bluuebunny/crossref_metadata_2025_split huggingface dataset:
Skin Delivery of the JVRS-100 Immunostimulant with and without Fluzone using a Protrusion Array Device (PAD) Results in Potent Adaptive Immune Responses (78.34)
Authors: Chang Stella , Gonzalez-Gonzalez Emilio , Contag Christopher H. , Fairman Jeff , Kaspar Roger , Speaker Tycho 
Published: January 2023

Abstract
Background: A protrusion array device (PAD) configured with hollow dissolvable microneedles was loaded with the JVRS-100 adjuvant or JVRS-100 combined with commercial Fluzone. The studies were designed to establish if adjuvant or adjuvant/vaccine could be administered via PAD patches. Methods: JVRS-100, Fluzone, or JVRS-100/Fluzone were loaded onto PAD patches and administered at day 0 and 14. At day 28 responses were assessed by HAI and ELISA (IgG, IgG1, and IgG2). Separate cohorts of mice were vaccinated intramuscularly (IM) with Fluzone and JVRS-100-containing patches were placed over the site of vaccination to see if intradermal administration of JVRS-100 would adjuvant IM vaccination. Results: The administration of JVRS-100 in addition to Fluzone stimulated a response greater than Fluzone alone patches as measured by geometric mean titer (EC50) for IgG (4.8x104 vs. 1.6x104) and IgG2 (4.5x103 vs 6.0x102). IgG1 titers were elevated in the Fluzone only group compared with JVRS-100/Fluzone (8.3x103 vs. 1.9x103). Administration of JVRS-100-containing patches over IM Fluzone vaccination increased IgG (3.7x105 vs. 2.2x105) and IgG2a (1.5x105 vs. 9.0x104). IgG1 titers were elevated in the Fluzone-only group compared with JVRS-100/Fluzone (1.5x105 vs. 2.9x105). HAI levels correlated with antibody titer levels for all groups. Conclusion: The antibody response from both studies indicate that the TH1 bias for JVRS-100 is retained as a dry PAD patch and intradermal administration over IM vaccination or combined for ID vaccination.
Tissue restricted activation of basophils mediates primary immunity against helminthes (79.22)
Authors: Sullivan Brandon M , Wu Davina , Liang Hong-Erh , Loke P'ng , McKerrow James H , Voehringer David , Locksley Richard M 
Published: January 2023

Abstract
CD4 T cells and the cytokines IL-4 and IL-13 are required for primary immunity to helminthes and for allergic responses. Although T-helper-2 (Th2) cells are believed to provide protection, genetic deletion of IL-4/13 from CD4 T cells revealed that cytokines from innate cells mediate Nippostrongylus brasiliensis clearance and survival after Schistosoma mansoni infection. Basophils were the only IL-4-secreting innate cells identified. Basophils were recruited to numerous sites, but basophil cytokine secretion occurred only in affected tissues, and was independent of CD4 T cell-derived IL-4/13 and IgE. Depletion of basophils compromised the ability to expel intestinal helminthes. Thus, basophils serve as innate effector cells during primary helminth infection by secreting cytokines at sites of tissue involvement, a process amplified during adaptive immunity by IgE through IL-4-mediated T cell help.
Basophils play an important role in the resolution of inflammation by inducing alternative activation of macrophages through Th2 cytokine release (80.4)
Authors: Karasuyama Hajime , Mukai Kaori 
Published: January 2023

Abstract
Basophils have long been thought to be an initiator or effector of allergic inflammation. Here we report that basophils can function as a terminator of inflammation in IgE-mediated chronic allergic inflammation. In this allergy model, mice were passively sensitized with hapten TNP-specific IgE and then challenged with subcutaneous injection of TNP-OVA in the ear. The allergen challenge induced not only immediate-type ear swelling (mast cell-dependent) but also delayed-onset ear swelling (basophil-dependent). Analyses with flow cytometry and RT-PCR revealed that alternatively activated macrophages (AAMs) appeared in the skin lesions as the inflammation proceeded. The AAMs were induced in T cell-deficient but not IL-4/IL-13-deficient mice, indicating that AAM-inducing Th2 cytokines were produced by non-T cells. Basophil depletion abolished the development of AAMs. The cell transfer experiment revealed that basophil-derived IL-4 was essential for the alternative activation of macrophages. Depletion or developmental block of AAMs resulted in deterioration and prolongation of inflammation with massive accumulation of inflammatory cells. Taken together, basophils induce alternative activation of macrophages through Th2 cytokine release, leading to the resolution of inflammation.
Nitration of aldolase: a critical post-translational modification in nitric oxide-mediated regulation of mast cell function (80.6)
Authors: Sekar Yokananth , Moon Tae Chul , Slupsky Carolyn M , Befus A. Dean 
Published: January 2023

Abstract
Nitric oxide (NO) is a short-lived free radical that regulates mast cell (MC) activities including inhibition of MC degranulation. To elucidate the molecular mechanisms underlying the effects of NO in MC, we investigated protein tyrosine nitration in human mast cell lines treated with the NO donor S-Nitrosoglutathione. Using 2D gel western blot analysis with an anti-nitrotyrosine antibody together with mass spectroscopy, we identified aldolase A, an enzyme of the glycolytic pathway as a target for tyrosine nitration in MC. Aldolase A nitration was associated with reduction in the Km and Vmax of aldolase in HMC-1 and LAD-2. Nuclear Magnetic Resonance analysis showed that despite these changes in activity of a critical enzyme in glycolysis, there was no significant change in total cellular ATP content, although the AMP/ATP ratio was increased. Elevated levels of lactate and pyruvate suggested that NO treatment enhanced glycolysis in MC. Reduced MC aldolase activity was associated with increased intracellular levels of its substrate, fructose 1,6 bisphosphate (FBP). Interestingly, FBP inhibited IgE-mediated MC degranulation in LAD-2 cells. Thus for the first time we report evidence of protein tyrosine nitration in two human MC lines and identify aldolase A as a prominent target that may regulate MC phenotype and function.
Funded by: Canadian Institutes of Health Research, The Lung Association, Alberta & NWT and 75th Anniversary Award, University of Alberta.
Imbalance of effector and regulatory T cells in the pathogenesis of experimental eosinophilic esophagitis (80.8)
Authors: Mishra Anil , Rothenberg Marc 
Published: January 2023

Abstract
RATIONALE: Eosinophilic esophagitis (EE) is an emerging disease and we previously demonstrated that adaptive T cell immunity is critical in the disease pathogenesis. However, the specific subsets of T cells that promote esophageal eosinophilia are not yet understood. Therefore, we focused our studies to examine the role of CD45RB T subsets, as the pathogenic and regulatory activity of T cells depends on the expression levels of CD45RB. METHODS: A flow cytometric analysis was performed to examine the frequency of CD45RBhigh and CD45RBlow T cell subsets in the mouse esophagus. A real time PCR analysis was performed to examine the pathogenic and anti-inflammatory characteristics of both subsets of T cells. In addition, we adoptively transferred these purified cell subsets in mice with experimental EE and examined their role in the disease pathogenesis.
RESULTS: An increase of CD4+CD45RBhigh and a decrease of CD4+CD45RBlow cells were observed in mouse esophagus following the induction of experimental EE. A relatively high transcript level of IL-5 and IL-13 in CD4+CD45RBhigh and IL-10 and TGF-β1 in CD4+CD45RBlow spleen purified cells was observed. Further, our initial experimentation revealed that mice adoptively transferred with 4X105 CD4+CD45RBlow T cells/week were protected from the induction of esophageal eosinophilia compared to control mice that did not received adoptively transferred cells. Mice receiving CD4+CD45RBhigh T cell had no significant change in the number of eosinophils in the esophagus compared to control mice following allergen challenge. CONCLUSIONS: CD4+CD45RBlow T cells have a critical role in down-regulating allergen-induced eosinophil recruitment to the mouse esophagus.
Evaluation of granulysin and perforin as candidate biomarkers for protection following vaccination with Mycobacterium bovis BCG or M. bovis delta-RD1 (81.10)
Authors: Scherer Charles C , Endsley Janice J , deAguiar Juliana , Jacobs Jr. William R , Larsen Michelle H , Palmer Mitchell V , Nonnecke Brian J , Golde William T , Waters W Ray , Estes D Mark 
Published: January 2023

Abstract
Development of improved vaccines against tuberculosis (TB) is directly correlated to the investigation of new and better correlates of protection after vaccination. We examined the kinetics of granulysin, perforin, IFN-gamma and Fas-L responses to M. bovis purified protein derivative (PPD) stimulation by peripheral blood mononuclear cells from BCG- and non-vaccinated cattle. Gene expression profiles following PPD stimulation showed significant increases in transcripts for granulysin and IFN-gamma in both CD4+ and CD8+ T cells in BCG-vaccinated as compared to non-vaccinated animals. Perforin & IFN-gamma examined by flow cytometry, showed a difference of 1-2% more PPD-specific cells in BCG-vaccinated than non-vaccinated animals. In a second trial, M. bovis BCG vaccinated animals had lower disease and pathological scores as compared to M. bovis Delta-RD1(a new vaccinate candidate) and the control group. Gene expression of granulysin and perforin were significantly increased in both vaccine groups (M. bovis BCG and M. bovis Delta-RD1 vaccinated) as compared to control after vaccination and challenge. IFN-gamma expression was increased only after vaccination and secretion was higher in the control, as compared to both vaccine groups demonstrating no correlation with protection upon vaccination.
Chinook Salmon (Oncorhynchus tshawytscha) Have Multiple Interferon-gamma Genes

(81.7)
Authors: Rhodes Linda D , Demlow S. Ellen 
Published: January 2023

Abstract
Interferon-gamma (IFNg) is a central cytokine in modulating innate and adaptive immunity in a wide range of vertebrates. A study of genes associated with an effective host response by Chinook salmon (Oncorhynchus tshawytscha) to a persistent Gram-positive bacterial pathogen (Renibacterium salmoninarum) suggested that an interferon-inducible signal was abrogated during early stages of exposure. Splenic DNA was amplified with IFNg primers from a related species, rainbow trout (Oncorhynchus mykiss), and multiple clones from individual fish were sequenced. Up to five genotypes per fish were found, and sequence variations included microsatellites within introns and single nucleotide polymorphisms (SNPs) within exons. Southern hybridizations supported the prediction that Chinook salmon possess at least three related IFNg loci. Multiple clones of cDNA, synthesized from RNA isolated from pronephros of fish exposed to an attenuated strain of R. salmoninarum, displayed a surprising abundance of SNPs. These findings are consistent with the phenomenon of genome duplication known to have occurred in teleosts. Although the presence of multiple loci complicates the analysis of IFNg gene expression, it may also offer a mode of immune modulation not found among mammals.
Research supported by National Marine Fisheries Service, NOAA Fisheries, U.S. Department of Commerce.
Peripheral blood and jejunal intraepithelial immune cell phenotypes associated with body weight of newly weaned pigs (81.9)
Authors: Davis Ellen , Rehberger Joshua , King Michael , Brown Dari C. , Maxwell Charles V. , Rehberger Tom 
Published: January 2023

Abstract
Early immune development has been purported to have ramifications on metabolic functions related to subsequent growth. To determine if specific immune cell populations were associated with pigs with heavy and light body weight (BW), piglets were weaned to an on-site conventional or a segregated weaning facility and immune cell phenotypes were characterized and correlated to piglet BW. Pig BW was measured and flow cytometry analyses were performed on PBMC and jejunal intraepithelial lymphocytes (IEL) on day 11, 18, and 25 after weaning. Ordination methods were performed to determine the relationship of specific immune cell phenotypes to piglet BW. Cell phenotypes consistent with γδ (CD4+γδ+CD8-, CD8+γδ+CD4-) and activated (CD25+) IEL were positively correlated to pig BW on day 11, whereas PBMC expressing MHC2 (MHC2+CD3-) resulted in a negative correlation. On day 18, CD25+CD8+CD4+ IEL were positively correlated and PBMC expressing CD8+CD4+ were negatively correlated to pig BW. CD25+ and CD4+CD8+ IEL populations were positively correlated to pig BW on day 25, whereas IEL γδ populations (γδ+CD8+CD4-, CD4+CD8+γδ+) and PBMC expressing MHC2 (MHC2+CD3-) were negatively correlated to BW. These data indicate that specific immune cell phenotypes correlate to pig BW after weaning. Specifically, IEL expressing CD25 are associated with pigs with heavier BW after weaning, whereas IEL expressing the γδ T cell receptor are associated with heavy BW pigs early after weaning but are correlated to low BW in older pigs.
CD4+ recent thymic emigrants are biased to the TH2 effector lineage (82.15)
Authors: Hendricks Deborah W , Boursalian Tamar , Fink Pamela J 
Published: January 2023

Abstract
Following intra-thymic development, T cells exit the thymus and join the peripheral T cell pool. These newly generated cells, termed recent thymic emigrants (RTEs), undergo phenotypic and functional maturation for the first 3 weeks they reside in the periphery. Using mice transgenic (Tg) for green fluorescent protein (GFP) driven by the Rag2 promoter (RAG2p-GFP Tg), in which GFP reliably distinguishes RTEs from their more mature naïve (MN) counterparts, our lab is able to isolate untouched RTEs for functional and phenotypic analysis.
CD4+ RTEs exhibit defects in proliferation, IL-2Rα upregulation, and IL-2 production when stimulated through the TCR. We now show that CD4+ RTEs also exhibit defects in Th0, Th1, Th17 and Treg lineage commitment, with diminished cytokine production and transcription factor expression. In contrast to these diminished functions, RTEs are enriched for IL-4 producers when differentiated under Th2, but not Th0, conditions. This bias to the Th2 lineage may be due to regulated expression of T-bet, a key Th1 transcription factor.
These data show that CD4+ RTEs represent a unique subset of the peripheral T cell pool, defined by a distinct interpretation of and response to immunological cues. These characteristics may be beneficial during the post-thymic maturation period to avoid inappropriate immune responses.
This work was supported by R01AI064318 from the NIAID (with a supplement to DWH).
Memory-like CD8+ T cells generated during homeostatic proliferation defer to antigen-experienced memory cells (82.18)
Authors: Cheung Kitty P , Yang Edward , Goldrath Ananda W 
Published: January 2023

Abstract
Naïve T cells proliferate in response to lymphopenia and acquire the phenotypic and functional qualities of memory T cells, providing enhanced protection against infection. How well 'memory-like' T cells generated during lymphopenia-induced homeostatic proliferation (HP-memory) differentiate into secondary memory cells and compete with antigen-experienced 'true-memory' cells is unknown. We found that CD8+ HP-memory T cells generated robust responses upon infection and produced a secondary memory population comparable to true-memory cells in the absence of competition. However, when true-memory and HP-memory T cells competed during infection, HP-memory cells contributed less to the effector population, contracted earlier and formed fewer secondary memory cells. Furthermore, HP- and true-memory cells demonstrated aberrant chemokine receptor expression and distinct localization within the spleen during infection, indicating differential access to signals necessary for secondary memory formation. Thus, HP-memory T cells provide protection without compromising the true-memory population. Differences in HP-and true-memory T cells may reveal the basis for competition for limited resources within the memory T cell compartment.
Role of TOSO in activation induced cell death of human T- and NK cells (82.2)
Authors: Narayanan Sriram , Borrego Francisco , Coligan John E. 
Published: January 2023

Abstract
Homeostasis of both NK and T cells in the periphery is regulated by activation induced cell death (AICD) mediated by signaling through Fas, and other death receptors. It has been shown that IL-2 is required for priming cells for this type of apoptosis. Cytokine primed NK cells, in addition to killing their target cells, also undergo AICD upon activation by activating ligands expressed by the target cells. TOSO is an anti-apoptotic molecule known to inhibit apoptosis mediated by the death receptor pathway and we investigated the role of TOSO in regulating AICD in NK and T cells. We observed by realtime PCR that stimulation of human NK cells and T cells in vitro with IL-2 resulted in down regulation of TOSO. We further analyzed TOSO expression levels on sorted human CD4 T cells and found that cells with an effector/memory phenotype have markedly lower levels of TOSO than naïve cells, which suggests that activation in vivo correlates with down-regulation of TOSO. In agreement, we found that activation of CD45RO+ cells in vitro dramatically down regulated TOSO compared to naïve CD4 T cells. Further, overexpression of Toso in Jurkat cells was able to inhibit Fas-mediated cell death. We hypothesize that IL-2 has a dual action: first, it activates and induces the proliferation of NK and T-cells and second, it down-regulates TOSO, thereby sensitizing or priming them for apoptosis upon re-stimulation. Considering that IL-2 is being used as a therapeutic agent for cancer treatment and the recent study showing that HIV envelope protein can down regulate TOSO levels on CD4 T cells, unraveling how TOSO functions and is regulated might have potential for treating cancer and other diseases.
Advanced age does not impair the development of functional memory CD8+ T cells following West Nile virus infection (83.4)
Authors: Lelic Alina , Parsons Robin , Hayes Lisa , Bramson Jonathan L 
Published: January 2023

Abstract
West Nile virus (WNV) infection of aged patients typically leads to the development of severe neuroencephalitis. Studies in mice have elucidated an important role for CD8+ T cell function in controlling WNV infection and we hypothesized that the increased severity of WNV illness in the aged results from impaired CD8+ T cell function due to immunosenescence. We examined the functional profile of WNV-specific CD8+ T cell memory responses from a cohort of young and aged WNV infected patients using polychromatic flow cytometry. We quantified production of IFN-γ, TNF-α, IL-2 and CD107a mobilization by WNV-specific CD8+ T cells. We did not observe any relationship between advanced age and either the breadth or the magnitude of CD8+ T cell function. The magnitude of IFN-γ and TNF-α responses to WNV epitope stimulation did not decline with advanced age. Production of IL-2 was not detected from either young or aged CD8+ T cells. The ability to degranulate in response to virus peptide was also the same in young and aged patients. Collectively these data fail to support our hypothesis and suggest that, contrary to current models of immunosenescence, aged patients do develop functional CD8+ T cell immunity following exposure to a novel virus infection. This work was funded by a contract from NIAID (N01-AI-40066).
Extrafollicular foci-derived B cells provide long-lived mucosal immunity (84.16)
Authors: Baumgarth Nicole , Rothaeusler Kristina 
Published: January 2023

Abstract
Antibodies to influenza are contributed by extrafollicular foci and germinal centers. The latter induce long-lived immunity via bone marrow plasma cells and memory B cells. Extrafollicular foci are thought to only provide early, short-lived responses generated from high-affinity B cells. The selection of the highest affine B cells into the early response might ensure survival from acute infection, but it seems counterintuitive that they would be excluded from the long-lived response. Here we assessed the contributions of the extrafollicular foci response to long-term immunity. We tracked a prototypic early B cell response to hemagglutinin (HA) of influenza A/PR8 in BALB/c mice by multicolor flow cytometry using an idiotype-specific mAb to the C12 idiotype (Id) together with fluorescent-labeled HA. C12Id+ HA-specific antibodies were measured by ELISA and ELISPOT. The results demonstrate the extrafollicular nature of the C12Id response and the lack of C12Id+ bone marrow plasma cells. Nonetheless, following infection HA-specific serum C12Id+ antibodies were measurable for the life of mice, secreted presumably by antibody-secreting C12Id+ plasma cells present in the lung. BrDU-labeling experiments suggested continuous turnover of these cells. Thus, extrafollicular foci induce long-term mucosal antibody responses via short-lived plasma cells. Supported by NIH AI051354 (N.B) and T32 HL007013 (K.R.).
Characterization of CD44-high memory phenotype T cells induced by enforced expression of the transcription factor PLZF (85.12)
Authors: Raberger Julia , Grabner Beatrice , Sakaguchi Shinya , Schebesta Alexandra , Boucheron Nicole , Blomberg K. Emelie M. , Berglöf Anna , Kolbe Thomas , Smith Edvard , Rülicke Thomas , Ellmeier Wilfried 
Published: January 2023

Abstract
We and others have shown that transgenic expression of the transcription factor PLZF in the T cell lineage induces the appearance of CD44-high memory-phenotype (MP) T-cells (Savage et al. Immunity 2008; Raberger et al. PNAS, 2008). These MP T cells arise already in the thymus and produce IFNã upon PMA/ionomycin stimulation, thus sharing certain features with innate-like T cells. Furthermore, we observed that PLZF expression is up-regulated in Itk-deficient T cells, which are enriched with innate-like T cell subsets. To study the PLZF-induced MP T cell population in more detail, we are investigating whether these cells, like other innate-like T cells, are selected on classical or non-classical MHC molecules. Furthermore, PLZF transgenic mice are intercrossed with several knockout mice (e.g. Itk-/- or IL-15-/- mice) to determine signaling pathways/molecules required for their development and maintenance. Our studies will provide more insight into transcription pathways controlling the development of MP T cells with "innate-like" functions.
Supported by the Austrian Science Fund FWF (SFB-F23, P19930).