bluuebunny/crossref_metadata_2025_split huggingface dataset:
Engineering filamentous phage as carriers that focus antibody responses against a peptide (47.11)
Authors: van Houten Nienke Elizabeth , Smith George P. , Scott Jamie K. 
Published: January 2023

Abstract
Antibody (Ab) responses to a B-cell epitope can be produced by immunization with a peptide bearing the epitope of interest. Such peptides are often poor immunogens, requiring conjugation or fusion to carrier proteins that provide T-cell help. However, carrier proteins may also divert the Ab response away from the targeted peptide via its own B-cell epitopes. Thus, an ideal peptide carrier should provide T-cell help to drive B-cell responses but contain relatively few B-cell epitopes.
The Ab response to filamentous phage is restricted to the 12 N-terminal residues of the major coat protein (pVIII) and the external domains of the minor coat protein, pIII. Previously, we reported that the Ab response against a synthetic peptide conjugated to phage was more focused on the peptide compared to ovalbumin as the carrier. In some cases, the anti-peptide Ab titer exceeded the anti-carrier Ab titer by &gt; 20 fold. The reduced complexity of the phage surface compared to ovalbumin’s may be responsible for this.
Here, we tested the hypothesis that Ab responses to weakly immunogenic peptides would be improved by removing immunodominant epitopes from the phage surface; immunogenic residues on pVIII were altered, and the external domains of pIII were truncated. The phage were tested alone as immunogens, and as carriers for pVIII-displayed peptides. The results and the implication for vaccine design are discussed.
Supported by NIH R01AI-49111
Complete Protective Immunity induced by Cottontail Rabbit Papillomavirus E1 Epitope DNA Vaccination in HLA-A2.1 transgenic rabbits (47.13)
Authors: Hu Jiafen , Schell Todd , Peng Xuwen , Cladel Nancy , Balogh Karla , Christensen Neil 
Published: January 2023

Abstract
Cottontail rabbit papillomavirus (CRPV)/HLA-A2.1 transgenic rabbit infection model has been reported previously. Because the CRPV genome can be modified extensively without loss of its function, this transgenic rabbit model can be used for testing the specificity of host immunity to specific HLA-A2.1 restricted papillomavirus protein epitopes in vivo. In this study, we incorporated online MHCI epitope prediction software, HLA-A2.1 transgenic mouse and rabbit systems together to demonstrate an efficient way to identify and test immunogenicity of HLA-A2.1 restricted epitopes from CRPVE1 (161–169 LLFRQAHSV and 303–311 MLQEKPFQL). Both epitopes were able to induce specific cytotoxic CD8 T cells in HLA-A2.1 transgenic mice immunized with peptides. Using an epitope DNA vaccination method, complete and partial protection against CRPV infection was achieved by CRPVE1/303–311 and CRPVE1/161–169 in HLA-A2.1 transgenic rabbits respectively. Our data further demonstrated the application of HLA-A2.1 transgenic rabbits as a potential in vivo model system to test papillomavirus therapeutic vaccines.
Passive immunity against infection with Trypanosoma brucei subsp. brucei in mice is conveyed by an antibody blocking the ligand-binding extracellular domain 1 (ed1) of the C2 receptor CRIT (47.20)
Authors: Inal Jameel Malhador , Rock Peter 
Published: January 2023

Abstract
Anti-CRIT-ed1 (recognising the first extracellular domain of CRIT) was passively transferred into mice which were subsequently challenged with 500 trypomastigotes of T. brucei subsp. brucei. At a peak of infection, eight days after challenge infection, mice that had received 0.5 mg anti-CRIT-ed1 (ascites) had significantly reduced levels of parasitaemia compared to control mice. This inhibiton was also found to be dose-dependent. As passive transfer of anti-CRIT-ed2, the antibody recognising the second extracellular domain did not reduce parasitaemia, the protection was deemed specific. The protection afforded by passive transfer of anti-CRIT-ed1 also delayed mortality. To exclude the possibility that the reduction in parasitaemia was due to intact IgG inducing complement-mediated lysis of trypomastigotes we found that whereas anti-CRIT-ed1 was unable to induce complement-mediated lysis of cultured trypomastigotes, the sera from T. brucei subsp brucei-infected mice induced lysis.
Detection of HBV Antigen-Specific Cellular Immune Responses after PrimeBoost Therapeutic Vaccination in Chronic Hepatitis B Patients (47.26)
Authors: Brown Dean , Bourne Victoria , Cripps Martin , Anderson Richard , Schneider Joerg 
Published: January 2023

Abstract
Hepatitis B virus (HBV)-specific T cell responses play an important role in the clearance of HBV infection. Heterologous Hi-8™ PrimeBoost vaccination using HBV surface antigen (HBsAg) is a promising approach for inducing T cell responses in chronic hepatitis B patients and results in viral suppression and liver function normalisation. Cellular immune responses against HBV antigens were determined in PBMC isolated from patients treated with Hi-8™ HBV therapy. Samples were tested by ex vivo IFN-γ ELISPOT assay and after 14 days of culture in the presence of overlapping peptides spanning the HBsAg and HBV core antigen (HBcAg) sequences. Ex vivo HBsAg-specific T cell responses were of very low frequency and transient. However, higher frequency T cell responses that could be correlated with clinical outcome were measured after culture. Notably, HBsAg but not HBcAg-specific responses increased after treatment, indicating that responses were driven by vaccination. Further experiments are being conducted to determine the relative contribution of CD4 and CD8 T cells to the immune response. In conclusion, HBsAg specific IFN-γ producing T cells were detected in PBMC from chronically infected patients and increased in those receiving Hi-8™ HBV therapy. Since the T cell immune responses could only be detected following in vitro culture, they are either of very low frequency in the peripheral blood or of a memory phenotype.
Comprehensive Analysis of the Antibody Response to Yersinia pestis by use of a Novel Whole Proteome Microarray (47.3)
Authors: Keasey Sarah L , Schmid Kara , Purcell Bret K , Morefield Garry L , Meegan James , Tikhonov Alex , Schweitzer Barry , Ulrich Robert G 
Published: January 2023

Abstract
A novel proteome microarray representing the majority of the 4086 Yersinia pestis KIM proteins was produced by high-throughput expression and purification. Rabbit hyper-immune sera were produced against proteomes extracted from several pathogenic gram-negative bacteria for use in validation assays. The antibody profile from each of the rabbits enabled detection of:
shared cross-reactive proteinssignature proteins common for two or more bacteria, andfingerprint proteins specific to each pathogen.
Unique proteins were recognized by convalescence sera from mice that survived plague following immunization with the experimental F1-V vaccine. Several new antigens were discovered that were recognized by antibody from rhesus that survived plague, whereas these Y. pestis proteins were not recognized by sera from animals surviving challenge with spores of the gram-positive Bacillus anthracis. Finally, analysis of sera from cynomolgus macaques acutely infected with Y. pestis or Bacillus anthracis produced antibody-binding patterns that were unique biomarkers for each disease. Our results demonstrate that the whole proteome microarray platform is very useful for establishing diagnostic biomarkers, detecting antibody responses to proteins that could potentially show promise in vaccine development, and also for determining the degree of antigen-cross reactivity between different strains or species of bacteria.
Antiviral effect of The ImmunomodulatorAS101 on West Nile Virus by AlphaVBeta3 integrin inhibition (47.33)
Authors: Sredni Benjamin , Indenbaum Victoria , Makarovsky Daria , Bin Hanna , Albeck Michael , Lustig Shlomo , Mendelson Ela 
Published: January 2023

Abstract
The identification of integrin αVβ3 as a receptor for West Nile virus (WNV) in recent years, created opportunities in the development of antiviral strategist against WNV infection. The immunomodulator AS101 [ammonium trichloro (dioxyethylene 0-0′)tellurate] has an antiviral effect in vivo and in vitro. Our study aimed at determining whether AS101 has an antiviral effect against WNV and its mechanism of work. Cell viability assay for Vero cells treated with AS101 before WNV infection, revealed up to 90% survival of the treated cells. More over, plaque forming unit (PFU) assay showed 83% inhibition at WNV-infected Vero cells after AS101 treatment. Significant decrease in the concentration of viral infectious particles was observed in supernatants of Vero cells infected with WNV at high MOI (5) after AS101 addition, and the viral envelope protein (E) expression decreased in Vero cells infected with WNV (MOI=5) and treated with AS101. In supernatants collected from virus infected and AS101 treated Vero cells immediately after inoculation were more virus particles, than in not treated cells. A novel AS101 effect was seen at attachment assay were AS101 inhibited Vero cells attachment to human integrin αVβ3 specific antibody in dose dependent manner. Thus, the results presented here shows that AS101 has an antiviral effect against WNV, and suggests that AS101 prevents WNV entry to the cells by inhibiting integrin αVβ3.
Directing the natural catalytic activity of antibodies to hepatitis C virus E2 protein (47.36)
Authors: Sapparapu Gopal , Planque Stephanie A , Wang Runsheng , Nishiyama Yasuhiro , Keck Zhen-Yong , Foung Steven , Paul Sudhir 
Published: January 2023

Abstract
The envelope protein E2 is involved in hepatitis C virus (HCV) entry into host cells. The proteolytic activity of antibodies (Abs) offers a novel neutralizing mechanism by permanent inactivation of multiple epitopes by a single Ab molecule. We studied the proteolytic cleavage of E2 linked to a glutathione S-transferase tag (E2-GST) by Abs purified from human sera. IgM but not IgG class Abs hydrolyzed recombinant E2 determined by an electrophoresis assay, consistent with previous observations that Ab catalysis is an innate immune function subject to deterioration over the course of adaptive B lymphocyte differentiation. To determine the feasibility of engineering HCV E2-specific proteolytic Abs, we cloned hybrid human IgGs composed of light chains with promiscuous proteolytic activity (clones GG63, HK13 and HK14; isolated from a lupus phage library) and the heavy chain of an HCV E2-specific Ab (clone CBH-7; isolated from an HCV infected subject). The hybrid Abs hydrolyzed E2-GST with little or no activity in the wildtype control Ab. The hybrid Abs displayed the ability to bind 125I-E2, confirming that the heavy chain subunit can imparts specificity for E2. These observations suggest the feasibility of harnessing the natural proteolytic activity of Abs to developing catalytic reagents with potential immunotherapeutic utility.
The neonatal calf tuberculosis vaccine model: immune responses to protective and non-protective vaccines after aerosol challenge with virulent Mycobacterium bovis (47.39)
Authors: Waters Ray , Palmer Mitchell , Nonnecke Brian , Thacker Tyler , Estes Mark , Jacobs William R. , Larsen Michelle 
Published: January 2023

Abstract
An attenuated Mycobacterium tuberculosis RD1 knockout and pantothenate auxotroph (mc26030) vaccine failed to protect neonatal calves from a low dose, aerosol M. bovis challenge. In contrast, M. bovis bacille Calmette Guerin (BCG)-vaccinates had reduced tuberculosis-associated pathology as compared to non- and mc26030-vaccinates. Mycobacterial colonization was not impacted by vaccination. Positive prognostic indicators associated with reduced pathology in the BCG-vaccinated group included: decreased antigen induced IFN-gamma, iNOS, IL-4, and MIP1-alpha responses and a diminished activation phenotype (i.e., ↓CD25+ and CD44+ cells and ↑CD62L+ cells) in mycobacterial-stimulated mononuclear cell cultures. The calf sensitization and challenge model provides an informative screen for candidate tuberculosis vaccines before their evaluation in costly non-human, primates.
Active Aβ vaccination-induced Aβ clearance enhanced by suppression of the CD40-CD40L interaction (48.11)
Authors: Bai Yun , Obregon Demian , Hou Huayan , Nikolic William , Mori Takashi , Zeng Jin , Ehrhart Jared , Giunta Brian , Fernandez Frank , Morgan Dave , Town Terrance 
Published: January 2023

Abstract
Active and passive Aβ immunization efficiently reduces amyloid plaque load and memory impairment in transgenic mouse models of Alzheimer’s disease. Immunization has also yielded favorable results for many patients; but a significant percentage also developed severe meningioencephalitis. Prior studies by our group suggest CD40 receptor-CD40 ligand interaction between T-cells and microglia initiates release of proinflammatory factors, which drive phenotypic switching from a phagocytic to an antigen presenting cell phenotype thus impairing the clearance of Aβ. We therefore tested whether a CD40 blockade could enhance Aβ vaccination-mediated Aβ clearance in vivo. Importantly in our present study, Aβ vaccinated CD40 deficient heterozygous transgenic mice over-expressing mutant amyloid precursor protein and presenilin-1 demonstrated enhanced Aβ clearance; compared with vaccinated PSAPP mice. Similar results were obtained with Aβ vaccinated PSAPP mice passively immunized with CD40L antibody injections. Taken together with our previous studies these data suggest that CD40 blockade, in the context of active Aβ vaccination, acts to enhance CNS Aβ clearance by maintaining microglia in a phagocytic phenotype. These data also may provide the basis for a method whereby enhancing Aβ vaccination-mediated Aβ clearance strategies.
4-1BB is Superior to CD28 Costimulation for Generating Effector CD8+ T Cells for Adoptive Immunotherapy (48.12)
Authors: zhang hua , Snyder Kristen , Suhoski Megan , Maus Marcela , Kapoor Veena , June Carl H , Mackall Crystal L 
Published: January 2023

Abstract
Artificial APCs (aAPCs) provide a new approach to expand cells for adoptive cell therapy, but optimal requirements for CD8+ expansion have not been defined. Beads expressing αCD3/αCD28 efficiently expand CD4+ cells, but not cytolytic CD8+ T cells. 4-1BB, a costimulatory molecule expressed by a minority of resting CD8+ T cells, is transiently upregulated by all CD8+ T cells following activation. We compared the capacity for aAPCs providing costimulation via 4-1BB vs. CD28 to expand cytolytic CD8+ T cell populations. Whereas αCD3/αCD28 aAPCs preferentially expand naive cells, αCD3/4-1BBL aAPCs preferentially expand memory cells, resulting in cultures with enhanced reactivity to previously primed viral antigens and leading to efficient expansion of electronically sorted CD8+ populations reactive toward viral or tumor-associated antigens. Moreover, using HLA-A2-Fc fusion proteins, 4-1BBL aAPCs selectively induced a 3 log expansion of antigen-specific CTL from polyclonal CD8+ PBL over 14 days; similar expansions were not induced using HLA-A2-Fc/αCD28 aAPCs. Compared to αCD3/αCD28 expanded CD8+ CTL, αCD3/4-1BBL expanded CD8+ T cells also mediate enhanced cytolytic capacity as a result of 4-1BB signaling itself and NKG2D upregulation. These results demonstrate that 4-1BB costimulation is superior to CD28 costimulation for expanding antigen specific CD8+ CTL for use in adoptive immunotherapy.
Genetic immunizations in the presence of a specific neurokinin 1 receptor agonist triggers Th1-Tc1 biased immune responses that protect against B16 melanomas (48.13)
Authors: Mathers Alicia R , Tkacheva Olga A , Janelsins Brian , Schuffeski William , Morelli Adrian E , Larregina Adriana T 
Published: January 2023

Abstract
Antigen (Ag) specific Th1-Tc1 cellular immunity induced by genetic immunizations is critical to prevent / eradicate tumors. The high number of resident dendritic cells (DCs) makes the skin an ideal microenvironment for genetic immunizations against tumors. Skin DCs become potent stimulators of Ag specific T cells by signals triggered during acute inflammation in peripheral tissues. The secretion of the pro-inflammatory neuropeptide substance P signaling via the neurokinin 1 receptor (NK1R) represents an early danger signal that stimulates APC functions of DCs. Here we tested whether signaling via the NK1R during skin genetic immunizations favors Th1-Tc1 immunity in C57BL/6 mice injected with B16 melanoma cells. We utilized the gene gun to deliver transgenic (tg) OVA or TRP-2 Ag to mouse skin and the NK1R agonist Sar9Met (O2)11-SP as a potential Th1 adjuvant. Our strategy expressed Ag exclusively in the epidermis and induced substantial migration of activated Langerhans cells (LCs) transporting tg Ag to skin draining lymph nodes, which resulted in generation of Th1-Tc1 immunity. Importantly delivery of tg TRP-2 plus NK1R agonist was able to protect mice against B16 melanoma tumor cell growth. Our data demonstrate that NK1R signaling provides the Th1 adjuvant effect that positively modulates the immunostimulatory functions of LCs during skin genetic immunizations leading to productive immune responses.
Form of antigen dictates immunity: Irradiated cell vs. whole cell lysate vaccination (48.16)
Authors: Buckwalter Matthew , Srivastava Pramod 
Published: January 2023

Abstract
The context in which antigen is perceived by the immune system dictates the quality of the ensuing immune response, which can range from tolerance to lasting immunity. In the current report we investigated the effects of vaccinating mice with antigen in two separate contexts, namely: Irradiated cells, or whole cell lysates. Using the MethA tumor model we observed that although a single vaccination with irradiated MethA cells leads to immunity in BALB/c mice, vaccination with the same cell equivalents of whole cell lysate does not. These results were surprising in light of the substantial body of literature demonstrating the immunostimulatory effects of cell lysates. We hypothesize that although each vaccination contains the same potential antigens, due to the different context in which they are delivered, the immune system will react to them differently. To test this we analyzed multiple characteristics of the immune response following vaccination with irradiated cells or whole cell lysates including: Tumor protection, the quantity and quality of an antigen specific T cell response, stability and persistence of antigen, and the effects of multiple vaccinations. We report dramatic differences between the two immunogens at each of the aspects tested. Our data suggests that although whole cell lysates contain antigen and the same potentially immunostimulatory components as irradiated cells, vaccinating mice with irradiated tumor cells leads to a much greater T cell response and tumor protection than does vaccinating with whole cell lysates.
Induction of Antigen-Specific CD8+ T cells in Melanoma Patients using a PrimeBoost Therapuetic Vaccine (48.18)
Authors: Anderson Richard John , Whale Kevin , Brown Dean , Scheibenbogen Carmen , Asemissen Anne Marie , Cripps Martin , Schneider Joerg ,   The Hi-8 MEL Phase I/II Study Group
Published: January 2023

Abstract
Induction of tumour-specific CD8+ T cell responses in melanoma patients is the major aim of antigen-specific immunotherapy. This study evaluated a prime-boost immunotherapeutic vaccine (Hi-8™ MEL) in stage III/IV melanoma patients using vectors encoding a polyepitope string of seven CD8+ T cell epitopes from five melanoma antigens. T cell responses were evaluated in cryopreserved PBMC samples taken before and during treatment. CD8+ T cells specific for the immunodominant Melan-A epitope were detected by ex vivo tetramer assay in 24/36 (67%) of all patients and in 10/11 (91%) of patients that received the highest treatment dose. Eleven of 36 (31%) patients mounted an ex vivo IFN-γ ELISPOT response: 9/36 (25%) to Melan-A and 3/36 (8%) to other epitopes in the string (NY-ESO-1 and MAGE-3). Notably, the Melan-A-tetramer+ CD8+ T cells in one patient reached 7.25% and acquired a CD45RA−CCR7− effector/memory phenotype. This patient also exhibited an ELISPOT response and a partial clinical response (&gt;12 months). Across the study, tetramer responders showed a median survival of 86 weeks versus 37 weeks for non-responders (p&lt;0.001), indicating that Hi-8™ MEL immunotherapy can elicit melanoma-specific CD8+ T cell responses that are associated with tumour control and increased survival. Additional immune assays are underway to further elucidate the nature of these efficacious T cell responses.
HLA-B-Associated Transcript 3 (BAT3) is an activating ligand for NKp30 and released from dendritic cells (48.2)
Authors: Simhadri Venkateswara Rao , Reiners Katrin S , McKinnon Peter J , Hallek Michael , Engert Andreas , von Strandmann Elke Pogge 
Published: January 2023

Abstract
Natural-killer (NK)-cells, originally defined as innate lymphocytes are regulators of adaptive immune responses via cytokine secretion and cross-talk with dendritic cells (DCs).
Both killing and maturation of DCs by NK cells is dependent on NKp30, a Natural Cytotoxicity Receptor (NCR) expressed on NK cells. However, the NKp30-specific ligands expressed on dendritic cells have remained elusive.
We recently identified BAT3 as the tumor associated ligand for NKp30. BAT3 is a nuclear protein characterized by the conserved BAG domain (Bcl-2 associated anthogene), that interacts with HSP70. Here we show that immature and mature DCs express BAT3 mRNA and protein. The protein is released from DCs into the extracellular environment upon exposure to stress signals such as serum starvation and heat shock. Moreover, the purified, recombinant BAT3 is functional as it binds to NKp30 and induces NK cell-mediated cytokine secretion. The inhibition of BAT3 using polyclonal antibodies prevents tumor rejection in vivo, demonstrating that BAT3 is necessary for the tumor rejection in a xenograft tumor model.
Thus, BAT3 is a novel ligand for the NK cell-receptor NKp30 and its role in the reciprocal activation of NK-DC cross-talk will be discussed. We present evidence that BAT3 is, together with HSP70 and HMGB1, a member of the emerging group of immune regulators that we call “Nucleokines”.
Support- Koeln Fortune Programme (89/2004).
Biodegradable Artificial Antigen Presenting Cells for ex vivo stimulation of antigen-specific T Lymphocytes (48.9)
Authors: Fahmy Tarek M , Steenblock Erin R 
Published: January 2023

Abstract
While dendritic cells (DCs) are the most potent professional antigen-presenting cells, their widespread utility in therapeutic approaches involving ex vivo stimulation of T lymphocytes is hindered by several practical factors such as generation of autologuous DCs, labor and isolation and preparation for clinical applications.
Thus, artificial antigen-presenting cells (aAPC) have been proposed and tested in the expansion of a number of specific T cells for the treatment of a variety of disease states. Here we have developed a novel physiologically compatible microparticulate system for ex vivo T cell stimulation using biodegradable polymers. Our approach fabricates FDA approved polymers into microparticles presenting T cell antigens, thus providing the multivalent interactions necessary for efficient stimulation of T cells. A key feature of this technology is the ability to co-encapsulate cytokines and program their release during and after activation and in a sustained fashion, facilitating long-term bio- availability and efficient stimulation of lymphocytes.