bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of Cytokines and Chemokines Circulating in Plasma Obtained from a Phase I Prime-Boost Dengue Vaccine Trial
Authors: Fessehazion Arsiema , Currier Jeffrey R , Koren Michael , Keiser Paul , Lin Leyi , Jarman Richard G , Friberg Heather 
Published: January 2023

Abstract
A safe and efficacious vaccine for dengue is seen as the best rationale for decreasing the burden of morbidity and mortality associated with dengue infection. Identification of acute phase soluble factors as biomarkers of successful vaccination would has ten the development of a dengue vaccine. The Walter Reed Army Institute Research (WRAIR) is exploring the potential of a Prime-Boost vaccine strategy using purified inactivated virus (PIV) and live attenuated virus (LAV) tetravalent dengue vaccine candidates. A phase 1 clinical trial was performed testing different dosing schedules, PIV followed by LAV or vice versa at 0–1 or 0–6 months. Plasma samples were collected pre-vaccination and at multiple time points immediately post-vaccination to assess the kinetics of the acute phase soluble factor response to vaccination. We tested the plasma samples using Luminex-based magnetic bead kits measuring 59 different chemokines and cytokines. Our results reveal that different cytokine profiles were induced in subjects depending on the vaccine modality and whether it was given as the prime or boost dose. LAV appeared to be most immunogenic, particularly when received as a booster dose after PIV priming. Production of cytokines, including sCD30, APRIL, sTNF-R2, and IL-1RA, occurred in the majority of subjects tested approximately 10–14days post-vaccination. This study supports other data from this trial demonstrating that the type of vaccine and the order in which it is received impacts immunogenicity. Correlations of soluble factor responses with traditional adaptive immune responses, such as neutralizing antibody and IFN-γ ELISpot assays, will be presented.
Paracrine rather than systemic glucocorticoids are biologically active in the thymus
Authors: Taves Matthew Donald , Mittelstadt Paul R , Korol Anastasia M , Hamden Jordan E , Abraham Ninan , Soma Kiran K , Ashwell Jonathan D 
Published: January 2023

Abstract
Thymocyte positive and negative selection are critical for generation of a competent and self-tolerant T cell repertoire. Glucocorticoids (GCs) protect thymocytes from T cell receptor (TCR)-induced death, and thymocyte-specific GC receptor (GR) deletion amplifies negative selection, weakening the TCR repertoire. Circulating GCs are secreted by the adrenals, but levels fluctuate widely with time of day and in response to stressors. To avoid such variation, thymus GCs might be regulated independently of the adrenals, as thymic epithelial cells (TECs) and possibly thymocytes express GC-synthetic enzymes. Whether local GC production is sufficient to affect thymocyte development in the presence of adrenal GCs, however, is unknown. Here, we have found that corticosterone, the major mouse GC, was locally elevated in the thymus compared to the blood, and that cultured thymus produced corticosterone from endogenous substrates via GC-synthetic enzyme activity. To test the source and importance of local GC synthesis in vivo, we generated mice with targeted deletion of the GC-synthetic enzyme Cyp11b1 in TECs (Cyp11b1foxn1-Cre) or thymocytes (Cyp11b1lck-Cre). As a measure of GC signaling we quantified thymocyte expression of the GC-responsive gene Gilz. Gilz mRNA was normal in Cyp11b1lck-Cre but reduced in Cyp11b1foxn1-Cre thymocytes, with a reduction equivalent to that in GR-deficient thymocytes. Basal GR signaling is thus driven overwhelmingly by TEC-rather than adrenal-derived corticosterone. These findings demonstrate the importance of paracrine GC function in vivo, and are consistent with a role for paracrine GCs in thymocyte selection.
Analysis of expression of genes encoding neutrophil and eosinophil granule proteins during in vitro eosinophilopoiesis
Authors: Chung Il Yup , Kim Sung Min , Hwang Sae Mi , Kim Karam 
Published: January 2023

Abstract
Neutrophils and eosinophils, the two prominent granulocytes in circulation, are characteristically distinguished by neutrophil primary granule proteins (NPGPs) and eosinophil specific granule proteins (ESGPs). They differentiate from myeloid progenitors in the bone marrow commonly through a successive stage encompassing promyelocyte, myelocyte, metamyelocyte, and band form. The purpose of this study is to analyze bifurcation of the two granulocytes by examining expression of the NPGP- and ESGP-encoding genes during eosinophilic differentiation of cord blood (CB) CD34+ cells for 24 days. All transcripts of the NPGP-encoding genes, including PRTN3, MPO, CTSG, AZU1, and ELANE, reach peaks at day 12 and steeply decrease thereafter, while the transcripts of ESGP-encoding genes, such as MBP and ECP, reached peaks at day 18. Similarly, upon differentiation of immature eosinophilic EoL-1 cells, the NPGP transcripts are markedly downregulated accompanying by upregulated ESGP transcripts. Thus, the NPGP-encoding genes appear to be transcriptionally active even at eosinophilic promyelocyte and myelocyte stages. As these cells undergo maturation through metamyelocyte and band form stages, a transcription network come into a play that favorably shifts expression the ESGP-encoding genes.
The human CD34(−)CD117(+)CD56(−)NKp44(−) population in secondary lymphoid tissue serves as a common progenitor to NK cells and ILC2s
Authors: Chen Luxi , Carson Mary , Nalin Ansel , Young Karen A , Scoville Steven D , Mundy-Bosse Bethany L , Caligiuri Michael A , Freud Aharon G 
Published: January 2023

Abstract
The developmental pathways of human innate lymphoid cells (ILCs) have not been fully elucidated. We reported that human natural killer (NK) cells can derive from progenitor cells naturally residing in tonsils. We hypothesized that Group 2 ILCs (ILC2s) also develop in tonsils through a shared pathway. In support of this, ex vivo flow cytometry analyses of enriched human tonsil ILCs revealed a putative pathway of human CD294(+) ILC2 development stemming from a lineage (Lin)(−)CD34(−)CD117(+) population from which non-overlapping CD94(+) NK cells also emerged. Furthermore, when purified tonsil Lin(−)CD34(−)CD117(+) cells were cultured for 28 days with murine OP9 stroma expressing the human Notch ligand, Delta-like 1, in the presence of human interleukin-7 and Flt3 ligand, they produced mutually distinct and functionally mature populations of CD94(+) NK cells and CD294(+) ILC2s. The tonsil Lin(−)CD34(−)CD117(+) population is heterogeneous for CD56 and NKp44. Evaluation of the in vitro lineage differentiation potentials of subsets within the Lin(−)CD34(−)CD117(+) population according to NKp44 and CD56 revealed that NKp44(−)CD56(−) cells gave rise to both NK cells and ILC2s, whereas fractions expressing NKp44 and/or CD56 gave rise to NK cells but only to negligible ILC2s. In fact, the NKp44(−)CD56(−) population gave rise to a significantly higher percentage of ILC2s (7.50 ± 0.6) compared to the NKp44(+)CD56(+/−) populations (1.22 ± 0.4) (n = 5, p &lt; 0.001). These data support a model whereby human NK cells and ILC2s can develop from a common Lin(−)CD34(−)CD117(+)CD56(−)NKp44(−) progenitor in tonsils and that the acquisition of NKp44 and/or CD56 restricts lineage differentiation potential towards the NK cell lineage.
3D visualization of sex differences in the immune response to influenza infection in the mouse lung
Authors: Condiff Emily K , Milanez-Almeida Pedro , Li Weizhe , Gerner Michael Y. , Germain Ronald N 
Published: January 2023

Abstract
Recent developments of imaging techniques, including histo-cytometry and a new tissue clearing and staining protocol called Clearing-enhanced 3D (Ce3D), allow for high-resolution, 3D multiplex quantitative imaging and analysis of cell populations in intact organ material. We have established a panel of 10–14 markers for use in Ce3D-cleared 250μm-thick sections of lung tissue to visualize both innate and adaptive aspects of the immune response as comprehensively as possible. This panel, combined with quantitative analysis provided by histo-cytometry, allows for the collection of larger, more informative imaging datasets than typically generated by conventional histochemistry methods. In our study, we applied these techniques to the mouse model of influenza A virus (IAV) infection to investigate the patterns of immune cell distribution and viral spread in the lungs of male and female mice. In C57BL/6J mice, we observed increased mortality in female mice compared to males when infected with the same dose of the lethal PR8 strain of IAV. Literature research as well as data from the infection of different transgenic mouse lines suggest that higher levels of innate inflammation in female mice may lead to the observed difference in mortality. Ce3D multiplex images show multiple differences in the distribution of both innate immune cells and infected areas in lungs from male and female mice. This study will contribute to a better understanding of both the immune response to IAV infection and sex differences in immunity in general, potentially leading to better design of tailored vaccines and treatments in the future.
Quantifying the relative impact of influenza virus mutations on viral fitness and escape from cellular immunity
Authors: Li Zheng-Rong Tiger , Zarnitsyna Veronika I , Lowen Anice C , Kohlmeier Jacob E , Antia Rustom 
Published: January 2023

Abstract
The relative conservation of T cell epitopes in human influenza viruses compared to antibody epitopes may be explained by two mechanisms: (1) mutations of the T cell epitopes are constrained by their potential to decrease viral fitness by altering protein function, or (2) the selection benefits gained by ‘escaping’ mutations are small due to the breadth of MHC polymorphisms among human population. The level of influenza-specific cellular immunity may further complicate the relative contribution of these two mechanisms. We developed a mathematical model incorporating the mutation cost, selection benefit, and allele frequency of ‘escapable’ MHC that indicated certain combinations of these three parameters determined the fate of influenza viruses harboring mutations in T cell epitopes. To measure the mutation cost and selection benefit, we developed a novel digital droplet PCR method for the simultaneous detection of wild-type and mutant influenza viruses harboring mutations in the immunodominant NP366–374 epitope. Primary infections with WT, NP-mutated, and 1:1 mixture of WT and mutated x31 influenza viruses showed no significant difference between the two viruses, while the secondary infection of PR8-primed B6 mice with the 1:1 mixture resulted in a 10-fold decrease in the peak of viral load of WT compared to NP-mutated viruses. Together, these data show that the mutation of a single immunodominant T cell epitope provides significant selection benefit with minimal fitness cost to the virus. Future studies in mice with diverse MHC alleles are underway to quantify the impact of varying levels of cellular immune pressure on the in vivo ability of mutant influenza viruses to escape T cell immunity.
Metabolic subsets of plasma cells and their turnover in the marrow
Authors: Wols Heather A Minges , Aguilera Alexya N , Witte Pamela L 
Published: January 2023

Abstract
The ability of long-lived plasma cells (LLPC) to maintain longevity and avoid metabolic burnout has been largely unexplored until recently. Our lab has shown that LLPC reside in hypoxic niches and are divided into two distinct subsets based on fluorescence following MitoTracker Red CM H2Xros staining: a predominant MitoHI subset, suggestive of aerobic respiration, and a rarer MitoLO subset, suggestive of glycolysis. These MitoHI and LO PC are comparable in function as detected by ELISPOT and ELISA following 24h ex vivo culture at oxygen levels as low as 0.5% (severe hypoxia). Taken together, these data suggest LLPC have the flexibility to use glycolysis when needed, although respiration may be the preferable metabolic strategy. To determine whether the MitoHI and LO PC subsets differed in turnover, we looked at 5-bromo-2′-deoxyuridine (BrdU) incorporation after ten days of labeling post antigen-boost to label PC forming from memory cells. Mice were sacrificed 13, 20, and 43 days following boost, and the frequency of BrdU+ PCs in the MitoHI and LO marrow compartments determined. The proportions of BrdU+ MitoHI PCs remained relatively stable from days 13 to 20 and slightly declined by day 43. Frequency of BrdU+ MitoLO PCs was initially greater than that in the MitoHI PC compartment, but rapidly diminished at subsequent time points. These data suggest that the turnover in the MitoLO compartment is much higher compared to the MitoHI compartment, which is more stable. Tracking antigen-specific PCs supported this conclusion. Thus, although marrow PC reside in hypoxic niches, they may prefer to use aerobic respiration and employ glycolysis only as an ‘as needed’ basis.
Development of an experimental model of Campylobacter jejuni intestinal colonization in commercial turkeys
Authors: Sylte Matthew Jon , Looft Torey , Johnson Timothy A. , Wu Zuowei , Zhang Qijing 
Published: January 2023

Abstract
The cecum is the main site where poultry are asymptomatically colonized by thermophillic Campylobacter spp. Consumption of contaminated poultry products is one of the main sources of human campylobacteriosis, for which Campylobacter jejuni is responsible for approximately 90% of the cases. In chickens, experimental models of C. jejuni colonization to examine the host-response have been established, but a model remains to be developed for commercial turkeys. We sought to develop a model to characterize intestinal colonization and host response in commercial turkeys using mutants of C. jejuni (strain NCTC 11168) constructed to express different antibiotic resistance markers (chloramphenicol (CjCm) or kanamycin (CjK)). Turkey poults were colonized by oral gavage at 21 days of age with up to 5×108 cfu of either CjCm, CjK or sterile PBS. Up to 8 poults/treatment were euthanized at days 2, 7 and 14 post-exposure and intestinal samples were assessed for colonization and host-response. Both mutants were consistently recovered from cecum, but not ileum, throughout the study, with ~107 cfu/g detected from cecal contents at day 14 post-exposure. Mild inflammatory changes were histologically noted in the cecum and cecal tonsil of CjCm or CjK colonized poults at days 7 and 14 post-exposure. Using IHC, Campylobacter antigen was detected between cecal villi by day 7, indicating persistent colonization. Host-response for cytokines (IL-6, -8, -10, -17A, -22 and others) and β-defensins was analyzed by qRT-PCR from intestinal tissues. This study demonstrates that CjCm and CjK persistently colonize the cecum of commercial turkeys and will be useful to evaluate strategies to reduce intestinal colonization by C. jejuni in turkeys and promote food safety.
Composition of the gut microbiota influences infection severity and pregnancy outcome in Plasmodium chabaudi-infected pregnant mice
Authors: Smith Catherine Diane , Cooper Caitlin A. , Moore Julie M. 
Published: January 2023

Abstract
Placental malaria, a severe clinical manifestation of Plasmodium falciparum infection, is a major cause of pregnancy loss, neonatal mortality, and severe maternal illness. We have developed a novel mouse model for pregnancy maintenance during maternal malaria infection utilizing outbred Swiss Webster mice. When infected with Plasmodium chabaudi in early gestation, several inbred mouse strains will abort their pregnancies at midgestation. However, outbred Swiss Webster mice infected with P. chabaudi in early gestation carry their pregnancies to term, allowing us to explore the immunological balance between parasite clearance and pregnancy success. The composition of the gut microbiota may alter this balance. As described by Villarino et al., C57BL/6 mice sourced from different vendors display microbiota-dependent differences in Plasmodium infection severity resulting in a susceptible or resistant phenotype. Similarly, pregnant Swiss Webster mice with susceptible gut microbes develop higher parasite burdens than mice with resistant gut microbes, although both cohorts produce live pups at term. Despite the lower parasite burdens observed in resistant mice, these dams tend to produce fewer viable pups per litter, suggesting a trade-off between accelerated parasite clearance and pregnancy success. We hypothesize that the difference in litter size between susceptible and resistant dams is a result of the elevated levels of pro-inflammatory cytokines observed in resistant dams. The levels of systemic TNF and IFN-γ, which have previously been associated with the clearance of blood-stage malaria infection and pregnancy loss in P. chabaudi-infected pregnant C57BL/6 mice, are expected to be higher in resistant mice.
Vitamin A deficiency in mice results in a hypermetabolic state, dysbiosis and alterations in bacterial metabolism
Authors: Tian Yuan , Nichols Robert G , Snyer Lindsay , Weaver Veronika , Cai Jingwei , Allen Joselyn Natasha , Vijay Anitha , Patterson Andrew D , Cantorna Margherita T 
Published: January 2023

Abstract
Vitamin A deficiency (A−) is the leading cause of preventable childhood blindness in developing countries and is associated with decreased ability to fight infection. A− mice are susceptible to Citrobacter rodentium infection and surviving mice fail to clear the infection. To better understand how vitamin A status influences the metabolome and gut microbiota, we systematically analyzed biological samples from vitamin A sufficient (A+, n = 6) and vitamin A deficient (A−, n = 6) mice using 1H NMR-based metabolomics and 16S rRNA gene sequencing. 1H NMR analyses showed that A− mice had disturbances in multiple metabolic pathways including alterations in energy metabolism (hyperglycemia, glycogenesis, TCA cycle, and lipoprotein biosynthesis) and the A− host showed overall signs of a hypermetabolic state. 16S rRNA gene sequencing indicated that the microbiota in A− mice had phenotypic as well as functional shifts (based on PICRUSt). Evaluation of bacterial metabolism showed enhanced amino acid metabolism, carbohydrate metabolism, DNA replication and repair, and signal transduction in A− mice. In addition, the A− microbiota had lower amino acid biosynthesis, vitamin metabolism, and cell motility. These findings suggest that vitamin A deficiency is associated with hyperglycemia, liver disease, and dysbiosis of the gut microbial communities. Bacteria do not express receptors for vitamin A and therefore it seems likely that the changes in bacterial communities and bacterial metabolism are indirectly regulated by vitamin A. How these changes might compromise the ability of the A− mice to clear a gastrointestinal infection with C. rodentium is an area of continuing investigation.
Role of Acinetobacter baumannii thioredoxin in bacterial dissemination by modulation of mucosal oxidative homeostasis
Authors: May Holly , Yu Jieh-Juen , Gupta Rishein , Guentzel M. Neal , Arulanandam Bernard 
Published: January 2023

Abstract
Acinetobacter baumannii is an increasing cause of hospital-acquired infections and a prominent cause of combat-related infections in the Middle East. Infections with this bacterium lead to a TLR4 response that initiates a potent innate immune response. Bacterial coinfection may occur in hospitalized patients, and endotoxin (LPS) released from one Gram-negative bacterium may have profound effects on pathogenesis of other co-infecting bacteria. Specifically, LPS induced oxidative stress has a significant impact on mucosal barrier function, leading to enhanced permeability and bacterial translocation. To delineate the importance of LPS in E. coli and Ab coinfection, we utilized a murine model of pulmonary Ab infection with oxidative stress induced via LPS injection. While the bacterial factors involved in translocation under oxidative stress are largely unknown, one plausible factor, based on previous research in our lab, is thioredoxin. Thioredoxin-1 (Trx1/TrxA) is a member of the thioredoxin protein superfamily that can be reversibly oxidized and reduced to facilitate reduction of disulfide bonds, as well as for protecting against free-radical damage. Our lab has created a mutant of Ab Clinical isolate 79 (Ci79) that lacks thioredoxin-1 (ΔTrxA). Using this mutant, the effects of thioredoxin on bacterial translocation during oxidative stress can be determined. Mice given E. coli LPS showed significantly increased translocation of Ci79 via organ burden. Mice infected with ΔTrxA showed significantly less translocation than the WT strains. Additionally, when Ci79 was treated with a TrxA blocker before infection, survival was increased and organ burden decreased.
Δ9-tetrahydrocannabinol (THC) activation of cannabinoid receptors induces unique changes in the murine gut microbiome and associated induction of myeloid-derived suppressor cells and Th17 cells.
Authors: Becker William J , Nagarkatti Mitzi , Nagarkatti Prakash S 
Published: January 2023

Abstract
Δ9-tetrahydrocannabinol (THC) is a partial agonist for cannabinoid receptors (CB1 & CB2), is the main psychoactive ingredient found in the Cannabis plant, and can modulate the nervous and immune system. Our intestine harbors a diverse array of bacteria, referred to as the microbiome, which is known to impact many bodily functions, including the immune system. In the current study, we investigated the effect of THC on the gut microbiome and the naïve immune response. We tested the effect of acute or chronic exposure of C57BL/6 mice to THC on the immune system, and correlated said changes with a flux in intestinal bacteria. Intraperitoneal exposure to THC resulted in migration of CD11b+Gr-1+ myeloid-derived suppressor cells (MDSCs) from the bone marrow to the peritoneal cavity. These MDSCs remained in the peritoneum throughout chronic THC administration, where they proliferated and produced IL-6. The peritoneal IL-6 led to an increase in T helper 17 (Th17) cells in the mesenteric lymph node (mLN), as well as increases of IL-17A in the serum and mLN supernatant of THC-treated mice. Indeed, an adoptive transfer of THC MDSCs increased the number of Th17 cells compared to bone marrow (BM) MDSCS, and in-vitro polarization experiments revealed THC MDSCs to be more potent inducers of Th17 cells than BM MDSCs. The alterations in the gut immune cells occurred in tandem with an increase in the number of Alphaproteobacteria in the cecum and feces of mice treated with THC compared to vehicle and naïve mice, and this effect was replicated after an adoptive transfer of THC MDSCS, but not BM MDSCs. Together, these data demonstrated that marijuana use can alter the gut microbiota as well as cause significant changes in the naïve immune system.