Immunologic Research最新文献

We present a series of preclinical studies focused on developing in vitro 2D and 3D models for assessing immunogenic factors in preventing infectious diseases. Human peripheral blood mononuclear cells (PBMC) and Calu-3 cell lines (bronchial epithelial cells) were used to develop 2D and 3D models. Peptides: Spike-S1-His (S-His), nucleocapsid-His and adjuvants: human adenovirus five serotype-based viral vector (AdV-D24-ICOSL-CD40L), armed with inducible co-stimulator (ICOSL) and CD40 ligand (CD40L), and a vector lacking these transgenes (AdV5/3) were used due to their effective initial interaction with antigen-presenting cells (APC). Studying the potency of biologics in vitro revealed a significant increase in the percentage of CD4⁺ TCM, CD4⁺ TEMRA, and CD4⁺ TEM lymphocyte subpopulations involved in memory cell generation after 24 h of treatment. Prolonging the exposure for 7 days, a significant increase in CD4⁺ cells was observed when PBMCs were treated with AdV1 (56.00 ± 0.26% vs. 48.17 ± 1.10%). In contrast, a decrease in CD8⁺ cells was observed in those treated with AdV1 (37.93 ± 0.35%) compared to AdV1 + S-His + N-His (38.47 ± 0.38%) versus the untreated group (44.63 ± 1.07%). A decrease in EMRA was noted when PBMCs were treated with AdV1 + S-His + N-His (2.97 ± 0.23% vs. 4.50 ± 0.35%). Moreover, it was pointed out that PBMCs treated with AdV1 alone or in combination with S-His and N-His showed an elevated number of naïve CD4⁺/CD8⁺ and SCM CD4⁺/CD8⁺ cells. No changes in the number of EMRA CD4⁺ subpopulations were detected when PBMCs were treated with AdV2 compared with untreated ones (4.27 ± 0.06% vs. 4.50 ± 0.35%). Analysis of the humoral response induced by AdV1, AdV2, S-His, N-His, AdV1 + S-His + N-His, and AdV2 + S-His + N-His showed that AdV1 alone (4.17 ± 0.25% vs. 3.17 ± 0.06%) and in combination with S-His and N-His (3.87 ± 0.25 vs. 3.17 ± 0.06%) slightly increased the number of CD19⁺ cells. RNA-Seq analysis of PBMC cells in the 3D model revealed gene overexpression, including FGFR4, associated with the Rap1 pathway in samples exposed to AdV1 + S-His + N-His. Thus, the proposed platform's impact on lymphocyte differentiation was confirmed, and cytokine profile analysis in this sample revealed elevated levels of IL-10, IL-12p70, and IL-8. All samples exposed to AdV showed increased levels of IFN-γ. The safety and biodistribution studies of the vaccine platform demonstrated that a 30-day exposure did not impact mice's survival or organ morphology. Exploring the CD40 pathway notably reveals its significant impact on immune cell populations, suggesting potential therapeutic avenues.

Since the beginning of the COVID-19 pandemic, various groups around the world have intensively worked in the development of vaccine candidates against SARS-CoV-2. Several vaccines have been approved in the past years; the majority is based on the Spike or RBD proteins and employs parenteral administration routes. Considering the recent history of coronavirus zoonotic events, which are known to have caused serious human health problems, the development of vaccines with a broad scope of protection and the potential to cut/reduce the transmission remains in the spotlight. The current global pandemic preparedness initiatives have also promoted the preclinical evaluation of a new group of coronavirus vaccines. In line with current needs, the goal of the present work is the preclinical evaluation, in two different mice strains, of a novel nasal vaccine candidate based on two highly conserved sarbecovirus proteins, S2 and nucleocapsid (N). The vaccine preparation, containing a CpG ODN as adjuvant, was able to generate high antibody titers against both antigens, in sera and bronchoalveolar lavages. This humoral response results cross-reactive to SARS-CoV-1 and MERS-CoV. In addition, the preparation induces IFNγ secretion, and a marked IgG2a response, against both proteins at the systemic compartment, consistent with the development of a Th1 pattern. Although further evaluations should be done, the level of cross-reactivity and the mucosal response obtained constitute promising features of this vaccine candidate.

Cell-free DNA (cfDNA) is emerging as a promising biomarker in liquid biopsy applications for cancer, with growing interest in its potential utility also for the diagnosis, monitoring of treatment response, and detection of relapse in hematologic malignancies. However, the precise origin and clinical relevance of cfDNA in these disorders remain to be fully clarified. In this study, we analyzed plasma samples from 98 patients with hematologic malignancies and 80 healthy donors using quantitative PCR (qPCR) to quantify cfDNA concentrations. We further examined associations between cfDNA levels and clinical parameters, including sex and measurable residual disease (MRD). Our results demonstrated significantly elevated cfDNA levels in patients compared to healthy individuals, with a strong correlation between cfDNA concentration and presence/MRD of the disease. We also identified sex-specific differences in cfDNA levels. Notably, our findings suggest that neutrophils, through the release and degradation of neutrophil extracellular traps (NETs), may constitute a relevant source of circulating cfDNA. In addition, cfDNA concentrations were significantly associated with MRD status, supporting the potential role of cfDNA as a non-invasive biomarker for disease monitoring. Overall, our data highlight the clinical relevance of cfDNA in hematologic malignancies, both as a surrogate for disease burden and as a possible player in disease pathophysiology, thus offering a promising avenue for improved diagnostic and therapeutic strategies.

Psoriasis is one of the most common chronic inflammatory skin diseases and is characterised by the uncontrolled proliferation of keratinocytes and their abnormal differentiation. Sirtuins are a group of enzymes that play an important role in post-translational modifications of proteins, such as deacetylation, poly-ADP-ribosylation, demalonylation and lipoamidation. They are found in various cell types and are involved in ribosomal DNA recombination, gene silencing and DNA repair. This study aimed to examine the plasma levels of sirtuin 3, 4 and 5 in patients with psoriasis and correlate these levels with clinical parameters. The study included 43 patients with plaque-type psoriasis and 28 healthy controls. The plasma concentrations of sirtuin 3 were statistically significantly increased in patients with psoriasis compared to the control subjects. The plasma concentrations of sirtuin 4 and 5 were statistically significantly lower in patients with psoriasis than in the control group. No statistically significant correlations were found between the plasma levels of sirtuin 3 and 4 and the psoriasis activity tools of PASI, DLQI and the BSA index or the selected clinical parameters in patients with psoriasis. Plasma concentrations of sirtuin 5 correlated statistically significantly with the BSA index, haemoglobin and leukocytes. The results of this study suggest the involvement of sirtuin 3, 4 and 5 in the pathogenesis of psoriasis. However, an explanation of the role of sirtuins in psoriasis requires further research.

Single-cell transcriptome analysis has made outstanding contributions to the identification of new cell lineages and the study of cancer immune microenvironment. Yet, the characterization of human liver type 1 innate lymphoid cells (ILC1s) and their dynamic changes in the tumor microenvironment have not been thoroughly studied at this detailed level. Here, we performed an integrated analysis of mouse and human liver immune cells to identify human liver ILC1s based on identified mouse liver ILC1s and to verify its functional similarity. Additionally, our findings highlighted the different expression patterns of the transcription factor EOMES in human versus mouse liver ILC1s, suggesting its reduced regulatory significance in human liver nature killer (NK) cells and ILC1s compared to murine models. A unique subset of intermediate innate lymphoid cells (intILCs) was identified, exhibiting traits of both human liver NK cells and ILC1s. Single-cell RNA sequencing (scRNA-seq) data analysis and TCGA dataset were utilized to characterize the distinct alterations in the genes and functions of NK cells, ILC1s, and intILCs in human hepatocellular carcinoma (HCC). It was found that the dynamic changes of liver ILC1s and intILCs, along with some of their subpopulations, may be key factors in tumor progression. This study provided new insights into the identification of ILC1s in human liver and the immunologic changes and mechanism of innate lymphoid cells (ILCs) in the tumor microenvironment, and these findings may be applicable to improving the diagnosis and treatment of hepatocellular carcinoma.

The field of epigenetics has significantly advanced our understanding of gene regulation in cancer, revealing dynamic modifications that do not alter the DNA sequence yet profoundly influence gene expression. These include DNA methylation, histone modifications, non-coding RNAs, chromatin remodeling, and RNA modifications. In malignancies such as colorectal, breast, lung, glioblastoma, and hematologic cancers, these epigenetic alterations contribute to tumor initiation, progression, and immune evasion. Emerging evidence reveals that such modifications shape the tumor-immune interface by influencing antigen presentation, immune cell infiltration, and cytokine signaling. This review explores the interplay between key epigenetic modifications and cancer immunity, emphasizing how these mechanisms contribute to immune escape and therapeutic resistance. We also examine the emergence of epigenetic therapies-particularly DNMT inhibitors, HDAC inhibitors, and BET inhibitors-as promising tools to reprogram immune responses and restore anti-tumor immunity. Furthermore, we discuss combinatorial approaches integrating epigenetic modulators with immune checkpoint inhibitors, underscoring their potential to enhance treatment efficacy. By outlining current challenges and emerging strategies, this review underscores the need for personalized epigenetic interventions and biomarker-driven approaches to improve outcomes in cancer immunotherapy. These insights establish epigenetic regulation as a critical frontier in next-generation cancer immunotherapy.

Current therapies for Systemic lupus erythematosus (SLE), such as immunosuppressants and glucocorticoids, are associated with significant side effects, necessitating alternative treatment approaches. Extracellular vesicles (EVs) have emerged as a potential therapeutic option due to their immunomodulatory properties and ability to regulate innate and adaptive immune responses. Thus, the objective of the present study was to systematically analyze the therapeutic potential of EVs for treating SLE. A systematic review was conducted according to PRISMA guidelines. An electronic search was performed in the PubMed/ MEDLINE, Scopus, Cochrane Library and Open Gray databases covering the period up to September 2024 to respond to the PICO answer "Would EVs have therapeutic potential in SLE?" Control of Systemic lupus erythematosus progression and the cellular and molecular mechanisms present were considered the primary and secondary outcome, respectively. The bias risk of studies was evaluated by SYRCLE's RoB. A total of 7 studies met the inclusion criteria, and the data exhibited that EVs reduced disease severity, improved survival rates and ameliorated organ-specific damage in SLE models. It was seen that EVs reduction of autoantibody, Th17 and Tfh cells, type-1 macrophage, neutrophils and pro-inflammatory cytokines, alongside an increase in Tregs, immunosuppressive cytokines and type-2 macrophage. The studies showed high scientific evidence. EV therapy exhibits potential in mitigating SLE progression by modulating immune responses and reducing inflammation. Further research is required to confirm these datas in humans, facilitating advancement for future clinical applications.

Different mechanisms are involved in migraine pathogenesis, including neurogenic inflammation, neurodegenerative processes, and a potential role of microglia. The aim of this study was to assess axonal and glial damage measuring serum levels of neurofilament light chain (NfL) and glial fibrillary acidic protein (GFAP) in migraine patients. Serum samples of 25 patients with episodic migraine (EM), 25 with chronic migraine (CM) diagnosed in accordance with the International Classification of Headache Disorders, 3rd edition (ICHD-3), and 50 age-matched healthy controls were prospectively collected. NfL and GFAP levels were assessed using ultrasensitive paramagnetic bead-based ELISA (SIMOA). Non-parametric tests were used for group comparison and 2-tailed Spearman analysis to assess correlations. GFAP levels were significantly increased in migraine patients (median 103.15 pg/mL [IQR 70.98-146.34] vs. 69.43 pg/mL [IQR 53.04-91.85], p < 0.001), particularly in those with medication overuse (106.08 [IQR 87.94-159.07] vs. 71.38 [IQR 54.16-135.06], p = 0.007), without difference between EM and CM (p = 0.985). Although NfL levels were not increased (p = 0.387), they were higher in patients with a long migraine course (rho 0.519, p < 0.001). Attack at sampling/days from last attack, migraine frequency/attack severity did not influence NfL or GFAP levels. Our findings demonstrate the occurrence of glial damage, particularly correlated with medication overuse, and the presence of axonal damage in the later disease stage, providing potential novel cues for the migraine pathogenesis.

A newly identified cell subset within CD8⁺ T cells expressing CXCR5 of follicular cytotoxic T cells (T_FC) lyse the infected or tumor cells. Recent studies have suggested that some T_FC cell subsets may be involved in the regulation of antibody responses. We aimed to determine the subset of T_FC which differentiates in patients diagnosed with chronic lymphocytic leukemia (CLL) and their role in CLL immunopathogenesis. The peripheral blood mononuclear cells were isolated from 29 CLL patients and 19 healthy subjects. Intracellular IL-4, IL-17, IL-21, IFN-γ, perforin, and granzyme-B levels were investigated in T_FC subsets. Increased levels of IL-4, IL-17, IL-21, IFN-γ and perforin, and decreased granzyme B expression levels were observed in CD40L⁺ T_FC cells compared to the levels in CD40L^- T_FC cells in the analysis of healthy individuals. T_FC and its subsets were analyzed in CLL patients and healthy individuals, T_FC and CD40L⁺ T_FC cells were increased in CLL patients and there was a positive correlation between T_FC and CD5⁺CD19⁺ cells. Moreover, increased number of T_FC cells were detected in CLL patients with more progressive disease. Higher expression level of IL-4, IL-17, IL-21, and IFN-γ was observed in CD40L⁺ T_FC cells of patients compared to the levels in healthy controls. Our findings might indicate that CD40L⁺ T_FC cells may have a B cell activating role rather than exhibiting a cytotoxic role. Considering the effects of CD40L⁺ T_FC cells on B cells, determining subsets of T_FC cells which differentiate and understanding the functions of these subsets is crucial to elucidate their roles in the pathogenesis of B cell malignancies.

Immunoglobulin G4-related disease (IgG4-RD) is a rare immune-mediated fibro-inflammatory condition that can affect nearly any organ, with breast involvement (IgG4-related sclerosing mastitis, IgG4-RM) being exceptionally uncommon. This disease primarily affects middle-aged to elderly women and is frequently misdiagnosed as malignancy due to non-specific clinical and imaging features. We report a case of a 44-year-old woman with a painless right breast mass showing hypoechoic ultrasound findings and magnetic resonance imaging (MRI) characteristics, including a high ADC value and homogeneous enhancement with a persistent time-intensity curve (TIC). Histopathology confirmed IgG4-RM, with no recurrence after 12 months. A review of 24 published lectures highlights the diagnostic challenges and imaging variability of IgG4-RM.