bluuebunny/crossref_metadata_2025_split huggingface dataset:
Developing an antibody targeting CεmX of mIgE for the treatment of allergic and other IgE-mediated diseases (VAC6P.954)
Authors: Chan Yueh-Hsuan , Lee Yu-Tzu , Chou Hung-Wen , Wu Pheidias C , Chen Jiun-Bo , Li Chau-Hong , Cheng Tien-Tien , Chen Nien-Yi , Lue Ko-Haung , Chang Tse-Wen 
Published: January 2023

Abstract
CεmX (also referred to as M1’) is a discrete domain of 52 a.a. residues, located between the CH4 domain and the C-terminal membrane anchor peptide of the ε heavy chain of membrane-bound IgE (mIgE) on human B lymphocytes. Antibodies that target CεmX are potentially useful in controlling IgE production for treating allergic and other IgE-mediated diseases. Based on its abilities to bind to mIgE with high affinity and to lyse mIgE-expressing B lymphocytes by apoptosis, ADCC, and other cytolytic mechanisms, humanized 4B12 monoclonal antibody (h4B12 mAb) is now under preclinical development. In an allergic asthma animal model employing genetically modified mice that express mIgE containing human CεmX domain, we have demonstrated that mAb 4B12 was effective in reducing antigen-specific IgE, but not immunoglobulins of other isotypes. 4B12 could also markedly alleviate airway hyperresponsiveness (AHR) to inhaled methacholine in those mice. Furthermore, we have demonstrated that h4B12 could inhibit the production of human IgE in mice that had been reconstituted with human PBMCs. We have now developed a transfected CHO cell line capable of producing h4B12 at a high yield in preparation for human clinical trials.
Therapeutic clearance of the virally infected nervous system via noncytopathic T cell interactions with resident myeloid cells (VAC6P.945)
Authors: Herz Jasmin , McGavern Dorian 
Published: January 2023

Abstract
The goal of anti-viral therapy is to promote pathogen clearance without causing tissue injury. Therapeutic administration of anti-viral T cells (adoptive immunotherapy) has shown promise in clinical studies for the treatment of CMV, EBV, and adenovirus. Our laboratory models adoptive immunotherapy by transferring anti-viral memory T cells into mice persistently infected from birth with lymphocytic choriomeningitis virus (LCMV). Here we demonstrate that brain-resident myeloid cells (microglia), in addition to neurons, are persistently infected with LCMV in carrier mice. Therapeutic memory T cells can completely purge virus from persistently infected microglia and neurons without causing blood brain barrier breakdown or cellular injury. This is accomplished through a tailored release of chemoattractants that recruit adaptive immune cells, but few pathogenic innate immune cells. In addition, memory T cells enlist the support of nearly all microglia by converting them into CD11c-expressing antigen-presenting cells (APCs). We observed similarly converted myeloid cells in the brains of humans infected with JC virus and HIV-1. Using two-photon microscopy, we revealed that during clearance of the LCMV-infected brain anti-viral CD8 T cells form stable, noncytopathic interactions with CD11c+ microglia, whereas CD4 T cells interact transiently. These data demonstrate that brain-resident APCs can thwart the cytolytic functions of anti-viral T cells while allowing viral clearance.
Use of biologic nanolipoprotein particles containing monophosphoryl lipid A as a novel vaccine platform against inhalational pathogens (VAC7P.956)
Authors: Dunkle Alexis , Blanchette Craig , Boone Tyler , Corzett Michele , Hwang Mona , Fischer Nicholas , Lehmann Doerte , Lychak Cheri , Hoeprich Paul , Driks Adam , Rasley Amy 
Published: January 2023

Abstract
Subunit vaccines are theoretically safe and easy to manufacture but require effective adjuvants and delivery systems to yield protective immunity, particularly at critical mucosal sites such as the lung. We investigated nanolipoprotein particles (NLPs) containing the Toll-like receptor 4 agonist monophosphoryl lipid A (MPLA) as a platform for intranasal vaccination against Bacillus anthracis. Modified lipids enabled attachment of disparate spore and toxin protein antigens. Intranasal vaccination of mice with B. anthracis antigen-MPLA-NLP constructs vs. delivery with free MPLA induced robust IgG and IgA responses in serum and in bronchoalveolar and nasal lavage. Typically, a single dose sufficed to induce sustained antibody titers over time. When multiple immunizations were required, specific antibodies were detected earlier in the boost schedule with MPLA-NLP-mediated delivery than with free MPLA. Administering combinations of constructs induced responses to multiple antigens, indicating potential for a multivalent vaccine preparation. No off-target responses to the species-matched NLP scaffold protein were detected. Using the ovalbumin/TCR transgenic model, MPLA-NLPs enhanced antigen-specific T cell expansion in lung-draining lymph nodes. In summary, the NLP platform enhances cellular, humoral, and mucosal responses to intranasal immunization, indicating promise for NLPs as a flexible, robust vaccine platform against B. anthracis and potentially other inhalational pathogens.
Recombinant outer membrane protein: a potential candidate for Th17 based vaccine against Klebsiella pneumoniae. (VAC7P.967)
Authors: Chen Kong , Goodwin Meagan , McAleer Jeremy , Nguyen Nikki , Way Emily , Kolls Jay 
Published: January 2023

Abstract
Bacterial pneumonia is a leading cause of mortality and one major pathogen associated with this disease is Klebsiella pneumoniae. Recently, the emergence of antibiotic resistant strains demands an effective vaccine against these bacteria. We have previously shown that intranasal immunization with heat-killed K. pneumoniae or crude outer membrane proteins isolated from K. pneumoniae induced antigen specific Th17 responses and these Th17 cells conferred serotype independent protection against various clinical isolates of K. pneumoniae including the recently described multidrug resistant New Delhi Metallo-beta-lactamase-1 strain. To develop a clinically relevant Klebsiella vaccine, we cloned single OMP genes by PCR from K. pneumoniae and successfully expressed one recombinant outer membrane protein, OmpX, in the BL21 E. coli strain. The purified recombinant OmpX was recognized by Klebsiella immune serum by direct ELISA and also recognized by Th17 cells from Klebsiella immunized mice. In vivo, intranasal immunization of the purified OmpX induced robust mucosal Th17 responses and left IL-17 producing gamma-delta T cells unaffected. Ongoing work will determine whether these Th17 responses will result in serotype independent protection against live bacterial challenge including the multidrug resistant strains.
Vaccination with a Hybrid 1 (H1) fusion protein combined with a liposomal adjuvant (CAF01) induced antigen specific T-cells 3 years post vaccination in a human clinical trial. (VAC7P.971)
Authors: Henson Dawn , van Dissel Jaap , Joosten Simone , Graves Andrew , Hoff Søren , Soonawala Darius , Prines Corine , Thierry-Carstensen Birgit , Andreasen Lars , de Visser Adriette , Agger Else , Ottenhoff Tom , Kromann Ingrid , Andersen Peter , Hokey David 
Published: January 2023

Abstract
Approximately one third of the world’s population is infected with Mycobacterium tuberculosis. Developing novel vaccines to protect against pulmonary tuberculosis is a public health priority. In this study, a Hybrid 1 (H1) subunit vaccine containing a recombinant fusion protein of Ag85B and ESAT-6 was paired with a two-component CAF01 liposomal adjuvant system developed and manufactured by Statens Serum Institut. A phase I clinical study was performed to evaluate H1:CAF01 in healthy non-BCG vaccinated adult male and female subjects between the ages of 18 and 55 years old. The subjects were randomized into four groups including H1 alone or H1 with 125/25µg, 313/125µg or 625/125µg CAF01 and were vaccinated on study days 0 and 56. PBMCs harvested approximately 150 weeks post vaccination were used to assess antigen specific responses by a 13-color intracellular cytokine staining assay. Vaccination with H1:CAF01 resulted in statistically significant Ag85B-specific CD4 polyfunctional CD154+ T cells compared to H1 alone. ESAT-6 stimulation resulted in detection of CD4 polyfunctional CD154+ T cells responses that were not elevated to a statistically significant extent compared to H1 alone. This is the first demonstration of the persistence of an antigen-specific cellular immune response up to 3 years after vaccination in a clinical trial using H1:CAF01 vaccination.
CCR10 expression is required for immunogenicity of a HIV-1env DNA vaccine encoding CCL28 to enhance HIV-1env-specific IgG and IgA at relevant mucosal sites (VAC7P.986)
Authors: Kutzler Michele , Kathuria Noshin , Curatola Ashley , Makurumidze Getrude , Myles Devon , Muthumani Kar , Sylvester Albert , Ginsberg Arielle , Hutnick Natalie , Moldoveanu Zina , Hunter Meredith , Khan Amir , Sardesai Niranjan , Mestecky Jiri , Marx Preston , Weiner David 
Published: January 2023

Abstract
Development of a vaccine that drives anti-viral mucosal B cell responses is critical for protection against HIV-1 infection. It is currently unknown whether triggering CCR10/CCL28 pathways in a DNA-based vaccine results in induction of HIV-1env specific B cell immunity at mucosal sites of infection. We hypothesized that co-immunization with HIV-1env/CCL28 molecular adjuvant would augment B cell responses at gastrointestinal and vaginal sites and require CCR10. CCL28 co-immunized WT mice displayed a significant enhancement of HIV-1env specific antibody titers in serum, feces and vaginal washes, and enhanced HIV-1 specific IgA responses were abrogated in CCR10KO mice. CCL28 co-immunization did not increase the breadth of linear B cell epitopes in WT mice, but augmented the dominant epitopes elicited by antigen immunization alone. The frequency of splenic and intestinal IgA+CD19+B220+CD138+CCR10+ plasmablasts was augmented in the CCL28 co-immunized WT mice over antigen-only immunized WT controls. The physiological relevance of these findings was confirmed in a NHP model of intravaginal SIVsmE660 challenge in which CCL28 co-immunization resulted in significant increases in serum/vaginal IgG/IgA, decrease in peak viral loads, significant suppression of viral titers over 120 days, and recovery of CD4 T cells. These data support a role for CCL28 in targeting protective anti-viral B cells to mucosal sites when delivered as molecular adjuvants for HIV-1env DNA-based vaccines.
High-affinity TCRs confer antigen sensitivity in HIV gag-specific effector CD8 T cells (VAC7P.991)
Authors: Wilson Caleph , Robles-Oteiza Camila , Riley James 
Published: January 2023

Abstract
Virus infected cells are eliminated by CTLs when TCR recognizes viral Ag on MHC. Previously, we demonstrated that HIV-1-GAG specific high-affinity (ha) TCR enhanced CD8 T cell killing of HIV-1-infected cells. To elucidate why haTCR expressing T cells were better at controlling HIV-1 replication, we asked whether WT or haTCRs expressing primary human T cells had a distinct cytokine profile when stimulated with monomeric pMHC attached to magnetic beads. In this minimalistic T cell activation system, we observed robust IFNγ, TNFα, IL-2 and MIP1β induction in the presence or absence of CD28 stimulation. Interestingly, αCD3 coated beads do not induce cytokine expression. Previously, we and others demonstrated that αCD3 stimulation failed to induce AKT activation, which has a prominent role in cytokine production. Here, TCR signaling alone activates AKT. Next, we addressed whether T cells expressing haTCRs were able to respond to less Ag, and thereby control HIV-1 replication better by destroying HIV-1 infected cells before they could effectively spread the virus. By mixing various ratios of cognate pMHC with non-cognate pMHC on magnetic beads, we show that CD8 T cells, and not CD4 T cells, expressing haTCRs are able to induce Ag specific activation of phospho-ERK1/2 and -AKT with a ~10 fold increased sensitivity when compared with WT TCR expressing T cells. Thus, implicating CD8 as playing an important role in regulating how a T cell perceives differences in TCR affinity.
Novel vaccination strategy: Francisella tularensis vaccines based on functionalized catanionic vesicles (VAC7P.963)
Authors: Richard Katharina , Mann Barbara , Stocker Lenea , Barry Eileen , Qin Aiping , Cole Leah , Hurley Matthew , Ernst Robert , Michalek Suzanne , Stein Daniel , DeShong Philip , Vogel Stefanie 
Published: January 2023

Abstract
Francisella tularensis (Ft) is a Gram-negative, immune-evasive coccobacillus that causes tularemia for which there is no FDA-approved vaccine. We have utilized a novel vaccine approach using synthetic nanoparticles made from catanionic surfactant vesicles (V), functionalized by incorporation of either Ft type B Live Vaccine Strain (Ft LVS) or Ft type A Schu S4 strain (Ft Schu S4) components (i.e., LVS-V and Schu S4-V, respectively). Immunization of C57BL/6 mice with bare V partially protected against Ft LVS, presumably through activation of an innate immune response, yet failed to protect against Ft Schu S4. In contrast, immunization with LVS-V fully protected mice against intraperitoneal (i.p.) Ft LVS challenge, while immunization of mice with either LVS-V or Schu S4-V partially protected C57BL/6 mice against an intranasal (i.n.) Ft Schu S4 challenge. LVS-V-immunization, but not immunization with V, elicited high levels of IgG against non-LPS epitopes and these antisera conferred passive protection against challenge with Ft LVS. Our recently published and ongoing studies aim to identify the protein targets of mouse antisera, study the mechanism of non-specific protection gained by immunization with this nanoparticle vaccine platform (adjuvant effect) in Ft LVS challenge, and enhance protection in Ft Schu S4 challenge. Our data extend the utility of functionalized catanionic surfactant vesicles as a vaccine platform for pathogens.
Poxviral infection elicits human neutralizing antibodies recognizing diverse epitopes of the major vaccinia virus surface protein D8 (VIR4P.1016)
Authors: Gilchuk Iuliia , Blum David , Fritz Gloria , Singh Vidisha , Kose Nurgun , Yu Yingchun , Sapparapu Gopal , Matho M. , Zajonc Dirk , Xiang Y. , Hammarlund Erika , Slifka Mark , Crowe James 
Published: January 2023

Abstract
Immunization with vaccinia virus (VACV) confers cross-protective immunity to variola, the causative agent of smallpox, and other clinically important poxviral species, such as cowpox and monkeypox viruses. Vaccine-mediated protection has been correlated with elicitation of broad neutralizing antibody (Ab) responses, however the specificity of human neutralizing antibodies to poxviruses is poorly understood. We used a highly-optimized human hybridoma technology to generate large panels of anti-VACV monoclonal Abs (mAbs) from vaccinia-immunized subjects or from subjects who had naturally acquired monkeypox infection. Immunologic characterization and DNA sequencing of the panel revealed 16 individual mAbs that recognized the immunodominant surface protein antigen D8. Several of anti-D8 mAbs possessed neutralizing activity against VACV, cowpox and monkeypox viruses. These Abs exhibited mainly a complement-dependent pattern of neutralization and they formed at least four cross-blocking groups, suggesting the existence of several non-overlapping neutralizing epitopes for D8. The results suggest that D8 is an important target for human neutralizing Abs generated following poxviral infection or vaccination, and suggest the existence of novel epitopes targeted by anti-D8 Abs that contribute to cross-protective immunity following smallpox vaccination.
Despite large clonal expansions of cytotoxic T cells with age and chronic CMV exposure, the underlying repertoire appears completely normal, suggesting that holes in the T cell repertoire are not responsible for immunosenesence (VIR4P.1012)
Authors: Mukherjee Rithun , Lindau Paul , Desmarais Cindy , DeGloria Jeanne , Utsugi Heidi , Warren Edus , Riddell Stanley , Makar Karen , Turtle Cameron , Robins Harlan 
Published: January 2023

Abstract
Age is thought to restrict the cytotoxic immune repertoire, both in terms of number and diversity of naïve T cells available for responding to novel pathogens and vaccinogens. Studies have shown that CMV-induced chronic stimulation exacerbates this immune restriction. CMV infection leads to CD8 T cell clonal expansions (TCEs), bearing classical memory as well as late-differentiated memory phenotypes. Such TCEs become huge in the elderly, where they have been held responsible for constriction of the remainder of the repertoire. In turn, this has been linked with the CMV-associated immune risk profile and increased mortality in the elderly, although it is not known how TCEs affect immune competence. Here we address the premise that immune competence with age could be compromised because of repertoire restriction. We do so by describing the structure of the elderly repertoire at an unprecedented resolution in terms of both clonal origin and broad CD8 T cell phenotypes. We then proceed to exhaustively identify CMV-specific clones. This presents us with a close look at the rest of the repertoire, which does not respond to CMV. We compare this with the full repertoire in CMV- individuals. Our observations suggest that CMV-specific TCEs are restricted to components of the repertoire that expand without affecting the absolute size or diversity elsewhere in the repertoire, so “holes” are unlikely to develop in the T cell repertoire.
Mechanisms of enhanced TRAIL, CD86, and CD40 expression by plasmacytoid dendritic cells from HIV-infected subjects (VIR5P.1027)
Authors: Serei Virian , Dai Jihong , Hodder Sally , Fitzgerald-Bocarsly Patricia 
Published: January 2023

Abstract
Plasmacytoid dendritic cells (pDC) are critical in viral immunity via their production of IFN-α and presentation of antigen. In HIV+ subjects, pDC decline impairs immune defense, while chronic in vivo stimulation of pDC contributes to immune activation. Here, we aimed to further explore pDC dysfunction in HIV+ subjects. We confirmed that pDC from HIV+ subjects have increased TRAIL, CD86, and CD40 expression compared to healthy donors. pDC from both HIV+ and healthy subjects upregulated TRAIL, CD86, and CD40 in response to overnight HIV- and HSV-stimulation. However, pDC from HIV+ subjects with CD4 &gt;200 showed increased TRAIL, CD86, and CD40 upregulation compared to healthy donors and HIV+ subjects with CD4 &lt;200. pDC from healthy donors were then used to explore how expression of these molecules occurs. Inhibition of endosomal acidification blocked TRAIL, CD86, and CD40 in virus-stimulated pDC. TLR7 inhibition in HIV-stimulated pDC partially inhibited this expression. While TLR9 blockage also inhibited CD40 expression by HSV-stimulated pDC, there was no effect on CD86 and TRAIL. Pro-inflammatory cytokines IL-6, TNF-α, and IFN-α were able to enhance TRAIL expression by HIV-stimulated pDC. Despite upregulation of CD86, HIV- stimulated pDC from healthy donors were poor inducers of allogeneic CD4 T cell expansion. We conclude that pro-inflammatory cytokines, along with TLR signaling, “prime” pDC in HIV-infected individuals to express TRAIL, CD86, and CD40.
Latent viral infection alters the immune architecture of the lung (VIR5P.1030)
Authors: Dwivedi Varun , Trgovcich Joanne , Mansfield Sara , Cook Charles 
Published: January 2023

Abstract
Human cytomegalovirus (HCMV) is ubiquitous virus infecting most humans and developing persistent latent infections. HCMV reactivation occurs in ~33% of immune-competent patients during critical illness associated with doubled risk of mortality. In many persistent infections Myeloid derived suppressor cells (MDSC) cause T cell dysfunction, creating an immunosuppressive microenvironment. We hypothesized that latent CMV infection induces MDSC, perhaps contributing the impairment of host resolution of infection and thus latency. BALB/c mice were infected with 1×106 PFU of Smith strain murine CMV and lung mononuclear cells (LMNC) were cultured ex vivo or stained for surface markers and analyzed by ELISA or FACS. During latency, we observed a change in frequency (million/gm of lung tissue) of cell subsets such as marked reduction of CD4+T and B cells, increased CD8+T cells and significant increase in NK cells compared to naïve mice. Interestingly, the percentage of MDSC was unchanged at 2 weeks, but was significantly reduced at12 weeks post infection (p.i.). Both IL-6 and TNF-α were elevated in unstimulated or LPS, ConA stimulated LMNC at 2 and 12 weeks p.i. conferring the risk of lung pathology under critical illness. Our findings suggest that during the transition from acute to latent infection, persistent pro-inflammatory lung immune environment is associated with reduced MDSC. Longitudinal studies are underway to explore the role of MDSC in lungs of latently infected mice.
Dysregulated host inflammatory response in rats with fatal encephalitic Rift Valley Fever (VIR5P.1021)
Authors: Hartman Amy , Caroline Amy , Bethel Laura , Powell Diana , Oury Tim , Reed Douglas 
Published: January 2023

Abstract
Rift Valley Fever virus (RVFV) is a zoonotic arbovirus capable of causing widespread outbreaks with severe morbidity, mortality, and socioeconomic consequences. Most patients develop an acute febrile disease but recover, while a small subset of infected humans develop more severe disease including hemorrhagic manifestations, meningoencephilitis, or retinitis resulting in blindness. Inbred rats have been shown to be a good model of severe human RVF. Lewis rats are highly resistant to subcutaneous inoculation with RVFV, but develop uniformly fatal encephalitis after exposure to low doses of RVFV by inhalation. Elevated levels of M-CSF and MCP-1 are seen in the blood on day 4 post-exposure concurrent with granulocytosis and thrombocytopenia that continues until death. IL-1β and GRO/KC, the rat homolog of IL-8, peaked slightly on day 4 p.i., decreased briefly, and were highly elevated in the blood of moribund rats. In the brain, many pro-inflammatory cytokine levels, including IL-1α, IFN-γ, and IL-18, were found to be greatly elevated at day 5 post-exposure, which coincides with the earliest detection of virus in the brain. Pathological evaluation of brain tissue indicates an influx of lymphocytes and neutrophils that corresponds with the onset of neurological signs and severe disease in the Lewis rats. These data suggest that the host immunological response may be contributing to the pathogenesis of RVFV in the brain and the outcome of infection.
Human cytomegalovirus microRNA miR-UL148D facilitates HCMV latency (VIR5P.1024)
Authors: Liu Yuan , Pan Chaoyun , Li Donghai , Liu Fenyong , Zen Ke 
Published: January 2023

Abstract
Human cytomegalovirus (HCMV), infects a majority of population and can undergo life-long, latent infection, is a leading cause of opportunistic and congenital disease. HCMV can encode 17 microRNAs (miRNAs) but the roles of these viral miRNAs during viral infection are unclear. Here, we infected human bone marrow and CD34+ hematopoietic progenitor Kasumi-3 cells with HCMV (strain NR-1) and determined the kinetics of cellular levels of all hcmv miRNAs by qRT-PCR. We found that, different from most other hcmv miRNAs which had high level at early days of infection, hcmv-miR-UL148D was highly expressed at the latent infection (10days or longer). Bioinformatics and experimental validation showed that hcmv-miR-UL148D targets the 3’UTR of host cell immediately-early response gene 5 (IER5). Functional assay indicated that downregulation of IER5 by hcmv-miR-UL148D transcriptionally upregulated CDC25B, leading to lower number of Kasumi-3 cells in G2/M phase. As G2/M phase is a cell cycle of large viral replication, decrease the number of cells in G2/M phase by hcmv-miR-UL148D would suppress viral replication and facilitate hcmv entering in latent infection. In contrast, NR1-Δhcmv-miR-UL148D, an hcmv mutant deleting hcmv-miR-UL148D, was lack of ability to target IER5, increase CDC25B, control the cell cycle of host cells, and achieve latent infection. Our results collectively demonstrate that hcmv-miR-UL148D facilitates hcmv latency through modulating host cell IER5-CDC25B axis.