediators. However, the molecular basis for glial activation and the signaling pathways regulating glial/innate immune responses remains poorly understood. Here, we examine the activation of retinoic acid inducible gene-like (RIG) receptors (RLRs) and the involvement of RLR signaling in the regulation of type I IFNs following spinal cord injury (SCI). Immunohistochemical analysis followed by confocal microscopy procedures reveals that astrocytes express two intracellular RLRs, RIG-I and melanoma-associated gene 5 (MDA5). Immunoblotting of spinal cord protein lysates after SCI and stretch injury of culture astrocytes indicates that injury to the CNS activates RLR signaling as determined by phosphorylation of IRF3 leading to production of type I IFNs. Stimulation of RIG-I and MDA5 with synthetic RNA (poly(I:C)) results in RLR signaling activation, phosphorylation of IRF3, and increased expression of glial fibrillary acidic protein and the intermediate filament protein vimentin, two hallmarks of reactive astrocytes. To the best of our knowledge this is the first time that the RLR signaling pathway is described as being involved in regulating astrocyte activation and modulation of the innate immune response after SCI.