Peripheral Blood Of Patients Research Articles

Abstract 6031: CD4+CD103+ tissue-resident memory T cells correlate with unfavorable prognosis in patients with head and neck cancer

Abstract T cell memory is an important mechanism for long-term protection against diverse pathogens such as cancers. Tissue-resident memory T cell (TRM) is a subset of memory T cells that stably resides in tissues and controls local immune homeostasis. In the present study, we investigated the immunological profile and clinical significance of TRM in patients with head and neck squamous cell carcinoma (HNSCC). We analyzed RNA-sequencing data and clinical data obtained from The Cancer Genome Atlas (TCGA) database. In total, 520 cases, including 97 human papillomavirus (HPV)-positive HNSCCs and 423 HPV-negative HNSCCs, were analyzed. TRM-enriched tumors were identified based on the expression of both CD4/CD8A and ITGAE (CD103). TRM-enriched tumors tended to correlate with HPV-positive status and early T factor. Gene Set Enrichment Analysis revealed the enrichment of pathways relating to cytotoxic immune response in TRM-enriched tumors. Moreover, the expressions of immunostimulatory and immune checkpoint-related genes were elevated in TRM-enriched tumors. Notably, the enrichment of CD4+TRMs in tumors was an independent prognostic factor for shorter progression-free survivals (PFS). We also analyzed peripheral blood mononuclear cells obtained from 77 patients with HNSCC. The increased CD8+CD103+TRMs correlated with HPV-positive status and primary lesion of oropharynx. The proportion of CD4+CD103+TRMs negatively correlated with that of CD4+ naïve T cells, while that of CD8+CD103+TRMs negatively correlated with that of effector T cells. The proportion of CD4+CD103+TRMs positively correlated with Ki-67 expression in CD4+ T cells. Moreover, the elevated CD4+CD103+TRMs was an independent prognostic factor for shorter PFS. In conclusion, we highlighted the clinical and immunological significance of CD103+TRMs in both tumor tissues and peripheral blood of patients with HNSCC. Further characterization of TRMs could lead to the development of novel biomarkers for patients with HNSCC. Citation Format: Hideyuki Takahashi, Shota Ida, Hiroe Tada, Kazuaki Chikamatsu. CD4+CD103+ tissue-resident memory T cells correlate with unfavorable prognosis in patients with head and neck cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 6031.

Pharmacogenomic analysis and clinical annotation of 635 patients.

In this study, we aimed to reveal the sequence analysis of 69 pharmacogenes in 635 patients and the clinical explanation of variants. Genomic DNA was isolated from peripheral blood of patients. Next-Generation Sequence analysis and bioinformatic analysis were performed to identify 69 pharmacogene variants. Variants with clinical annotation were identified. Analysis of 69 pharmacogenes in a total of 635 patients identified 9485 variants. The number of distinct variants identified in each gene was 1409. Of these 1409 variants, the number of variants registered in PharmGKB was 126. Among the 126 variants registered in PharmGKB, 26 variants were identified that had a direct association with clinical annotation and drug efficacy or toxicity. The most common variant genes were DPYD, CYP2C19, VKORC1,UGT1A1, RYR1 and MTHFR. These 26 variants with clinical annotation were identified in 327 (51%) different individuals. Such a high frequency of critical variants (51%) points to the need for pharmacogenetic studies. The results are extremely important in terms of determining the drug dose according to the genomic status of individuals receiving drug therapy and preventing unnecessary health expenditures.

Detecting plasma SHOX2, HOXA9, SEPTIN9, and RASSF1A methylation and circulating cancer cells for cholangiocarcinoma clinical diagnosis and monitoring.

Cholangiocarcinoma (CCA), also known as bile duct cancer, is a devastating malignancy primarily affecting the biliary tract. To assess their performance in clinical diagnosis and monitoring of CCA, plasma methylation and circulating tumor cells were detected. Plasma samples were collected from Hubei Cancer Hospital (n = 156). Plasma DNA was tested to detect SHOX2, HOXA9, SEPTIN9, and RASSF1A methylation using TaqMan PCR. Circulating tumor cells (CTCs) were detected in the peripheral blood of patients using the United States Food and Drug Administration-approved cell search system before and after clinical therapy. The CCA diagnostic value was estimated using the area under the curve. The independent prognosis risk factors for patients with CCA were estimated using Cox and logistic regression analyses. The sensitivity and specificity of the four DNA plasma methylations exhibited 64.74% sensitivity and 93.88% specificity for detecting CCA. The receiver operating characteristic curve of the combined value for CCA diagnosis in plasma was 0.828 ± 0.032. RASSF1A plasma methylation was related to the prognosis of patients with CCA. We determined the prognostic hazard ratio for CCA using CTC count, tumor stage, methylation, and carbohydrate antigen 19-9 levels as key factors. Our overall survival nomogram achieved a C-index of 0.705 (0.605-0.805). SHOX2, HOXA9, SEPTIN9, and RASSF1A plasma methylation demonstrated increased sensitivity for diagnosing CCA. RASSF1A plasma methylation and CTCs were valuable predictors to assess CCA prognosis and recurrence.

Impairment of platelet mitochondrial respiration in patients with chronic kidney disease with and without diabetes.

Chronic kidney disease (CKD) and diabetic kidney disease (DKD) are major public health problems, and their burden is growing relentlessly with the aging of the global population. Their early recognition is now a public health priority, and there is an unmet need for the identification of specific biomarkers in minimally invasive or non-invasive samples. Mitochondrial dysfunction plays a pivotal role in the development and progression of both CKD and DKD and circulating platelets have emerged as an ideal candidate for the assessment of the respiratory function. The present study assessed mitochondrial respiration in platelets isolated from the peripheral blood of patients with DKD and CKD compared to healthy controls. The study included a total number of 89 subjects, as follows: 30 DKD patients divided into three subgroups based on the urinary albumin-to-creatinine ratio (uACR): 20 normoalbuminuric, 10 microalbuminuric, and 10 macroalbuminuric, 29 CKD patients (positive controls) and 20 healthy individuals (negative controls). Platelets were isolated by differential centrifugations and a high-resolution respirometry protocol was adapted to assess mitochondrial respiration dependent on complex I (CI) and complex II (CII). A significant reduction of the CI-supported active respiration was found in the normoalbuminuric DKD patients and further decreased in the microalbuminuric DKD subgroup. Both CI and CII-dependent coupled respiration and the maximal uncoupled respiration were significantly reduced in the macroalbuminuric DKD subgroup. In conclusion, mitochondrial respiration impairment in peripheral platelets is evident from the early stages of DKD. Moreover, platelet mitochondrial respiration was more severely impaired in patients with macroalbuminuric DKD as compared to those with CKD. Further, more extensive follow-up studies are warranted to determine whether platelet respiratory mitochondrial dysfunction could serve as a peripheral biomarker for kidney mitochondrial dysfunction and/or as a prognostic tool in DKD.

Expression and clinical significance of CXCR3 on effector T cells in the peripheral blood of patients with Alzheimer's disease

Objective: This study investigated the expression of C-X-C motif chemokine receptor 3 (CXCR3) on CD45RO⁺ T cells in the peripheral blood of patients with Alzheimer's disease (AD) and its association with clinical features. Methods: A total of 41 AD patients and 30 age-and sex-matched healthy controls (HCs) were recruited from the Department of Neurology at the Medical Division of PLA General Hospital between September 2022 and March 2024. Flow cytometry was used to quantify CXCR3 expression on CD45RO⁺ T cell subsets in peripheral blood. Dementia severity in AD patients was assessed using the Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA). Spearman correlation analysis examined the relationship between CD45RO⁺CXCR3⁺ T cell levels and cognitive function in the AD group. Receiver operating characteristic (ROC) curve analysis determined the predictive utility of CD45RO⁺CXCR3⁺ T cells for AD, quantified by the area under the curve (AUC). Results: Compared to healthy controls, AD patients exhibited significantly elevated levels of CD8⁺CD45RO⁺CXCR3⁺ T cells [17.8% (7.2%, 40.3%) vs. 8.2% (5.1%, 12.3%), Z=-2.59, P<0.05]. However, no significant differences were observed for CD4⁺CD45RO⁺CXCR3⁺ T cells, CD4⁺CD45RO⁻CXCR3⁺ T cells, or CD8⁺CD45RO⁻CXCR3⁺ T cells (P>0.05). Spearman correlation analysis revealed a negative correlation between CD8⁺CD45RO⁺CXCR3⁺ T cell levels and cognitive scores (MMSE: r=-0.72, P<0.05; MoCA: r=-0.70, P<0.05). ROC analysis demonstrated an AUC of 0.81 for CD8⁺CD45RO⁺CXCR3⁺ T cells in predicting AD, with a sensitivity of 59.0% and specificity of 93.3%. Conclusions: CXCR3 expression is significantly upregulated on CD8⁺CD45RO⁺ T cells in AD patients, and its levels correlate with cognitive impairment severity. These findings suggest that CD8⁺CD45RO⁺CXCR3⁺ T cells may serve as a potential biomarker for AD diagnosis and progression monitoring.

Peripheral blood HIF-1α levels: a study on their predictive ability for early-stage diabetic retinopathy.

This study aims to investigate the correlation between the levels of hypoxia-inducible factor-1α (HIF-1α) in the peripheral blood of patients with type 2 diabetes（T2D）and early-stage diabetic retinopathy (DR), and to evaluate its potential as a predictor of early DR. From November 2021 to December 2023, 70 patients who underwent fundus photography at Zhangzhou Second Hospital were recruited. Based on the results, patients were categorized into T2D group comprising 25 patients and DR group comprising 45 patients. Additionaly,18 healthy individuals who underwent routine physical examinations during the same period were included as a control group. Serum levels of the HIF-1α protein were measured in all three groups. The results indicated that patients in the early-stage DR group had significantly higher levels of HIF-1α, Albumin (Alb), Ca, and Blood Urea Nitrogen (BUN)) compared to the T2D group (). Additionally, patients in the DR group exhibited higher levels of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), fasting plasma glucose (FPG), and glycated hemoglobin (HbA1C) than the control group. The incidence rates of DR in groups A, B,C,and D were 18.18%, 31.82%, 59.09%, and 95.45%, respectively, with statistically significant differences.Spearman's correlation analysis revealed significant positive correlations between HIF-1α and age, Disease Duration (years), Systolic Blood Pressure (SBP), Cr, FPG, and HbA1c, with a strong positive correlation between HIF-1α and HbA1c optimal cutoff value of 2.3855 ng/mL Conclusion: Increased levels of HIF-1α in peripheral blood are closely linked to the development of diabetic retinopathy, suggesting that HIF-1α may act as a potential biomarker for the early detection and prediction of DR risk.

Cell-Type-Specific mRNA N6-Methyladenosine Landscape and Regulatory Mechanisms Underlying Immune Dysregulation of Sleep-Deprived.

Sleep deprivation impairs daytime cognitive functioning and is a risk factor for various diseases related to immune dysregulation. N6-methyladenosine (m6A) is a common epigenetic RNA modification with essential roles in regulating neurogenesis and circadian rhythms. m6A dysregulation resulting in immune imbalance has received much attention. In this study, we elucidated the landscape and specific mechanisms of m6A regulators in the peripheral blood of patients with sleep deprivation through RNA sequencing and single-cell transcriptomics data sets. We observed that m6A regulator upregulation aggravated sleep loss and immune disorders. Women were more sensitive to sleep deprivation. Notably, m6A regulator ALKBH5 was downregulated in peripheral blood mononuclear cells (PBMC) of patients with sleep deprivation at the transcriptome level. However, ALKBH5 was cell-type specific upregulated in T cells (TC), B cells (BC), and natural killer (NK) cells, involving the dysregulation of acquired immune mechanisms by aberrant cell-cell communication that mediated ligand-receptor interactions across diverse cell types. Furthermore, the immune dysregulation of sleep loss could be regulated by a potential pathway between ALKBH5 and CD99 in SH-SY5Y and HT22 cells. Sleep deprivation group CD3+/CD45+ T cells had higher levels of ALKBH5 mRNA than the control group. This study demonstrated that abnormal m6A modification patterns caused by m6A regulators play a key role in the dysregulation of innate and acquired immunity in sleep deprivation.

The Role of Tim-3+T cell subsets in the peripheral blood of patients with COVID-19 and diabetes

The Role of Tim-3+T cell subsets in the peripheral blood of patients with COVID-19 and diabetes

Colorectal Cancer-Derived Exosomes Impair CD4+ T Cell Function and Accelerate Cancer Progression via Macrophage Activation.

Background: Exosomal programmed death ligand 1 (PD-L1), an exosomal membrane protein found in many tumor types, is consider to aid in regulation of the immune microenvironment. However, the functions and the mechanisms underlying the exosome-mediated regulation of the immune microenvironment in colorectal cancer (CRC) remain unknown. Methods: Western blotting was used to investigate the levels of exosomal PD-L1 in the peripheral blood of patients with CRC and healthy controls. A CRC mouse model was constructed by administering 10 mg/kg azoxymethane (AOM) and dextrane sodium sulfate (DSS) intraperitoneally. The mice were then administered the control or CRC-derived exosomes to examine the regulatory effect of the exosomes on macrophage infiltration and CRC development. In vitro studies, using a coculture system, and flow cytometry analysis were conducted to examine the relationship between the regulatory effect of CRC-derived exosomes on CD4+ T cells and tumor-associated macrophages. RNA-seq and reverse transcription-quantitative polymerase chain reaction assays were used to investigate the mechanisms underlying the regulatory effect of the CRC-derived exosomes on macrophage proliferation and the regulation of the immune microenvironment during CRC development. Results: In patients with CRC, higher levels of exosomal PD-L1 were associated with a more severe form of disease. The treatment of mice with AOM/DSS-induced CRC with CRC-derived exosomes resulted in high levels of macrophage proliferation, increased PD-L1 levels in macrophages, and accelerated CRC progression. Importantly, analysis of an in vitro coculture system and flow cytometry analysis showed that the CRC-derived exosomes transported PD-L1 into macrophages and impaired CD4+ T cell function. Preliminary data suggest that the NF-κb signaling pathway regulates the function of CRC-derived exosomal PD-L1-dependent macrophages. Conclusion: CRC-derived exosomes induce the proliferation of macrophages and increase their PD-L1 levels. They also impair CD4+ T cell function and promote CRC progression. Our findings reveal a novel exosomal PD-L1-mediated crosstalk between the CRC cells and immune cells in the CRC microenvironment.

CD121b-positive neutrophils predict immunosuppression in septic shock.

Septic shock is linked with high mortality and significant long-term morbidity in survivors. However, the specific role of neutrophils in septic shock pathophysiology remains scarce in recent research. Peripheral blood immune cells from healthy donors and patients with septic shock were analyzed using single-cell RNA sequencing and batch RNA sequencing. We measured serum CD121b in both patients and healthy donors. Peripheral immune cells were isolated and exposed to either a CD121b recombinant protein or a CD121b blocking antibody to evaluate the expression of inflammatory factors. Additionally, in a humanized mouse sepsis model, the expression of CD121b in neutrophils across different tissues was assessed following treatment with all-trans retinoic acid (ATRA). This study identified a subset of CD10-CD121b+ neutrophils in the peripheral blood of patients with septic shock. These patients exhibited elevated concentrations of soluble CD121b in serum and urine. Furthermore, outcomes revealed that the presence of CD121b+ neutrophils positively correlated with the severity of septic shock. These cells displayed immunosuppressive characteristics; after blocking CD121b, proinflammatory cytokines increased in peripheral immune cells. Additionally, we found that treatment with ATRA down-regulated the expression of CD121b. CD121b is closely associated with the progression of septic shock and may serve as a potential predictor indicator of immunosuppression for the condition.

Biomarker Identification in Patients with Multiple Sclerosis Treated with Autologous Hematopoietic Stem Cell Transplantation

Introduction: Approximately 80% of individuals with multiple sclerosis (MS) have a positive response to autologous hematopoietic stem cell transplantation (aHSCT). Markers that may predict the transplant outcome are necessary. The objective of this work is to identify markers that may refine the selection of patients with multiple sclerosis who could benefit from aHSCT. Methods: We evaluated the levels of six biomarkers in the peripheral blood of patients with MS before aHSCT. The design of this study is cross-sectional; patients were divided into two transplant-responses-at-12-months groups, responders (ΔEDSS &lt; 0) and non-responders (ΔEDSS &gt; 0). Pre-transplant samples were used to assess the different markers. Results: Thirty-four patients were enrolled: fourteen were non-responders and twenty were responders to aHSCT. Among the evaluated biomarkers, a significant difference was only detected in miR-146a levels, with increased values in the non-responder group. Conclusions: The biomarker miR146a could be useful to evaluate the response to aHSCT in patients with MS.

Personalized tumor-specific amplified DNA junctions in peripheral blood of patients with glioblastoma.

Monitoring disease progression in patients with high-grade gliomas (HGGs) is challenging due to treatment-related changes on imaging and the requirement for neurosurgical intervention to obtain diagnostic tissue. DNA junctions in HGGs often amplify oncogenes, making these DNA fragments potentially more abundant in blood than monoallelic mutations. Herein, we piloted a cell-free DNA approach for disease detection in plasma of patients with HGGs by leveraging patient-specific DNA junctions associated with oncogene amplifications. Whole genome sequencing of grade 3 or 4 IDH-mutant or wild-type astrocytomas was utilized to identify amplified junctions. Individualized qPCR assays were developed using patient-specific primers designed for the amplified junction. ctDNA levels containing these junctions were measured in patient plasma samples. Unique amplified junctions were evaluated by individualized semi-quantitative PCR assays in presurgical plasma of 18 patients, 15 with tumor-associated focal amplifications and three without. High-copy number junctions were robustly detected in plasma of 14/15 patients (93.3%) with amplified junctions and none of the controls. Changes in junction abundance correlated with disease trajectory in serial plasma samples from five patients, including increased abundance of amplified junctions preceding radiographic disease progression. In patients with grade 3 or 4 astrocytomas who had tumor-associated amplifications, patient-specific amplified junctions were successfully detected in assayed plasma from most patients. Longitudinal analysis of plasma samples correlated with disease trajectory, including cytoreduction and progression.

HMGB1 promotes immune abnormalities in preeclampsia by recruiting monocyte/decidual macrophages and inducing M1 polarization.

Preeclampsia (PE) is a severe pregnancy complication associated with immune imbalance and placental hypoxia stress. Decidua macrophages (dMφ) are essential at maternal fetal interface, which is abnormally activated and excessively transformed to the M1-phenotype in PE. High mobility group box 1(HMGB1), released from necrotic cells after injury, accumulates in the placenta and peripheral blood of patients with PE. Therefore, this study aims to investigate the interaction between macrophages and trophoblasts, exploring how HMGB1 affects macrophage functions and its potential involvement in the pathophysiology of PE. Decidua tissue was obtained from 11 women with severe pre-eclampsia (sPE), 13 women with pre-eclampsia (PE), and 13 women with normal pregnancies. HMGB1 levels in decidua were evaluated by immunohistochemistry (IHC), western blot, and qPCR. Additionally, primary dMφ and peripheral blood monocytes (pMo) were isolated to develop a co-culture model simulating maternal fetal interface cell model of PE. Flow cytometric analysis and in vitro cell migration assay investigated the interaction between macrophages and HMGB1. This study identified elevated HMGB expression in PE patients. HMGB1 expressed widely in trophoblast, decidual stromal cells, and the extracellular matrix. Furthermore, hypoxia induced trophoblast to express and secrete HMGB1, promoting pMo and dMφ migration. Additionally, HMGB1 recruited monocyte-induced macrophages (pMφ) and modulated M1 macrophage polarization. HMGB1 is increased at the maternal-fetal interface of PE, which recruits monocytes and macrophages and induces their polarization to M1. This study offers insights into the suppressive effects of the crosstalk between dMφ and trophoblasts at the maternal-fetal interface, lending credence to macrophage-targeted interventions of PE as a potential therapeutic strategy.

Unbalanced CD4+IL-10+ and more activated and exhausted CD8+ lymphocytes characterized patients with a major depressive episode.

Depression is a highly prevalent psychiatric condition referred to as a major depressive episode (MDE). The underlying causes and mechanisms of depression remain poorly elucidated and, in consequence, the efficacy of the current therapies. Inflammation has been proposed to contribute to the pathophysiology of MDE with altered innate and adaptive immune responses. The present work aimed to shed light on the effector CD4+ T helper (Th) cell subsets response and the activation and exhaustion condition of CD8+ lymphocytes in the peripheral blood of patients with MDE. Patients with MDE (n= 57) and healthy controls (HCs) (n= 34) were recruited. The intracellular production of interferon (IFN) γ (Th1), interleukin (IL)-17 (Th17), and IL-10 (CD4+ IL-10+) as the activation and exhausted status of CD8+ lymphocytes were evaluated by flow cytometry. The frequencies of Th1 and Th17 cells did not show statistical differences, but a lower percentage of CD4+IL-10+ cells (P < 0.05) and CD4+IFNγ+IL-10+ cells (P < .01) were found in patients with MDE compared with HCs. The reduction in the CD4+IL-10+ and CD4+IFNγ+IL-10+ population negatively correlated with the number of depressive episodes (P < .05 and P < .01) and duration of illness (P < .01). A higher proportion of activated CD8+CD69+ (P < .0001) with potentially exhausted phenotype CD8+PD1+LAG3+ (P < .0001) were observed, which positively correlated with the severity, number of episodes, and duration of illness in MDE compared with HCs. Altogether, patients with MDE are characterized by unbalanced Th1/Th17/CD4+IL-10+ cells and increased activation and exhaustion of cytotoxic CD8+ T cells. Therapeutic strategies restoring this balance and effective control of inflammatory responses may be beneficial in MDE. SIGNIFICANCE STATEMENT: The pathophysiology of major depressive episode (MDE) remains poorly defined, hampering the development of more tailored therapeutic interventions. Inflammation has been proposed to initiate or maintain the MDE. This study demonstrates that effector CD4+ T helper cells are altered in MDE, showing a reduction in CD4+IL-10+ cells; meanwhile, CD8+ lymphocytes are more activated but with an exhausted phenotype. This study provides valuable insights into the lymphocyte response, which should lead to advances in the therapeutics of mood disorders.

A combination of hepatic leukemia factor and circulating tumor cells serve as effective biomarkers for lung adenocarcinoma prognosis.

Lung adenocarcinoma (LUAD) is a highly malignant tumor with the highest mortality rate among all cancers. Early diagnosis and prognosis are important factors in treatment. Hepatic leukemia factor (HLF) is thought to be closely associated with lung cancer metastasis. It is downregulated in lung cancer tissues and negatively correlated with the number of metastasis-activating circulating tumor cells (CTCs) in the peripheral blood of patients. In this study, we analyzed data from LUAD samples in TCGA and found that HLF was significantly upregulated in samples with EGFR mutations. Immunohistochemical (IHC) staining of 343 clinical samples also revealed a trend of HLF upregulation in patients with EGFR mutations. EGFR is one of the driver genes in non-small cell lung cancer (NSCLC), and the proportion in LUAD is as high as 50% in the East Asian population. In this study, EGFR mutation was not significantly correlated with the prognosis of LUAD patients and the number of CTC was also not related to EGFR mutation, but was closely related to HLF expression, with more CTCs being captured in the peripheral blood of patients with low expression of HLF (SI ≤ 4). By following up these 343 LUAD patients, high HLF expression (SI > 4) was found to be an independent protective factor for progression-free survival regardless of EGFR status (P<0.001), whereas high CTC count (> 3) was an independent risk factor for recurrence or death in LUAD patients (P<0.001). When low HLF and high CTCs coexisted, patients had the shortest median survival time. Patients with low HLF or high CTCs appeared alone had a moderate median survival time. Patients had the longest median survival time when HLF was high and CTCs were low. In summary, we believe that HLF expression in cancer tissues and the number of CTCs can be used as effective biomarkers for predicting the prognosis of LUAD, which plays an important role in clinical diagnosis and prognosis judgment.

MicroRNA-24-3p targeting Top1 in perirenal fat is involved in circulating inflammation and high cardiovascular disease risk in patients with primary aldosteronism

ContextPatients with primary aldosteronism (PA) are at a high risk of cardiovascular diseases (CVD) and metabolic syndrome. Notable inflammatory and fibrotic changes and differential microRNA (miRNA) expression profiles in the perirenal fat observed in PA may contribute to this increased risk, however, which has not been fully elucidated.ObjectiveThis study aimed to explore the role of high expression of miR-24-3p in perirenal fat in circulating inflammation and its correlation with a high risk of CVD in patients with PA.MethodsPerirenal fat thickness (PRFT) measured by computed tomography (CT), miR-24-3p expression in perirenal fat, circulating inflammatory factors from adrenal veins and peripheral blood in patients with PA were analyzed. In vitro, white and brown adipocytes with miR-24-3p overexpression or inhibition respectively were stimulated with aldosterone and a unidirectional co-culture model of adipocytes and HUVEC was established. The target genes of miR-24-3p were identified.ResultsPatients with PA and CVD have significantly higher PRFT than those without CVD. The expression level of miR-24-3p in perirenal fat was significantly positively correlated with PRFT. MiR-24-3p was significantly upregulated in the perirenal fat of PA and was associated with increased adipogenesis, inflammation, and oxidative stress, correlating with plasma aldosterone concentration (PAC), PRFT, cardiac remodeling, and weight gain. The IL-6 level in the peripheral blood was elevated in patients with PA and CVD, and the affected adrenal vein had the highest IL-6 level. Targeting Top1, miR-24-3p modulated aldosterone-induced effects in adipocytes and influenced IL-6 secretion, thereby affecting HUVEC.ConclusionThe upregulation of miR-24-3p in the perirenal fat induced inflammation and oxidative stress by targeting Top1, which may contribute to a high risk of CVD in patients with PA.

Assessment of Immune Cell Populations in the Peripheral Blood of Patients With Metastatic Prostate Cancer.

Background Prostate cancer (PCa) encompasses a heterogeneous spectrum, ranging from indolent to highly aggressive forms, with approximately 10-20% of patients with initially localized disease later becoming metastatic. Oligometastatic PCa (OMPC) represents an intermediate state between locally advanced and high-volume metastatic disease. Understanding the immune landscape of OMPC and plurimetastatic PCa (PMPC) can provide valuable insights into disease biology, with potential implications for treatment strategies and prognosis. Aim and objective This study aimed to evaluate alterations in circulating immune cell subsets between OMPC and PMPC to identify potential immune biomarkers and therapeutic targets. Methods We conducted a retrospective cohort study of 43 mPC patients. Patients were stratified into two groups based on metastatic spread: OMPC (≤5 metastatic lesions in bone or lymph nodes) and PMPC (>5 lesions and/or visceral involvement). Peripheral blood mononuclear cells (PBMCs) were isolated and analyzed via flow cytometry for key immune subsets, including γδ T cells, αβ T cells, and regulatory T cells, with functional assessments performed using cytokine stimulation. Statistical analysis used the Mann-Whitney-Wilcoxon test, with p ≤ 0.05 considered significant. Results OMPC patients exhibited significantly increased γδ2+ T cells compared to PMPC, suggesting enhanced immune surveillance in low metastatic burden. A trend toward elevated γδ2+ T cells expressing interferon-gamma (IFN-γ) was observed in PMPC. No significant differences were observed in other immune subsets. Conclusions γδ2+ T cells represent a distinct immune subset in PCa, potentially influencing disease progression. Despite the small sample size, these findings highlight γδ2+ T cells as promising biomarkers and therapeutic targets. Prospective studies with a larger sample size are warranted to confirm the significance of these findings and explore the mechanistic roles of these cells and possible clinical applications in metastatic PCa (mPC).

Aberrant Inflammatory Cells of Peripheral Blood in Nasal Inverted Papilloma

Introduction: Nasal inverted papilloma (NIP) is a common nasal benign tumor, with a complex pathogenesis closely linked to inflammatory response. It can be classified into Grade-I and Grade-II phenotypes based on nasal epithelium abnormality. Our objective is to investigate potential inflammatory biomarkers in peripheral blood of NIP patients for preoperative assessment of disease severity. Methods: We retrospectively analyzed inflammatory cells in the peripheral blood of 63 healthy controls and 82 NIP patients, while distinguishing different phenotypes of NIP through immunofluorescent and hematoxylin and eosin staining. Using receiver operating characteristic curve analysis, we assessed the predictive value of the monocytes%-to-lymphocytes% ratio (M%L%R) for peripheral blood abnormalities in NIP and explored its correlation with clinical data by Spearman r characteristic. Results: We observed an increase in monocytes% and decrease in lymphocytes% proportion, while the level of M%L%R was upregulated in the peripheral blood of NIP patients (all p < 0.05). Immunofluorescent staining revealed basal cell proliferation and squamous metaplasia, along with inhibited differentiation of ciliated and goblet cells in the nasal epithelium of NIP. Correlation analysis demonstrated a positive correlation between M%L%R and degree of nasal epithelial deterioration (r = 0.2999, p = 0.0062). Finally, M%L%R level was significantly increased in both Grade-I and Grade-II patients (all p < 0.05). Conclusion: The expression level of M%L%R in the peripheral blood of patients with NIP can serve as a potential biomarker for preoperative evaluation of the severity of nasal epithelial lesions.

Delineation of monocytic and early-stage myeloid-derived suppressor cells in the peripheral blood of patients with hepatocarcinoma.

In patients with hepatocellular carcinoma (HCC), increased myeloid-derived suppressor cells (MDSC) relate to aggressiveness and poor prognosis. Favorable responses with immune checkpoint inhibitors demonstrate that HCC is susceptible to immune activation, suggesting that the elimination of MDSC would provide therapeutic benefits. However, a global analysis of the different MDSC subsets in HCC is still missing. Here we phenotyped circulating myeloid cell subsets (monocytes, M-MDSC, PMN-MDSC and eMDSC) by flow cytometry in HCC and hepatocholangiocarcinoma patients and in healthy donors (HD). Isolated myeloid CD33+ cells were analyzed in immunosuppression assays, and cytokines were quantified in the supernatants. Arginase-1 activity (Arg-1) was analyzed in serum samples. All three proportions of MDSC, together with the immunosuppressive Arg-1, were significantly increased in HCC compared with HD. An important proportion of eMDSC expressed CD25, the IL-2 receptor α chain, and CD25+ eMDSC were also significantly expanded in HCC patients. HCC-CD33+ cells, enriched in M-MDSC and eMDSC, in vitro inhibited both CD4+ and CD8+ T cell proliferation and IL-2 production, and augmented IL-10, IL-6, and TNF-α. The correlation between the inhibition of T lymphocyte proliferation and M-MDSC was the strongest, while eMDSC or CD25+ eMDSC did not show antiproliferative capacity. Despite this functional difference, M-MDSC, CD25+ eMDSC, and CD25 expression in eMDSC were more prominent in advanced HCC as defined by a higher number of nodules, TNM stage, and alpha-fetoprotein level. This better delineation of M-MDSC and eMDSC phenotype and function in HCC could help to design therapies more likely to succeed in clinical trials.

Lymphocyte phenotyping in untreated children patients with chronic allergic asthma

This study aimed to isolate and phenotype lymphocytes in untreated children patients with chronic allergic asthma. To reach such aim the study involved (25) patients from children (17 male and 9 female) whom their ages where between (3-10) years, in addition to (15) apparently healthy children (9 male and 6 female) in the same ages involved as control group. The data demonstrated that there was a significant increase in the mean percentages of T-lymphocytes (CD3+ cells) in the peripheral blood of patients (66.75±0.29)**, in comparison with control group (43.58±0.19), a significant increase in the mean percentages of T-helper lymphocytes (CD4+ cells) in the peripheral blood of patients (51.14±0.55), in comparison with control group (39.17±0.23) and the mean percentages of B-lymphocytes (CD20+ cells) was also increased significantly in the peripheral blood of patients (29.63±0.20) when it compared with the mean percentages of the same cells in control group (18.60±0.80). Besides a significant decrease in the mean percentages of T-suppressor lymphocytes (CD8+ cells) in the peripheral blood of patients (11.31±0.05), in comparison with control group (16.42±0.15). Finally the results of this study showed a significant increase in the mean percentages of the ratio of (CD4+ cells/CD8+ cells) in the peripheral blood of patients (55.34±0.41), in comparison with control group (31.25±0.09).