Sialic Acid-Siglec-E Interactions Regulate the Response of Neonatal Macrophages to Group B Streptococcus.

The mammalian Siglec receptor sialoadhesin (Siglec1, CD169) confers innate immunity against the encapsulated pathogen group B Streptococcus (GBS). Newborn lung macrophages have lower expression levels of sialoadhesin at birth compared with the postnatal period, increasing their susceptibility to GBS infection. In this study, we investigate the mechanisms regulating sialoadhesin expression in the newborn mouse lung. In both neonatal and adult mice, GBS lung infection reduced Siglec1 expression, potentially delaying acquisition of immunity in neonates. Suppression of Siglec1 expression required interactions between sialic acid on the GBS capsule and the inhibitory host receptor Siglec-E. The Siglec1 gene contains multiple STAT binding motifs, which could regulate expression of sialoadhesin downstream of innate immune signals. Although GBS infection reduced STAT1 expression in the lungs of wild-type newborn mice, we observed increased numbers of STAT1+ cells in Siglece-/- lungs. To test if innate immune activation could increase sialoadhesin at birth, we first demonstrated that treatment of neonatal lung macrophages ex vivo with inflammatory activators increased sialoadhesin expression. However, overcoming the low sialoadhesin expression at birth using in vivo prenatal exposures or treatments with inflammatory stimuli were not successful. The suppression of sialoadhesin expression by GBS-Siglec-E engagement may therefore contribute to disease pathogenesis in newborns and represent a challenging but potentially appealing therapeutic opportunity to augment immunity at birth.

Corrigendum: Advances and potential of omics studies for understanding the development of food allergy.

[This corrects the article DOI: 10.3389/falgy.2023.1149008.].

Macrophage Polarizations in the Placenta and Lung are Associated with Bronchopulmonary Dysplasia.

The intricate interplay between macrophage polarization and placenta vascular dysfunction has garnered increasing attention in the context of placental inflammatory diseases. This study delves into the complex relationship between macrophage polarization within the placenta and its potential impact on the development of vascular dysfunction and inflammatory conditions. The placenta, a crucial organ in fetal development, relies on a finely tuned balance of immune responses for proper functioning. Disruptions in this delicate equilibrium can lead to pathological conditions, including inflammatory diseases affecting the fetus and newborn infant. We explored the interconnectedness between placental macrophage polarization and its relevance to lung macrophages, particularly in the context of early life lung development. Bronchopulmonary dysplasia (BPD), the most common chronic lung disease of prematurity, has been associated with abnormal immune responses, and understanding the role of macrophages in this context is pivotal. The investigation aims to shed light on how alterations in placental macrophage polarization may contribute to lung macrophage behavior and, consequently, influence the development of BPD. By unraveling the intricate mechanisms linking macrophage polarization, placental dysfunction and BPD, this research seeks to provide insights that could pave the way for targeted therapeutic interventions. The findings may offer novel perspectives on preventing and managing placental and lung-related pathologies, ultimately contributing to improved maternal and neonatal health outcomes.

Advances and potential of omics studies for understanding the development of food allergy.

The prevalence of food allergy continues to rise globally, carrying with it substantial safety, economic, and emotional burdens. Although preventative strategies do exist, the heterogeneity of allergy trajectories and clinical phenotypes has made it difficult to identify patients who would benefit from these strategies. Therefore, further studies investigating the molecular mechanisms that differentiate these trajectories are needed. Large-scale omics studies have identified key insights into the molecular mechanisms for many different diseases, however the application of these technologies to uncover the drivers of food allergy development is in its infancy. Here we review the use of omics approaches in food allergy and highlight key gaps in knowledge for applying these technologies for the characterization of food allergy development.

Sp3 is essential for normal lung morphogenesis and cell cycle progression during mouse embryonic development.

Members of the Sp family of transcription factors regulate gene expression via binding GC boxes within promoter regions. Unlike Sp1, which stimulates transcription, the closely related Sp3 can either repress or activate gene expression and is required for perinatal survival in mice. Here, we use RNA-seq and cellular phenotyping to show how Sp3 regulates murine fetal cell differentiation and proliferation. Homozygous Sp3-/- mice were smaller than wild-type and Sp+/- littermates, died soon after birth and had abnormal lung morphogenesis. RNA-seq of Sp3-/- fetal lung mesenchymal cells identified alterations in extracellular matrix production, developmental signaling pathways and myofibroblast/lipofibroblast differentiation. The lungs of Sp3-/- mice contained multiple structural defects, with abnormal endothelial cell morphology, lack of elastic fiber formation, and accumulation of lipid droplets within mesenchymal lipofibroblasts. Sp3-/- cells and mice also displayed cell cycle arrest, with accumulation in G0/G1 and reduced expression of numerous cell cycle regulators including Ccne1. These data detail the global impact of Sp3 on in vivo mouse gene expression and development.

Data-driven longitudinal characterization of neonatal health and morbidity.

Although prematurity is the single largest cause of death in children under 5 years of age, the current definition of prematurity, based on gestational age, lacks the precision needed for guiding care decisions. Here, we propose a longitudinal risk assessment for adverse neonatal outcomes in newborns based on a deep learning model that uses electronic health records (EHRs) to predict a wide range of outcomes over a period starting shortly before conception and ending months after birth. By linking the EHRs of the Lucile Packard Children's Hospital and the Stanford Healthcare Adult Hospital, we developed a cohort of 22,104 mother-newborn dyads delivered between 2014 and 2018. Maternal and newborn EHRs were extracted and used to train a multi-input multitask deep learning model, featuring a long short-term memory neural network, to predict 24 different neonatal outcomes. An additional cohort of 10,250 mother-newborn dyads delivered at the same Stanford Hospitals from 2019 to September 2020 was used to validate the model. Areas under the receiver operating characteristic curve at delivery exceeded 0.9 for 10 of the 24 neonatal outcomes considered and were between 0.8 and 0.9 for 7 additional outcomes. Moreover, comprehensive association analysis identified multiple known associations between various maternal and neonatal features and specific neonatal outcomes. This study used linked EHRs from more than 30,000 mother-newborn dyads and would serve as a resource for the investigation and prediction of neonatal outcomes. An interactive website is available for independent investigators to leverage this unique dataset: https://maternal-child-health-associations.shinyapps.io/shiny_app/.

Carcass Characteristics, Meat Quality and Nutritional Composition of Kadaknath, a Native Chicken Breed of India.

The study was carried out to investigate the carcass and meat quality traits and nutritional profile of the meat of the Kadaknath, a unique native chicken breed in comparison with commercial broilers. The yield of the carcass, breast and giblets of the Kadaknath was lesser (p < 0.01), while that of the legs, wings, back, and neck was higher (p < 0.01) than broilers. The meat of the Kadaknath was significantly (p < 0.0001) darker (42.44, 50.92) and more yellow (6.23, 8.99) than broilers. The decline in pH of the meat was lower (p < 0.001) in the Kadaknath compared to broilers. Kadaknath meat had more protein and less fat, moisture and ash content than broilers (p < 0.01). Furthermore, it was richer (p < 0.01) in 11 amino acids, including those which are known to impart a sweet and umami taste, than the meat of broilers (3 amino acids). Both genotypes were almost similar in meeting the daily requirements of indispensable amino acids of adult human. The study concluded that the Kadaknath differed in carcass and meat quality characteristics from the broilers, and the nutritional quality of Kadaknath meat in terms of high protein and less fat and higher content of amino acids (tasty type) was better in Kadaknath meat as compared to broiler meat.

Progressive Metabolic Abnormalities Associated with the Development of Neonatal Bronchopulmonary Dysplasia.

Objective: To assess the longitudinal metabolic patterns during the evolution of bronchopulmonary dysplasia (BPD) development. Methods: A case-control dataset of preterm infants (<32-week gestation) was obtained from a multicenter database, including 355 BPD cases and 395 controls. A total of 72 amino acid (AA) and acylcarnitine (AC) variables, along with infants' calorie intake and growth outcomes, were measured on day of life 1, 7, 28, and 42. Logistic regression, clustering methods, and random forest statistical modeling were utilized to identify metabolic variables significantly associated with BPD development and to investigate their longitudinal patterns that are associated with BPD development. Results: A panel of 27 metabolic variables were observed to be longitudinally associated with BPD development. The involved metabolites increased from 1 predominant different AC by day 7 to 19 associated AA and AC compounds by day 28 and 16 metabolic features by day 42. Citrulline, alanine, glutamate, tyrosine, propionylcarnitine, free carnitine, acetylcarnitine, hydroxybutyrylcarnitine, and most median-chain ACs (C5:C10) were the most associated metabolites down-regulated in BPD babies over the early days of life, whereas phenylalanine, methionine, and hydroxypalmitoylcarnitine were observed to be up-regulated in BPD babies. Most calorie intake and growth outcomes revealed similar longitudinal patterns between BPD cases and controls over the first 6 weeks of life, after gestational adjustment. When combining with birth weight, the derived metabolic-based discriminative model observed some differences between those with and without BPD development, with c-statistics of 0.869 and 0.841 at day 7 and 28 of life on the test data. Conclusions: The metabolic panel we describe identified some metabolic differences in the blood associated with BPD pathogenesis. Further work is needed to determine whether these compounds could facilitate the monitoring and/or investigation of early-life metabolic status in the lung and other tissues for the prevention and management of BPD.

The Lung Microenvironment Instructs Gene Transcription in Neonatal and Adult Alveolar Macrophages.

Immaturity of alveolar macrophages (AMs) around birth contributes to the susceptibility of newborns to lung disease. However, the molecular features differentiating neonatal and mature, adult AMs are poorly understood. In this study, we identify the unique transcriptomes and enhancer landscapes of neonatal and adult AMs in mice. Although the core AM signature was similar, murine adult AMs expressed higher levels of genes involved in lipid metabolism, whereas neonatal AMs expressed a largely proinflammatory gene profile. Open enhancer regions identified by an assay for transposase-accessible chromatin followed by high-throughput sequencing (ATAC-seq) contained motifs for nuclear receptors, MITF, and STAT in adult AMs and AP-1 and NF-κB in neonatal AMs. Intranasal LPS activated a similar innate immune response in both neonatal and adult mice, with higher basal expression of inflammatory genes in neonates. The lung microenvironment drove many of the distinguishing gene expression and open chromatin characteristics of neonatal and adult AMs. Neonatal mouse AMs retained high expression of some proinflammatory genes, suggesting that the differences in neonatal AMs result from both inherent cell properties and environmental influences.

Antenatal Mesenchymal Stromal Cell Extracellular Vesicle Therapy Prevents Preeclamptic Lung Injury in Mice.

In preeclamptic pregnancies, a variety of intrauterine alterations lead to abnormal placentation, release of inflammatory and/or antiangiogenic factors, and subsequent fetal growth restriction with significant potential to cause a primary insult to the developing fetal lung. Thus, modulation of the maternal intrauterine environment may be a key therapeutic avenue to prevent preeclampsia-associated developmental lung injury. A biologic therapy of interest is mesenchymal stromal cell-derived extracellular vesicles (MEx), which we have previously shown to ameliorate preeclamptic physiology through intrauterine immunomodulation. To evaluate the therapeutic potential of MEx to improve developmental lung injury in experimental preeclampsia, using the heme oxygenase-1-null mouse (Hmox1-/-) model, preeclamptic pregnant dams were administered intravenous antenatal MEx treatment during each week of pregnancy followed by analysis of fetal and postnatal lung tissues, amniotic fluid protein profiles, and lung explant and amniotic fluid cocultures in comparison with control and untreated preeclamptic pregnancies. We first identified that a preeclamptic intrauterine environment had a significant adverse impact on fetal lung development, including alterations in fetal lung developmental gene profiles in addition to postnatal alveolar and bronchial changes. Amniotic fluid proteomic analysis and fetal lung explant and amniotic fluid cocultures further demonstrated that maternally administered MEx altered the expression of multiple inflammatory mediators in the preeclamptic intrauterine compartment, resulting in the normalization of fetal lung branching morphogenesis and developmental gene expression. Our evaluation of fetal and postnatal parameters overall suggests that antenatal MEx treatment may provide a highly valuable preventative therapeutic modality for amelioration of lung development in preeclamptic disease.

Effect on physiological and production parameters upon supplementation of fermented yeast culture to Nicobari chickens during and post summer.

Nicobari is an indigenous bird reared for meat and eggs. This study evaluated the effect of heat stress on plasma levels of leptin, growth hormone and their receptors, liver AMP kinase, plasma cholesterol and lipid peroxide (MDA). The laying period coincided with the post summer period. The birds were equally divided into three groups, control group was offered ad libitum feed and treatment groups were supplemented with fermented yeast culture at 700 mg (T1) and 1.4 g/kg (T2) of feed/day. The levels of plasma Leptin and GH hormones were higher (p < 0.05) in the control group when compared to the treatment groups. The expression of the hormone receptors was higher in the brain, and MMP3 gene expression in the magnum was lower in the treatment group. Plasma cholesterol, MDA and AMP kinase were significantly higher (p < 0.05) in the control group. Fermented yeast culture supplementation decreased feed intake and increased egg production parameters, which indicates a greater efficiency of supplementation. Supplementation reduced the severity of necrosis of villi in the jejunum when compared to control. In conclusion, higher ambient temperature during summer had negative effect on production parameters through modulation of physiological parameters which could be ameliorated by supplementation of FYC.

Effect of Supplementation of Fermented Yeast Culture on Hormones and Their Receptors on Exposure to Higher Temperature and on Production Performance after Exposure in Nicobari Chickens.

Heat stress (HS) affects the production performance in chickens and causes economic loss to the producers. Most of the studies have been conducted on and for the welfare of broilers. We still lack information on the physiological parameters being affected during chronic heat stress in layers. To fill this gap, the present study evaluated the effect of heat stress (induced in the chamber) during the prelaying period (21-23 weeks) on plasma levels of the hormones leptin and ghrelin and GH and expression of the respective receptors and heat stress markers. Three groups were considered, one at room temperature (CR) and the other two groups (SH and CH) subjected to heat stress at 39°C for four hours for three weeks (21-23 weeks of age). The SH group (SH) feed was supplemented with fermented yeast culture (FYC, 700 mg/kg), whereas the CH group was devoid of it. After that, all the groups were shifted to shed under natural ambient conditions till 31 weeks of age. Studies were restricted to production performance only. Feed offered without yeast culture (CH group) had a smaller concentration of plasma hormones (P < 0.01) and increased expression fold of the hormone receptors (P < 0.01). Further, the group also presented higher liver AMP kinase enzyme, plasma MDA (malondialdehyde), and cholesterol concentrations. These changes likely explained the decrease in feed intake and the CH group's body weight and further reduced the production performance during the laying period. Supplementation with FYC to birds had an opposite effect on the above-mentioned parameters, reducing HS effects. In summary, supplementation with FYC (700 mg/kg) maintained physiological parameters as in the CR group under HS conditions and negated adverse effects on parameters both before and during laying periods.

Estimation of breeding value, genetic parameters and maternal effects of economic traits in rural male parent line chicken using pedigree relationships in an animal model.

Breeding value (BV), genetic parameters and additive genetic, and maternal effects were evaluated on growth and production traits utilizing data from eight generations employing animal model in a rural male parent line (PD-6) chicken at ICAR-Directorate of Poultry Research, Hyderabad, India. The least squares means (LSM) for body weight (BW) and shank length (SL) up to 6 weeks of age varied significantly (p ≤ .01) among the generations and hatches. BW increased significantly (p ≤ .01) over the generations and decreased with the hatches. Sex also had a significant effect on BW and shank length except for BW at 0 day (BW0). LSM for BW (BW6) and Shank length (SL6) at 6 weeks of age were 598.84 ± 0.79 g and 74.57 ± 0.04 mm, respectively. Males recorded significantly (p ≤ .01) higher BWs and shank length. All the production traits were significantly (p ≤ .01) influenced by the generation effect. The overall LSM for age at sexual maturity (ASM), egg production at 40 weeks (EP40) and egg weight at 40 weeks (EW40) were 164.93 ± 0.23 days, 74.66 ± 0.40 eggs and 54.79 ± 0.08 g, respectively. Model 3 with additive, maternal permanent environmental and residual effects was the appropriate model for BW2, BW4, BW6, SL4 and SL6, whereas Model 4 with maternal effects was the best for BW0. The heritability estimates for BW6 and SL6 were 0.22 ± 0.02 and 0.18 ± 0.02, respectively. Model 1 with additive direct and residual effects was the best appropriate model for all the production traits. The heritability estimates of EP40 and EW40 were 0.16 ± 0.04 and 0.34 ± 0.05, respectively. BW and shank length were highly correlated with significant (p ≤ .05) positive association from different components. The correlation coefficient from direct additive component between egg production and BW40 was negative, while it was positive with less magnitude between egg production and BW20. The egg production and egg weights had a negative association at different ages. BV of SL6, the primary trait of selection, was significant (p ≤ .05) across the generations and increased linearly with an average genetic gain of 1.05 mm per generation. BV of BW6 was also significant (p ≤ .05) and increased linearly as correlated response with an average genetic response of 22.34 g per generation. BV of EP40 showed an increasing trend with a genetic gain of 0.02 eggs per generation. The EW 40 also increased linearly with an average genetic gain of 0.06 g. The average inbreeding coefficient of the population was 0.015. The study concluded that the population was in ideal status with a linearly increasing trend of average BV with negligible inbreeding over the eight generations of selection.

Developmental Immaturity of Siglec Receptor Expression on Neonatal Alveolar Macrophages Predisposes to Severe Group B Streptococcal Infection.

Streptococcus agalactiae (Group B Streptococcus, GBS) is the most common neonatal pathogen. However, the cellular and molecular mechanisms for neonatal susceptibility to GBS pneumonia and sepsis are incompletely understood. Here we optimized a mouse model of GBS pneumonia to test the role of alveolar macrophage (ΑΜΦ) maturation in host vulnerability to disease. Compared with juvenile and adult mice, neonatal mice infected with GBS had increased mortality and persistence of lung injury. In addition, neonatal mice were defective in GBS phagocytosis and killing. ΑΜΦ depletion and disruption of ΑΜΦ differentiation in Csf2-/- mice both impaired GBS clearance. AMΦ engage the heavily sialylated GBS capsule via the cell surface Siglec receptors Sn and Siglec-E. Although both newborn and adult ΑΜΦ expressed Siglec-E, newborn ΑΜΦ expressed significantly lower levels of Sn. We propose that a developmental delay in Sn expression on ΑΜΦ may prevent effective killing and clearing of GBS from the newborn lung.

Transcriptional profiling of lung macrophages identifies a predictive signature for inflammatory lung disease in preterm infants.

Lung macrophages mature after birth, placing newborn infants, particularly those born preterm, within a unique window of susceptibility to disease. We hypothesized that in preterm infants, lung macrophage immaturity contributes to the development of bronchopulmonary dysplasia (BPD), the most common serious complication of prematurity. By measuring changes in lung macrophage gene expression in preterm patients at risk of BPD, we show here that patients eventually developing BPD had higher inflammatory mediator expression even on the first day of life. Surprisingly, the ex vivo response to LPS was similar across all samples. Our analysis did however uncover macrophage signature genes whose expression increased in the first week of life specifically in patients resilient to disease. We propose that these changes describe the dynamics of human lung macrophage differentiation. Our study therefore provides new mechanistic insight into both neonatal lung disease and human developmental immunology.

Computational Approach to Identifying Universal Macrophage Biomarkers.

Macrophages engulf and digest microbes, cellular debris, and various disease-associated cells throughout the body. Understanding the dynamics of macrophage gene expression is crucial for studying human diseases. As both bulk RNAseq and single cell RNAseq datasets become more numerous and complex, identifying a universal and reliable marker of macrophage cell becomes paramount. Traditional approaches have relied upon tissue specific expression patterns. To identify universal biomarkers of macrophage, we used a previously published computational approach called BECC (Boolean Equivalent Correlated Clusters) that was originally used to identify conserved cell cycle genes. We performed BECC analysis using the known macrophage marker CD14 as a seed gene. The main idea behind BECC is that it uses massive database of public gene expression dataset to establish robust co-expression patterns identified using a combination of correlation, linear regression and Boolean equivalences. Our analysis identified and validated FCER1G and TYROBP as novel universal biomarkers for macrophages in human and mouse tissues.

Transcriptomic and epigenetic mechanisms underlying myeloid diversity in the lung.

The lung is inhabited by resident alveolar and interstitial macrophages as well as monocytic cells that survey lung tissues. Each cell type plays distinct functional roles under homeostatic and inflammatory conditions, but mechanisms establishing their molecular identities and functional potential remain poorly understood. In the present study, systematic evaluation of transcriptomes and open chromatin of alveolar macrophages (AMs), interstitial macrophages (IMs) and lung monocytes from two mouse strains enabled inference of common and cell-specific transcriptional regulators. We provide evidence that these factors drive selection of regulatory landscapes that specify distinct phenotypes of AMs and IMs and entrain qualitatively different responses to toll-like receptor 4 signaling in vivo. These studies reveal a striking divergence in a fundamental innate immune response pathway in AMs and establish a framework for further understanding macrophage diversity in the lung.

Differential Immune Activation in Fetal Macrophage Populations.

Distinct macrophage subsets populate the developing embryo and fetus in distinct waves. However little is known about the functional differences between in utero macrophage populations or how they might contribute to fetal and neonatal immunity. Here we tested the innate immune response of mouse macrophages derived from the embryonic yolk sac and from fetal liver. When isolated from liver or lung, CD11bHI fetal liver derived macrophages responded to the TLR4 agonist LPS by expressing and releasing inflammatory cytokines. However F4/80HI macrophages from the yolk sac did not respond to LPS treatment. While differences in TLR4 expression did not appear to explain these data, F4/80HI macrophages had much lower NLRP3 inflammasome expression compared to CD11bHI macrophages. Gene expression profiling also demonstrated LPS-induced expression of inflammatory genes in CD11bHI macrophages, but not in F4/80HI cells. Genes expressed in LPS-treated CD11bHI macrophages were more likely to contain predicted NF-κB binding sites in their promoter regions. Our data show that CD11bHI macrophages derived from fetal liver are the major pro-inflammatory cells in the developing fetus. These findings could have important implications in better understanding the fetal inflammatory response and the unique features of neonatal immunity.