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Allergy May 2022Changes in immune cell composition during the immunological window within the first years after birth are not fully understood, especially the effect that different...
BACKGROUND
Changes in immune cell composition during the immunological window within the first years after birth are not fully understood, especially the effect that different lifestyles might have on immune cell functionality.
METHODS
Peripheral blood mononuclear cells from mothers and their children at birth and at two anvd five years were analyzed by mass cytometry. Immune cell composition and functionality was analyzed according to family lifestyle (anthroposophic and non-anthroposophic).
RESULTS
We found no significant differences in the proportions of major immune lineages between anthroposophic and non-anthroposophic children at each time point, but there were clear changes over time in the proportions of mononuclear leukocytes, especially in B-cells and T lymphocytes. Phenotypic distances between cord blood and maternal blood were high at birth but decreased sharply the first two years, indicating strong phenotypic convergence with maternal cells. We found that children exhibited similar stimulation responses at birth, but subsequently segregated into two discrete functional trajectories. Trajectory 1 was associated with a decrease in tumor necrosis factor alpha (TNFa) production by CD4 T- and NK-cells, while Trajectory 2 depicted an increase in the production of IL-2 and interferon gamma (INFg) by T-cells. In both trajectories, there was an increase in IL-17A production by T-cells resulting in prominent differences at five years of age.
CONCLUSIONS
This exploratory study suggests that leukocyte frequencies and cell phenotypes change with age in the same way across all children, while functional development follows one of two discrete trajectories that largely segregate by family lifestyle, supporting the hypothesis that early environmental exposures imprint immune cell function which may contribute to IgE sensitization. Our results also support that the first two years are critical for the environmental exposures to imprint the immune cells. Further studies with larger sample sizes are required to validate our findings.
Topics: Fetal Blood; Humans; Interferon-gamma; Killer Cells, Natural; Leukocytes, Mononuclear; Life Style
PubMed: 35094423
DOI: 10.1111/all.15232 -
Immunobiology May 2020Crotalus neutralising factor (CNF) is an endogenous γ-type phospholipase A (PLA) inhibitor that inhibits the toxic action of crotoxin, a neurotoxin present in Crotalus...
Crotalus neutralising factor (CNF) is an endogenous γ-type phospholipase A (PLA) inhibitor that inhibits the toxic action of crotoxin, a neurotoxin present in Crotalus durissus terrificus venom. However, its effects on the activation and modulation of immune cells, which play a major role in the development of inflammation, is not known. The objective of the present study was to assess the effects of CNF on human leukocyte modulation in vitro by analysing the following parameters: cell viability, phagocytic capacity, lipid droplet formation, reactive oxygen species production, nitric oxide production, p38 MAPK activation, and cytosolic PLA (cPLA) gene expression. Neutrophils and peripheral blood mononuclear cells from healthy donors were isolated via the density gradient method, resuspended in RPMI medium, and incubated with RPMI (negative control), LPS, or PMA (positive control) or CNF (sample test) at a concentration of 50 μg/mL. Results showed that CNF was not toxic to human neutrophils after 48 and 72 h of incubation. CNF treatment induced an increase in PBMCs and neutrophil phagocytic capacity, as well as the formation of lipid droplets within these cells after 1 h of incubation. However, CNF did not induce the formation of reactive oxygen and nitric oxide species. Moreover, CNF induced p38 MAPK protein phosphorylation and cPLA gene expression in neutrophils. The data obtained herein showed that CNF action modulates human leukocytes, CNF activates important signalling pathways for human leukocytes, and it is pro-inflammatory. These findings also complement previous studies on CNF action on human peripheral blood leukocyte function.
Topics: Biomarkers; Glycoproteins; Humans; Immunomodulation; Leukocytes; Leukocytes, Mononuclear; Neutrophils; Oxidative Stress; Phospholipases A2; Reactive Oxygen Species; Reptilian Proteins; p38 Mitogen-Activated Protein Kinases
PubMed: 32183984
DOI: 10.1016/j.imbio.2020.151932 -
Scientific Reports May 2021Early life stress increases one's risk for health problems later in life, and many studies find that these effects are sex-differentiated. Here, we examined... (Observational Study)
Observational Study
Early life stress increases one's risk for health problems later in life, and many studies find that these effects are sex-differentiated. Here, we examined relationships between multiple sources of early life stress and adult immune function in humans across several functional assays. Adult participants provided retrospective information about their childhood (a) socioeconomic status, (b) household unpredictability, and (c) exposure to adverse experiences. Participants' peripheral blood mononuclear cells (PBMCs) were then isolated for use in functional assays of immune performance: (a) tumor cell lysis by natural killer cells, (b) phagocytosis of Escherichia coli bioparticles, and (c) mitogen-induced leukocyte proliferation and cytokine release. In men, lower childhood socioeconomic status predicted decrements in immunological performance across functional assays, along with greater spontaneous cytokine release from PBMCs. These changes co-occurred with elevations in plasma testosterone levels. Similar effects were not observed for other sources of stress, nor were they found in women (with the exception of spontaneous cytokine release). These findings provide evidence that low childhood socioeconomic status has a lasting negative impact on multiple aspects of immune function, particularly in men.
Topics: Adolescent; Adverse Childhood Experiences; Cell Proliferation; Cytokines; Female; Humans; Immunity; Immunoassay; Killer Cells, Natural; Leukocytes, Mononuclear; Male; Sex Factors; Social Class; Young Adult
PubMed: 33972662
DOI: 10.1038/s41598-021-89413-y -
STAR Protocols Dec 2021Deep immune profiling is essential for understanding the human immune system in health and disease. Successful biological interpretation of this data requires consistent...
Deep immune profiling is essential for understanding the human immune system in health and disease. Successful biological interpretation of this data requires consistent laboratory processing with minimal batch-to-batch variation. Here, we detail a robust pipeline for the profiling of human peripheral blood mononuclear cells by both high-dimensional flow cytometry and single-cell RNA-seq. These protocols reduce batch effects, generate reproducible data, and increase throughput. For complete details on the use and execution of this protocol, please refer to Savage et al. (2021).
Topics: Computational Biology; Flow Cytometry; Humans; Leukocytes, Mononuclear; Monitoring, Immunologic; Sequence Analysis, RNA; Single-Cell Analysis
PubMed: 34806044
DOI: 10.1016/j.xpro.2021.100900 -
Journal of Affective Disorders Dec 2014Adverse childhood experiences (ACEs) are associated with poor physical and mental health outcomes in adulthood. Adverse childhood experiences are also associated with...
BACKGROUND
Adverse childhood experiences (ACEs) are associated with poor physical and mental health outcomes in adulthood. Adverse childhood experiences are also associated with shortened leukocyte telomere length (LTL) in adults, suggesting accelerated cell aging. No studies have yet assessed the relationship of ACEs to LTL in individuals with major depressive disorder (MDD), despite the high incidence of antecedent ACEs in individuals with MDD. Further, no studies in any population have assessed the relationship of ACEs to the activity of telomerase, the major enzyme responsible for maintaining LTL, or the relationship between telomerase and LTL in individuals with ACEs.
METHODS
Twenty healthy, unmedicated adults with MDD and 20 healthy age-, sex- and ethnicity-matched controls had ACEs assessed and had blood drawn for LTL and peripheral blood mononuclear cell (PBMC) resting telomerase activity.
RESULTS
In healthy controls, greater ACE exposure was associated with shorter LTL (p<.05) but was unassociated with telomerase activity. In MDD, however, the opposite pattern was seen: greater ACE exposure was unrelated to LTL but was associated with increased telomerase activity (p<.05) and with a higher telomerase:LTL ratio (p=.022).
LIMITATIONS
Study limitations include the small sample size, a single timepoint assessment of telomerase activity, and the use of retrospective self-report to assess ACEs.
CONCLUSIONS
These results replicate prior findings of shortened LTL in healthy adults with histories of multiple ACEs. However, in MDD, this relationship was substantially altered, raising the possibility that activation of telomerase in ACE-exposed individuals with MDD could represent a compensatory response to endangered telomeres.
Topics: Adult; Case-Control Studies; Child; Depressive Disorder, Major; Female; Humans; Leukocytes; Leukocytes, Mononuclear; Life Change Events; Male; Mental Disorders; Retrospective Studies; Telomerase; Telomere; Telomere Homeostasis; Telomere Shortening
PubMed: 25173430
DOI: 10.1016/j.jad.2014.07.035 -
ACS Biomaterials Science & Engineering Aug 2023The manufacturing process of chimeric antigen receptor T cell therapies includes isolation systems that provide pure T cells. Current magnetic-activated cell sorting and...
The manufacturing process of chimeric antigen receptor T cell therapies includes isolation systems that provide pure T cells. Current magnetic-activated cell sorting and immunoaffinity chromatography methods produce desired cells with high purity and yield but require expensive equipment and reagents and involve time-consuming incubation steps. Here, we demonstrate that aptamers can be employed in a continuous-flow resin platform for both depletion of monocytes and selection of CD8 T cells from peripheral blood mononuclear cells at low cost with high purity and throughput. Aptamer-mediated cell selection could potentially enable fully synthetic, traceless isolations of leukocyte subsets from a single isolation system.
Topics: Leukocytes, Mononuclear; CD8-Positive T-Lymphocytes; Leukocytes; Chromatography
PubMed: 37467493
DOI: 10.1021/acsbiomaterials.3c00651 -
Lab on a Chip Mar 2021In molecular and cellular biological research, cell isolation and sorting are required for accurate investigation of cell populations of specific physical or biological...
In molecular and cellular biological research, cell isolation and sorting are required for accurate investigation of cell populations of specific physical or biological characteristics. By employing unique cell properties to distinguish between heterogeneous cell populations, rapid and accurate sorting with high efficiency is possible. Dielectrophoresis-based cell manipulation has significant promise for separation of cells based on their physical properties and is used in diverse areas ranging from cellular diagnostics to therapeutic applications. In this study, we present a microfluidic device that can achieve label-free and size-based cell separation with high size differential resolution from a mono-cellular population or complex sample matrices. It was realized by using the tunnel dielectrophoresis (TDEP) technique to manipulate the spatial position of individual cells three dimensionally with high resolution. Cells were processed in high speed flows in high ionic strength buffers. A mixture of different sizes of polystyrene micro-particles with a size difference as small as 1 μm can be separated with high purity (>90%). For the first time, high-pass, low-pass, and band-pass filtering within a mono-cellular mammalian cell population were demonstrated with a tunable bandwidth as small as 3 μm. In addition, leukocyte subtype separation was demonstrated by sorting monocytes out of peripheral blood mononuclear cells (PBMCs) from whole blood with high purity (>85%). Its ability to deliver real-time adjustable cut-off threshold size-based cell sorting and its capability to provide an arbitrary cell size pick-up band could potentially enable many research and clinical applications.
Topics: Animals; Cell Separation; Lab-On-A-Chip Devices; Leukocytes, Mononuclear; Monocytes; Polystyrenes
PubMed: 33313615
DOI: 10.1039/d0lc00853b -
Cytokine Dec 2021Interleukin-15 (IL-15) is a pleiotropic cytokine that plays pivotal roles in innate and adaptive immunity. It is also a promising cytokine for treating cancer. Despite...
Interleukin-15 (IL-15) is a pleiotropic cytokine that plays pivotal roles in innate and adaptive immunity. It is also a promising cytokine for treating cancer. Despite growing interest in its use as an immunotherapeutic, its safety and immunological effects in dogs have not been reported. In this study, healthy dogs were given recombinant canine IL-15 (rcIL-15) intravenously at a daily dose of 20 μg/kg for 8 days and monitored for 32 days to determine the safety and immunological effects of rcIL-15. The repeated administration of rcIL-15 was well tolerated, did not cause any serious side effects, and promoted the selective proliferation and activation of canine anti-cancer effector cells, including CD3CD8 cytotoxic T lymphocytes, CD3CD5CD21, and non-B/non-T NK cell populations, without stimulating T lymphocytes. The rcIL-15 injections also stimulated the expression of molecules and transcription factors associated with the activation and effector functions of NK cells, including CD16, NKG2D, NKp30, NKp44, NKp46, perforin, granzyme B, Ly49, T-bet, and Eomes. These results suggest that rcIL-15 might be a valuable therapeutic adjuvant to improve immunity against cancer in dogs.
Topics: Animals; Antigens, CD; Cell Proliferation; Cytotoxicity, Immunologic; Dogs; Forkhead Transcription Factors; Gene Expression Regulation; Granzymes; Humans; Interleukin-15; K562 Cells; Killer Cells, Natural; Leukocyte Count; Leukocytes, Mononuclear; Lymphocyte Subsets; RNA, Messenger; Recombinant Proteins; T-Box Domain Proteins
PubMed: 34103211
DOI: 10.1016/j.cyto.2021.155599 -
Psychoneuroendocrinology Oct 2022Mitochondria are multifunctional energy-producing and signaling organelles that support life and contribute to stress adaptation. There is a growing understanding of the...
BACKGROUND
Mitochondria are multifunctional energy-producing and signaling organelles that support life and contribute to stress adaptation. There is a growing understanding of the dynamic relationship between stress exposure and mitochondrial biology; however, the influence of stress on key domains of mitochondrial biology during early-life, particularly the earliest phases of intra-uterine/prenatal period remains largely unknown. Thus, the goal of this study was to examine the impact of fetal exposure to stress (modeled as the biological construct allostatic load) upon mitochondrial biology in early childhood.
METHODS
In n = 30 children (range: 3.5-6 years, 53% male), we quantified mitochondrial content via citrate synthase (CS) activity and mtDNA copy number (mtDNAcn), and measured mitochondrial bioenergetic capacity via respiratory chain enzyme activities (complexes I (CI), II (CII), and IV (CIV)) in platelet-depleted peripheral blood mononuclear cells (PBMCs). In a cohort of healthy pregnant women, maternal allostatic load was operationalized as a latent variable (sum of z-scores) representing an aggregation of early-, mid- and late-gestation measures of neuroendocrine (cortisol), immune (interleukin-6, C-reactive protein), metabolic (homeostasis model assessment of insulin resistance, free fatty acids), and cardiovascular (aggregate systolic and diastolic blood pressure) systems, as well as an anthropometric indicator (pre-pregnancy body mass index [BMI]).
RESULTS
An interquartile increase in maternal allostatic load during pregnancy was associated with higher mitochondrial content (24% and 15% higher CS and mtDNAcn), and a higher mitochondrial bioenergetic capacity (16%, 23%, and 25% higher CI, CII and CIV enzymatic activities) in child leukocytes. The positive association between maternal allostatic load during pregnancy and child mitochondrial content and bioenergetic capacity remained significant after accounting for the effects of key pre- and post-natal maternal and child covariates (p's < 0.05, except CI p = 0.073).
CONCLUSION
We report evidence that prenatal biological stress exposure, modeled as allostatic load, was associated with elevated child mitochondrial content and bioenergetic capacity in early childhood. This higher mitochondrial content and bioenergetic capacity (per leukocyte) may reflect increased energetic demands at the immune or organism level, and thus contribute to wear-and-tear and pathophysiology, and/or programmed pro-inflammatory phenotypes. These findings provide potential mechanistic insight into the cellular processes underlying developmental programming, and support the potential role that changes in mitochondrial content and bioenergetic functional capacity may play in altering life-long susceptibility for health and disease.
Topics: Allostasis; DNA, Mitochondrial; Energy Metabolism; Female; Humans; Leukocytes, Mononuclear; Male; Mitochondria; Pregnancy
PubMed: 35853381
DOI: 10.1016/j.psyneuen.2022.105868 -
Mitochondrion May 2020Changes in whole blood (WB) mitochondrial DNA (mtDNA) content are associated with health and disease. Platelet-derived mtDNA can confound WB mtDNA content measurements....
Changes in whole blood (WB) mitochondrial DNA (mtDNA) content are associated with health and disease. Platelet-derived mtDNA can confound WB mtDNA content measurements. From a sample of 44 volunteers, we show that platelet mtDNA content and platelet:leukocyte ratio are both dependent predictors of WB mtDNA content, but that platelet count itself is not. Furthermore, when platelet:leukocyte ratio increased by <2-fold ex vivo, the effect on WB mtDNA content was minimal. Altogether, this study clarifies the contribution of platelet mtDNA content rather than platelet count on WB mtDNA content measurements, and identifies defined parameters for future research on WB mtDNA content.
Topics: Adult; DNA, Mitochondrial; Female; Healthy Volunteers; Humans; Leukocyte Count; Leukocytes, Mononuclear; Male; Mitochondria; Platelet Count; Real-Time Polymerase Chain Reaction; Young Adult
PubMed: 32156645
DOI: 10.1016/j.mito.2020.03.001