bluuebunny/crossref_metadata_2025_split huggingface dataset:
A replicating Adenovirus-SIV recombinant prime/protein boost regimen induces SIVspecific T and B cell responses in lymph nodes associated with humoral immunity in the genital mucosa of female Rhesus macaques
Authors: Hait Sabrina Helmold , Hogge Christopher James , Mushtaq Zuena , Hoang Tanya , Robert-Guroff Marjorie 
Published: January 2023

Abstract
The efficacy of HIV vaccines relies on potent antibody (Ab) responses at major transmission sites such as the female genital tract (FGT). CD4+ T follicular helper (TFH) cells, specialized in providing B cell help in germinal centers (GCs) of secondary lymphoid organs, play an important role in initial stages of HIV-specific Ab responses. Here we investigated SIV-specific T and B cell responses and FGT immunity in Rhesus macaques following mucosal priming with Ad5hr-SIV(Env/gag/nef) vectors at wks 0 & 13 and intramuscular boosting with SIV gp120 in alum at wks 26 & 38. Lymph nodes (LNs) were collected pre-vaccination and 3–4 days post-2nd prime and post-2nd boost. Vaginal washes were obtained prior to and 3 wks post-immunizations. GC TFH and B cell frequencies positively correlated post-2nd prime (p=0.04; r=0.6) suggesting an effect of early TFH cells in GCs. GC SIV-specific TFH cells were significantly enhanced post-2nd boost (p=0.01). BCL6 upregulation was also seen in GC-TFH cells at this time, confirming the GC phenotype of TFH cells. GC SIV-specific memory B cell levels were higher post-2nd boost compared to post-prime (p=0.03) indicating efficient SIV-specific B cell generation. GC B cells post-2nd prime and boost strongly associated with SIV-specific IgG levels in vaginal washes (p=0.03, r=0.7; p=0.002, r=0.8). GC centrocytes are critical for long-lasting Ab secreting cells. GC centrocytes post 2nd prime and boost tended to correlate with vaginal wash SIV-specific IgG levels (p=0.06, p=0.05; r=0.6 for both). Overall, our vaccine regimen elicited GC T and B cell responses that supported SIV-specific humoral immunity in the FGT suggesting further development of replicating mucosal Ad-vaccines is warranted for females as well as males.
Weaning and postnatal age influence the early time course and nature of intestinal mast cell activation and mucosal inflammation in a porcine model of early life adversity
Authors: Wilson Neco , Li Yihang , Rajput Mrigendra , Thelen Kyan , Kerr Katie , Moeser Adam J. 
Published: January 2023

Abstract
Early life adversity (ELA) is a risk factor for later life emergence of functional and inflammatory GI disorders in people and animals. In this study, we compared the early GI immune responses in male piglets exposed to early weaning (EW) at 16–18 d of age, a form of ELA we’ve previously shown result in altered GI developmental and health trajectories similar to ELA in humans, with that of late weaned piglets (LW: 28 d of wean age). RNA transcriptome analysis of ileal mucosa at 24h post-weaning revealed that EW pigs exhibited a greater number of differentially expressed genes (765 vs 110 genes, for EW and LW piglets, respectively), characterized by a large number of upregulated genes associated with immune cell trafficking and inflammation. Mast cells (MC), stress-sensitive innate immune cells which orchestrate the immune response, were acutely activated in EW and LW pigs but in a differential manner. Compared with LW piglets, EW piglets exhibited higher levels of serum histamine which coincided with a downregulation in the gene expression of histamine degrading enzymes, DAO and HNMT in intestinal mucosa. Mast cell tryptase (MCT7) and chymase (CMA1) gene expression were upregulated in ileal and colonic mucosa (within 3h post-weaning) in both EW and LW piglets; however, this response was greater in LW pigs. Together, these data show that weaning stress in piglets induces rapid GI MC activation which precedes immune cell recruitment and intestinal inflammation and that postnatal age has a significant influence on the level and nature of this response. Further investigation of how EW and LW piglets differentially regulate early GI immune responses is expected to provide new insight into the mechanisms of GI and immune disorders associated with ELA.
Characterization of bovine γδ T cells phenotype following Mycobacterium bovis vaccination or virulent infection
Authors: Guerra-Maupome Mariana , Palmer Mitchell V , Waters Ray , McGill Jodi L. 
Published: January 2023

Abstract
Bovine tuberculosis caused by Mycobacterium bovis is a significant veterinary health problem. γδ T cells participate in the control of mycobacterial infections. Accumulating data suggest that mycobacterial infection regulates memory/effector phenotype and immune functions of mycobacterium-responsive γδ T cells. To date, the impact of both virulent M. bovis infection and Bacille Calmette-Guerin (BCG) vaccination on bovine γδ T cells memory/effector phenotype remains unknown. In this study, we addressed the functional and phenotypical differences in memory/activation marker, and chemokine receptor expression of M. bovis-specific γδ T cells following BCG vaccination or virulent M. bovis infection. Analysis of γδ T cells in blood, bronchoalveolar lavage fluid, and regional lymph nodes indicated that γδ T cells have phenotypic differences based on the expression of CD27 that suggest distinct functional properties. Here, we show that most peripheral γδ T cells display a CD27+ phenotype. Antigen recall responses showed that expression of CD27 correlates with the expansion of M. bovis-specific γδ T-cells. Thus, the CD27+ subset may comprise the adaptive γδ T cell compartment responding to virulent M. bovis infection and vaccination. Furthermore, we show that γδ T cells from neonatal calves are activated by aerosol BCG vaccination and adopt a lung-homing phenotype that is similar to that observed during virulent M. bovis infection, suggesting an important role for this T cell subset in the response to mucosal vaccination. The ability of γδ T cells to mount a robust local immune response supports the hypothesis that vaccine-elicited γδ T cell immunity might prove beneficial.
Disease tolerance and immune response in bovine trypanosomiasis is mediated by CD14 promoter gene
Authors: Morenikeji Olanrewaju , Thomas Bolaji 
Published: January 2023

Abstract
Bovine trypanosomiasis is a major drawback for cattle production in sub-Saharan Africa. Immune response to infection is thought to be mediated by genetic diversity in pattern recognition receptors, leading to disease tolerance or susceptibility. We elucidated naturally occurring variations within the bovine CD14 gene promoter in trypanotolerant (N’dama) and trypanosusceptible (White Fulani) cattle, using genomic and transcriptomic methods. We compared our gene sequences with reference bovine genome UMD 3.1 assembly from the Ensembl database. In all, we identified a total of 54 and 41 SNPs at the CD14 promoter for Ndama and White Fulani respectively, including 10 (rs210109064, rs473185091, rs521360374, rs454858160, rs432371570, rs446947915, rs442402639 and rs456854916) common to both. Interestingly, we found 36 SNP’s in 9 SNP regions that were unique to Ndama that were either functional or transcript overlap variants. The significantly higher SNP variation and density at the CD14 promoter region in Ndama could be responsible for its trypanotolerance, potentially an evolutionary adaptation to disease. To further validate this, we examined the expression of CD14 in tissue samples, with our results showing a significant upregulation and higher expression in Ndama compared to White Fulani (p&lt;0.05). These results confirm that trypanotolerant Ndama significantly expressed CD14 and the presence of a higher SNP variation at the CD14 gene promoter plays an important role in pathogen recognition and initiation of immune response, which is absent in White Fulani (trypanosusceptible). This is a major player in host defense mechanism against trypanosomiasis that could be harnessed for breeding and selection purposes.
Transcriptome analysis of distinct coelomocyte populations in Strongylocentrotus purpuratus
Authors: Fugmann Sebastian D , Cheng Jun Fang , Huang Hsiao Han 
Published: January 2023

Abstract
Coelomocytes are important cellular mediators of immunity in echinoderms. In the purple sea urchin (Strongylocentrotus purpuratus) these cells can readily be divided into phagocytes, vibratile cells, red spherule cells, and colorless spherule cells based on their morphology. While some information has been reported about genes that are expressed in specific types of coelomocytes, a comprehensive analysis of the gene expression pattern in each cell type was not available. Using a density gradient coelomocytes from individual adult urchins were separated into three populations (phagocytes, a mixture of vibratile and colorless spherule cells, and red spherule cells), RNA was isolated, and next generation sequencing was employed to determine the transcriptome of each cell population. All three cell populations showed clear differences in their gene expression patterns, and the differences in transcript levels for selected genes that were found to be uniquely expressed in one cell type were confirmed by qRT-PCR using cDNA obtained from additional sea urchins. This bulk population data was then complemented by single-cell RNAseq to obtain information about suitable population and sub-population markers for subsequent studies of coelomocyte biology.
Annotation of the WC1 gene family in Sus scrofa and evaluation of individual SRCR domain affinity for Mycobacterium bovis and Leptospira spp
Authors: Le Page Lauren Ann , Buck Jessica , Boisvert Natalie , Gillespie Alexandria , Yirsaw Alehegne , Hudgens Edward , Hsu Haoting , Baldwin Cynthia L , Telfer Janice C 
Published: January 2023

Abstract
γδ T cells respond to a variety of proteins and non-proteinaceous molecules independent of MHC presentation, making them less susceptible to immune evasion and thus; an attractive target for next-generation vaccines. Unfortunately, definitive methods to prime these cells remain elusive. WC1, a member of the group B Scavenger Receptor Cysteine-Rich (SRCR) superfamily, is expressed exclusively on the γδ T cells of swine and ruminants and functions as hybrid co-receptor and pattern recognition receptor for the γδ TCR. It is known that there are thirteen unique WC1 genes expressed in cattle, each containing up to eleven extracellular SRCR domains. Bovine WC1+ γδ T cells share restriction in their TCR gene usage, yet respond to different pathogens based on which WC1 molecule(s) they express. This can be attributed to the ability of the expressed WC1 molecule to directly bind whole pathogens via its SRCR domains. Because WC1 is expressed as a multigene array, we hypothesize that each WC1 gene has co-evolved with a different set of pathogens, and may be exploited to generate a protective immune response from WC1+ γδ T-cells. We sought to identify and characterize WC1 genes expressed in swine, and evaluate their pathogen binding potential. We have obtained cDNA evidence for ten swine WC1 genes, containing up to six extracellular SRCR domains. Through annotation with MAKER, we have confirmed the presence of seven of these genes in the current assembly, Sus scrofa 11.1, and identified four unique exon-intron gene structures. We have shown that SRCR domains from multiple WC1 genes differentially bind Leptospira spp, and Mycobacterium bovis BCG Danish and Pasteur strains.
Isolation and characterization of acute phase protein C-reactive protein in shark serum: a potential biomarker for assessing health in wild sharks
Authors: Orndorff Samantha Mae , Merly Liza 
Published: January 2023

Abstract
The acute phase protein, C-reactive protein (CRP), has become an established clinical biomarker in both animal and human health, where it is used to assess the degree of acute and/or chronic inflammation. CRP has also been proposed as a potential biomarker for measuring health in sharks by indicating when a shark is experiencing an inflammatory state or is under a chronic state of stress that may have significant physiological impact. Despite the promise in using increased levels of CRP as well as other acute phase proteins as indicators of compromised health, there has not to date been a developed shark-specific assay to accurately measure CRP. An initial step in the development of a specific bioassay for CRP is isolation and characterization of the protein. The purpose of the present study is to use both field and laboratory techniques to obtain serum samples and isolate CRP from nurse sharks, Ginglymostoma cirratum. A secondary aim is to characterize the molecule and produce shark CRP specific antisera. A two step isolation procedure is now being refined to positively identify the pentraxins present and characterize them further. Protein affinity chromatography was used to identify serum amyloid P component (SAP) in addition to CRP given that they are both pentraxin proteins displaying similar binding properties. Isolated protein bands resulting from concentrated chromatographic peaks were analyzed by LC-MS/MS. Results thus far include the isolation of a pentraxin domain-containing protein within shark serum.
Inflammatory changes and melanization in the Atlantic salmon (Salmo salar) targeted by In situ hybridization
Authors: Bjoergen Haavard , Hordvik Ivar , Koppang Erling Olaf 
Published: January 2023

Abstract
Research on the immune system of salmonid fish is attracting increased attention, however, progress within this field has been hampered by the general lack of cell markers targeting different immune molecules. Here, we circumvent these challenges by applying in situ hybridization (ISH) to reveal basic cellular responses in salmon immune reactions.
Using RNAscope® (ACD, Newark, CA), we have established ISH targeting immunoglobulin (Ig) M and T RNA. Further, we also target other immune genes where we previously had no markers, and tyrosinase, thought to be important for the function of the melano-macrophages which are special, melanin-producing immune cells found in ectothermic vertebrates. We provided target gene sequences to the manufacturer, which supplied us with probes, and we performed ISH according to the manufacturer’s instructions. Different inflammatory conditions in farmed salmon, including cancerous changes from the gut and muscle tissue with chronic myositis and melanization due to the presence of melano-macrophages, were screened by standard histology and subsequently subjected to ISH.
For the IgM results, we compared IHC and ISH results, as we have antisera targeting IgM. The results were highly similar. ISH thus seems as a good method that may be used in salmon when applicable and especially to target transcripts where no antibody recognizing the resulting protein is available. Further, we reveal hitherto non-described cellular compositions of inflammatory changes in salmon using probes detecting several different immune cell transcripts. Our results show that ISH using RNAscope® works in Atlantic salmon and is highly reproducible.
Evaluation of epitope specificity in the immune response to PR8 H1N1 influenza virus infection in neonatal and adult African Green monkeys
Authors: Clemens Elene A , Angeletti Davide , Holbrook Beth C , Aycock Scott Tyler , Yewdell Jonathan W. , Alexander-Miller Martha A 
Published: January 2023

Abstract
Generation of effective antibody responses following virus infection is challenging in neonates as a result of impairments in the innate and adaptive immune systems that characterize early life. This leaves them particularly susceptible to severe disease following virus infection, e.g. influenza A virus. A potential contributor to the efficacy of the immune response generated following infection is the epitope specificity of the elicited antibodies. Previous studies in adult mice have reported a defined and reproducible pattern of immunodominance among antibodies directed to the neutralizing epitopes on the head of the influenza hemagglutinin (HA) molecule. The impact of age on the immunodominance pattern of HA-specific antibodies has not been explored. To address this, epitope recognition was quantified in plasma collected from newborn and adult African Green monkeys on d14 following PR8 influenza virus infection. For these analyses we used HA molecules engineered to singly express each of the neutralizing epitopes identified on the HA head. In addition, we used a construct that restricts recognition to the stem region of PR8 HA. Our analyses revealed a similar pattern of recognition by HA-specific IgG between infants and adults in the 14 day period following infection. In contrast, the HA-specific IgM pools exhibited distinct binding patterns in these two groups. The most striking of these differences was in the production of antibodies capable of recognizing the stem portion of HA. These data suggest antibody immunodominance patterns may be modulated with age and sheds new light on the regulation of potentially broadly protective stem responses to influenza virus.
The immune responses to influenza vaccine in aged and metabolic disease occurred cynomolgus macaques
Authors: Urano Emiko , Murakata Yoshiko , Okamura Tomotaka , Yasutomi Yasuhiro 
Published: January 2023

Abstract
The analysis of age-related immune senescent is extremely meaningful to understand the vital homeostasis in individuals among the aging society. Influenza virus (FluV) infection is a major public health concern due to its high infectivity and ability to severe respiratory illness in elderly people. The reduced immune responses after Flu vaccination in humans are recognized in aging and also in lifestyle diseases such as diabetes mellitus (DM). Understanding the hypofunction in aging and lifestyle diseases is necessary to optimize vaccination resume and the comprehensive analysis of that will provide a great impact on public health burden. However, several factors affect the biofunctional analysis in human.
Cynomolgus macaques (CMs) show human-like characteristics such as higher brain functions, long life span, single pregnancy. In the present study, several groups of CMs, including DM and hyperlipidemia (HL) occurred CMs, were subcutaneously immunized with seasonal trivalent Flu vaccine and assessed immune responses. The reduced humoral and cellular immune responses to Flu vaccine were observed in the elderly CMs group compare to the young group and induced anti-HA antibody titers were negatively correlated with age. In particular, Flu-specific immune responses were considerably decreased in DM group despite their youth. However, the reduced humoral and cellular immune responses were not observed in HL group, unlike DM CMs. These results suggested that elderly or DM and HL occurred CMs should be a powerful tool to reveal the comprehensive mechanisms for hypofunction in aging and lifestyle diseases through a long-life span. This study will help to enhance the immune response to vaccine in metabolic disease and aged person.
Mechanism of Influenza Virus Induced Inflammatory Cell Death
Authors: Sannula Kesavardana , Kanneganti Thirumala-Devi 
Published: January 2023

Abstract
RNA viruses and host cell death responses are co-evolved which prompt the fate of immune responses. Inflammatory cell death is a form of programmed signaling cascade which lead to the loss of cell integrity and secretion of leaderless pro-inflammatory cytokines. Host innate immune system exerts inflammatory cell death to curtail virus replication and initiate protective inflammatory and adaptive responses. This also causes host tissue damage if uncontrolled. Elevated inflammatory responses and lung damage are primary causes for morbidity and mortality during pathogenic influenza infections. However, little is known about key mechanisms regulating influenza A virus (IAV) induced cell death and inflammation.
We identified Z-DNA Binding Protein 1 (ZBP1) as a sensor of IAV infection to trigger inflammatory cell death in in vitro and in vivo. ZBP1 induced inflammatory cell death is specific to nuclear replicating IAV but not the other RNA viruses those replicate in the cytosol. We found that the ZBP1 translocate to the nucleus prior to the cell death execution. Unlike other innate receptors, ZBP1 recognizes unique patterns of RNA in ribonucleoprotein complexes (vRNPs) of IAV. This nucleate RHIM domain driven oligomeric RIPK3 signaling scaffolds to facilitate RIPK3-Caspase-8 dependent inflammatory cell death (both necroptosis and pyroptosis). Our biochemical studies identified RIG-I dependent activation, ubiquitination of ZBP1 and loss of mitochondrial integrity precedes inflammatory cell death which facilitate the assembly of cell death signaling complexes proximal to the cellular membrane. Overall, our work demonstrates the mechanism of sensing of nuclear replicating IAV vRNPs to trigger programmed inflammatory cell death.
Fibroblasts from individuals with Trisomy 21 have increased Type-I Interferon responses to HSV-1 compared to typical adults
Authors: Royzman Kristina , Drews Caitlyn , Maxwell Tyler , Singh Gurkirat , Keefe Caitlin , James Stephanie 
Published: January 2023

Abstract
Down syndrome (Trisomy 21; T21) results from an extra copy of chromosome 21. This results in a myriad of abnormalities, including immune dysfunction. Previous research has focused only on children with T21. However, improved medical interventions and treatments have led to a rapid increase in the lifespan of people with T21. Thus, there is now a population of adults with T21 and a critical gap in our understanding of immunity in these adults. It is well known that adults with T21 are far more likely to die from respiratory infections compared to their disomic counterparts. This is despite recent published reports that individuals with T21 have an increased interferon response, and the Interferon-α receptor is located on q21. Our objective was to determine if a.) IFNα stimulated fibroblasts with trisomy 21 had greater interferon stimulated gene (ISG) and protein expression compared to disomic fibroblasts; and b.) If virally infected fibroblast cells with trisomy 21 had an increased type I IFN response after viral infection compared to disomic fibroblasts. Cells were treated with IFNα or virally infected with Herpes Simplex Virus-1 (HSV) for 6 or 24 hours. Gene expression and viral replication was measured by qPCR and protein expression measured by Western blot. Our results indicate that fibroblast cells with trisomy 21 have a much greater expression of the ISG15 and Mx1 proteins compared to disomic fibroblasts when stimulated with IFNα. During HSV infection, fibroblasts with trisomy 21 had a significantly greater gene expression of ISGs compared to disomic fibroblasts although viral replication rates were similar between the two groups. This data suggests that individuals with T21 have a greater type I IFN response compared to typical adults.