bluuebunny/crossref_metadata_2025_split huggingface dataset:
The VSVΔG/ZEBOV GP vaccine protects against an Ebola infection and induces humoral and cellular immunity (128.16)
Authors: Alimonti Judie B. , Qiu Xiangguo , Fernando Lisa , Melito Leno , Feldmann Friedericke , Stroher Ute , Feldmann Heinz , Jones Steven 
Published: January 2023

Abstract
Ebola virus (EBOV) is a highly infectious filovirus causing a severe hemorrhagic fever in humans with fatality rates approaching 90%. We used the live attenuated recombinant vaccine vector vesicular stomatitis virus (VSV) to create a vaccine for EBOV. Ten cynomolgus macaques were immunized either intramuscularly (IM), orally (OR), or intranasally (IN) with VSVΔG/ZEBOVGP 28 days before receiving a lethal challenge of Zaire ebolavirus (ZEBOV). There were no signs of illness in the vaccinated animals, and no loss of lymphocytes typically seen during infection. ZEBOVGP-specific humoral and cell mediated immune responses (CMIR) were generated by all immunization routes. Surprisingly, in general, the IN route generated more potent humoral responses, while the IM route generated early IL-2 and IFN-γ CMIR. We demonstrate that a single dose of the VSVΔG/ZEBOVGP vaccine delivered by any of the three routes induces cellular and humoral immune responses and protects non-human primates against a lethal challenge of ZEBOV.
Tegument- and glycoprotein-reactive HSV-1-specific CD4 and CD8 T-cells localize to human trigeminal ganglia (128.24)
Authors: Jing Lichen , Verjans G.M.G.M , Haas Juergan , Bailer Susanne , McClurkan Christopher L , Chong Tiana M , Koelle David M 
Published: January 2023

Abstract
Chronic HSV infections of the natural human host are characterized by lifelong latency in dorsal root ganglia. It is not clear which viral proteins can access antigen presentation pathways in the human ganglia. HSV-1-infected ganglia showed increased inflammation compared to virus-negative ganglia. The leukocyte infiltrate in human HSV-infected TG was shown to include HSV-specific CD4 and CD8 T-cells using interferon-gamma (IFN-g) readouts. By cloning HSV-1 ORFeome in a large set of eGFP fusion plasmids, We have interrogated the fine specificity of TG-derived T-cell lines. Antigen presenting cells for CD8 assays were HLA class I heavy chain /HSV-1 ORF co-transfectants, while HLA class II-matched PBMC were used as APC to stimulate CD4 responses. CD8 recognition of discrete HLA class I-restricted epitopes was documented for two distinct HSV-1 glycoproteins, gL and gK, expressed with late kinetics, as well as for the major transactivator tegument protein, VP16. Tetramer and IFN-g staining showed that up to 20% of TG CD8 cells recognized single HSV-1 epitopes. Glycoprotein-specific CD8 T-cells displayed nanomolar avidity and cross-recognized the homologous HSV-2 nonamer peptides. Interestingly, VP16 was also recognized by TG-infiltrating CD4 T-cells. HSV-1 lytic cycle proteins can access ganglionic HLA class I and II antigen presentation pathways during chronic TG infection, implying that local APC somehow sample viral proteins from neuronal reactivation events. Local antigen-specific T-cells may contribute to the life-long equilibrium between latency and clinical reactivation, and influence disease phenotype. Supported by NIH grant AI50132.
The contributions of glycosylation and GPI-linkage of the murine cytomegalovirus immune evasin, m157, to functional interactions with its lectin-like Ly49 receptors (128.25)
Authors: Carlin-Brown Lindsey E. , Guseva Natalya V. , Heusel Jonathan W. 
Published: January 2023

Abstract
The MCMV-encoded glycoprotein, m157, is recognized by the activating Ly49HC57Bl/6 and inhibitory Ly49I129S lectin-like NK cell receptors. We and others have shown that Ly49H is tolerant to multiple amino acid changes in m157 with the exception of K161, which is predicted to anchor a unique N-terminal α0-helix to the main m157 chain. We undertook site-directed mutagenesis to address the contributions of N-linked glycosylation and the GPI anchor of m157 to Ly49H and Ly49I recognition. Variants of m157 containing single or multiple N-to-Q substitutions at positions N178, N187, N213, and N267, or fusions between the m157 extracellular domain and the transmembrane + cytoplasmic domains of DAP12 or CD4, were expressed in myeloid or fibroblast cell lines. We find that 1) only fully deglycosylated m157 has reduced capacity to activate Ly49H or chimeric Ly49HI reporter cells; 2) m157-CD4 transmembrane versions of m157 fully activate Ly49H and chimeric Ly49HI reporter lines; and 3) Ly49H+ NK cells are activated by all N-glycosylation and transmembrane variants of m157. Thus, neither N-glycosylation nor GPI anchoring of m157 is required for functional trans interactions with lectin-like Ly49 receptors.
Lethal poxvirus infection can be treated at later stages of infection using Abs to the non-structural type I IFN binding protein (128.30)
Authors: Xu Renhuan , Sigal Luis 
Published: January 2023

Abstract
Ectromelia virus (ECTV) is an Orthopoxvirus (OPV) that causes mousepox, the murine equivalent of human smallpox caused by the OPV Variola virus (VARV). Both ECTV and VARV encode a Type I IFN binding protein (EVM166 in ECTV) that inactivates IFNƒÑ. We have previously shown that ECTV deficient in EVM166 is avirulent and that immunization of mice with recombinant EVM166 prevented mousepox in BALB/c mice challenged with WT ECTV. Here, we tested whether EVM166 protein can also be used as a target for post-exposure therapy. Strikingly, we found that rabbit polyclonal or mouse mAb to EVM166 given in a single dose 1, 3 or even 5 days post infection (PI) were highly protective. Moreover, treatment with anti-EVM 166 significantly reduced virus loads and liver pathology. Furthermore, we found that EVM166 protein is present in the sera of mice from day 5 PI and that this was significantly reduced following anti-EVM 166 treatment. Analysis of gene expression demonstrated that mice treated with anti-EVM166 Ab produced ~5-fold less Type I IFN in the liver but had ~5-fold higher expression of the IFN inducible genes such as Mx1. This suggests that the amount of type I IFN production in the liver is dependent on virus loads and that anti-EVM 166 Ab treatment decreases virus loads in the liver by permitting IFN signaling and an anti-viral state of the organ. Our study provides a new promising strategy for post-exposure treatment of poxvirus diseases and yields important basic information about the mechanisms whereby Type I IFNs protect in vivo and the host-pathogen interaction.
This work was supported by NIH grant R01AI065544 to LJS
Inhibition of HIV-1 infectivity across subtypes by the green tea catechin, epigallocatechin gallate, without altered immune function (128.6)
Authors: Nance Christina L , Siwak Edward B , D'Souza Melinda , Songa Sonia , McMullen Ashley M , Shearer William T 
Published: January 2023

Abstract
Previously, we presented evidence that the green tea catechin, epigallocatechin gallate (EGCG), binds to the CD4 molecule at the gp120 attachment site on T cells. CD4+ T cells, B cells and macrophages were isolated from HIV-1 uninfected donors. HIV-1 infectivity by HIV-1 p24 ELISA on M-tropic (R5) subtypes B, C and G; T-tropic (X4) subtypes B and D; and dual tropic (R5/X4) subtype B. Immune function studies: cytotoxicity by ViCell, apoptosis by flow cytometry, immunoglobulin (Ig) production by ELISA and cytokine profiling by multiplex platform. Statistical significance by Student's t test. EGCG significantly inhibited HIV-1 infectivity on human immune cells in a dose-dependent manner (6-100μM) (p&lt;0.01-0.001). At physiologic concentration of 6μM, EGCG significantly inhibited HIV-1 p24 antigen production across HIV-1 subtypes [B (p&lt;0.001), C, D, and G (p&lt;0.01)]. The specificity of the EGCG-induced inhibition was substantiated by the failure of EGCG derivatives lacking galloyl and/or pyrogallol side groups to alter HIV-1 p24 levels. EGCG-induced inhibition of HIV-1 infectivity was not due to cytotoxcity, cell growth inhibition, nor apoptosis. Neither Ig nor cytokine production were significantly altered. EGCG inhibits HIV-1 infectivity across a broad spectrum of HIV-1 subtypes without significant effects on immune function, thereby supporting EGCG as a candidate for HIV-1 therapy.
(NIH AT003084)
Identification of a Mamu Class I-restricted listeriolysin T-cell epitope in rhesus macaques infected with Listeria monocytogenes (LM) (129.13)
Authors: Williams Kristina M. , Bigley Elmer C. , Babu Uma S. 
Published: January 2023

Abstract
LM is a gram positive foodborne pathogen that causes a flu-like illness in immunocompetent patients, but immunocompromised patients can develop septicemia and meningitis resulting in a high mortality rate. The primate model of LM infection is being used to investigate both innate and adaptive components of immunity in an animal model that is more relevant to human disease. Cytotoxic T lymphocytes were isolated from rhesus monkeys infected with LM by nasogastric intubation and used to identify Class I-restricted LM epitopes. Peptides were eluted from the surface of syngeneic B lymphocytes that were incubated overnight with heat-killed LM, and then peptides were separated by reversed-phase HPLC. An 11-mer peptide was identified that induced significant target cell lysis. The amino acid sequence of this peptide was determined by Edman degradation and confirmed by mass spectrometry. The sequence of this peptide encompassed amino acids 220-230 of the listeriolysin protein. Synthetic peptides were synthesized according to this sequence. The CTL responses induced by these peptides were comparable to the eluted native peptide.
Immunoglobulin G and M Detection for Leprosy Diagnosis (129.18)
Authors: Duthie Malcolm S , Goulart Isabela M B , Reed Steven G , Lobato Janaina , Goulart Luiz R 
Published: January 2023

Abstract
Leprosy is a curable disease that requires better diagnostic and prognostic tools to accompany preventive and therapeutic strategies. Circulating IgM antibodies against the M. leprae phenolic glycolipid-I (PGL-I) antigen have previously been used to assess humoral immunity. The presence of elevated titers of anti-PGL-I IgM reflects total bacterial load in the body; these antibodies, however, are generally low or absent in paucibacillary patients. Our objective was to compare 3 new recombinant antigens, ML0405, ML2331, and LID1 (comprising critical regions from ML0405 and ML2331), which recognize serum IgG, in comparison and in combination with PGL-I IgM. Serum samples were collected from 105 patients across the leprosy spectrum at the initial diagnosis and again after treatment. Patient positivity for LID1, ML0405, ML2331 and PGL-I tests was 67%, 62%, 65%, and 76%, respectively. A combination of the LID1 and PGL-1 antigens gave a positivity of 80%. After treatment, the ML0405 and PGL-I assays decreased their ELISA index values for all clinical forms, except for the LL form. Importantly, both antigens have been positively correlated with the bacillary load, clinical forms and the operational classification at diagnosis. Our results suggest that the combination of LID1 and PGL-I antigens, recognizing the IgG and IgM response, respectively, could be employed as an auxiliary tool for leprosy diagnosis.
This work was funded in part by American Leprosy Mission.
TgMAPK1 is an important virulence determinant in Toxoplasma gondii affecting stage differentiation. (129.23)
Authors: Brumlik Michael James , Ludwig Sara M , Pandeswara Srilakshmi , Jeansonne Duane P , Livi Carolina B , Daniel Benjamin J , Church Kristina M , Kious Mark J , Curiel Tyler J 
Published: January 2023

Abstract
Toxoplasma gondii is a medically important, obligate intracellular protozoan parasite that causes significant morbidity and mortality in hosts with defective cellular immunity. Using antisense RNA knockdown, we generated T. gondii tachyzoites expressing 5 - 10-fold less TgMAPK1, a stress-response mitogen-activated protein kinase. In infection, T. gondii converts from the rapidly-dividing tachyzoite to the latent bradyzoite form. We show here that T. gondii tachyzoites with reduced TgMAPK1 display distinctive bradyzoite characteristics, including expressing Bag1 (a bradyzoite-specific antigen) and significantly reducing Sag1 (a tachyzoite-specific antigen). Although knockdown (KD) tachyzoites were dramatically less sensitive to the anti-proliferative effects of IFN-γ than wildtype (WT), these parasites were nonetheless significantly less virulent than WT in BL6 mice, based on: (i) KD parasites achieved only a fraction of the parasite burden in tissues of infected mice relative to control parasites, (ii) mice infected with KD produced substantially less IFN-γ, and (iii) the majority of mice challenged with KD were able to survive lethal challenge. These results show that TgMAPK1 is an important virulence determinant affecting stage conversion, proliferation, and replication. This work was supported by funds from NIH AI060424 and Johnson & Johnson Focused Giving.
Characterization of immune responses to Yersinia pestis infection: resistant versus susceptible mice(129.29)
Authors: Lambert Nathaniel D , Nilles Matthew L , Bradley David S 
Published: January 2023

Abstract
We have previously reported that inbred B10.T(6R) mice are resistant to Yersinia pestis infection, compared to susceptible strains, e.g. Swiss Webster mice; the mean LD50's for B10.T(6R) females is 14000 CFU's, compared to Swiss Webster females, at 22 CFU's. Interestingly, B10.T(6R) mice demonstrated a more robust pro-inflammatory response early in infection, and a lesser anti-inflammatory response later on, when compared to susceptible Swiss Webster mice. Significantly higher levels of TNF-alpha were detected in B10.T(6R) on day 1 while TNF-alpha responses were delayed in Swiss Webster mice. Conversely, IL-10 levels were not changed in B10.T(6R) mice while IL-10 levels increased significantly in Swiss Webster mice. Yersinia outer membrane proteins (YOP) are translocated into the host cell's cytoplasm via a type three secretion system. After contact with a eukaryotic cell, YOPs then interact with cellular components to disrupt the host's immune response and induce the production of suppressive cytokines, e.g. IL-10. We postulate that the resistant B10.T(6R) strain of mice are able to overcome initial infection with Y. pestis through higher levels of pro-inflammatory cytokines and decreased levels of immune suppressive cytokines later in infection.
Influenza-specific memory CD4 T cells regulate acute inflammation independent of innate immune recognition (130.10)
Authors: Strutt Tara  M. , McKinstry K. Kai , Dutton Richard W. , Swain Susan L. 
Published: January 2023

Abstract
It is generally accepted that the flow of immunologically relevant information during the early stages of responses against pathogens is one-way, - that inflammation induced upon pattern recognition by highly conserved receptors of the innate immune dramatically impacts subsequent antigen-specific T and B cell responses. We asked if the reverse occurs, and if cells of the adaptive immune system can influence the character and magnitude of innate inflammatory responses. We show here that resting, antigen-specific memory CD4 T cells can dramatically alter innate inflammatory responses within 36 hours of viral infection in a manner independent of other T cells and TLR signaling. Virus-specific memory CD4 T cells transferred to naïve mice that are then challenged with influenza induce greater expression of multiple inflammatory mediators both at the site of infection and systemically upon cognate recognition of antigen in an IFN-gamma independent fashion. Our results show that the adaptive immune system can profoundly influence the character of inflammation following pathogen challenge, demonstrating a new role for memory CD4 T cells in controlling virus titers during protective immune responses.
Coxsackievirus B3 has profoundly different inhibitory effects on the MHC class I and class II antigen presentation pathways (131.2)
Authors: Kemball Christopher C , Harkins Stephanie , Whitmire Jason K , Feuer Ralph , Flynn Claudia T , Whitton J Lindsay 
Published: January 2023

Abstract
Coxsackievirus B (CVB) infections cause considerable morbidity and mortality, and strategies to treat or prevent CVB-associated disease may benefit from a clearer understanding of the host immune response to virus infection. CVB3 induces minimal endogenous naïve CD8 and CD4 T cell responses, possibly because viral antigen (Ag) presentation is extremely limited. Several CVB3 proteins are known to cooperatively downregulate MHC class I on infected cells in vitro. In this study, we used transgenic (Tg) CD4 and CD8 T cells as sensors to evaluate viral Ag presentation by the MHC class I and class II pathways in vivo. Our analysis revealed a striking difference in Tg T cell responses: CD4 Tg T cells proliferated in CVB3-infected mice whereas CD8 Tg T cells failed to divide. Moreover, virus infection generated multi-functional memory CD4 T cells, which expanded dramatically following challenge infection, and rapidly differentiated into secondary effector Th1 cells. Although naïve CD8 Tg T cells were unresponsive to CVB3, they persisted in the host and responded vigorously when stimulated by a different virus encoding their cognate Ag. These data suggest that CVB3 utilizes a powerful immune evasion strategy in vivo that differentially affects the two Ag presentation pathways, and that this difference regulates the host's capacity to mount CD4 and CD8 T cell responses. This work was supported by NIH R01 AI42314 and T32 NS41219.
Engineering Immunity against HIV - a Stem Cell Vaccine (132.12)
Authors: Yu Kenneth K , Tsai Jonathan , Yang Lili , Baltimore David 
Published: January 2023

Abstract
An effective vaccine against HIV is urgently needed. We are pursuing a novel gene therapy approach to combat HIV by engineering HSCs to produce human B cells programmed to make anti-HIV broadly neutralizing antibodies (bNAbs). We accomplish this by using lentiviral vectors to modify the long-lived HSCs with engineered bNAb genes. We hypothesize the transplantation of the engineered HSCs will confer life-long HIV-specific immunity; in effect, a "Stem Cell Vaccine" for HIV. In nature the soluble Ab and membrane BCR are produced by complex alternative splicing events difficult to implement in the context of a lentiviral vector. We have overcome this obstacle by developing a novel "Synthetic-Switch" antibody gene that mimics the function of the membrane-bound BCR and secretory antibody. Furthermore, the Synthetic-Switch approach enables us to produce "isotype-switched" anti-HIV B cells without the need for complex DNA rearrangement. The Synthetic-Switch approach thus successfully captures the essential functions of the roughly 2-Mbp immunoglobulin loci in a compact 10kb lentiviral vector. Herein we present this successful first step toward an anti-HIV stem cell vaccine.
How the strength of peptide-MHC and TCR interaction impacts CD8 T cell responses (132.24)
Authors: Zehn Dietmar , Bevan Michael J. 
Published: January 2023

Abstract
The pool of naïve T cells specific for any foreign peptide/MHC-I epitope is diverse and contains cells that differ in their functional avidity. Theoretical considerations imply that there are likely to be more low than high avidity T cells specific for any foreign antigen. We studied to what extent lower avidity T cells participate in an immune response and we investigated how the dominance of high avidity T cells evolves over the course of an infection. To control the strength of stimulation that CD8 T cells receive during an immune response, we used a model system in which OT-1 TCR transgenic CD8 T cells were stimulated during a Listeria monocytogenes infection either by their native ligand SIINFEKL or by different SIINFEKL-derived altered peptide ligands. Using this system we observed that even very weak TCR-ligand interactions are sufficient to induce rapid T cell proliferation and to generate functional effector and memory CD8 T cells. However, the strength of the TCR-ligand interaction impacts when expansion stops and contraction begins and when the cells exit lymphoid organs, i.e. strongly stimulated T cells contract and exit lymphoid organs later than do weakly stimulated cells. Our data challenges the prevailing view that strong TCR ligation is a prerequisite for CD8+ T cell activation. Instead, very weak interactions are sufficient for activation, but strong TCR ligation is required to sustain T cell expansion.
Pre-clinical testing of T cell responses to influenza vaccine/adjuvant combinations in older adults (132.5)
Authors: Ma Kewei , Haynes Laura , Wilschut Jan , McElhaney Janet E. 
Published: January 2023

Abstract
Objective: The purpose of this experiment was to determine whether inflammatory cytokines or TLR ligands could be combined with influenza vaccine in older adult PBMC to enhance the response to live influenza virus challenge.
Methods: PBMC from five older adults (&gt;65 years old) and six young (20-38 years old) adults were obtained at six months post-vaccination and cultured for five days with split-virus vaccine (SVV) and the TLR3 ligand, poly I:C, or a cocktail of pro-inflammatory cytokines (TNF-α, IL-1, IL-6; T/1/6). Stimulated PBMC were then challenged with live influenza virus for 20 hours and supernatants and lysates prepared for assays of IL-1, IL-6, TNF-α in 5-day supernatants, and the IFN-γ:IL-10 ratio and granzyme B activity in 20-hour virus-challenged PBMC.
Results: We observed an increased response to SVV in younger compared to older adults, but a dose-response relationship between poly I:C (added to a fixed dose of SVV) and the IFN-γ:IL-10 ratio and granzyme B levels improved the response in older adult PBMC. This response was associated with a dose-response increase in pro-inflammatory cytokine levels in SVV/poly I:C-stimulated PBMC. However, a cocktail of these inflammatory cytokines (T/1/6) added to the PBMC culture suppressed rather than enhanced the T cell response in older adults.
Conclusion: Poly I:C combined with influenza vaccine was shown to enhance the T cell response to influenza in aged PBMC by mechanisms that cannot be replicated by the simple addition of inflammatory cytokines to the vaccine.
Characterization of the diversity of the CD8+ T cell repertoire against the same epitope induced by bacterial or viral immunization (132.9)
Authors: Rudd Brian D , Venturi Vanessa , Davenport Miles P , Smithey Megan , Alzhanova Dina , Way Sing Sing , Nikolich-Zugich Janko 
Published: January 2023

Abstract
Prevention of infectious diseases is most efficiently accomplished by vaccination, and most vaccines rely on generation of protective antibodies. On the other hand, elicitation of CD8+ T cells plays a key role in immune protection against many intracellular pathogens. Despite the widespread success of many vaccines, the underlying basis of these protective immune responses is still poorly understood. While vaccine development has historically focused on the magnitude of the response, recent advances in immunology suggest that sufficient diversity of CD8+ T cell immune responses is also critical in controlling various infections and such information may be especially critical in the rational design of vaccines against chronic and persistent infections. To address these issues, we are currently investigating whether CD8+ T cell repertoire against a protective epitope can be influenced by the infectious/inflammatory context in which the epitope is encountered. In an effort to assess the extent of TCR diversity that results from different infectious vectors, CD8+ T cells specific for the immunodomant epitope (gB 498-505) were sequenced from mice infected with HSV-1 or with Vaccinia virus (VV) or Listeria monocytogenes (LM) expressing the immunodominant peptide. By comparing CD8+ T cell repertoires induced by different pathogens, we hope to understand how and to what extent the CD8+ T cell repertoire is able to shaped.
NIH grant #7R01AI066096-05