bluuebunny/crossref_metadata_2025_split huggingface dataset:
Eliciting mucosal and systemic CD8+ T-cell responses via pulmonary vaccination with synthetic nanoparticles (113.21)
Authors: Li Adrienne , Moon James , Elkhader Jamal , Yen Mimi , Abraham Wuhbet , Suh Heikyung , Irvine Darrell 
Published: January 2023

Abstract
Mucosal vaccines offer needle-free administration and can stimulate immune responses systemically and at mucosal sites. “Vectorless” vaccines based on antigens packaged in synthetic nanoparticles (NPs) offer potential advantages in manufacturing, enhanced safety, and lack of anti-vector immune responses. To this end, we tested the ability of lipid NPs encapsulating ovalbumin or SIV-gag epitopes with Toll-like receptor agonists to stimulate mucosal immunity following pulmonary vaccination in mice. CD11c+B220+MHC-II+CD205+ DCs and CD11c+MHC-IIloF4/80+CD11b+ macrophages were the major cell types acquiring antigen in the lungs. Following prime and boost, a dramatic expansion of endogenous antigen-specific CD8+ T-cells to ~15% of CD8+ cells in blood was observed-twice the level seen with soluble antigen vaccines and 3 times the level achieved by s.c. immunization. Antigen-specific T-cell expansion was detected in the local lung tissue (~60% of CD8 cells), as well as in the gut and vaginal tract (~40% CD8 cells). These T-cells produced effector cytokines, and lasting CD127hi KLRG1lo effector memory cells were detected at 11wks after priming in both systemic and mucosal compartments. ELISA analysis of serum, fecal pellets and vaginal washes demonstrated that humoral responses were also stimulated. This data provides evidence that mucosal delivery of synthetic NPs can safely and effectively stimulate CD8+ T cells at sites relevant for protection against mucosal pathogens such as HIV.
Identification and surface display on liposomes of neutralizing linear B-cell epitopes as a strategy for biodefense vaccine development (113.25)
Authors: Vance David , Yermakova Anastasiya , Mantis Nicholas 
Published: January 2023

Abstract
The design of subunit vaccines for Category A-B toxins, including anthrax and ricin, has proven largely unsuccessful. Recombinant subunit antigens are minimally protective, difficult to manufacture and/or highly unstable. Thus, there is an urgent need to develop alternative, more effective approaches to vaccine design. With ricin toxin as a model system, we are using a two-tiered approach to address this issue. First, we have identified and validated continuous epitopes on ricin’s A (RTA) and B (RTB) subunits that are recognized by toxin neutralizing monoclonal antibodies (mAbs). In the case of mAb CMA2, pepscan analysis identified residues 190-CLTSDSNIRETVVKI-204 on RTB as being the target of antibody binding. Screening of a phage-displayed peptide library revealed that residues DxNxR are the critical contact points between CMA2 and ricin. An RTA-specific mAb, PB10, bound to peptide 97-NQEDAEAITHLF-108 in pepscan analysis, and phage display identified residues QExA as the likely contact points. The second tier of this project involves attaching these specific peptide epitopes polyvalently to the surface of liposomes (100 nm), with or without the TLR4 agonist MPLA present in the liposome bilayer, and then testing whether these liposomes, when injected into mice, are capable of eliciting protective anti-ricin antibodies. In preliminary studies, we have successfully produced liposomes displaying peptide epitopes from RTA and RTB, and have initiated mouse immunization studies.
Stable incorporation of functionally active purified GPI-anchored immunostimulatory molecules on H5 influenza VLP surfaces by protein transfer. (113.33)
Authors: Patel Jaina , Song Jae Min , Shashidharamurthy Rangaiah , Compans Richard , Kang Sang-Moo , Selvaraj Periasamy 
Published: January 2023

Abstract
H5N1 influenza, a bilayered lipid enveloped virus, is highly pathogenic and can lead to a pandemic threat; therefore, a potent vaccine is required. The use of virus-like particles (VLPs) as a vaccine has recently gained importance as they are safe and highly immunogenic. VLPs are similar in size and structure to their viral counterparts but do not contain the viral genome required for replication. Although VLPs are immunogenic, full protection and cross-reactivity to other influenza strains does not always occur; therefore, we aim to modify H5 influenza VLPs to incorporate immunostimulatory molecules (ISMs) by using a protein transfer method. Previously, we have shown that GPI-anchored ISMs (ICAM-1 and IL-12) are able to incorporate onto H5 influenza VLPs by a simple and easy protein transfer method. In this study, we show that purification of GPI-IL-12 does not alter the functionality of the protein in stimulating proliferation of ConA-activated splenocytes suggesting that purified GPI-ISMs remain functionally active. Furthermore, the detection of incorporated GPI-ISMs onto the surface of H5 influenza VLPs by protein transfer was conducted by using immunogold staining electron microscopy and incorporation occurred uniformly as detected by flow cytometry analysis. Moreover, these GPI-ISMs remain stably expressed on the VLP surface for at least a week at 4oC. This study suggests that H5 influenza VLPs can be modified to stably express ISMs on its surface by protein transfer.
IL-2 and IL-6 cooperate to enhance the generation of effectors from aged human CD8 T cells to live influenza virus (113.34)
Authors: Zhou Xin , Behzad Hayedeh , Haynes Laura , Reed Steven , Swain Susan , McElhaney Janet 
Published: January 2023

Abstract
An aAge-related decline in cytolytic activity has been mainly described in CD8+ T cells and , with relative sparing of CD4+ T cells. wWe have shown that the poor CD8+ effector T cell response to influenza A/H3N2 challenge is due to a decline in the proportion and function of these cytolytic T lymphocytes (CTL). Hee we report In this study, we showed that therethis response corresponds to a significant decline in IL-2 levels but increased IL-6 levels in influenza-stimulated PBMCs from vaccinated older compared to young adults. We find that addition of exogenous Supplementation of PBMC with IL-2 and IL-6 to PBMC cultures with restoresd the proliferative response and reducesd the proportion of apoptotic effector CD8+ T cells in older adult PBMC to resemble that of younger adults. To further elucidate the mechanism, Wwe finshowed that exogenous IL-2 and IL-6 resulted in the differentiation of naïve CD4+ T cells into effector T helper cells which express IL-2 in response to influenza virus stimulation. Further, use of a TLR4 ligand combined with influenza vaccine ,whichto respectively stimulate IL-6 and IL-2 production, resultsed in an enhanced granzyme B response of aged CD8 T cells to a live influenza virus challenge. These results suggest provide a mechanism that could be targeted in the development of influenza vaccines to potentially with enhance theird efficacy in older adults.
NLRC5 dependent activation of the inflammasome in response to bacteria (114.8)
Authors: Davis Beckley , Roberts Reid , Barker Brianne , Duncan Joseph , Ting Jenny 
Published: January 2023

Abstract
The nucleotide-binding domain leucine-rich repeat-containing proteins, NLRs, are intracellular sensors of pathogen-associated molecular patterns and damage-associated molecular patterns. A subgroup of NLRs can form inflammasome complexes, which facilitate the maturation of procaspase 1 to caspase 1, leading to IL-1β and IL-18 cleavage and secretion. NLRC5 is predominantly expressed in hematopoietic cells and has not been studied for inflammasome function. RNA interference-mediated knockdown of NLRC5 nearly eliminated caspase 1, IL-1β, and IL-18 processing in response to bacterial infection, pathogen-associated molecular patterns, and damage-associated molecular patterns. This was confirmed in primary human monocytic cells. NLRC5, together with procaspase 1, pro-IL-1β, and the inflammasome adaptor ASC, reconstituted inflammasome activity that showed cooperativity with NLRP3. The range of pathogens that activate NLRC5 inflammasome overlaps with those that activate NLRP3. Furthermore, NLRC5 biochemically associates with NLRP3 in a nucleotide-binding domain-dependent but leucine-rich repeat-inhibitory fashion. These results invoke a model in which NLRC5 interacts with NLRP3 to cooperatively activate the inflammasome
mTORC2 controls T-cell receptor processing and surface expression during thymocyte development (115.18)
Authors: Werlen Guy , Chou Po-Chien , Oh Won , Wu Chang-Chih , Ruegg Markus , Hall Michael , Jacinto Estela 
Published: January 2023

Abstract
An efficient immune response relies on the presence of T-cells expressing a functional T-cell receptor (TCR). While the mechanisms generating TCR diversity for antigenic recognition are well defined, what controls its surface expression is less known. The mammalian target of rapamycin (mTOR) forms two structurally and functionally distinct protein complexes, mTOR complex 1 (mTORC1) and mTORC2. While mTORCs were found to regulate T lymphocyte activation and lineage commitment, their role in early thymocyte development remains to be defined. Here we found that mTORC2 modulates T-cell ontogeny and activation by controlling the quantity of TCR that reaches the cell surface of developing thymocytes as well as transformed T-cells. Deletion of the mTORC2 component rictor at early stages of T cell ontogeny (DN2 and onward) led to aberrant maturation and increased proteasomal degradation of nascent TCR polypeptides. Consequently, the levels of TCR as well as other receptors, including CD4, CD8 and CD147 were significantly attenuated on the surface of rictor-deficient thymocytes. Since CD147 was also defectively glycosylated and expressed in SIN1-deficient fibroblasts, our findings suggest that mTORC2 is involved in the co/post-translational processing of membrane receptors. Thus, we uncover a novel function for mTORC2 that provides rationale to target this complex in disorders caused by deregulated expression of surface receptors.
Post-translational regulation of RhoH via the proteasomal pathway (115.2)
Authors: Tamehiro Norimasa , Oda Hiroyo , Suzuki Harumi 
Published: January 2023

Abstract
RhoH, an atypical GTPase-deficient Rho GTPase, is exclusively expressed in hematopoietic lineage cells and is required for preTCR- and TCR-dependent signal transduction during T cell development in the thymus. In the current study, we defined tyrosine residues critical for stability of RhoH protein. HA-tagged mutant RhoH with the substitution of all five tyrosines to phenylalanine was not stable when expressed in 293 cells. The mutant RhoH degraded more quickly and was more highly ubiquitinated than wildtype RhoH, and also its degradation was proteasome dependent. We also found that anti-CD3 plus -CD28 stimulation in T lymphocytes enhanced ubiquitination of RhoH in primary lymphocytes, indicating that TCR-stimulation could induce active RhoH degradation. From analysis of each single mutant of five tyrosines, we determined that three tyrosine residues (Y33, Y73 and Y148) were critical for proteasome-dependent protein degradation, whereas the other two (Y38 and Y83) were not. Furthermore, both Y38 and Y83 mutants showed association with ZAP-70, like wildtype RhoH, suggesting that phosphorylation of these two tyrosines were not important for the binding of RhoH to ZAP-70. From these results, we propose that stability of RhoH protein is regulated by tyrosine phospholyation-dependent ubiquitin-proteasomal degradation, as well as the lysosomal degradation pathway reported before.
Eri1 regulates microRNA homeostasis and mouse natural killer cell development and anti-viral function (115.9)
Authors: Ansel K Mark , Thomas Molly , Abdul-Wajid Sarah , Panduro Marisella , Babiarz Joshua , Rajaram Misha , Woodruff Prescott , Lanier Lewis , Heissmeyer Vigo 
Published: January 2023

Abstract
Natural killer (NK) cells play a critical role in early host defense to infected and transformed cells. Here we show that mice deficient in Eri1, a conserved 3’-to-5’ exoribonuclease that represses RNA interference, have a cell-intrinsic defect in NK cell development and maturation. Eri1-/- NK cells displayed delayed acquisition of Ly49 receptors in the bone marrow and a selective reduction in Ly49D and Ly49H activating receptors in the periphery. Furthermore, Ly49H+ NK cells deficient in Eri1 failed to expand efficiently during mouse cytomegalovirus (MCMV) infection. Consequently, Eri1 was required for immune-mediated control of MCMV. We identified miRNAs as the major endogenous small RNA target of Eri1 in mouse lymphocytes. Both NK and T cells deficient in Eri1 displayed a global, sequence-independent increase in miRNA abundance. Ectopic Eri1 expression rescued defective miRNA expression in mature Eri1-/- T cells. Thus mouse Eri1 regulates miRNA homeostasis in lymphocytes and is required for normal NK cell development and anti-viral immunity.
Fatal pneumococcal infection in mice deficient for the actin-bundling protein L-plastin (117.26)
Authors: Morley Sharon Celeste , Todd Elizabeth , Deady Lauren , Davis Chris , Muenzer Jared 
Published: January 2023

Abstract
L-plastin (LPL) is a leukocyte-specific actin-bundling protein that increases the stability of actin-based structures such as podosomes. Fewer lymphocytes and dendritic cells populate the lungs of LPL-/- mice, presumably due to defective motility of these cell types. To determine whether defective immunity in LPL-/- mice would result in increased susceptibility to infection, we challenged mice intra-tracheally with S. pneumoniae. Less than 20% of LPL-/- mice survived pneumococcal infection, compared to 80% of wild-type mice. Rapid mortality of LPL-/- mice correlated with a 100-fold higher bacterial burden in the blood and bronchoalveolar lavage fluid 24 hours following infection, indicating a significant failure to control and limit bacterial growth in vivo. Defective clearance of pneumococcus was apparent as early as six hours after intra-tracheal challenge, and comparison to Rag1-/- mice indicated defective innate immunity. Preliminary experiments demonstrated defective phagocytosis of pneumococci by LPL-/- alveolar macrophages, providing an explanation for the early clearance defect observed. No requirement for LPL in macrophage function has been previously reported. Delineation of the molecular pathways critical to early innate pulmonary defense against pneumococci through the use of mice deficient for LPL will support research efforts aimed at reducing the global health burden of pneumococcal pneumonia.
Genetic polymorphism in the human LILRB3/LILRA6 locus (118.13)
Authors: Bashirova Arman , Apps Richard , Vince Nicolas , Yu Xu , Carringotn Mary 
Published: January 2023

Abstract
LILRB3 is an orphan receptor expressed on the surface of myeloid cells. It is structurally distinct from HLA class I binding LILRs and extremely polymorphic in the extracellular domain. LILRB3 can mediate negative signaling through its ITIM-bearing cytoplasmic tail. LILRA6 is identical to LILRB3 in the extracellular part but is potentially an activating receptor. The molecules are encoded by two neighboring genes on human chromosome 19. We have analyzed polymorphism in the LILRB3/A6 locus and found that LILRA6 exhibits a level of polymorphism comparable to LILRB3 with similar polymorphic sites. Moreover, we identified copy number variation (CNV) in the LILRA6 locus with 33% of healthy individuals possessing duplication of LILRA6 and 4% bearing deletion of the gene. Duplications and deletions of LILRA6 and a constant number of LILRB3 seem to be a consequence of unequal crossing-over between the two genes resulting in exchange of the extracellular domains between the receptors. Such exchange explains the maintenance of structural similarity of the two receptors and identity of polymorphic sites. Analysis of mRNA expression in the major PBMC fractions shows that LILRA6 is primarily expressed on monocytes, similarly to LILRB3, and its expression level correlates with the CNV. We suggest that the LILRA6 CNV may influence the ratio of inhibitory vs activating versions of this receptor on the cell surface, potentially affecting the biological consequence of LILRB3/A6 ligation.
IgE antibody-mediated degranulation of cutaneous mast cells induces thermal pain in mice (118.8)
Authors: Chatterjea Devavani , Mack Madison , Wetzel Abigail , Shabani Estela , Allen Juliann 
Published: January 2023

Abstract
Pain is a key component of inflammation and a public health issue of significance. Mast cell activity has been implicated in pain responses associated with irritable bowel syndrome, migraine headaches, interstitial cystitis, postoperative pain, and endometriosis, but the mechanisms underlying mast cell involvement in pain pathologies are not clear. We show that degranulation of plantar mast cells caused by passive local antibody (IgE anti-DNP/HSA) sensitization and systemic antigen (DNP/HSA) challenge, induces thermal hyperalgesia as well as significant edema, neutrophil influx, and upregulation of IL-1beta and IL-6 proteins in the plantar tissue. Pre-treatment with histamine receptor H2 and H3/4 antagonists significantly abrogates pain, but not edema, associated with IgE/antigen-mediated mast cell degranulation This suggests that while pain has a multifactorial pathophysiology, tissue-resident sentinel mast cells may be a critical initiators of allergen-invoked hyperalgesia in mice.
Increased bacterial colonization and delayed wound closure in aged mice is associated with altered leukocyte recruitment (118.9)
Authors: Brubaker Aleah , Rendon Juan , Choudhry Mashkoor , Kovacs Elizabeth 
Published: January 2023

Abstract
We developed a murine model of cutaneous wound infection to elucidate the role of innate immune dysfunction in aberrant responses to wound infection with advanced age. Young (10-14 weeks) and aged (18-20 months) BALB/c mice were subjected to 3mm dorsal punch wounds inoculated with ~10^3 CFU Staphylococcus aureus (S.aureus) per wound. At days 1, 3, 7 and 10, aged mice exhibited heightened bacterial colonization (F=0.0648; Age, p=0.0002; Time, p=0.002 by two-way ANOVA) and increased wound size compared to young mice (F=0.7666; Age, p&lt;0.0001; Time, p&lt;0.0001 by two-way ANOVA). At day 1, there was no difference in wound neutrophil (PMN) or macrophage (MΦ) recruitment between young and aged mice despite higher levels of the chemokines KC, MIP-2 and MCP-1 in aged animals (p&lt;0.05). Conversely, at day 3, fewer numbers of PMN and MΦ were recruited to wounds in aged mice (p&lt;0.05) though chemokine levels in aged mice were comparable to young. The frequency or expression of TLR2+ wound PMN or MΦ was not affected by age. No differences in S. aureus phagocytosis by wound PMN or MΦ were observed. Interestingly, in circulation, aged mice had a higher frequency of CXCR2+ PMN and elevated CXCR2 expression compared to young mice at early time points (p&lt;0.05). Together, these data suggest that early dysregulation of leukocyte recruitment impairs resolution of cutaneous wound infection with advanced age.
Administration of anti-FcER1 antibody injections daily for 10 consecutive days delays type 1 diabetes onset in NOD mice (119.11)
Authors: Torrero Marina , Hübner Marc , Shi Yinghui , Mitre Edward 
Published: January 2023

Abstract
Helminth infections protect against autoimmune diseases. In this study, we investigated whether mimicking the immune response induced by helminths could protect against autoimmunity in the absence of actual infection. As helminths cause chronic IgE-mediated activation of basophils and mast cells, we tested the hypothesis that continuous activation of these cells would delay the onset of diabetes in NOD mice. Twelve week old NOD mice were injected i.p. daily with 5ug of anti-FcER1 antibody or isotype control for 10 days and onset of diabetes assessed. In vitro effects of anti-FcER1 were studied by measuring IL-4 production in basophils by flow cytometry and in vivo effects were assessed by measuring plasma histamine and IL-4 levels 30 min and 18 h after i.p. injection of antibody. Cellular immune responses were evaluated by measuring cytokine production from splenocytes by ELISA and assessing frequencies of natural T and B regulatory cells by multicolor flow cytometry. Injections of anti-FcER1 resulted in release of IL-4 and histamine into the bloodstream, consistent with in vivo basophil and mast cell activation. Anti-FcER1 or isotype treatment increased production of IL-10 from splenocytes. By 20 weeks of age, 80% of mice in the control and isotype groups had developed diabetes, compared to only 20% of mice treated with anti-FcER1. These results demonstrate that anti-FcER1 therapy can delay the onset of Type I diabetes in NOD mice.
Vitamin C insufficiency aggravates Dextran Sodium Sulfate (DSS)-induced colitis in Gulo(-/-) mice (120.20)
Authors: Kim Hyemin , Bae Seyeon , Yu Yeonsil , Kim Yejin , Kim Hang-Rae , Hwang Young Il , Kang Jae Seung , Lee Wang Jae 
Published: January 2023

Abstract
Intestinal mucosal damage in the inflammatory bowel diseases (IBD) involves the dysfunctional immunoregulation of the gut. Among the immunoregulatory factors, reactive oxygen species (ROS) are produced in abnormally high levels in IBD and, their destructive effects may contribute to the initiation or propagation of the disease. Vitamin C not only scavenges free radicals as an antioxidant but also has anti-inflammatory effects. Therefore, we investigated the effect of vitamin C on DSS-induced colitis in gulo(-/-) mice which cannot synthesize vitamin C like human. Vitamin C-insufficient gulo(-/-) mice showed decreased survival and lowering recovery efficacy. It accompanied with more severe colitis such as epithelial erosion, infiltration of inflammatory cells and contraction of colon. The production of pro-inflammatory cytokine, IL-6 was remarkably higher in DSS-treated vitamin C-insufficient gulo(-/-) mice than vitamin C-sufficient gulo(-/-) mice and wild type mice. In contrast, the production of anti-inflammatory cytokine, IL-22 was significantly decreased and IL-22 producing NKp46(+) cells were lower in vitamin C-insufficient gulo(-/-) mice. The phosphorylation of STAT3, a downstream signaling of IL-6 and IL-22, was increased in both epithelial cells and lamina propria cells. Taken together, it suggests that vitamin C represents a protective effect on DSS-induced colitis by regulating the production of cytokine and the induction of inflammation.
Effect of spa spring water on cytokine expression in human keratinocyte HaCaT cells and on differentiation of CD4+ T cells (120.26)
Authors: Lee Hye Jin , Cho Kyung-Ah , Woo So-Youn , Lee Kyung-Ho , Kim Jin-Wou 
Published: January 2023

Abstract
Skin contributes to the first line of defense against foreign materials outside of our body. In inflammatory skin disease, the pathogenesis is due to an immune reaction in the keratinocytes, immune cells and soluble mediators. Spa therapy is widely used for the treatment of inflammatory skin disease but the mechanisms are only partly understood by immune regulation. Spa therapy or balneotherapy in dermatologic disease can affect the secretion of pro-inflammatory cytokines, IL-1alpha and tumor-necrosis factor (TNF) from keratinocytes and possibly affect T cell differentiation. In this study, we evaluated the effect of spa spring water from Yong-Goong-Oun-Chun on cells and investigated the skin immune reaction. The treatment of spa spring water from Yong-Goong-Oun-Chun decreased the expression of pro-inflammatory cytokines under TLR stimulation to HaCaT cells and APCs. In addition, spa spring water attenuated the differentiation process of subsets of CD4+ T cells, i.e., Th1, Th2 and Th17 cells. All these immune parameters can be used to evaluate the efficacy of spa spring water in Korea in terms of immune modulatory effect.