Pattern Of Cellular Response Research Articles

A novel multi-omics data analysis of dose-dependent and temporal changes in regulatory pathways due to chemical perturbation: a case study on caffeine

Comprehensive analysis of multi-omics data can reveal alterations in regulatory pathways induced by cellular exposure to chemicals by characterizing biological processes at the molecular level. Data-driven omics analysis, conducted in a dose-dependent or dynamic manner, can facilitate comprehending toxicity mechanisms. This study introduces a novel multi-omics data analysis designed to concurrently examine dose-dependent and temporal patterns of cellular responses to chemical perturbations. This analysis, encompassing preliminary exploration, pattern deconstruction, and network reconstruction of multi-omics data, provides a comprehensive perspective on the dynamic behaviors of cells exposed to varying levels of chemical stimuli. Importantly, this analysis is adaptable to any number of omics layers, including site-specific phosphoproteomics. We implemented this analysis on multi-omics data obtained from HepG2 cells exposed to a range of caffeine doses over varying durations and identified six response patterns, along with their associated biomolecules and pathways. Our study demonstrates the effectiveness of the proposed multi-omics data analysis in capturing multidimensional patterns of cellular response to chemical perturbation, enhancing understanding of pathway regulation for chemical risk assessment.

Diagnostic Value of Immunological Biomarkers in Children with Asthmatic Bronchitis and Asthma.

Background and Objectives: This study aimed to investigate the diagnostic value of immunological biomarkers in children with asthmatic bronchitis and asthma and to develop a machine learning (ML) model for rapid differential diagnosis of these two diseases. Materials and Methods: Immunological biomarkers in peripheral blood were detected using flow cytometry and immunoturbidimetry. The importance of characteristic variables was ranked and screened using random forest and extra trees algorithms. Models were constructed and tested using the Scikit-learn ML library. K-fold cross-validation and Brier scores were used to evaluate and screen models. Results: Children with asthmatic bronchitis and asthma exhibit distinct degrees of immune dysregulation characterized by divergent patterns of humoral and cellular immune responses. CD8+ T cells and B cells were more dominant in differentiating the two diseases among many immunological biomarkers. Random forest showed a comprehensive high performance compared with other models in learning and training the dataset of immunological biomarkers. Conclusions: This study developed a prediction model for early differential diagnosis of asthmatic bronchitis and asthma using immunological biomarkers. Evaluation of the immune status of patients may provide additional clinical information for those children transforming from asthmatic bronchitis to asthma under recurrent attacks.

89 Lung Single Cell Transcriptomics to Guide the Development of AOP Anchored Cell-Based Assays in Response to Nanoparticle Exposure

Abstract Inhaled nanoparticles (NP) can cause acute and chronic inflammation. Here we seek to identify NP-specific cellular perturbation pathways and the underlying key cell types to inform adverse outcome pathway (AOP) anchored in vitro studies, by single cell transcriptomics. We exposed mice intratracheally to carbon black (CNP), tangled double-walled (DWCNT) and rigid, multi-walled carbon nanotubes (MWCNT). All lungs underwent single-cell RNA sequencing (scRNA-seq), histology and BAL analysis. At the chosen doses all NPs caused comparable airspace neutrophilia at 12h, which increased until d6 for the CNTs but remained elevated until d28 only for MWCNT, indicating the key event (KE) acute and chronic inflammation. Comparing scRNA-seq and BAL cytokine levels at 12h, differentiated the fate of inflammation NP specifically. In agreement with airspace neutrophilia, CNP uniquely triggered GM-CSF and CXCL1 release, with Csf2 and Cxcl1 mRNAs being expressed mainly in alveolar epithelial cells. DWCNT caused CCL2, -3 and -4 release involving interstitial macrophages and monocytes, associated with acute high lung monocyte and at d6 high BAL macrophage numbers. MWCNT in contrast caused ample BAL cytokine increases involving different cell types and specifically caused a Th2 cytokine response (CCL11, IL10), a crucial KE of fibrosis AOPs. MWCNT triggered cell communication analysis uncovered NP-specific networks including epithelial cells, fibroblasts, macrophages, dendritic cells and endothelial cells. Our study uncovers early NP-specific cell perturbations and identified first specific cellular response pattern to guide AOP predictive cell-based testing strategies. Further analysis of the cellular dynamics shall enhance our understanding of NP specific cell circuits and related pathologies.

Cellular and molecular effects of non-ionizing electromagnetic fields.

The way that living cells respond to non-ionizing electromagnetic fields (EMF), including static/extremely-low frequency and radiofrequency electromagnetic fields, fits the pattern of 'cellular stress response'- a mechanism manifest at the cellular level intended to preserve the entire organism. It is a set pattern of cellular and molecular responses to environmental stressors, such as heat, ionizing radiation, oxidation, etc. It is triggered by cellular macromolecular damage (in proteins, lipids, and DNA) with the goal of repairing and returning cell functions to homeostasis. The pattern is independent of the type of stressor encountered. It involves cell cycle arrest, induction of specific molecular mechanisms for repair, damage removal, cell proliferation, and cell death if damage is too great. This response could be triggered by EMF-induced alternation in oxidative processes in cells. The concept that biological response to EMF is a 'cellular stress response' explains many observed effects of EMF, such as nonlinear dose- and time-dependency, increased and decreased risks of cancer and neurodegenerative diseases, enhanced nerve regeneration, and bone healing. These responses could be either detrimental or beneficial to health, depending on the duration and intensity of the exposure, as well as specific aspects ofthe living organism being exposed. A corollary to electromagnetic hypersensitivity syndrome (EHS) could be an inappropriate response of the hippocampus/limbic system to EMF, involving glucocorticoids on the hypothalamic-pituitary-adrenal axis.

The role of multi-low molecular weight organic acids on phenanthrene biodegradation: Insight from cellular characteristics and proteomics

Polycyclic aromatic hydrocarbons (PAHs) are carcinogenic and ubiquitous pollutants that need to be solved. The low-molecular-weight organic acid (LMWOA) holds the promise to accelerate the capacity of microbes to degrade PAHs. However, the degradation mechanism(s) with multi-LMWOAs has not been understood yet, which is closer to the complex environmental biodegradation in nature. Here, we demonstrated a comprehensive cellular and proteomic response pattern by investigating the relationship between a model PAH degrading strain, B. subtilis ZL09-26, and the mixture LMWOAs (citric acid, glutaric acid, and oxalic acid). As a result, multi-LMWOAs introduced a highly enhanced phenanthrene (PHE) degradation efficiency with up to 3.1-fold improvement at 72 h, which is accompanied by the enhancement of strain growth and activity, but the releasement of membrane damages and oxidative stresses. Moreover, a detailed proteomic analysis revealed that the synergistic perturbation of various metabolic pathways jointly governed the change of cellular behaviors and improved PHE degradation in a network manner. The obtained knowledge provides a foundation for designing the artificial LMWOAs mixtures and guides the rational remediation of contaminated soils using bio-stimulation techniques.

Generalized Modules for Membrane Antigens (GMMA), an outer membrane vesicle-based vaccine platform, for efficient viral antigen delivery.

Vaccine platforms enable fast development, testing, and manufacture of more affordable vaccines. Here, we evaluated Generalized Modules for Membrane Antigens (GMMA), outer membrane vesicles (OMVs) generated by genetically modified Gram-negative bacteria, as a vaccine platform for viral pathogens. Influenza A virus hemagglutinin (HA), either physically mixed with GMMA (HA+STmGMMA mix), or covalently linked to GMMA surface (HA-STmGMMA conjugate), significantly increased antigen-specific humoral and cellular responses, with HA-STmGMMA conjugate inducing further enhancement than HA+STmGMMA mix. HA-STmGMMA conjugate protected mice from lethal challenge. The versatility for this platform was confirmed by conjugation of rabies glycoprotein (RABVG) onto GMMA through the same method. RABVG+STmGMMA mix and RABVG-STmGMMA conjugate exhibited similar humoral and cellular response patterns and protection efficacy as the HA formulations, indicating relatively consistent responses for different vaccines based on the GMMA platform. Comparing to soluble protein, GMMA was more efficiently taken up in vivo and exhibited a B-cell preferential uptake in the draining lymph nodes (LNs). Together, GMMA enhances immunity against viral antigens, and the platform works well with different antigens while retaining similar immunomodulatory patterns. The findings of our study imply the great potential of GMMA-based vaccine platform also against viral infectious diseases.

P06.01.A Deconvolution of immunotherapy-treated glioblastoma identifies cellular heterogeneity and plasticity at the single-cell level

Abstract Background Glioblastoma is the most aggressive cancer originating in the brain with an average survival of 15 months. One of the characteristics of glioblastoma is the high level of intra-tumour heterogeneity (ITH), but the composition and complexity at the single-cell level is poorly understood. Here, we aimed to assess the effects and consequences of immune checkpoint inhibitor (ICI) on the cellular and molecular heterogeneity of glioblastoma tumours using at the single cell level. Material and Methods In collaboration with the phase I trials unit at Rigshospitalet, we performed paired molecular analysis of glioma cells from primary and relapse surgery after ICI treatment. Samples were analysed using single-cell RNA sequencing (scRNA-seq) as well as bulk RNA sequencing and whole exome DNA sequencing. Results In an effort to trace cellular lineages we developed and refined methods to a identify copy number changes using scRNA-seq. To this end, we identified clonal and subclonal tumour cell populations in each sample. We found high levels of ITH prior to treatment, both with respect to the glioblastoma subtype enrichment and the cell type-specific gene expression. Using expression-based cell-type classification, we found defined recurrent cell-type populations present at both surgery time points. The immune checkpoint treatment had consequences on the cellular phenotypes and proportions of tumour cells, suggesting a level of plasticity in the neoplastic cells. Moreover, we identified examples of clonal dynamics and sweeps following ICI treatment, pointing to potential treatment response and resistance in these population. Conclusion In summary, we pursued single cell-focused analysis of ICI treated glioblastoma patients to study the cellular and molecular heterogeneity within and between glioblastoma patients, which pointed to recurrent patterns of cellular responses following ICI treatment.

Synchrotron Fourier Transform Infrared Microscopy Spectra in Cellular Effects of Janus Kinase Inhibitors on Myelofibrosis Cancer Cells

Janus kinase (JAK) deregulation of the JAK/signal transducers and activators of transcription pathway leads to myelofibrosis that can be treated by JAK inhibitors including Ruxolitinib and Tofacitinib. Even though both inhibitors are effective against myelofibrosis, each of them has a different mode of action in the cells. Ruxolitinib is an inhibitor for selective JAK1/2, and Tofacitinib is an inhibitor for JAK3. This study evaluated the chemical fingerprints of TF-1 cells after JAK inhibitor treatments by the synchrotron Fourier transform infrared microspectroscopy (S-FTIR) spectrum. Tofacitinib and Ruxolitinib treatments in TF-1 cells were applied with a chemical fingerprint approach in S-FTIR spectroscopy and in vitro cytotoxicity in a cell-based assay. Principal component analysis or PCA was utilized to classify three cell treatments with three biochemical alteration absorbances of lipid vibration by the C–H stretching, protein amide I that appeared from the C=O stretching, and a P=O phosphodiester bond from nucleic acids. The results showed that the inhibition effect of Ruxolitinib on the TF-1 cell lines was two-fold higher than Tofacitinib. PCA distinguishes untreated and drug-treated cells by detecting cellular biochemical alteration. The loading plots identify that proteins and nucleic acids were the different main components in disparate cell treatments. Tofacitinib was distinct from the others in lipid and nucleic acid. The second derivative spectra of the three molecular components had decreased lipid production and accumulation, changes in secondary structures in proteins, and a high level of RNA overexpression in cell treatment. The JAK inhibitors caused different spectroscopic biomarkers of the modifications of secondary protein conformation, stimulated cell lipid accumulation, and phosphorylation from untreated cells. The alteration of cellular biochemical components suggests that FTIR is a potential tool to analyze specific patterns of drug cellular responses at the molecular level.

Time-resolved quantitative phosphoproteomics reveals cellular responses induced by caffeine and coumarin

Protein phosphorylation is a critical way that cells respond to external signals and environmental stresses. However, the patterns of cellular response to chemicals at different times were largely unknown. Here, we used quantitative phosphoproteomics to analyze the cellular response of kinases and signaling pathways, as well as pattern change of phosphorylated substrates in HepG2 cells that were exposed to caffeine and coumarin for 10 min and 24 h. Comparing the 10 min and 24 h groups, 33 kinases were co-responded and 32 signaling pathways were co-enriched in caffeine treated samples, while 48 kinases and 34 signaling pathways were co-identified in coumarin treated samples. Instead, the percentage of co-identified phosphorylated substrates only accounted for 4.31% and 9.57% between 10 min and 24 h in caffeine and coumarin treated samples, respectively. The results showed that specific chemical exposure led to a bunch of the same kinases and signaling pathways changed in HepG2 cells, while the phosphorylated substrates were different. In addition, it was found that insulin signaling pathway was significantly enriched by both the caffeine and coumarin treatment. The pattern changes in phosphorylation of protein substrates, kinases and signaling pathways with varied chemicals and different time course shed light on the potential mechanism of cellular responses to endless chemical stimulation.

The presence of serum anti-SARS-CoV-2 IgA appears to protect primary health care workers from COVID-19.

The patterns of humoral and cellular responses to SARS‐CoV‐2 were studied in Swedish primary health care workers (n = 156) for 6 months during the Covid‐19 pandemic. Serum IgA and IgG to SARS‐CoV‐2, T‐cell proliferation and cytokine secretion, demographic and clinical data, PCR‐verified infection, and self‐reported symptoms were monitored. The multivariate method OPLS‐DA was used to identify immune response patterns coupled to protection from Covid‐19. Contracting Covid‐19 was associated with SARS‐CoV‐2‐specific neutralizing serum IgG, T cell, IFN‐γ, and granzyme B responses to SARS‐CoV‐2, self‐reported typical Covid‐19 symptoms, male sex, higher BMI, and hypertension. Not contracting Covid‐19 was associated with female sex, IgA‐dominated, or no antibody responses to SARS‐CoV‐2, airborne allergy, and smoking. The IgG‐responders had SARS‐CoV‐2‐specific T‐cell responses including a cytotoxic CD4+ T‐cell population expressing CD25, CD38, CD69, CD194, CD279, CTLA‐4, and granzyme B. IgA‐responders with no IgG response to SARS‐CoV‐2 constituted 10% of the study population. The IgA responses were partially neutralizing and only seen in individuals who did not succumb to Covid‐19. To conclude, serum IgG‐dominated responses correlated with T‐cell responses to SARS‐CoV‐2 and PCR‐confirmed Covid‐19, whereas IgA‐dominated responses correlated with not contracting the infection.

TAMI-68. DECONVOLUTION OF IMMUNOTHERAPY-TREATED GLIOBLASTOMA IDENTIFIES CELLULAR HETEROGENEITY AND PLASTICITY AT THE SINGLE-CELL LEVEL

Abstract Glioblastoma is the most aggressive cancer originating in the brain with an average survival of 15 months. One of the characteristics of glioblastoma is the high level of intra-tumor heterogeneity (ITH), but the composition and complexity at the single-cell level is poorly understood. Here, we aimed to assess the effects and consequences of immune checkpoint inhibitor (ICI) on the cellular and molecular heterogeneity of glioblastoma tumors at the single cell level. In collaboration with the phase I trials unit at Rigshospitalet, we performed paired molecular analysis of glioma cells from primary and relapse surgery after ICI treatment. Samples were analyzed using single-cell RNA sequencing (scRNA-seq) as well as bulk RNA sequencing and whole exome DNA sequencing. In an effort to trace cellular lineages we developed and refined methods to a identify copy number changes using scRNA-seq. To this end, we identified clonal and subclonal tumor cell populations in each sample. We found high levels of ITH prior to treatment, both with respect to the glioblastoma subtype enrichment and the cell type-specific gene expression. Using expression-based cell-type classification, we found defined recurrent cell-type populations present at both surgery time points. The immune checkpoint treatment had consequences on the cellular phenotypes and proportions of tumor cells, suggesting a level of plasticity in the neoplastic cells. Moreover, we identified examples of clonal dynamics and sweeps following ICI treatment, pointing to potential treatment response and resistance in these populations. We additionally found a recurrent pattern of a small population showing high levels of cell cycle activation and simultaneously expressing markers of stem cell potential. In summary, we pursued single cell-focused analysis of ICI treated glioblastoma patients to study the cellular and molecular heterogeneity within and between glioblastoma patients, which pointed to recurrent patterns of cellular responses following ICI treatment.

Tunable heat shock protein-mediated NK cell responses are orchestrated by STAT1 in Antigen Presenting Cells

The release of Heat Shock Proteins (HSPs) from aberrant cells can initiate immune responses following engagement of the HSPs with antigen presenting cells (APCs). This is an important mechanism for cancer immunosurveillance and can also be modeled by vaccination with HSPs through various routes, targeting specific APCs expressing the HSP receptor CD91. Immunological outcomes can be varied as a result of the broad expression of CD91 in different dendritic cell and macrophage populations. We investigated the cellular response of different APCs to the prototypical immunogenic HSP, gp96, in the context of Th1 immunity. Although APCs generally express similar levels of the HSP receptor CD91, we uncovered APC-distinct, downstream signaling pathways activating STAT1, and differential STAT1 induced genes. As a result of this differential and unique signaling we determined that gp96-activated macrophages, but not DCs are capable of activating NK cells to produce IFN-gamma. These data demonstrate that different APC subsets elicit unique intracellular signaling responses to HSPs which result in different patterns of downstream cellular activation and immune responses. Collectively this provides a novel tunable and autochthonous immune response to extracellular HSPs which has important implications on the development of immunity to cancer and infectious disease, as well as homeostasis.

How humic acid and Tween80 improve the phenanthrene biodegradation efficiency: Insight from cellular characteristics and quantitative proteomics

Polycyclic aromatic hydrocarbons (PAHs) are toxic and recalcitrant pollutants, with an urgent need for bioremediation. Systematic biodegradation studies show that surfactant-mediated bioremediation is still poorly understood. Here, we investigated a comprehensive cellular response pattern of the PAH degrading strain B. subtilis ZL09–26 to (non-)green surfactants at the cellular and proteomic levels. Eight characteristic cellular factor investigations and detailed quantitative proteomics analyses were performed to understand the highly enhanced phenanthrene (PHE) degradation efficiency (2.8- to 3-fold improvement) of ZL09–26 by humic acid (HA) or Tween80. The commonly upregulated pathway and proteins (Arginine generation, LacI-family transcriptional regulator, and Lactate dehydrogenase) and various metabolic pathways (such as phenanthrene degradation upstream pathway and central carbon metabolism) jointly govern the change of cellular behaviors and improvement of PHE transport, emulsification, and degradation in a network manner. The obtained molecular knowledge empowers engineers to expand the application of surfactants in the biodegradation of PAHs and other pollutants.

Cytokine Response Profiles of School-Aged Children Infected with Schistosomiasis before and after Praziquantel Treatment.

Schistosomiasis is a parasitic disease that affects millions of people in 78 countries globally. Children under the age of 14, who have the chronic disease may suffer from anemia and malnutrition that contribute to lost days at school and pervasive learning disabilities. The infection is prevalent in Kenya, especially in endemic areas, contributing to significant morbidity. The cellular response pattern is associated with both the acute and chronic phases of the disease, in which cytokines play a critical role. The objective of this study was to evaluate the cytokine profiles of IL-4, IL-2, IL-10, IL-5, IFN-γ, and TNF in serum samples of infected school-aged children by using flow cytometry before and after treatment. The analysis indicated a shift in the expression of the cytokines after treatment with all the cytokines being downregulated, except TNF. There was a general trend of decrease in the expression of the cytokines at six and twelve weeks after treatment as compared to the pretreatment levels. There were statistically significant differences in the expression in IL-2 (P=0.001∗∗), IL-4 (P=0.033∗), IL-10 (P=0.001∗∗∗), IFN-γ (P=0.023∗), and IL-5 (P=0.0001∗∗∗), except in TNF (P=0.095). The reduction in the cytokine levels can be directly related to the influence of the drug praziquantel, modulating the cytokine response by elimination of adult worms, decline in parasitic load, and reduction of morbidity. Therefore, cytokine response is directly related with the influence of treatment in the variation of the immune response.

Using a big data approach to explore in situ chondrocyte response to impact loading in articular cartilage

Using a big data approach to explore in situ chondrocyte response to impact loading in articular cartilage

Distinct Patterns of Humoral and Cellular Immune Responses Following SARS-CoV-2 mRNA Vaccination in Patients With Immune-Mediated Neurological Disorders on Anti-CD20 Therapy: A Prospective Cohort Study

Background: Patients with neuroimmunological disorders on anti-CD20 therapy are at increased risk of severe COVID-19. However, little is known about SARS-CoV-2 vaccine efficacy in this cohort. In particular, the impact of B cell depletion on humoral and cellular immune responses to SARS-CoV-2 vaccination remains poorly defined. No study has addressed vaccine responses to the SARS-CoV-2 delta variant in immunocompromised individuals so far.Methods: In this prospective cohort study we investigated humoral and cellular responses in serial samples from 164 individuals after SARS-CoV-2 mRNA vaccination (82 patients with neuroimmunological disorders on anti-CD20 therapy and 82 healthy controls). Antibodies were quantified using the Elecsys anti-SARS-CoV-2 S immunoassay against the receptor-binding (RBD) domain. T cell responses against the SARS-CoV-2 Wuhan strain and the delta variant were assessed by IFN-g enzyme-linked immunosorbent spot assays. Findings: Following vaccination, SARS-CoV-2 –specific antibodies were detected in 57/82 (70%) patients compared to 82/82 (100%) healthy controls (pInterpretation: Humoral immunity following SARS-CoV-2 mRNA vaccination can be achieved in patients on anti-CD20 therapy once B cells start to repopulate. In the absence of B cells, a robust and stable T cell response is generated which may provide a potent defense against severe COVID-19 in this high-risk patient population. Funding Information: Funded partly by a Medical-Scientific fund of the Major of the federal capital of Vienna (grant Covid003).Declaration of Interests: BK has received honoraria for speaking and for consulting from Biogen, BMS-Celgene, Johnson&Johnson, Merck, Novartis, Roche, Teva and Sanofi-Genzyme outside of the submitted work. No conflict of interest with respect to the present study. FL has received honoraria for speaking and consulting from Almirall, Biogen-Idec, Celgene BMS, MedDay, Merck-Serono, Novartis, Pfizer, Roche, Sanofi-Genzyme, Santhera; Schering, Teva-Ratiopharm. There are no conflicts of interest with respect to the present study, PR has received honoraria for speaking and for consulting from Alexion, Almirall, Amicus, Biogen, Merck, Novartis, Roche, Teva, Sandoz, and Sanofi Genzyme outside of the submitted work. He has received research grants from Amicus, Biogen, Merck and Roche outside the submitted work. GZ recieved speaking honoria from biogen and has participated in meetings sponsored by or received travel funding from Biogen, Merck, Novartis, Roche, Sanofi-Genzyme and Teva. No conflict of interest regarding this study, GB has participated in meetings sponsored by, received speaker honoraria or travel funding from Biogen, Celgene/BMS, Lilly, Merck, Novartis, Roche, Sanofi-Genzyme and Teva, and received honoraria for consulting Biogen, Celgene/BMS, Novartis, Roche, Sanofi-Genzyme and Teva. He has received financial support in the past 12 months by unrestricted research grants (Celgene/BMS, Novartis). No conflict of interest with respect to the present study, ADB declares no conflict of interest related to the content of this article. Independent of this study, ADB has participated in meetings sponsored by or received speaker honoraria or travel funding from Biogen, Merck, Novartis, Roche, Celgene (BMS), and Sanofi. She has received an unrestricted grant from Merck GmbH, an affiliate of Merck KGaA and is currently supported by a research grant from Biogen. TB has no particular conflicts of interest regarding the present study. In general, he has participated in meetings sponsored by and received honoraria (lectures, advisory boards, consultations) from pharmaceutical companies marketing treatments for multiple sclerosis: Almirall, Bayer, Biogen, Biologix, Bionorica, Genzyme, GSK, Janssen-Cilag, MedDay, Merck, Novartis, Octapharma, Roche, Sanofi/Genzyme, TG Pharmaceuticals, TEVA ratiopharm and UCB. His institution has received financial support in the last 12 months by unrestricted research grants (Biogen, Bayer, Merck, Novartis, Sanofi/Genzyme, and TEVA ratiopharm) and for participation in clinical trials in multiple sclerosis sponsored by Alexion, Bayer, Biogen, Merck, Novartis, Octapharma, Roche, Sanofi/Genzyme, and TEVA. LS, HH, RT, FZ, WR, KZ, IW, AS, MG, MM, KR, SW, JHA, ST report no conflicts of interest.Ethics Approval Statement: Subjects gave written informed consent. The study was approved by the ethics committee of the Medical University of Vienna, Austria, EK Nr. 1073/2021.

Coxsackievirus B3 Infection of Human Neural Progenitor Cells Results in Distinct Expression Patterns of Innate Immune Genes.

Coxsackievirus B3 (CVB3), a member of Picornaviridae family, is an important human pathogen that causes a wide range of diseases, including myocarditis, pancreatitis, and meningitis. Although CVB3 has been well demonstrated to target murine neural progenitor cells (NPCs), gene expression profiles of CVB3-infected human NPCs (hNPCs) has not been fully explored. To characterize the molecular signatures and complexity of CVB3-mediated host cellular responses in hNPCs, we performed QuantSeq 3′ mRNA sequencing. Increased expression levels of viral RNA sensors (RIG-I, MDA5) and interferon-stimulated genes, such as IFN-β, IP-10, ISG15, OAS1, OAS2, Mx2, were detected in response to CVB3 infection, while IFN-γ expression level was significantly downregulated in hNPCs. Consistent with the gene expression profile, CVB3 infection led to enhanced secretion of inflammatory cytokines and chemokines, such as interleukin-6 (IL-6), interleukin-8 (IL-8), and monocyte chemoattractant protein-1 (MCP-1). Furthermore, we show that type I interferon (IFN) treatment in hNPCs leads to significant attenuation of CVB3 RNA copy numbers, whereas, type II IFN (IFN-γ) treatment enhances CVB3 replication and upregulates suppressor of cytokine signaling 1/3 (SOCS) expression levels. Taken together, our results demonstrate the distinct molecular patterns of cellular responses to CVB3 infection in hNPCs and the pro-viral function of IFN-γ via the modulation of SOCS expression.

A stereo-selective growth inhibition profile of ginsenoside Rh2 on human colon cancer cells

ABSTRACTWe compared the stereo-selective growth inhibition effects of purified enantiomeric pairs of ginsenoside Rh2 (20(S)-ginsenoside Rh2 and 20(R)-ginsenoside Rh2) on human colon cancer cells in vitro with non-invasive real-time cellular analysis. When 50 μM, respectively, of 20(S)-ginsenoside Rh2 and 20(R)-ginsenoside Rh2 were administered to the colon cancer cell line HT-29, only the S-type ginsenoside Rh2 lowered cancer cell viability. Similarly, when the same experiment was conducted on cancer cells with different levels of ginsenoside treatments (20, 40, 60, and 80 μM), cancer cell viability was decreased in a dose-dependent manner only in the 20(S)-ginsenoside Rh2 treatment groups. Cytotoxicity of cancer cells was observed only in the S form-treated group when cellular response and growth pattern were analyzed via real-time cellular analysis. These findings suggest that the anti-tumor effect of ginsenoside Rh2 on colon cancer cells is S-form selective.

Brucella canis induces canine CD4+ T cells multi-cytokine Th1/Th17 production via dendritic cell activation.

Brucella canis is a small intracellular Gram-negative bacterium that frequently leads to chronic infections highly resistant to antibiotic therapy in dogs. Also, it causes mild human brucellosis compared to other zoonotic Brucella spp. Herein we characterize the cellular immune response elicited by B. canis by analysing human and canine CD4+ T cells after stimulation with autologous monocyte-derived dendritic cells (MoDCs). Human and canine B. canis-primed MoDCs stimulated autologous CD4+ T cells; however, a Th1 response was triggered by human MoDCs, whereas canine MoDCs induced Th1/Th17 responses, with increased CD4+ T cells producing IFN-γ and IL-17A simultaneously. Each pattern of cellular response may contribute to host susceptibility, helping to understand the differences in B. canis virulence between these two hosts. In addition, other aspects of canine immunology are unveiled by highlighting the participation of IL-17A-producing canine MoDCs and CD4+ T cells producing IFN-γ and IL-17A.

TLR4-mediated activation of the ERK pathway following UVA irradiation contributes to increased cytokine and MMP expression in senescent human dermal fibroblasts.

Exposure to ultraviolet (UV) radiation is a major contributing factor to premature aging (photoaging) and skin cancer. In vitro models of cellular senescence have proven to be very useful for the study of slow and progressive accumulation of damage resulting in the growth arrest of aging skin cells. In this study, we compared UVA-induced cellular responses in non-senescent (NS) vs. senescent (S) human dermal fibroblasts (HDFs). HDFs were irradiated with a single dose of UVA (7.5 J/cm2) and QuantSeq 3' mRNA sequencing was performed to assess differential gene expression. Both NS and S HDFs expressed similar numbers of differentially expressed genes, although distinct sets of genes were differentially expressed between the two groups. Higher expression of matrix metalloproteinases (MMPs) and Toll-like receptor (TLR) pathway genes, such as TLR4, MyD88, and CXCL-8, was detected in S HDFs as compared with NS HDFs, and UVA exposure led to a downregulation of collagen genes, such as COL8A2 and COL5A3. Consistent with gene expression profiling, enhanced IL-6 and IL-8 secretion was observed in S HDFs compared with NS HDFs, in response to UVA. Furthermore, we show that TLR4-mediated ERK pathway is responsible for the UVA-mediated mitochondrial dysfunction as well as increased secretion of MMP-1 and IL-8 in S HDFs. Taken together, our results demonstrate the UVA-induced common and distinct molecular patterns of cellular responses between NS and S HDFs and suggest TLR4/ERK pathways as candidate targets to reduce senescent phenotypes.