bluuebunny/crossref_metadata_2025_split huggingface dataset:
Minipigs as neonatal animal model for tuberculosis vaccine efficacy testing
Authors: Juarrero Mercedes Gonzalez , Ramos Laylaa , Obregon-Henao Andres , Henao-Tamayo Marcela , Bowen Richard , Izzo Angelo A. , Lunney Joan 
Published: January 2023

Abstract
Many vaccines against childhood diseases are administered early after birth. Despite the fact that neonatal-infant immunity differs from adult immunity, vaccine development studies frequently test efficacy in adult rather than in neonatal animal models. A good example of this scenario is Bacillus Calmette-Guerin (BCG). BCG is the only vaccine available against tuberculosis (TB). The vaccine lacks efficacy against child and adulthood pulmonary disease or reactivation of latent TB but prevents disseminated TB in children and is thus widely used in countries with endemic TB as part of the neonatal vaccine regimen. There are several new vaccines that have shown efficacy against TB in adult animal models yet fail to protect infants from TB disease in clinical trials. In this novel approach, we used the minipig as a neonatal animal model for evaluation of immune responses to BCG vaccination. The longitudinal immune responses of neonatal mini-piglets vaccinated with BCG were followed until adolescence, with the same monitoring applied to a group of unvaccinated mini-piglets. We characterized the memory and activation phenotype of T cells, cytokine profile, and monocyte activation in response to BCG stimulation from 4 weeks of age into adolescence- age of 24 weeks. Further, we challenged both vaccinated and non-vaccinated animals via aerosol route with a low dose aerosol of Mycobacterium tuberculosis strain HN878 and we characterized changes between the two groups in the course of immune responses following challenge. Based on comparison of immune responses to BCG in minipigs with similar studies in infants, our findings suggest that minipigs have the potential to serve as an effective neonatal animal model for vaccine development.
Birth weight affects immune response of piglets during the peri-weaning period
Authors: Verso Luca Lo , Matte Jacques , Talbot Guylaine , Lapointe Jérôme , Bissonnette Nathalie , Guay Frédéric , Ouattara Bazoumana , Luna Uanderson Verissimo , Lessard Martin 
Published: January 2023

Abstract
Relationships between birth weight and a range of immune markers were studied in piglets during the peri-weaning period. A total of 47 sows and their respective litters adjusted to 12 piglets were used. Piglets were weighed after birth and within each litter 2 low weight (LW) and 2 high weight (HW) animals were identified: the average weight of the 2 groups was 1.12 and 1.81 ± 0.17 kg, respectively. Within litter, one couple of LW and HW piglets was sacrificed at 16 and 23 d of age, two days after weaning at day 21, to characterise different leukocyte subsets in mesenteric lymph nodes (MLN) and blood by flow cytometry. Jejunal mucosal and MLN samples were also collected to measure the expression of genes involved in immune response by qPCR. The percentage of blood γδT cells was higher in HW piglets before and after weaning (P=0.01 and P&lt;0.001, respectively). At 16 d, LW animals had higher amounts of CD4+CD8+ T cells and dendritic cells in the blood (P=0.01 and P=0.04, respectively) and in MLN (P=0.02 and P=0.009 respectively), and lower count of blood cytotoxic T cells (P=0.01) than HW. Moreover, jejunal expression of TNFα was reduced in LW piglets (P=0.02), while MLN expression of CCL23, BMP2 and SPP1 was increased (P=0.003, P=0.002 and P=0.02, respectively). Two days after weaning (d 23), HW piglets had higher percentages of T cells (P=0.001) and lower amount of non-T cells (P=0.03) and natural killer cells (P=0.04) in blood compared to LW animals. These data revealed that piglet birth weight affects the proportion of leukocyte subsets in different tissues and the expression of genes involved in the immunity. Globally, our results suggest that the development of the immune system after birth differed between LW and HW piglets.
β-glucan induced training and tolerance: alterations to primary monocytes
Authors: Byrne Kristen A. , Beiki Hamid , Tuggle Christopher K. , Loving Crystal L. 
Published: January 2023

Abstract
In mice and humans, exposure to β-glucan can induce epigenetic reprograming in monocytes resulting in enhanced responses to heterologous agonists. Epigenetic reprograming is the basis for innate memory, which includes both decreased immune responsiveness (innate tolerance) and increased responsiveness (innate training). As a valuable food animal and medically relevant species, we sought to understand the phenotypic and mechanistic alterations induced by β-glucan on swine cells to develop methods to enhance health while limiting antibiotic usage. Thus, primary porcine monocytes were stimulated with β-glucan from S. cerevisiae or C. albicans, rested for 5 d, and then restimulated with lipopolysaccharide (LPS; TLR 4 agonist) or Pam3CSK4 (synthetic triacylated lipopeptide; TLR 2 agonist) to determine trained or tolerant phenotype (increase or decrease in cytokine production relative to unstimulated controls). Zymosan (β-glucan from S. cerevisiae) primed monocytes exhibited a tolerant phenotype (decreased IL-1β and TNF-α production compared to controls) when restimulated with LPS or Pam3CSK4. However, β-glucan from C. albicans (the primary β-glucan used in mouse and human studies) primed porcine monocytes for increased cytokine production after LPS and Pam3CSK4 stimulation, an indication of trained immunity. Epigenetic analysis of the accessibility of the genome to transposase (ATAC-sequencing) showed changes in promoter peaks with treatment, providing insight into mechanisms of tolerance versus training. These data indicate that β-glucan can induce both training and tolerance in porcine monocytes, but the source and purity of β-glucan likely impacts innate memory.
Mucosal cellular immune responses following Escherichia coli O157:H7 bacterin vaccination associated with reduced fecal shedding
Authors: Schaut Robert G. , Boggiatto Paola M. , Palmer Mitchell , Loving Crystal L. , Sharma Vijay 
Published: January 2023

Abstract
Parenterally-delivered, bacterin-based vaccines can significantly limit shedding of Shiga-toxin producing Escherichia coli (STEC) O157:H7 from cattle, providing a potential intervention strategy to limit foodborne contamination. While the peripheral antibody response to STEC bacterin has been characterized, induction of STEC-specific cellular immunity, particularly at the site of colonization, remains poorly described. As STEC-specific antibodies likely provide some protection, cellular immune responses likely contribute to limiting colonization also. Following vaccination and challenge, lymphocytes were detected in the rectoanal junction (RAJ), a preferred colonization site for E. coli O157:H7. T cells present in the RAJ were positive for interferon (IFN)-γ mRNA, including a subset of γδ+ CD8+ T cells. Specifically within the RAJ, γδ+ T cells expressing IFN-γ mRNA were greater in number compared to classical αβ+ T cells. Also, distinct lymphoid follicles were present in greater number at the RAJ in vaccinated/challenged animals and fecal STEC-specific IgA was detected. These vaccine -primed responses likely support localized immune responses upon challenge. Local T cell-produced IFN-γ may impact epithelial cells, subsequently limiting E. coli adherence. Thus, distinct immune changes were induced at the mucosa of vaccinated and challenged animals, providing insight on mechanisms associated with limiting shedding. Enhancing mucosal immunity may be critical in the further development of efficacious vaccines for controlling STEC O157:H7 in ruminants, thus limiting human infection.
Somatic hypermutation of TCRα contributes to thymic positive selection in sharks
Authors: Ott Jeannine A. , Castro Caitlin , Deiss Thaddeus , Ota Yuko , Flajnik Martin F. , Criscitiello Michael F. 
Published: January 2023

Abstract
In mammals and probably all vertebrates, receptor editing of TCR alpha genes enhances immature T cell positive selection over a three-day interval in the thymic cortex. Surprisingly we found extensive somatic hypermutation (SHM) operating at the TCRα locus in the nurse shark thymus, implying that SHM contributes to receptor modifications that enhance positive selection. We analyzed mutation in TCRα families of clones with the same VJ rearrangement. Additionally, in situ hybridization showed the strongest activation-induced cytidine deaminase (AID) expression in the central thymic cortex and bordering the corticomedullary junction. The frequency of mutation at TCRα was as high as that seen at B cell receptor (BCR) loci in sharks and mammals. Complementarity determining regions (CDRs) accumulated significantly more mutations than framework regions (FWs), and significantly more mutations in CDRs resulted in amino acid replacement than in silent mutation. We saw a preference for transition mutations as well as a strong bias toward G:C substitutions within AID hotspots, especially within CDR regions. We suggest that TCRα utilizes SHM to boost positive selection and perhaps to broaden diversification of the αβ T cell repertoire in sharks, the first reported use of this process in thymic diversification in vertebrates.
Plasma surfactant proteins and lipid mediators in the airway associate with different stages of viral infection and resolution in a sub-lethal mouse Influenza A model
Authors: Nguyen Allen , Wong Kent , Chakrabarti Ari , Jiang Jenny , Ulufatu Sheila , Liu Shannon , Valle Nicole , Park Summer , Xu Min , Rosenberger Carrie , Anania Veronica , McBride Jacqueline 
Published: January 2023

Abstract
Viral quantification serves as a valid biomarker for detecting Influenza A infection however viral burden is not closely associated with disease severity or clinical outcomes. Biomarkers reflecting the status of the host immune response are needed to monitor and/or predict disease outcomes in a hospitalized setting.
Here, we utilized a sub-lethal mouse model of Influenza A infection to explore plasma and airway cytokines, plasma surfactant proteins (SP-D, SP-A and CC16), and airway bioactive lipid mediators as potential biomarkers of lung leakage and host response, respectively. This study aimed to assess the pharmacodynamic response of these biomarkers after treatment with an efficacious dose of an anti-HA stalk-binding antibody and to assess association of these biomarkers with disease severity metrics.
Plasma SP-D and IP-10 correlated with lung viral titers while SP-A and CC16 tracked more closely with changes in bronchial albumin levels and body weight. Bioactive lipid precursors Arachidonic Acid (AA), Docosahexaenoicacid (DHA) and Eicosapentaenoic acid (EPA), reported to have both pro- and anti-inflammatory properties, increase in the lung with infection. 17-HDHA, a DHA metabolite reported to be pro-resolving, was induced later during disease resolution. With anti-HA antibody treatment, all plasma surfactant proteins and precursor lipids in the airway were reduced and 17-HDHA production was accelerated.
In summary, plasma SP-D may serve as an early biomarker of host response and/or potentially indicate a decline in lung epithelium integrity. Plasma SP-A, CC16, and pro-inflammatory lipid mediators such as AA may reflect the ongoing host response and may therefore be more closely associated with disease outcomes.
Cytokines levels in nasal lavage in pediatric patients infected with respiratory viruses
Authors: Rosas-Taraco Adrian Geovanni , Leon-Icaza Stephen A. , Gonzalez-Chapa Jorge A. , Silva-Ramirez Beatriz , Martinez-Castilla Azalia M. , Treviño-Garza Consuelo , De la O-Cavazos Manuel E. , Montes-Tapia Fernando F. , Salinas-Carmona Mario C. 
Published: January 2023

Abstract
Respiratory viruses are responsible of acute respiratory infections that affect worldwide population causing morbi-mortality and children younger than 5 years are the most affected population. Cytokines can modulate the immune response but little is known about their levels in nasal mucosa and less if these would correlate with symptoms severity. One hundred twenty-one pediatric patients with clinically diagnosed acute respiratory infections were sequentially enrolled in the Department of Pediatrics of the University Hospital. Throat swabs were performed and nucleic acid was isolated to viral detection by Luminex xTAG RVP kit and qRT-PCR. Nasal lavage were performed in each patients to evaluate cytokines levels by multiplex immunoassay. Disease severity was evaluated according to clinical characteristics on the day of enrollment. The most frequently found viral respiratory infections were hRV, RSV, and hMPV. hMPV load correlated with disease severity (P&lt;0.05). Pro-inflammatory, Th1, Th2 and Th17 cytokines were detected in nasal lavage in those patients, however only pro-inflammatory cytokines were found higher in severe disease. Pro-inflammatory, Th1/Th2/Th17 cytokines correlated with symptoms, mainly with fever (P&lt;0.05). Our findings show that disease severity correlates with viral load and cytokines in nasal lavage may regulate local immune response against respiratory virus.
Exchange Proteins Directly Activated by cAMP and the Role of these Proteins in Respiratory Syncytial Virus Infection
Authors: Wu Wenzhe , Choi Eunjin , Ren Yuping , Chen Yu , Liu Shengxuan , Ren Junping , Garofalo Roberto P. , Zhou Jia , Bao Xiaoyong 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) is a single-stranded, negative-strand RNA virus belonging to Paramyxoviridae family, and is in the genus Pneumovirus. It was discovered in chimpanzees in 1955 and subsequently confirmed to be a human pathogen shortly after that. RSV is one of the most common viruses to infect young children and high-risk adults worldwide, and the leading cause of inflammation of the lungs (pneumonia) or the lungs’ airways (bronchiolitis) in infants. Due in part to the lack of long-term immunity after infection, making reinfection frequent. There is a single antiviral treatment for RSV currently approved, but its use is limited by questionable efficacy, side effects, and cost, and it is recommended that it be used only for patients at-risk for severe disease, on a case-by-case basis. In this study, we discovered an exchange protein directly activated by cyclic AMP (EPAC) that can serve as a potential therapeutic target for RSV. In both lower and upper epithelial cells, the treatment of EPAC inhibitor (ESI-09), but not PKA inhibitor (H89), significantly inhibits the RSV replication and pro-inflammatory cytokine/chemokine induction. In addition, RSV-activated transcriptional factors belonging to the NF-κB and IRF families are also suppressed by ESI-09. Through isoform-specific gene knock down, we found that EPAC2, but not EPAC1, plays a dominant role in controlling RSV replication and viral-induced host responses. Experiments using both EPAC2 knock out and EPAC2 specific inhibitor support such roles of EPAC2. Therefore, EPAC2 could be a promising therapeutic target to regulate RSV replication and associated inflammation.
Detrimental role of type I interferon signaling in respiratory syncytial virus infection
Authors: Ansar Maria , Komaravelli Narayana , Ivanciuc Teodora , Casola Antonella , Garofalo Roberto P. 
Published: January 2023

Abstract
Respiratory Syncytial Virus (RSV), the primary cause of lower respiratory tract infections in young children, is a poor inducer of type I interferons (IFN) and is resistant to its canonical antiviral activity. Paradoxically, increased type I IFN has been observed in some studies to correlate with more severe disease in RSV-infected children. Previously, we have shown that a pathogenic reactive oxygen species (ROS) production in the course of RSV infection is caused by a progressive reduction in nuclear and cellular levels of the transcription factor NF-E2-related factor 2 (NRF2), the primary regulator of antioxidant enzyme (AOE) gene expression. Thus, the goal of this study was to determine the contribution of type I IFN to lung innate/inflammatory responses, modulation of AOE expression, and airway disease in experimental RSV infection. Using either IFN alpha/beta receptor knockout mice or IFNAR neutralizing antibody, we found that IFN signaling blockade attenuated RSV-mediated NRF2 degradation and rescued AOE levels (i.e. SOD 1 and Catalase) in the lung, resulting in improved airway function and clinical disease. We also showed that NRF2/AOE rescue in mice lacking IFN signaling is due to the reduced expression of the promyelocytic leukemia protein (PML), a key component of PML nuclear bodies. In addition, levels of RSV-induced pro-inflammatory cytokines including IL-1α/β, IL-6, IL-12(p40), TNF-α, and the chemokines CXCL1, CCL2, 3, 4, and 5 were also reduced in IFN deficient mice. In conclusion, this data shows for the first time that IFN type I signaling plays a critical role in enhancing RSV-mediated airway disease through an innate immune pathways that involve the PML-NRF2-AOE axis.
Role of mucin 19 in the respiratory tract.
Authors: McBride Kaitlin E. , Cheemarla Nagarjuna R. , Guerrero-Plata Maria Antonieta 
Published: January 2023

Abstract
Human metapneumovirus (HMPV) is a negative sense, single stranded RNA virus that belongs to the Pneumovirdae family, and represents an important pathogen that causes severe bronchiolitis and pneumonia worldwide. There is currently no vaccine against this virus, so it is important to study the aspects of the immune response induced by HMPV in order to understand its mechanism of infection. Because infiltration of mucus is a hallmark of HMPV infection, it is warranted to study the role of mucus in the disease process. Mucin proteins make up the major component of mucus and can be found within the airway and lungs. Using an in vitro model of HMPV infection and human bronchial epithelial cells, we found that HMPV induced predominantly a mucin 19 expression. Those data were confirmed in mice, where HMPV induced predominant levels of muc19 protein. Mucin 19 is a secreted gel-forming mucin, whose function is largely understudied, particularly in the respiratory tract. Our findings led us to hypothesize that mucin 19 plays a role in HMPV infection. To test that, we used a muc19 knockout mouse model. Lung samples were collected at different time points. Sample analysis showed that the absence of muc19 led to an increased pulmonary inflammation and body weight loss. Interestingly, flow cytometry data indicate that the lack of Muc19 resulted in a decreased CD4+ T cell count in the lung, and reduced numbers of CD4+ and CD8+ T cells in the lymph nodes after HMPV infection. These novel findings yield relevant information regarding the contribution of the mucins, specifically Muc19, on the immune response induced by respiratory viral infections in the lung.
Genome-wide screens identify novel components of the MHC I pathway.
Authors: Kriegsman Barry A. , Cruz Freidrich M. , Merino-Rodriguez Elena , BelleIsle Janice , Rock Kenneth L. 
Published: January 2023

Abstract
Antigen presentation is the process by which cells generate and display peptides on major histocompatibility complex (MHC) molecules for T cell recognition. Although it is well known that antigen presentation is critical for effective adaptive immunity, not all of the machinery driving antigen presentation has been identified. Because there is significant potential to exploit antigen presentation to develop therapeutics against cancers, microbes, and autoimmune diseases, it is important that we advance our understanding of these pathways. To this end, our lab has conducted two independent genome-wide screens to identify novel genes important for MHC I presentation. In one screen, we silenced mouse dendritic cells (DC3.2 cell line) with siRNA and tested the ability of the silenced dendritic cells to cross-present (i.e., present peptides derived from exogenous antigen on MHC I). In another screen, we used a lentiviral sgRNA library to knock out human genes in HeLa-Cas9 cells and identify genes that positively regulate surface MHC I levels. We are currently validating several hits from these screens and characterizing the mechanisms by which they affect MHC I presentation.
A Treg-specific deletion of Helios causes autoimmune lipodystrophy and metabolic syndrome
Authors: Thornton Angela M , Gavrilova Oksana F , Penna Vinay R , Korty Patricia E , Shevach Ethan M 
Published: January 2023

Abstract
Regulatory T (Treg) cells suppress immune activation in a dominant manner and play a critical role in the maintenance of self-tolerance. A subpopulation (60–75%) of Foxp3+T regulatory (Treg) cells express the transcription factor Helios. To examine the function of Helios in Treg cells, we have generated Treg-specific Helios deficient mice (cKO, Heliosflxflx Foxp3cre). Although both Treg development in the cKO mice and their in vitro suppressor function are normal, the selective deletion of Helios in Tregs leads to slow, progressive systemic immune activation with a Th1 phenotype, hypergammaglobulinemia, and enhanced germinal center formation. Initially, we observed significant lymphocytic infiltrates only in the salivary gland and not in any other organs typically affected by Treg dysregulation. Strikingly, the mice developed lipodystrophy, hepatic steotosis and insulin resistance. Further analysis revealed a significant lymphocytic infiltrate in both the inguinal and perigonadal adipose tissue, indicating autoimmune mediated destruction of the white adipose tissue (WAT). We have further shown that the lymphocytic infiltrate specific to WAT can be transferred from mice with lipodystrophy to immunodeficient mice, confirming the autoimmune nature of the lipodystrophy. Thus, Helios deficiency in Treg disrupts immune homeostasis in the adipose tissue, leading to the destruction of WAT and redirection of lipids to the liver, and ultimately causes metabolic dysfunction.
Supported by the Intramural Research Program of the NIAID, NIH.
The absence of CD137 ligand upregulates the expression of the immunosuppressive molecule soluble CD137
Authors: Foda Bardees M. , Forsberg Matthew H , Ciecko Ashley E , Mueller Kevin W , Geurts Aron , Chen Yi-Guang 
Published: January 2023

Abstract
The gene encoding the co-stimulatory molecule CD137 is located within the Idd9.3 T1D susceptibility locus and contributes to diabetes progression in NOD mice. We have previously shown that CD137 expression in T cells has dual functions: CD4+CD137+ T cells inhibited T1D development while CD8+CD137+ T cells showed potent diabetogenecity. The protective function of CD137 in CD4+T cells is likely due to the significant amounts of soluble CD137 (sCD137) produced by Foxp3+ Tregs. sCD137 is an alternatively spliced form of CD137 lacking the transmembrane encoding exon. Treating NOD mice with recombinant sCD137 prevents T1D development. We aim to study the impact of CD137 ligand on the expression of sCD137. We used CRISPR/Cas9 to generate a strain of NOD depleted of CD137L (NOD.Tnfsf9−/−). Development of T1D was repressed in the newly generated strain. Deficiency of CD137L was associated with increased levels of serum sCD137. CD137-deficient-T cells contributed significantly to the elevated serum levels of sCD137. Ligand-deficient-Treg cells produced higher levels of sCD137 proteins compared to wild-type Tregs. Moreover, we detected increased levels of sCD137 transcripts in Tregs lacking the expression of CD137L. We direct our efforts to uncover the molecular mechanisms involved in the regulatory role of CD137L influencing the expression of sCD137.