Background:Streptococcus pneumoniae remains a leading cause of community-acquired pneumonia. Severe pulmonary inflammation and subsequent tissue damage are driven significantly by the overexpression of both the Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-κB) and Mitogen-Activated Protein Kinase (MAPK) signaling pathways. Objective:To investigate the immunomodulatory effects of rhein in a mouse model of Streptococcus pneumoniae-induced pneumonia. Methods:Forty male C57BL/6 mice were randomly assigned to four experimental groups: healthy control, S. pneumoniae infected control, and two rhein-treatment groups (10 mg/kg and 30 mg/kg). Expression levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, IL-10, transforming growth factor (TGF)-β, monocyte chemoattractant protein-1 (MCP-1), cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) in lung tissues and bronchoalveolar lavage fluid (BALF) were quantified using reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). Additionally, ELISA and Western blotting were employed to assess the phosphorylation profiles of MAPK and NF-κB. Results:Rhein significantly decreased the lung wet/dry weight ratio, reduced bacterial load, and downregulated COX-2 and iNOS expression. Cytokine analysis revealed marked reductions in pro-inflammatory mediators (IL-6, IL-1β, TNF-α, and MCP-1) alongside elevated levels of the anti-inflammatory cytokine IL-10 in both lung tissues and BALF. Furthermore, ELISA and Western blotting confirmed that rhein significantly inhibited the phosphorylation of MAPK and NF-κB. Conclusion:Rhein attenuates S. pneumoniae-induced pulmonary inflammation and injury by modulating the MAPK/NF-κB signaling pathways. These findings highlight Rhein as a promising potential therapeutic agent for pneumococcal pneumonia and related inflammatory lung diseases.
Background:Immunity protects organisms from pathogens, diseases, and external noxious agents. Naturally derived polysaccharides are well-documented to exhibit potent immunomodulatory effects. However, the exact immunostimulatory properties and underlying mechanisms of polysaccharides derived from Momordica charantia (MCPS) remain to be clearly elucidated. Objective:To investigate the immunomodulatory effects of MCPS on murine macrophages and to elucidate the underlying molecular mechanisms. Methods:MCPS was isolated from Momordica charantia. RAW 264.7 murine macrophages were treated with varying concentrations of MCPS, and its effects on cell viability, nitric oxide (NO) production, cytokine secretion, surface marker expression, and key intracellular signaling pathways were evaluated using standardized analytical assays. Potential endotoxin contamination was systematically excluded using a polymyxin B neutralization assay. Results:MCPS exhibited no cytotoxicity across all tested concentrations. Notably, MCPS significantly stimulated NO production and enhanced the secretion of pro-inflammatory cytokines, indicative of robust macrophage activation. This was accompanied by a marked upregulation of macrophage surface activation markers. Furthermore, MCPS activated the MAPK and NF-κB signaling pathways. This cytokine production was significantly attenuated by pathway-specific inhibitors, confirming the functional involvement of these cascades. Importantly, polymyxin B suppressed LPS-induced responses but did not alter MCPS- stimulated activation, indicating that the immunomodulatory effects were independent of endotoxin contamination. Conclusion:MCPS effectively promotes murine macrophage activation without cytotoxicity by stimulating NO production, cytokine secretion, and activation marker expression through the MAPK and NF-κB signaling pathways. These findings demonstrate that MCPS holds strong potential as a natural immunostimulatory agent capable of enhancing host immune responses.
Background:Natural Killer (NK) cells are innate lymphoid cells that eliminate malignant cells via perforin/granzyme-mediated cytotoxicity. This study investigates whether Olive Leaf Extract (OLE), rich in oleuropein and hydroxytyrosol, can enhance NK-cell cytotoxicity against colorectal cancer (CRC) cells. Although OLE exhibits direct anticancer effects, its therapeutic utility is constrained by poor bioavailability, requiring supraphysiological concentrations for direct cytotoxicity. Objective:To evaluate the effect of OLE on the cytotoxic activity of NK-92 cells against HT-29 colorectal cancer cells within a co-culture model. Methods:Olive leaves obtained from Balıkesir, Türkiye were dried and extracted with 70% methanol. The effects of OLE on the viability of HT-29 cells and NK-92 cells were evaluated using MTT and ATP assays. Additionally, the cytotoxic activity of NK-92 cells against HT-29 cells was assessed in a direct co-culture system. Granzyme B and perforin levels were measured using ELISA kits. Results:OLE inhibited the proliferation of HT-29 cells in a dose-dependent manner, with an IC₅₀ values of 548 µg/mL. In NK-92 cells, low concentrations of OLE (100-200 µg/mL) promoted cell proliferation, whereas higher concentrations exerted cytotoxic effects. In co-culture experiments, NK-92-mediated cytotoxicity against HT-29 cells was significantly enhanced by the addition of OLE at non-toxic concentrations (100 and 200 µg/mL). This enhanced cytotoxicity was further supported by a significant increase in granzyme B and perforin levels following OLE treatment. Conclusion:Our findings suggest that OLE can elicit a potent anticancer response via NK cells at lower, physiologically achievable doses. These results highlight a promising therapeutic strategy for CRC, leveraging OLE's immunomodulatory effects to enhance innate antitumor defenses.
Background:Patients with advanced non-small cell lung cancer (NSCLC) often experience poor clinical outcomes following chemotherapy. Heat-sensitive moxibustion (HSM), a traditional therapeutic intervention, has been reported to improve symptoms and quality of life in patients with cancer. Objective:To evaluate the effect of HSM in patients with advanced NSCLC following chemotherapy. Methods:Between January 2023 and February 2024, patients with advanced NSCLC were enrolled. Eligible patients were randomly assigned to either the chemotherapy (control) group or the chemotherapy plus HSM treatment (HSM) group. Following treatment, the percentages of CD4+ T cell subsets were evaluated, and the serum levels of CD4+ T cells-associated cytokines were measured using enzyme-linked immunosorbent assay (ELISA). The expression levels of CD4+ T cells-related factors were assessed by RT-qPCR and Western blot analysis. The overall response rate (OPR) and disease control rate (DCR), Karnofsky Performance Status (KPS) score, and incidence of adverse events were compared between the two groups. Results:A total of 90 patients with NSCLC were included with 45 patients in each group. The ratio of Th1/Th2 was significantly elevated, whereas the Treg/Th17 ratio was significantly decreased in the HSM group compared with the control group. The HSM group exhibited higher serum levels of IL-2, IL-17A and IFN-γ, and lower levels of IL-4 and IL-10 than the control group. HSM treatment significantly upregulated the expression of T-bet and IL-17A while downregulating GATA3 expression at both the mRNA and protein levels. The HSM group demonstrated superior clinical outcomes, as evidenced by higher KPS score, ORR, DCR, along with a lower incidence of adverse events. Conclusion:HSM may enhance immune function in patients with advanced NSCLC undergoing chemotherapy by modulating the balance of Th1/Th2 and Treg/Th17 cell subsets.
Background:Toxoplasma gondii is an obligate intracellular protozoan parasite with a unique global prevalence and is the causative agent of toxoplasmosis. MicroRNAs are key epigenetic elements crucial that modulate the immune response by influencing cytokine expression. Objective:To investigate serum levels of IL-37 alongside the expression of microRNAs miR-604 and miR-1302-3p in Iraqi patients with toxoplasmosis, and to evaluate their potential correlations and regulatory relationships. Methods:A total of 200 non-pregnant women were enrolled in the present study, consisting of 100 toxoplasmosis- positive and 100 healthy controls. Toxoplasmosis status was determined via serological screening using a rapid diagnostic test (TORCH) and ELISA for IgM, IgG antibodies. For the molecular analysis, total RNA was extracted from whole blood samples and reverse-transcribed into cDNA. Quantitative real-time PCR (RT-qPCR) was then performed on both groups to quantify the expression levels of microRNA-604 and microRNA-1302-3p. Results:The expression levels of both microRNA-604 and microRNA 1302-3p were significantly reduced in toxoplasmosis patients compared to the control group in patients (relative expression: 2.1896 ± 1.12221 ng/L vs. 1.6384 ± 1.09466 for microRNA-604; 4.0896 ± 1.42896vs. 2.5764 ± 1.16224 for microRNA 1302-3p). ). Conversely, serum levels of IL-37 were significantly higher in the patient group (295.0604 ± 45.79983 ng/L) compared to the control group (36.7183 ± 3.83299 ng/L). Conclusion:Our findings demonstrate that miRNA-604 and 1302-3p are downregulated in toxoplasmosis patients, a state associated with the marked induction of pro-inflammatory cytokine IL-37. These altered expression profiles suggest a coordinated role in the pathogenesis of toxoplasmosis, highlighting their potential utility as novel diagnostic biomarkers or therapeutic targets.
Background:Eosinophilic colitis (EC) is a rare eosinophilic gastrointestinal disorder with increasing frequency in recent years. Although normal eosinophil density varies substantially by colonic segment, several proposed eosinophil thresholds have been used in the literature to support the diagnosis in the appropriate clinical context. However, some patients with clinical features suggestive of eosinophilic colitis (EC) do not demonstrate increased tissue eosinophil counts on histopathologic examination. Objective:To compare eosinophil-derived cell-free granules based on expression of their surface markers between two groups of patients with symptoms consistent with EC: those with increased tissue eosinophils and those without tissue eosinophilia. Methods:This study included 10 pediatric patients with histologically confirmed EC and 13 patients with clinical suspicion of EC. Eosinophil-derived cell-free granules were evaluated in colonic tissue samples using immunohistochemistry (IHC), with antibodies against MBP and CCR3. Results:Eosinophil-derived cell-free granules were detected in all specimens from patients with suspected EC and in the majority of specimens from patients with confirmed EC. Degranulating eosinophils were identified in all specimens from both groups, although the proportion of degranulating eosinophils varied among patients. Furthermore, the abundance of granules was significantly associated with the percentage of degranulating eosinophils. Conclusion:Our findings indicate that IHC can be used to detect eosinophil-derived granules in colonic tissue. The presence of cell-free eosinophil granules with varying abundance, along with evidence of eosinophil degranulation in all specimens from patients with suspected EC, may provide supportive histopathological features that contribute to improving the diagnostic assessment of EC.
Type 1 diabetes (T1DM) is an autoimmune disorder marked by a characteristic dysfunction of insulin-secreting beta-cells in the pancreatic islets. This destructive process is not a single event but a chronic inflammatory cascade, propelled by immune cells-including T-lymphocytes and macrophages-that release potent pro-inflammatory cytokines. This review delves into the growing body of evidence implicating specific inflammation-promoting mediators-including Interleukin-1β (IL-1β), Tumor Necrosis Factor-α (TNF-α) and Interferon-γ (IFN-γ) -as master regulators of beta-cell death. We also explore how modern analytical techniques enable precise mapping of cytokine patterns in the blood, revealing a dynamic and ongoing disease process long that precedes clinical symptoms onset. By weaving together findings from clinical and preclinical studies, this article argues that profiling these inflammatory signals provides a crucial real-time, functional readout of autoimmune activity that complements traditional, static autoantibody measurements. We conclude by discussing the significant potential of integrating cytokine data into existing models to create more robust risk stratification tools and to pave the way for interventions that could intercept the disease pathway before critical beta-cell mass is lost.
Background: Rheumatoid arthritis (RA) and ankylosing spondylitis (AS) are chronic autoimmune inflammatory disorders in which TNF-alpha plays a keyl role. Anti-TNF agents are widely used in the management of these diseases. Since TNF-alpha is also involved in the pathogenesis of autoimmune thyroid disease (AITD), this study aimed to assess whether anti-TNF therapy influences thyroid function. Methods: This prospective study included 50 RA and 51 AS patients aged 18-85 years. Patients received either conventional treatment (DMARDs and NSAIDs/sulfasalazine) or anti-TNF agents. Serum TSH, free T4, and anti-TPO levels were measured at baseline and after 6 months. Thyroid dysfunction and AITD prevalence were compared between two treatment groups. Results: In the RA group, subclinical hyperthyroidism was observed in both arms; In the anti-TNF group, one patient had hypothyroidism and another had subclinical hypothyroidism. Improvement in subclinical hyperthyroidism was seen in one patient in the DMARD arm, while improvement occurred in the patient with hypothyroidism and in one patient with subclinical hyperthyroidism in the anti-TNF arm(p>0.05). In the AS group, central hyperthyroidism developed in one patient receiving conventional treatment. In anti-TNF group one patient with subclinical hypothyroidism improved to normal values, while another developed central hypothyroidism. Anti-TPO positivity was 18% in the conventional group and 3.4% in the anti-TNF group (p>0.05). A significant TSH change was observed only in the RA-DMARD group (p<0.05);While no significant changes in free T4 were detected in any group. Conclusion: Anti-TNF therapy showed no significant effect on thyroid function or autoimmune thyroid disease in patients with RA and AS during the six-month follow-up.
Background:Dendritic cells (DCs) are the most potent antigen-presenting cells, playing a central role in T cell activation and the stimulation of B cell antibody production. Owing to their immunological significance, DCs have been extensively explored for applications in cancer immunotherapy, dendritic cell-based vaccines, and strategies to overcome the immunosuppressive tumor microenvironment. Objective:Given the broad applications of DCs and the need for efficient generation protocols, the present study aimed to optimize key parameters influencing the production of bone marrow-derived dendritic cells (BMDCs). Methods:Variables such as mouse strain, cytokine concentrations, culture plate type, differentiation duration, incubation temperature, cell seeding density, and media replacement intervals were evaluated. CD11c expression was used as the primary marker to quantify BMDCs, while CD80 and CD86 were used as indicators of maturation status. Results:The optimized protocol yielded 60-70% CD11c+ BMDCs and 70-80% CD80/CD86 expression within 5 days. Conclusion:This robust and highly reproducible protocol offers valuable applications for in vitro dendritic cell generation studies.
Background:Recurrent pregnancy loss (RPL) is a global health challenge in women. It has been reported that Toxoplasma gondii (T. gondii) infection and interleukin-6 (IL-6) bioavailability contribute to RPL. Objective:This study investigated IL-6 and IL-6 receptor (IL-6R) gene polymorphisms and blood IL-6 levels in T. gondii-infected women with RPL. Methods:The study comprised 163 women, including 83 infected (Group A: subgroups A1, A2) and 80 healthy (Group B). Demographic data, blood, and placental tissue samples were collected. The blood samples were screened for IL-6 levels through ELISA. DNA was extracted, and T. gondii-DNA was identified using PCR. IL-6 and IL-6R genes were amplified by PCR and sequenced using the Sanger method. Results:The IL-6 gene -174G/C (p=0.005) and -597G/A (p=0.0012) polymorphisms were significantly associated, while the -572G/C polymorphism was insignificantly associated (p=0.143) with infection and RPL. The mean blood IL-6 level was significantly variable across all groups (p < 0.001). The IL-6 gene -174GG and -597GG genotypes, and the IL-6R gene AA genotype, were common in Group-A2 (47.5%, 37.5%, 50%) and Group-B (56.2%, 60%, 68.8%), respectively, and were also significantly associated with elevated blood IL-6 levels. The -572GG genotype was common and was not significantly associated with elevated blood IL-6 levels across all groups (p=0.143). The IL-6 gene -174CC and -597AA genotypes and IL-6R gene CC genotype were highly prevalent in Group-A1 (48.8%, 41.9%, and 51.1%, respectively) and associated with low blood IL-6 levels. Conclusion:The IL-6 gene -174G/C, -597G/A, and IL-6R gene A/C polymorphisms play significant roles in RPL in women infected with T. gondii, and change IL-6 bioavailability.
Background:Miscarriage occurs when a pregnancy ends spontaneously before the fetus is viable outside the uterus. Early miscarriage, sometimes referred to as first-trimester miscarriage, happens before 12 weeks. Vascular endothelial growth factor (VEGF), an angiogenic protein essential for blood vessel formation, and cluster of differentiation 68 (CD68), a marker of macrophages, are vital to the immune response. Objective:This research aimed to evaluate VEGF and CD68 expression patterns in placental tissues and decidua from first-trimester miscarriage, to ascertain their potential roles in immune response and angiogenesis in miscarriage. Methods:The study included 60 women who experienced spontaneous abortion during the first trimester, 30 with missed abortion, and 30 with incomplete abortion. Using immunohistochemical staining, the expression of CD68 and VEGF in decidua and placental tissues was investigated. Expression levels were scored semi-quantitatively by averaging data from five fields per slide and subjected to statistical analysis. Results:Building on these methods, immunohistochemical staining confirmed significantly elevated CD68 expression in both placental and decidua tissues from missed and incomplete abortions, indicating enhanced macrophage infiltration. Widespread staining (>50%) was seen in 50% of incomplete abortion decidua and 30% of missed abortion tissues. In contrast, VEGF expression was predominantly negative across all samples, with no case showing positive staining, indicating impaired angiogenesis. Conclusion:The findings highlight a dual pathological mechanism in early miscarriage characterized by enhanced inflammatory macrophage infiltration and deficient angiogenic support. This imbalance may contribute to placental dysfunction and pregnancy failure. These results underscore the potential diagnostic and therapeutic relevance of immune-angiogenic pathways in early miscarriage.
Background:Rheumatoid arthritis (RA) and ankylosing spondylitis (AS) are chronic autoimmune inflammatory disorders in which TNF-α plays a key role. Anti-TNF agents are widely used in the management of these diseases. Objective:Since TNF-α is also involved in the pathogenesis of autoimmune thyroid disease (AITD), this study aimed to assess whether anti-TNF therapy influences thyroid function. Methods:This prospective study included 50 RA and 51 AS patients aged 18-85 years. Patients received either conventional treatment (DMARDs and NSAIDs/sulfasalazine) or anti-TNF agents. Serum TSH, free T4, and anti-TPO levels were measured at baseline and after 6 months. Thyroid dysfunction and AITD prevalence were compared between the two treatment groups. Results:In the RA group, subclinical hyperthyroidism was observed in both arms; in the anti-TNF group, one patient had hypothyroidism, and another had subclinical hypothyroidism. Improvement in subclinical hyperthyroidism was observed in one patient in the DMARD arm, in one patient with hypothyroidism, and in one patient with subclinical hyperthyroidism in the anti-TNF arm (p>0.05). In the AS group, central hyperthyroidism developed in one patient receiving conventional treatment. In the anti-TNF group, one patient with subclinical hypothyroidism improved to normal values, while another developed central hypothyroidism. Anti-TPO positivity was 18% in the conventional group and 3.4% in the anti-TNF group (p>0.05). A significant TSH change was observed only in the RA-DMARD group (p < 0.05), while no significant changes in free T4 were detected in any group. Conclusion:Anti-TNF therapy showed no significant effect on thyroid function or autoimmune thyroid disease in patients with RA and AS during a six-month follow-up.
Background:Autophagy genes are essential for proper uterine function, reproductive physiology, and the maintenance of endometrial atrophy (EA). Objective:This study aims to clarify how these genes impact endometrial thickness and their significance in the apoptosis of peripheral blood mononuclear cells (PBMCs) from patients with thin endometria. Methods:Blood samples were collected from 40 patients with a thin endometrium and 40 healthy controls, all in the non-pregnancy stage. Real-Time PCR was used to measure the expression levels of autophagy genes ATG5, ATG7, LC3B, Beclin1, FOXO1, FOXO3a, FOXO4, and FOXO6 in 40 women with thin endometrium and 40 healthy women. Apoptosis was also assessed by flow cytometry. To further elucidate the biological pathways and processes associated with the differentially expressed autophagy genes, we conducted a KEGG pathway enrichment analysis using the EnrichR tool. Results:Evaluation of autophagy gene expression showed a significant difference between the studied groups, with higher expression of ATG5, ATG7, LC3B, Beclin1, FOXO1, FOXO3a, FOXO4, and FOXO6 in the patient group. Moreover, there was a positive correlation between LC3B expression and the frequency of apoptotic cells in the studied patients. The EnrichR tool identified the enriched pathways: "Shigellosis," "FOXO signaling," "Fruptosis," and, to a lesser extent, "Longevity regulating pathway," "Autophagy," and "Mitophagy." Conclusion:Our results showed that autophagy genes associated with apoptosis in PBMCs may be involved in endometrial thinning in EA patients.
Background:Atherosclerosis is a chronic inflammatory, immune-mediated disease that is a leading cause of global mortality and disability. Coronary artery calcification (CAC) is a key predictor of the severity of coronary artery disease (CAD). Interleukin-38 (IL-38), a newly identified anti-inflammatory cytokine, may modulate inflammation and prevent atherosclerosis progression. Objective:This study aimed to evaluate the association between serum IL-38 levels and CAC severity among patients referred to the CT angiography unit at Razieh Firooz Hospital in Kerman City. Methods:In this cross-sectional study, 151 patients aged 50-70 years were evaluated. The mean age of the participants was 60.1 ± 6.9 years. CAC severity was determined using the Agatston scoring method and multi-detector CT scanners. Serum IL-38 levels were measured via enzyme-linked immunosorbent assay (ELISA). Statistical analyses were performed using an independent T-test and multivariable logistic regression. Results:Comparing serum IL-38 levels across CAC severity categories showed a statistically significant difference (P = 0.039). Mean serum IL-38 in patients with non-severe and severe calcification were 16.8 ± 5.5 pg/mL and 19.4 ± 4.9 pg/mL, respectively. However, in the multivariable regression analysis, adjusted for major risk factors including sex, age, diabetes, hypertension, and smoking, the association between serum IL-38 levels and CAC severity was not significant (P>0.05). In subgroup analyses, the significant association between IL-38 and CAC severity was observed only in older participants and in patients with established cardiovascular risk factors. Conclusion:Although serum IL-38 levels were higher in patients with severe CAC, this association did not remain significant after adjustment for major cardiovascular risk factors. Therefore, the observed elevation may reflect age- or risk-related inflammatory changes rather than a direct role of IL-38 in calcification. So, this relationship remains unclear. Further investigation is needed to clarify the potential context-dependent function of IL-38 in atherosclerosis progression.
Background:Cervical cancer progression is driven by immune evasion mechanisms, including PD-L1-mediated suppression of T cell activity. BATF, a transcription factor, has been linked to tumor immunity; cancer remains incompletely understood. Objective:This study investigates the BATF-STAT1-PD-L1 axis in tumor progression and immune regulation. Methods:BATF expression was analyzed in cervical cancer tissues and cell lines. Functional assays assessed the impact of BATF knockdown on proliferation, apoptosis, autophagy, and migration. STAT1 regulation was examined via chromatin immunoprecipitation and Western blot. PD-L1 expression was measured by flow cytometry. In vivo xenograft models were used to evaluate tumor growth and response to PD-L1 blockade. Results:BATF was upregulated in cervical cancer and promoted tumor growth, metastasis, and immune evasion. BATF knockdown suppressed proliferation, enhanced apoptosis and autophagy, and reduced migration. Mechanistically, BATF transcriptionally activated STAT1, which induced PD-L1 expression. BATF suppression enhanced CD8⁺ T cell infiltration and improved the efficacy of PD-L1 blockade in vivo. Conclusion:BATF promotes cervical cancer progression by modulating STAT1 and PD-L1. Targeting BATF may enhance anti-tumor immunity and improve immunotherapy outcomes.
Background:Exosomes (EXOs) are small vesicles derived from endosomes and secreted by most living cells including tumor cells. In recent years, these vesicles have been recognized as key mediators of intercellular communication, playing essential roles in the regulation and orchestration of diverse physiological and pathological processes within the organism. Objective:To further investigate hepatocellular carcinoma (HCC)-derived exosomes containing tumor-associated antigens and to evaluate their immunostimulatory capacity and antitumor effects using in vitro and in vivo approaches. Methods:Following isolation from tumor cells, exosomes were characterized and subsequently co-cultured with dendritic cells (DCs). The expression of surface molecules associated with DC maturation was then assessed using flow cytometry. A mouse liver cancer model was established and animals were randomly assigned to three groups: a negative control group (treated with PBS), an iDC group, and a DC-TEXs (tumor-derived exosomes) group. Tumor volume was monitored in all groups, with a focus on changes in immune cell populations and cytokine levels. Results:Our in vitro studies showed that Hepa1-6 cell-derived EXOs dose-dependently enhanced dendritic cell (DC) maturation, as evidenced by increased expression of surface MHC-II molecules, co-stimulatory markers (CD40, CD80, CD86), and the maturation marker CD83. In vivo studies using subcutaneous HCC mouse models demonstrated that TEX administration significantly alters the tumor immune microenvironment, mainly through increased T lymphocyte infiltration and proliferation. Conclusion:Our results suggest that TEXs can serve as endogenous immunotherapeutic agents by eliciting tumor-specific T lymphocyte responses through DC activation cascades. These findings provide novel insights into the therapeutic exploitation of tumor-derived vesicles for the treatment of hepatocellular carcinoma.
Background:Placental tissue contains a variety of tumor-associated antigens. Antigen-specific T cells generated by expanded dendritic cells (DCs) loaded with placental peptides have the potential to exert antitumor effects both in vitro and in vivo. Objective:To investigate the immunotherapeutic potential of placental peptides as a novel source of tumor-associated antigens (TAAs). We hypothesized that DCs, expanded and matured in vitro and pulsed with placental peptide extracts, can effectively prime antigen-specific T cells (ASTs) with robust cytotoxic activity against various tumor cell lines and in vivo tumor models. Methods:Mass spectrometry was used to identify tumor-related peptides within the placental extract. In vitro assays were employed to assess the expansion of DCs, their maturation following exposure to placental peptide preparations, and the subsequent activation of T cells. Results:The cytotoxic activity of ASTs was assessed and showed strong tumor cell killing across three cancer cell lines, U87MG (glioblastoma), SH-SY5Y (neuroblastoma), and MCF-7 (breast cancer). In a neuroblastoma animal model, ASTs treatment significantly reduced tumor proliferation, indicating substantial therapeutic potential in vivo. Conclusion:Our findings suggest that DCs pulsed with placental peptides can generate ASTs with potent antitumor activity in vitro and in vivo. Additional studies are needed to determine applicability across other tumor types, refine therapeutic parameters, and assess clinical potential.
Background:Proper invasion and migration of trophoblasts are critical for successful human pregnancy. Tumor necrosis factor-α (TNF-α) has two biologically active forms: the membrane-bound TNF-α (mTNF) and the soluble TNF-α (sTNF). Objective:To investigate the role of both forms of TNF-α in trophoblasts invasion and migration and to elucidate their underlying mechanisms. Methods:We exposed HTR-8/SVneo trophoblasts to exogenous mTNF or sTNF and evaluated their cellular behaviors. Proliferation was assessed using a Cell Counting Kit-8, while invasion and migration were assessed through Transwell assays. Additionally, we measured mRNA levels of matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinases-1 (TIMP-1), and plasminogen activator inhibitor-1 (PAI-1) using qRT-PCR. The expression of IκB-α was determined by western blot analysis. Results:Unlike sTNF, mTNF enhances the invasion and migration of trophoblasts. Mechanistic analysis showed that mTNF increased MMP-9 expression while decreasing TIMP-1 and PAI-1 expression, and it inhibited the activation of NF-κB signaling pathway in HTR-8/SVneo cells with or without lipopolysaccharide (LPS) treatment. Conclusion:This research uncovered a new function of mTNF in regulating trophoblast invasion and migration, offering a new approach to treating pregnancy-related diseases associated with inadequate trophoblast invasion.
Background:Tumor-draining lymph nodes play a pivotal role in orchestrating immune cell trafficking and initiating antitumor responses. Among immunoregulatory molecules, programmed cell death protein 1 (PD-1) has emerged as a central mediator in tumor-induced immunosuppression. Objective:To investigate the expression patterns of PD-1 and its ligands (PD-L1, PD-L2) in the tumor-draining lymph nodes of patients with breast cancer (BC). Methods:Lymph node samples were freshly collected from BC patients undergoing surgery. Mononuclear cells were isolated and analyzed by flow cytometry for surface markers CD45, PD-1, PD-L1, and PD-L2. Data were analyzed using FlowJo v10.8.1. Results:PD-1 was detected on 9.48 ± 5.19% of CD45+ cells, whereas PD-L1 and PD-L2 were expressed at lower levels (1.73 ± 0.85% and 1.68 ± 0.84%, respectively). Despite a significant reduction in the percentage of CD45+ lymphocytes, the frequencies of PD-1+ and PD-L2+ subsets were significantly elevated in patients with poorly-differentiated and advanced-stage tumors (P<0.05). Additionally, the frequency of PD-1+ lymphocytes was significantly higher in patients with the triple-negative tumors (P=0.014) and in those negative for estrogen and progesterone receptor (P=0.001). Conclusion:Elevated expression of PD-1 and its ligands in BC-draining lymph nodes is associated with adverse clinical features, suggesting their role in immune evasion. These findings along with higher frequency of PD-1+ lymphocyte in triple-negative patients may inform subtype-specific therapeutic strategies and predict responsiveness to PD-1/PD-Ls blockade therapies. Future studies should include functional analyses with broader immunophenotyping to further elucidate these mechanisms.
Keloid, as a skin fibrotic proliferative disorder, have a complex pathogenesis that remains incompletely understood. It is characterized by abnormal and excessive scar formation following skin injury. The occurrence and development of keloids are closely associated with immune dysregulation. Immune cells, such as T cells, macrophages, mast cells, and Langerhans cells, play crucial roles in the formation of keloids. These immune cells contribute to keloid initiation and progression through mechanisms including cytokine secretion, promotion of inflammatory responses, and regulation of fibroblast proliferation and collagen synthesis. With advances in immunological research, the roles of fibroblasts, keratinocytes and melanocytes in the immunological dysregulation underlying keloids have received increasing attention. This paper aims to review recent progress on the abnormal immunological regulation involving these three epidermal cell types, in order to provide new insights and theoretical foundations for the treatment of this disease.
