bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunometabolic editing: a new type of immunoediting in tumorigenesis (TUM7P.1027)
Authors: Tyson Danielle , Ho Ping-Chih , Tsui Yao-Chen , Kaech Susan 
Published: January 2023

Abstract
Immune evasion and deregulated metabolic transformation are hallmarks of cancer that contribute to tumor aggressiveness and metastasis through different mechanisms. However, it has not been examined whether there is a connection between these two hallmarks. In this study, we approach this question by both melanoma engraftment and a genetically modified melanoma mouse model. First we investigated whether the presence of CD4 or CD8 T cells during tumorigenesis has any effect on a tumors metabolic profile by depleting these cells post-tumor induction. We found that T cell depletion in the early phase of tumorigenesis led to down regulation of hexokinase 2 and pyruvate kinase isozyme M2. Then we investigated whether the effector functions of T cells, especially CD40L and interferon gamma (IFNγ), promote tumor cells to acquire deregulated metabolic activity. It appears the IFNγ may be a major player in the metabolic selection process. Finally, we aim to examine whether the metabolic activity of tumor cells has an effect on the immune activity. In the pilot test, we identified three different melanoma cell lines derived from genetically modified melanomas that have distinct glycolytic activity and oxidative phosphorylation. These tools lay a foundation for future analysis to investigate whether the metabolic features of melanoma cells can play a regulatory role in controlling anti-tumor immunity and the formation of an immunosuppressive microenvironment.
Regulation of CTL trafficking into tumors and anti-tumor immunity by chemoattractant receptors BLT1 and CXCR3 (TUM7P.1015)
Authors: Chheda Zinal , Sharma Rajesh , Jala Venkatakrishna , Bodduluri Haribabu 
Published: January 2023

Abstract
A major hurdle limiting the success of tumor immunotherapies in the clinic is poor recruitment of CD8+ T cells (Cytotoxic T Lymphocytes-CTL) into tumors despite efficient CTL responses in the periphery. However, little is known about the mechanisms that regulate their infiltration into tumors. We investigated the role of leukotriene B4 receptor, BLT1 and CXCR3, the receptor for CXCL9 and CXCL10 in anti-tumor immunity using a syngeneic B16 melanoma mice model. Significant acceleration in tumor growth and reduced survival was observed in both BLT1-/- and CXCR3-/- mice as compared to WT mice. Tumor immune infiltration studies revealed significant reduction of CTLs in BLT1-/- and CXCR3-/- tumors as compared to WT tumors, their frequencies being similar in the periphery. Adoptive transfer of WT but not BLT1-/- or CXCR3-/- CTLs reduced tumor growth in Rag2-/- animals due to enhanced WT CTL infiltration to tumors. Furthermore, adoptive transfer of BLT1-/-, CXCR3-/- or BLT1-/-CXCR3-/- CTLs together with WT CTLs showed that initial migration of BLT1+/+ CTLs to tumors facilitates additional CTL recruitment in a CXCR3 dependent manner. An anti-PD-1 antibody based vaccine completely lost its efficacy in both BLT1-/- and CXCR3-/- mice suggesting a requirement for these receptors in generating sustained anti-tumor immune response. These results demonstrate a critical role for BLT1 and CXCR3 in CTL migration to tumors and thus can be targeted to elicit effective anti-tumor responses.
Rescuing CTLs from undergoing TCR restimulation induced necroptosis improves tumor control (VAC7P.1034)
Authors: Kesarwani Pravin , Gudi Radhika , Chatterjee Shilpak , Scurti Gina , Nishimura Michael , Mehrotra Shikhar 
Published: January 2023

Abstract
Engineering T cells with tumor epitope reactive T cell receptors (TCR) for adoptive cell transfer (ACT) therapy requires rapid proliferation and expansion, resulting in replicative senescent phenotype and increased susceptibility to cell death. Cell death may occur as apoptosis (programmed) or necrosis (un-programmed), or programmed necrosis (termed necroptosis). Our data shows that human T cells engineered with tumor epitope tyrosinase specific TIL1383I TCR upon restimulation with cognate antigen undergo caspase independent and reactive oxygen species mediated cell death cell death that involves JNK. Further, electron microscopic studies revealed that primarily effector-memory phenotype bearing T cells underwent antigen stimulated cell death resembling necroptosis, which was averted by pretreating with necroptosis inhibitor NecroX2 that inhibits RIPK1. Further, in vivo studies done to treat subcutaneously established B16-F10 tumors in C57BL/6 mice showed better tumor control when adoptively transferred tumor epitope gp100 reactive T cells were pretreated with NecroX2. Long-term tracking studies showed that NecroX2 pretreated T cells persisted better than the untreated T cells alone, and could account for differences in tumor growth. Our data highlights that understanding the morphology/type of cell death that a tumor reactive T cell may undergo upon TCR restimulation is important to effectively rescue anti-tumor T cells for increasing persistence and improving ACT.
Functional deficiencies in chimeric antigen receptor T cells from geriatric donors (VAC7P.1035)
Authors: Guha Prajna , Cunetta Marissa , Thorn Mitchell , Point Gary , Junghans Richard , Katz Steven 
Published: January 2023

Abstract
Age-related immune dysfunction poses unique and poorly defined challenges to the use of cancer immunotherapy in geriatric patients. We sought to test our hypothesis that CAR-T from geriatric patients (gCAR-T, n=3, &gt;65 years) are functionally impaired relative to CAR-T from younger patients (yCAR-T, n=3, 18-45 years). Donor T cells were genetically modified to express CAR specific for carcino-embryonic antigen (CEA), presently in use in our phase I trials. Untransduced and CAR-T cells were expanded ex vivo with IL2 (3000IU/ml) or IL15 (5ng/ml) for 20 days. A trend towards higher transduction efficiencies (35.2% yCAR-T vs 24.9% gCAR-T, p=0.09) and cell expansions of 2.5 fold, (p=0.01, two-tailed t-test) were observed in yCAR-T cells for both treatments as compared to gCAR-T. yCAR-T contained higher proportions of CD4 effector cells (EC) (64.3+7.6%,p=0.003), CD4 effector memory (EM) cells (43.2+6.8%,p=0.002) and CD8 effector memory cells (EM) (16.5+3.6%,p=0.002). In accordance with higher numbers of CD4 and CD8 TEM, yCAR-T demonstrated higher levels of (1.3-fold over gCAR-T, p=0.03) CEA specific cytotoxicity compared to gCAR-T, with maximum cytotoxicity observed in IL15 treated yCAR-T cells. Enhanced tumor killing by yCAR-T correlated with increased levels of perforin (1.3+0.1 ng/ml,p=0.01) and granzymeB (0.4+0.03 ng/ml, p=0.02). Our pilot study suggests gCAR-T demonstrate age-related deficiencies that may impact production and anti-tumor efficacy.
Evaluating use of the probiotic yeast Saccharomyces boulardii for the synthesis and delivery of oral therapeutics (VAC8P.1061)
Authors: Hudson Lauren , McDermott Courtney , Fasken Milo , Kuiper Emily , Guiliano David , McBride Shonna , Corbett Anita , Lamb Tracey 
Published: January 2023

Abstract
Recent studies have explored the possibility of using microorganisms to synthesize and deliver therapeutics for the treatment of gastrointestinal disorders. Probiotic organisms, particularly the eukaryotic probiotic yeast Saccharomyces boulardii, may be well suited for this task given their current clinical applications for the treatment of gastrointestinal infections and inflammatory bowel diseases. Saccharomyces is furthermore capable of performing eukaryotic post translational modifications and can thus be made to express a wide array of therapeutic proteins. In order to increase the ease with which S. boulardii can be transformed to express recombinant protein, we developed auxotrophic mutant strains of S. boulardii using UV mutagenesis. Unlike wild type S. boulardii, these mutant S. boulardii can be genetically manipulated without reliance on antibiotic selection. These mutants also show a similar profile of resistance to conditions found in the gut, such as low pH and bile acid salts, relative to wild type S. boulardii. Critically, we demonstrate through oral gavage experiments that viable mutant S. boulardii recovered from murine gastrointestinal immune tissues continue to express recombinant protein. These mutant S. boulardii will thus permit studies to evaluate the ability of eukaryotic probiotics to deliver oral recombinant therapeutics and provide adjunctive therapy for the treatment of gastrointestinal disorders.
Leukocyte immune-type receptors are markers of antiviral cytotoxic T lymphocytes in channel catfish, Ictalurus punctatus (VET1P.1125)
Authors: Taylor Erin , Wilson Melanie , Bengten Gertrud 
Published: January 2023

Abstract
LITRs are a multigene family in teleosts that encode immunoglobulin superfamily receptors with activating or inhibitory signaling capability, but their functional role in the immune response is not well understood. These immunoregulatory receptors exhibit sequence homology to both Fc receptors and molecules encoded in the leukocyte receptor complex. By using a combination of immunoprecipitation and protein identification by LC-MS/MS, we determined that the monoclonal antibody CC41 binds to LITRs on clonal cytotoxic T lymphocytes (CTL). Consequently, we hypothesize that this antibody can be used to monitor the in vivo antiviral CTL response. To test this hypothesis, we injected gynogenetic catfish with a channel catfish virus (CCV)-infected MHC-matched clonal T cell line (G14D-CCV) and collected peripheral blood lymphocytes (PBL) at 3, 5, and 9 days after injection for analyses by flow cytometry. The percentage of CC41+ cells was elevated five days after primary immunization and three days after booster immunization. When the CC41+ cell population was magnetically isolated from the PBL, they were found to specifically kill CCV-infected targets, as shown by 51Cr-release assays, and this killing is partially inhibited by the addition of CC41. When subjected to flow cytometric mRNA analyses, the CC41+ cells were found to express message for CD3γδ, perforin, and eomesodermin (EOMES), indicating that these cells are in fact CTL.
Diminished innate, but not adaptive, immune responses in migrating adult Chinook salmon, Oncorhynchus tshawytscha (VET1P.1127)
Authors: Dolan Brian , Fisher Kathleen , Colvin Michael , Benda Susan , Danley Courtney , Peterson James , Kent Michael , Zwollo Patty , Schreck Carl 
Published: January 2023

Abstract
In wild systems, animals have to balance the energy required to mount an immune response with other requirements necessary to survive and procreate. An extreme example is migrating adult Pacific salmon species, which cease feeding at the start of their migration from salt water to fresh water. During the migration, salmon must swim against strong currents, compete for mates, evade predation, and contend with freshwater pathogens. We quantified the immune response in migrating adult Chinook salmon, Oncorhynchus tshawytscha, at different times and locations during the run using several different measures. We measured the levels of immunoglobulin heavy chain mu and heavy chain tau mRNA transcript in the anterior kidney and found that levels of both secreted and membrane bound forms of the heavy chain did not diminish during migration, and importantly, that they remained unchanged even after spawning. We also measured the innate anti-bacterial ability of plasma isolated from fish to prevent the growth of a lab strain of E. coli. Contrary to the humoral immune response, the innate anti-bacterial components of plasma diminished at later time points in the run leading up to spawning. Diminished immune response was not due to IL-10 secretion, as transcripts for this cytokine were similar at all time points that were examined. These data suggest that in the face of limited energy resources, innate immune responses diminish while adaptive responses remain unchanged.
Characterizing the murine immune response to Corynebacterium pseudotuberculosis: approaches to developing a C. pseudotuberculosis vaccine (VET1P.1119)
Authors: Hino Christopher , Crowley Daniel , Chisholm Gregory , Geiger Kristina , Sumida Megan , Molinder Karen , Pollock Roberta 
Published: January 2023

Abstract
Pigeon Fever is a major infectious equine disease caused by the Gram-positive, intracellular bacterium Corynebacterium pseudotuberculosis. The disease manifests as external abscesses, internal infections, or ulcerative lymphangitis. Horses with external abscesses have a very low mortality rate, while horses with the less common internal infections have a high mortality rate of up to 40%. We have developed a mouse model for this disease to examine the roles of different T-cell and antibody responses to C. pseudotuberculosis infection. BALB/c and C57Bl/6 mice were challenged intradermally with varying doses of live bacteria following immunization with crude bacterial fractionations. Analysis of serum antibody levels of unvaccinated mice suggests that both mouse strains did not mount a significant humoral immune response against C. pseudotuberculosis infection. By contrast, mice mounted a high IgG response after being immunized with crude bacterial fractions, which providing partial protection against infection. We have generated monoclonal antibodies from immunized mice and used these antibodies to immunoprecipitate their respective antigens. Three proteins have been identified by mass spectrometry, and are being cloned so that the recombinant proteins can be tested for their effectiveness as vaccine components. These studies highlight the potential of a vaccine strategy based on the use of immunogenic bacterial proteins for use in a protective vaccine.
Th17 deviation under influence of Th1 response attenuates the inflammation in late phase of influenza virus infection (VIR5P.1138)
Authors: Huang Ching-Tai , Dutta Avijit , Chen Tse-Ching , Lin Chun-Yen , Lin Yung-Chang , Chang Chia-Shiang , He Yueh-Chia 
Published: January 2023

Abstract
Overwhelming inflammation with cytokine storm has been postulated as a major cause of severe influenza disease in human. In our influenza Ag- specific mouse experimental system, we discovered a Th17 deviation of CD4+ T cell response with resultant attenuation of inflammation in the late phase of influenza infection. Naïve hemagglutinin (HA)-specific CD4+ T cells respond to acute influenza virus infection with Th1 phenotype. In the presence of antigen specific Th1 CD4+ T cells, naïve CD4+ T cells adoptively transferred 4 days after the acute infection are activated with deviation to the Th17 phenotype. The second batch of naïve HA-specific CD4+ T cells is activated in an environment enriched with Th1 cytokines and active TGF-β. They proliferate, secrete IL-17 but not IFN-γ and express LAG-3 on the cell surface. They can suppress T cell activation in vitro and decrease lung inflammation in vivo. The 6.5 mice, with monotonous TCR repertoire responsive to influenza HA, reveal a Th17 deviated CD4+ T cell response and significant survival benefit in influenza virus infection. Consecutive waves of naïve CD4+ T cells from the thymus in 6.5 mice are probably activated as subsequent batches of naïve CD4+ T cells, with Th17 deviation which attenuates the inflammation in late phase of influenza virus infection.
Herpes simplex virus tegument protein VP11/12-derived epitopes preferentially recall polyfunctional effector memory CD8+ T cells from seropositive asymptomatic individuals and protect “humanized” HLA-A*02:01 transgenic mice against ocular herpes (VIR6P.1172)
Authors: Benmohamed Lbachir , Khan Arif , Srivastava Ruchi 
Published: January 2023

Abstract
In this study, we used multiple prediction computer-assisted algorithms to identify 10 potential HLA-A*02:01-restricted CD8+ T cell epitopes from the 716 amino acids sequence of Herpes Simplex Virus type 1 virion tegument phosphoprotein 11/12 (HSV-1 VP11/12). Three out of ten epitopes exhibited high to moderate binding affinity to HLA-A*02:01 molecules. In ten sequentially studied HLA-A*02:01 positive, HSV seropositive healthy asymptomatic (ASYMP) individuals (who have never had clinical herpes disease), the most frequent, robust and polyfunctional effector CD8+ T-cell responses, as assessed by a combination of tetramer frequency, granzyme B, granzyme K, perforin, CD107a/b cytotoxic degranulation, IFN-g and multiplex cytokines assays, were predominantly directed against three epitopes: VP11/1266-74, VP11/12220-228 and VP11/12702-710. Interestingly, ASYMP individuals had significantly higher proportion of CD45RAlowCCR7lowCD44highCD62LlowCD27lowCD28lowCD8+ effector memory T cells (TEM) specific to the three epitopes, compared to symptomatic (SYMP) individuals (with a history of numerous episodes of recurrent ocular herpetic disease). Moreover, immunization of HLA-A*02:01 transgenic mice with the three ASYMP CD8+ TEM cell epitopes induced a strong protective immunity against ocular herpes infection and disease. Our findings outline the features of protective HSV-specific CD8+ T cells that should guide the development of an effective T-cell-based herpes vaccine.
Mouse polyomavirus tropism alters magnitude and functionality of virus-specific CD8 T cells (VIR6P.1173)
Authors: Qin Qingsong , Jin Ge , Lukacher Aron 
Published: January 2023

Abstract
Mouse polyomavirus (MPyV) is a ubiquitous persistent natural mouse pathogen. Human JC polyomavirus variants associated with progressive multifocal leukoencephalopathy have discrete amino acid mutations in VP1, the major capsid protein, that affects tropism for glial cells. A glutamic acid (E) to glycine (G) difference at position 92 of VP1, shifts the profile to tumors induced by MPyV from epithelial to mesenchymal cell origin. Here, we asked if this shift in tropism affects the MPyV-specific CD8 T cell response, which controls MPyV infection/tumorigenesis. We found that the A2 (92E-VP1) and RA (92G-VP1) MPyV strains differ sharply in MPyV-specific CD8 T cell responses. A2 virus generates a robust, high magnitude virus-specific CD8 T cell response, whereas RA virus elicits a markedly smaller anti-MPyV CD8 T cell response. Infection by an A2 92G-VP1 mutant virus recapitulates the antiviral CD8 T cell response seen with RA virus. Notably, the antiviral CD8 T cell recall response during persistent infection by this A2 mutant significantly exceeded that of A2 virus. Using immunization with peptide-loaded dendritic cells in the setting of bystander inflammation by A2 or the mutant virus, we found that A2 virus stimulates a higher magnitude MPyV-specific CD8 T cell response; this difference is negated in DC-deficient Flt3-/- mice. Together, these data indicate that viral tropism for DCs is a major determinant of the quantity and quality of the anti-MPyV CD8 T cell response.
CD8+ T cells from elite controllers recognize and kill HIV abortively infected CD4+ T cells (VIR6P.1168)
Authors: Monel Blandine , Boucau Julie , McKeon Annmarie , LeGall Sylvie , Walker Bruce 
Published: January 2023

Abstract
Understanding the underlying immunological mechanisms of spontaneous HIV control by HIV+ elite controllers (EC) remains an important step in the development of efficient HIV vaccines and functional cures. To understand control mechanisms triggered by CD8+ T cell we established a system in non-activated peripheral blood mononuclear cells (PBMC) using infectious HIV (NL4.3 IRES GFP) carrying the fusion protein Vpr-Beta lactamase (to assess viral entry) and a staining strategy to study the CD8+ T cell response by flow cytometry. First we showed that non-activated CD4+ T cells are permissive for HIV entry but this leads mostly to abortive infection (b-lac+ GFP-). CD8+ T cells from EC were able to recognize these HIV+ cells, became activated (IFNg+, MIP1b+) and underwent degranulation (CD107a+). This activation was initiated by the presentation of HIV peptides from incoming viruses and HLA-I blockade almost abolished this response. We confirmed by mass spectrometry HIV peptides processed in the cytoplasm of CD4+ cells shortly after HIV entry. These results suggest that the ability of CD8+ T cells to form immunological synapses allows them to be activated by low levels of HIV antigens from incoming particles at the surface of early-infected CD4+ T cells. Killing of abortively HIV-infected cells early after HIV entry might avoid inflammation due to pyroptosis and therefore contribute to HIV control.
Antigen processing constraints on CD4+ T cell epitope dominance: an approach to improved epitope prediction (APP5P.113)
Authors: Landry Samuel , Charles Tysheena , Mettu Ramgopal 
Published: January 2023

Abstract
We present a novel method for CD4+ epitope prediction based on modeling antigen processing constraints. Given an antigen with known three-dimensional structure, our algorithm combines four types of data to construct a profile of conformational stability that allows us to identify regions that are most accessible to proteolysis. Using this profile, we construct a profile of epitope-likelihood based on the pattern of unstable-to-stable transitions. We validate our method using 23 datasets of experimentally measured CD4+ T-cell responses of mice bearing IAb or HLA-DR4 alleles as well as of human subjects. Overall, we show that an antigen processing-based approach has at least as much, and sometimes more, predictive power for epitope prediction than existing, sequence-based methods that rely on MHC class II binding affinity estimates.
CD4 T Cell reactivity to both native and citrullinated-MOG epitopes identified in HLA-DR4 mouse model Of demyelinating disease (BA11P.137)
Authors: Kersh Anna 
Published: January 2023

Abstract
Multiple sclerosis (MS) is an autoimmune disease characterized by inflammation and demyelination in the central nervous system (CNS). HLA-DR4 is an MHC class II gene associated with increased risk for MS. Humanized HLA-DR4-transgenic mice were used to probe T cell specificity and affinity during demyelinating disease progression in response to myelin oligodendrocyte glycoprotein (MOG). Using the micropipette adhesion frequency assay to obtain sensitive, physiologically relevant measurements, only 26% of CNS-infiltrating CD4 T cells are reactive to MOG in symptomatic HLA-DR4-expressing mice. Moreover T cells probed for specificity to a modified MOG peptide containing a citrulline residue in position 101 instead of arginine revealed T cells reactive to MOG alone, citrulline-modified MOG, or to both. Of importance, the citrullinated-MOG peptide does not induce paralytic disease on its own. We believe that these results are the first demonstration of T cell reactivity to citrullinated-MOG epitopes in the context of HLA-DR4 presentation to T cells. We suggest that T cell cross-reactivity to native and citrullinated epitopes of MOG provides a mechanism of disease progression and escape from immune tolerance. Further, reactivity to the citrullinated neoepitope has great implications for use as a potential biomarker of disease therapeutic efficacy since citrullination is the result of a deimination reaction that occurs during prolonged inflammation.