•The co-culture model of mouse PBMC and RAW264.7 macrophages was established.•PBMC inhibit M1 phenotypic differentiation in RAW264.7 cells.•PBMC promote M2 phenotypic differentiation in RAW264.7 ...cells.•Macrophages boost the polarization of monocytes within PBMC into M2 macrophages.•STAT pathways are involved in the regulation of macrophages by PBMC.
Peripheral blood mononuclear cells (PBMC), sourced autologously, offer numerous advantages when procured: easier acquisition process, no in vitro amplification needed, decreased intervention and overall increased acceptability make PBMC an attractive candidate for cell therapy treatment. However, the exact mechanism by which PBMC treat diseases remains poorly understood. Immune imbalance is the pathological basis of many diseases, with macrophages playing a crucial role in this process. However, research on the role and mechanisms of PBMC in regulating macrophages remains scarce. This study employed an in vitro co-culture model of PBMC and RAW264.7 macrophages to explore the role and mechanisms of PBMC in regulating macrophages. The results showed that the co-culturing led to decreased expression of inflammatory cytokines and increased expression of anti-inflammatory cytokines in RAW264.7 or in the culture supernatant. Additionally, the pro-inflammatory, tissue matrix-degrading M1 macrophages decreased, while the anti-inflammatory, matrix-synthesizing, regenerative M2 macrophages increased in both RAW264.7 and monocytes within PBMC. Moreover, co-cultured macrophages exhibited a significantly decreased p-STAT1/STAT1 ratio, while the p-STAT6/STAT6 ratio significantly increased. This suggests that PBMC may inhibit M1 macrophage polarization by blocking STAT1 signaling cascades and may promote M2 macrophage polarization through the activation of STAT6 signaling cascades. Overall, this study sheds light on the role and mechanism of PBMC in regulating macrophages. Moreover, it was found that monocytes within co-cultured PBMC differentiated into M2 macrophages in the presence of macrophages. This finding provides experimental evidence for the use of PBMC in treating inflammatory diseases, especially macrophage-depleting inflammatory diseases such as osteoarthritis.
•Small molecules in immunotherapy offer a promising frontier for treating immunological diseases.•Compound B, a derivative of Compound A, shows metabolic stability and safety with no PBMC ...toxicity.•Compound B boosts the immune system by promoting pro-inflammatory cytokines and T-cell proliferation.•RNA sequencing demonstrates upregulation of immune response-related genes.•Compound B holds potential as an immune-modulating therapeutic agent, with ongoing target identification and validation.
The use of small molecules in immunotherapy is a promising and rapidly growing field with the potential to revolutionize the way we treat a wide range of immunological diseases. The success stories of using small molecules in immunotherapy have been highlighted with many drugs, such as checkpoint inhibitors, which have achieved significant success in treating several types of tumors. However, despite this success, the development of small molecules for immunotherapy is still in its infancy, and there are many challenges that remain to be addressed. In a recent study, our laboratory reported the immunostimulatory effect of a small molecule (Compound A) (A). In this paper, we studied a derivative of this molecule (Compound B) (Fig. 1B). and analysed its physiochemical properties, immunological activities, and transcriptome profiling. Compound B exhibited acceptable metabolic stability and no toxicity against PBMCs. We also demonstrated that Compound B was capable of modulating the immune system by inducing pro-inflammatory cytokines and promoting T-cell proliferation. RNAseq results showed that Compound B was able to significantly upregulate genes involved in stimulating the immune response pathways. Our findings suggest that Compound B may serve as a promising therapeutic agent to modulate the immune system. The identification and validation of the molecular targets responsible for its immunological activities are underway.
The expression of interferon (IFN) stimulated genes (ISGs) in lymphocytes has been used for pregnancy diagnosis in cattle. However, among-cow variability has yielded sub-optimal predictive accuracy. ...We hypothesized that the expression of ISGs (ISG15, OAS1, RSAD2, CLEC3B, and AKR1B1) in early pregnancy varies according to the proportion of Bos indicus (B. indicus) genetics on females. Multiparous cows were classified in three genetic groups, High Angus (HA; n = 45 0–33% Brahman influence), Angus-Brahman (AB; n = 30 34–67%), and High Brahman (HB; n = 19 68–100%) and submitted to a Select-Synch + CIDR protocol. Cows that displayed estrus (n = 94) were artificially inseminated (Day0; D0). On D19, blood samples were collected to obtain peripheral blood mononuclear cells (PBMC) and measure progesterone (P4) concentrations. On D30, pregnancy diagnosis was performed. The expression of RSAD2 in PBMC of pregnant cows was positively related to the proportion of B. indicus genetics of the groups, but not the expression of ISG15 and OAS1. In pregnant cows, the proportion of B. indicus genetics was negatively associated to circulating levels of P4 concentrations. The P4 concentrations were related positively with RSAD2 expression. ROC curve results determined that for cattle with B. indicus genetics lower than 67%, the CLEC3B and AKR1B1 combination was the most accurate option to predict the outcome of pregnancy. In cows with more than 68% of B. indicus genetics, RSAD2 provided the best accuracy. In conclusion, there is a relationship between the proportion of B. indicus genetics and the ISGs gene expression in PBMC during pregnancy.
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•Expression of classical interferon-stimulated genes was not affected by the proportion of B. indicus genetics in pregnant cattle.•RSAD2 expression was influenced negatively by the plasmatic concentration of progesterone.•RSAD2 was the best predictor of pregnancy for cows with greater proportion of B. indicus genetics.
Nanoparticles are increasingly used in biomedical applications to influence the way the immune system reacts to tumors and infectious disease-causing agents. Nanoparticles not-intended for ...immunomodulation can also influence immune responses by affecting immune cell subsets' viability and/or activity. While immunophenotyping is commonly used to assess the effects of drugs and nanoparticles on immune cell subsets, no standardized approach exists due to the breadth of available cell models and instrumentation. In this chapter, we describe a protocol for flow cytometer calibration and reagent qualification prior to its use in the immunophenotyping experiment. The strategies described herein can be adapted to other instruments. The subsequent chapter-immunophenotyping part II (Chap. 25 )-provides detailed instructions for applying this methodology to analyze nanoparticle effects on subsets of immune cells present in peripheral blood.
Despite growing evidence that Long noncoding RNAs (lncRNAs) can regulate gene expression and widely take part in autoimmune and inflammatory diseases, our knowledge of systemic lupus erythematosus ...(SLE)-related lincRNAs remains limited. In this study, we aimed to explore the contribution of the lncRNA metastasis associated lung adenocarcinoma transcript 1 (MALAT1) to the pathogenesis of SLE. PBMCs were obtained from SLE patients and healthy donors. The expression levels of MALAT-1 were measured by quantitative PCR. Small interfering RNA (siRNA) was then used to knock down the expression of MALAT1 in order to determine the role of MALAT1 in the expression levels of IL-21 and SIRT1 signaling pathway in primary monocytes of SLE patients. Here, we found MALAT-1 expression was abnormally increased in SLE patients and predominantly expressed in human monocytes. Additionally, silencing MALAT-1 significantly reduced the expression of IL-21 in primary monocytes of SLE patients. Furthermore, MALAT-1 exerts its detrimental effects by regulating SIRT1 signaling. Our results demonstrate that MALAT-1 is the key regulatory factor in the pathogenesis of SLE and provides potentially novel target for therapeutic intervention.
Abstract
Background
The World Health Organization characterizes novel coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), as a ...pandemic. Here, we investigated the clinical, cytokine levels; T-cell proportion; and related gene expression occurring in patients with COVID-19 on admission and after initial treatment.
Methods
Eleven patients diagnosed with COVID-19 with similar initial treatment regimens were enrolled in the hospital. Plasma cytokine, peripheral T cell proportions, and microfluidic quantitative polymerase chain reaction analyses for gene expression were conducted.
Results
Five patients with mild and 6 with severe disease were included. Cough and fever were the primary symptoms in the 11 COVID-19 cases. Older age, higher neutrophil count, and higher C-reactive protein levels were found in severe cases. IL-10 level significantly varied with disease progression and treatment. Decreased T-cell proportions were observed in patients with COVID-19, especially in severe cases, and all were returned to normal in patients with mild disease after initial treatment, but only CD4+ T cells returned to normal in severe cases. The number of differentially expressed genes (DEGs) increased with the disease progression, and decreased after initial treatment. All downregulated DEGs in severe cases mainly involved Th17-cell differentiation, cytokine-mediated signaling pathways, and T-cell activation. After initial treatment in severe cases, MAP2K7 and SOS1 were upregulated relative to that on admission.
Conclusions
Our findings show that a decreased T-cell proportion with downregulated gene expression related to T-cell activation and differentiation occurred in patients with severe COVID-19, which may help to provide effective treatment strategies for COVID-19.
Suppressed T-cell immune response and decreased T cells occurred in patients with COVID-19 related to downregulated gene expression involved in T-cell activation and differentiation, especially in severe disease, against SARS-CoV-2 infection and at the early period of treatment.
Bovine herpesvirus-1 is highly contagious virus of cattle and buffaloes all over the world. It establishes lifelong latency in ganglionic neurons of the peripheral nervous system. Since, trigeminal ...ganglia are the main sites of latency, therefore, it is challenging to detect BoHV-1 in latently infected live animals. No research work has been done to correlate the sero-prevalence and latency in peripheral blood mononuclear cells (PBMC). The present study was designed to detect BoHV-1 latency related transcript or microRNA in peripheral blood mononuclear cells of sero-positive animals. The highly sensitive RT-qPCR assays based on TaqMan chemistry have been developed for the detection of transcripts of BoHV-1 latency. The limit of detection (LOD) of the assays for ORF-1 specific RT-qPCR and miRNA specific RT-qPCR was 460 copies and 117 copies respectively. The efficiency of the developed assays was 93.5% for ORF-1 and 97.93% for miRNA transcript. None of the PBMC samples of seropositive animals found positive for ORF-1 and miRNA transcripts in developed assays.The absence of latency specific transcripts in PBMC might be due to very low expression i.e. beyond the LOD of newly developed assays or absence of latency in PBMC of seropositive animal. However, further studies are required to establish the fact. To the best of authors knowledge, this is the first report of latency-specific RT-qPCR assay development and its application in PBMC of BoHV-1 seropositive cattle.
Prostate cancer is the second most common cancer and the fifth leading cause of cancer‐associated death in men. Previous studies have revealed a surprising ability for an immature population of ...myeloid cells called myeloid‐derived suppressor cells (MDSCs) in the commencement and development of many tumors, including those of prostate cancer. Herein, the molecular and cellular changes of MDSCs in prostate cancer in both human and nonhuman models are reviewed. The suppressive function of MDSCs are also discussed with a particular focus on the role of IL‐6 and JAK/STAT3 signaling pathways in the induction of their suppressive activity. Ultimately, a brief review of MDSC‐targeting approaches for potential cancer therapy is presented.
Review of the suppressive role of myeloid‐derived suppressor cells in the context of both human and clinical models of prostate cancer
Methicillin-resistant Staphylococcus aureus (MRSA) is a deleterious human pathogen responsible for severe morbidity and mortality worldwide. The pathogen has attained high priority in the World ...Health Organization (WHO) - Multidrug-resistant (MDR) pathogens list. Emerging MDR strains of S. aureus are clinically challenging due to failure in conventional antibiotic therapy. Biofilm formation is one of the underlying mechanisms behind the antibiotic resistance. Hence, attenuating biofilm formation has become an alternative strategy to control persistent infections. The current study is probably the first that focuses on the antibiofilm and antivirulence potential of myrtenol against MRSA and its clinical isolates. Myrtenol exhibited a concentration-dependent biofilm inhibition without causing any harmful effect on cell growth and viability. Further, microscopic analysis validated the biofilm inhibitory efficacy of myrtenol against MRSA. In addition, myrtenol inhibited the synthesis of major virulence factors including slime, lipase, α-hemolysin, staphyloxanthin and autolysin. Inhibition of staphyloxanthin in turn sensitized the MRSA cells to healthy human blood and hydrogen peroxide (H2O2). Notably, myrtenol treated cells were deficient in extracellular DNA (eDNA) mediated autoaggregation as eDNA releasing autolysis was impaired by myrtenol. Biofilm disruptive activity on preformed biofilms was observed at concentrations higher than minimum biofilm inhibitory concentration (MBIC) of myrtenol. Also, the non-cytotoxic effect of myrtenol on human peripheral blood mononuclear cell (PBMC) was evidenced by trypan blue and Alamar blue assays. Transcriptional analysis unveiled the down-regulation of global regulator sarA and sarA mediated virulence genes upon myrtenol treatment, which is well correlated with results of phenotypic assays. Thus, the results of the present study revealed the sarA mediated antibiofilm and antivirulence potential of myrtenol against MRSA.
Among the properties melatonin is claimed to possess, are the immuno-inflammation inductive capacities that would be responsible of some of the paramount of activities melatonin is reported to have ...in most of the human pathological conditions. In the present paper, we measured the effect of melatonin on established cellular models of immuno-inflammation, and found none. The discrepancies are discussed, especially because those properties are reported at pharmacological concentration (1 μM and beyond) at which the melatonin receptors are desensitized by internalization, leading to putative non-receptor-dependent mechanism of action.
•The melatonin enthusiasts are ruining Science reputation, by claiming facts that are often wishful thinking.•Melatonin is not able to boost most of the actors of the immune system, even at pharmacological concentrations.•We hope to be able to show that some of the activities of melatonin are pure fantasies.
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