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A major symptom of diabetes mellitus (DM) is unfit hyperglycemia, which leads to impaired wound healing. It has been reported that the migration of fibroblasts can be suppressed under high glucose (HG) conditions. In our previous study, we introduced a serum-free culture method for mononuclear cells (MNCs) called quantity and quality control culture (QQc), which could improve the vasculogenic and tissue regeneration ability of MNCs. In this study, we described a culture model in which we applied a high glucose condition in human dermal fibroblasts to simulate the hyperglycemia condition in diabetic patients. MNC-QQ cells were cocultured with fibroblasts in this model to evaluate its role in improving fibroblasts dysfunction induced by HG and investigate its molecular mechanism. It was proven in this study that the impaired migration of fibroblasts induced by high glucose could be remarkably enhanced by coculture with MNC-QQ cells. PDGF B is known to play important roles in fibroblasts migration. Quantitative PCR revealed that MNC-QQ cells enhanced the gene expressions of PDGF B in fibroblasts under HG. Taken with these results, our data suggested a possibility that MNC-QQ cells accelerate wound healing via improving the fibroblasts migration and promote the gene expressions of PDGF B under diabetic conditions.

Chronic obstructive pulmonary disease (COPD) is considered the fourth-leading causes of death worldwide; COPD is caused by inhalation of noxious indoor and outdoor particles, especially cigarette smoke that represents the first risk factor for this respiratory disorder. To mimic the effects of particulate matter on COPD, we isolated peripheral blood mononuclear cells (PBMCs) and treated them with combustion-generated ultrafine particles (UFPs) obtained from two different fuel mixtures, namely, pure ethylene and a mixture of ethylene and dimethylfuran (the latter mimicking the combustion of biofuels). UFPs were separated in two fractions: (1) sub-10 nm particles, named nano organic carbon (NOC) particles and (2) primarily soot particles of 20-40 nm and their agglomerates (200 nm). We found that both NOC and soot UFPs induced the release of IL-18 and IL-33 from unstable/exacerbated COPD-derived PBMCs. This effect was associated with higher levels of mitochondrial dysfunction and derived reactive oxygen species, which were higher in PBMCs from unstable COPD patients after combustion-generated UFP exposure. Moreover, lower mRNA expression of the repairing enzyme OGG1 was associated with the higher levels of 8-OH-dG compared with non-smoker and smokers. It was interesting that IL-18 and IL-33 release from PBMCs of unstable COPD patients was not NOD-like receptor 3/caspase-1 or caspase-8-dependent, but rather correlated to caspase-4 release. This effect was not evident in stable COPD-derived PBMCs. Our data suggest that combustion-generated UFPs induce the release of caspase-4-dependent inflammasome from PBMCs of COPD patients compared with healthy subjects, shedding new light into the biology of this key complex in COPD.

Research into the efficacy of photobiomodulation therapy (PBMT) in reducing inflammation has been ongoing for years, but standards for irradiation methodology still need to be developed. This study aimed to test whether PBMT stimulates in vitro human peripheral blood mononuclear cells (PBMCs) to synthesize pro-inflammatory cytokines, including chemokines. PBMCs were irradiated with laser radiation at two wavelengths simultaneously (λ = 808 nm in continuous emission and λ = 905 nm in pulsed emission). The laser radiation energy was dosed in one dose as a whole (5 J, 15 J, 20 J) or in a fractionated way (5 J + 15 J and 15 J + 5 J) with a frequency of 500, 1,500 and 2,000 Hz. The surface power densities were 177, 214 and 230 mW/cm 2 , respectively. A pro-inflammatory effect was observed at both the transcript and protein levels for IL-1β after PBMT at the energy doses 5 J and 20 J (ƒ=500 Hz) and only at the transcript level after application of PBMT at energy doses of 20 J (ƒ= 1,500; ƒ=2,000 Hz) and 5 + 15 J (ƒ=500 Hz). An increase in CCL2 and CCL3 mRNA expression was observed after PBMT at 5 + 15 J (ƒ=1,500 Hz) and 15 + 5 J (ƒ=2,000 Hz) and CCL3 concentration after application of an energy dose of 15 J (frequency of 500 Hz). Even though PBMT can induce mRNA synthesis and stimulate PBMCs to produce selected pro-inflammatory cytokines and chemokines, it is necessary to elucidate the impact of the simultaneous emission of two wavelengths on the inflammatory response mechanisms.

Cigarette filter microplastics are composed of cellulose acetate that does not undergo biological or photo-degradation. These microplastics are readily dispersed and can be found abundantly in water, soil, and air. These fibers possess high absorption capabilities, allowing them to collect and retain pollutants such as toxic elements. As a result, they are regarded as potential dangers to living organisms. The purpose of this study was to analyze the immune response of human peripheral blood mononuclear cells (PBMCs) when exposed to cigarette filter microfibers, measuring the secretion of the inflammatory cytokines TNFα (tumor necrosis factor-alpha) and IL-6 (interleukin-6). In this study, we examined how used cigarette cellulose acetate microfibers affect the viability of peripheral blood mononuclear cells in an appropriate culture medium at three concentrations: 50, 100, and 200 µg/ml. In addition, this study investigated the release of inflammatory cytokines TNFα and IL6 from PBMCs exposed to 200 µg/ml cigarette filter cellulose acetate. The results showed that increasing the concentration of cellulose acetate fibers of one of the brands in the culture medium has a significant effect on reducing cell viability. The 200 µg/ml in DW is more effective than 50 and 100 µg/ml in reducing cell viability. Peripheral blood mononuclear cells showed an inflammatory immune response when exposed to 200 µg/ml cellulose acetate from cigarette filters. They produced inflammatory cytokines that showed a significant increase compared to the control sample. In general, it can be concluded that cellulose acetate fibers in contact with body cells stimulate them and cause an inflammatory response.

Bromelain, a proteolytic enzyme from pineapple, is a promising anticancer candidate due to its ability to induce apoptosis in various malignant cell lines. While its antitumor efficacy is explored, the safety profile of bromelain in normal human cells requires further validation to support its clinical application. This study examines the cytotoxic, proapoptotic, and genotoxic effects of bromelain at concentrations ranging from 0 to 120 µg/mL on normal human peripheral blood mononuclear cells (PBMCs). The following assessments were conducted: cell survival, protein integrity (thiol and amino groups), mitochondrial membrane potential, types of cell death, and DNA damage. The results indicate that bromelain at the studied concentrations exhibits low overall cytotoxicity toward PBMCs. While it did not cause nonspecific proteolysis of cellular proteins, bromelain at concentrations of 60 and 120 µg/mL triggered apoptosis (9.8% and 15.08%, respectively). The proapoptotic effect observed at 120 µg/mL in PBMCs was associated with several factors: decreased mitochondrial membrane potential, increased expression of proapoptotic caspases-9 and − 3, the proapoptotic protein Bax, and subsequent PARP cleavage. Furthermore, bromelain demonstrated dose-dependent genotoxic effects in PBMCs across the entire tested range (30–120 µg/mL), with a maximum DNA damage of 16.0% in the comet tail. These findings highlight the concentration-dependent response of human PBMCs to bromelain and provide important safety data that should be considered when evaluating its potential as a therapeutic agent.

Background Type 1 diabetes mellitus (T1DM) is a prototypic endocrine autoimmune disease resulting from an immune-mediated destruction of pancreatic insulin-secreting β  cells. A comprehensive immune cell phenotype evaluation in T1DM has not been performed thus far at the single-cell level. Methods In this cross-sectional analysis, we generated a single-cell transcriptomic dataset of peripheral blood mononuclear cells (PBMCs) from 46 manifest T1DM (stage 3) cases and 31 matched controls. Results We surprisingly detected profound alterations in circulatory immune cells (1784 dysregulated genes in 13 immune cell types), far exceeding the count in the comparator systemic autoimmune disease SLE. Genes upregulated in T1DM were involved in WNT signaling, interferon signaling and migration of T/NK cells, antigen presentation by B cells, and monocyte activation. A significant fraction of these differentially expressed genes were also altered in T1DM pancreatic islets. We used the single-cell data to construct a T1DM metagene z-score (TMZ score) that distinguished cases and controls and classified patients into molecular subtypes. This score correlated with known prognostic immune markers of T1DM, as well as with drug response in clinical trials. Conclusions Our study reveals a surprisingly strong systemic dimension at the level of immune cell network in T1DM, defines disease-relevant molecular subtypes, and has the potential to guide non-invasive test development and patient stratification.

Human peripheral blood mononuclear cells (hPBMCs) are widely used in fundamental research and clinical applications as studying their responses to activation is an effective way to uncover functional alterations and disease associated phenotypes. However, the availability of samples in large numbers at a specific time and location remains challenging, hence they often might preferably be collected and cryopreserved for later analysis. While the effect of cryopreservation on viability and cell surface expression is well established, changes in activity and cytokine secretion still lead to conflicting results as it is often measured in bulk or within the cells. Here, we used our platform for dynamic single-cell multiplexed cytokine secretion measurement and compared it to a traditional intracellular cytokine staining to quantify the effect of cryopreservation on cytokine secretion and expression of individual hPBMCs. Following stimulation with LPS or anti-CD3/CD28 antibodies for up to 36 or 72 h incubation, we observed distinct alterations in cytokine responses due to cryopreservation when comparing to fresh samples, but also remarkable consistencies for some cytokines and parameters. In short, the frequencies of cytokine-secreting cells in cryopreserved samples were lower for IL-6 (LPS), IL1-β (CD3/CD28) and IFN-γ (CD3/CD28), while the frequency and dynamics of IL-8 secretion were strongly impacted in all cases. We observed a large disconnect between cytokine expression and secretion for TNF-α, where the expression dramatically increased after cryopreservation, but actual secretion was, in comparison, remarkably stable. The polyfunctionality of single cells was altered by cryopreservation in specific co-secreting populations led by the effects on IL-6 or IL-8 secretion. Among immune cells, cryopreservation seemed to affect lymphocytes and monocytes differently as effects appeared early on in lymphocytes while generally observed in later time points in monocytes. Together, this study offers an in-depth quantitative insight into the biological behavior of immune cells in response to cryopreservation and stimulation, further providing some insights into conflicting results in the literature as well as guidelines for researchers planning to assess cytokine-secreting from frozen hPBMCs in immunological research or clinical applications.

Susceptibility to pathogen infections and efficacy of vaccination highly depend on the immune status of the piglet. Here, we measured immunocytes in piglets from birth to weaning to elucidate how immunocyte populations change during development and are affected by weaning. Crossbred piglets were used. Suckling piglets were euthanized at 1, 7, 14, 21, 28 or 35 days old (3~4 piglets at each time point). In addition, seven piglets were weaned at 21 days old, with four being euthanized at 28 days old and the remaining at 35 days old. Piglet carcasses were dissected, and blood, mesenteric lymph nodes (MLN) and spleen were sampled. In total, seven antibodies were used to stain the immunocyte population. Dynamics of myeloid (CD3–SWC3+CD16+), natural killer (NK; CD3–SWC3–CD16+), killer T (CD3+CD8+), helper T (CD3+CD4+) and B (CD3–CD21+) cells were analyzed. Percentage of innate immunity cells such as myeloid cells declined (p < 0.05) from the first day after birth. In contrast, percentage of NK cells increased in piglets while they were still suckling. Killer T, helper T, and B cell populations increased around 2~3 weeks after birth. No significant differences in the populations of the evaluated cell types were observed between suckling and weaned piglets at least for 14 days post weaning.

Sepsis is characterized by a dynamic and heterogeneous immune response in which hyperinflammation and immunosuppression often coexist and change over time. Peripheral blood mononuclear cells (PBMCs), including monocytes and lymphocyte subsets, provide an accessible window into this evolving immune trajectory and can be repeatedly assessed during disease progression. In this narrative review, we synthesize recent evidence from single-cell and spatial omics, mechanistic studies, and PBMC-based biomarker research, with a focus on three connected themes: time-dependent PBMC remodeling, monocyte/macrophage dysfunction, and translational immune biomarkers. Current evidence indicates that PBMC remodeling in sepsis is marked by monocyte state transitions, HLA-DR downregulation, lymphocyte apoptosis and exhaustion, regulated cell death, and immunometabolic reprogramming. These changes help explain why some patients recover after the acute inflammatory phase, whereas others progress toward persistent immunoparalysis, secondary infection, prolonged critical illness, or poor outcomes. PBMC-derived biomarkers, including transcriptomic signatures, monocyte phenotypes, lymphocyte exhaustion markers, metabolic indicators, and functional immune assays, may improve immune phenotyping and risk stratification. However, clinical translation remains limited by cohort heterogeneity, age-related differences, timing-dependent immune states, assay standardization, and insufficient prospective validation. Future studies should prioritize longitudinal, age-stratified, and biomarker-guided designs to determine whether PBMC-based immune monitoring can support individualized immunomodulatory strategies in sepsis.

Background Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a severe autoimmune neuropsychiatric disease. Brain access of anti-NMDAR autoantibody through the blood–brain barrier (BBB) is essential for pathogenesis. Most previous animal models limit the investigation of etiologies of BBB damage in patients. Methods In this study, we established a novel humanized mouse model of anti-NMDAR encephalitis by intraperitoneal injection of patients’ peripheral blood mononuclear cells (PBMCs) into BALB/c Rag2 −/− Il2rg −/− Sirpα NOD Flk2 −/− mice. Results We found that engraftment of patients’ PBMCs not only produced potent anti-GluN1 autoantibodies, but also disrupted BBB integrity to allow brain access of autoantibodies, resulting in a hyperactive locomotor phenotype, anxiety- and depressive-like behaviors, cognitive deficits, as well as functional changes in corresponding brain regions. Transcriptome analysis suggested an exaggerated immune response and impaired neurotransmission in the mouse model and highlighted Il-1β as a hub gene implicated in pathological changes. We further demonstrated that Il-1β was produced by endothelial cells and disrupted BBB by repressing tight junction proteins. Treatment with Anakinra, an Il-1 receptor antagonist, ameliorated BBB damage and neuropsychiatric behaviors. Conclusions Our study provided a novel and clinically more relevant humanized mouse model of anti-NMDAR encephalitis and revealed an intrinsic pathogenic property of the patient’s lymphocytes.