Iranian Journal of Immunology最新文献

Background: Antitumor-targeting drugs can stimulate dendritic cells (DCs) indirectly through the shedding of dying tumor cells as part of what is referred to as a "danger signal". Although chemotherapeutic agents have been shown to kill dendritic cells (DCs), the effects of low, non-cytotoxic doses on DC function have not been studied.

Objective: To investigate the impact of various concentrations of mitomycin C at low, non-cytotoxic doses on the maturation of DCs.

Methods: THP-1 monocytes were differentiated into immature dendritic cells using IL-4 and GM-CSF. HCT116 colorectal cancer cells were treated with mitomycin C at concentrations ranging from 10 to 80 nM and co-cultured with undifferentiated dendritic cells. The expression of co-stimulatory molecules (CD11c, CD80, CD83, CD86, HLA-DR, CD14) and IL-12p70 secretion were measured.

Results: Different dosages of Mitomycin C-treated HCT116 cells enhanced the maturation of dendritic cell markers (CD80, CD83, CD86, HLA-DR), but reduced CD14 levels (p< 0.01). While increasing the Mitomycin C dose to 80 nM further upregulated HLA-DR and CD86 expression, the release of IL-12 was highest a 50 nM concentration of mitomycin C (686.7 ± 125.7 pg/mL compared to 263.8 ± 4.8 pg/mL in controls; p < 0.05). IL-12 levels were not significantly increased even with 30 nM Mitomycin C.

Conclusion: Low concentrations of Mitomycin C contributed to an increase in dendritic cellmaturation and an increase in the expression of co-stimulatory molecules (CD80, CD86, CD83, and HLA-DR), along with the secretion of cytokines such as IL-12p70, IL-2, and GM-CSF.

Background: Systemic Lupus Erythematosus (SLE) is a chronic inflammatory disease that affects multiple organ systems. The resulting inflammation disrupts adipocyte metabolism, thereby altering the levels of adipokines.

Objective: To assess e serum levels of Asprosin and CTRP-6 as novel adipokines in SLE patients compared to controls, and their association with lipid profiles, oxidative stress markers, and clinical parameters.

Methods: This case-control study involved 41 SLE patients and 41 healthy controls. Serum CTRP-6 and Asprosin levels were measured using ELISA, while total antioxidant capacity (TAC) and malondialdehyde levels were measured using a colorimetric assay.

Results: The mean serum CTRP-6 level was significantly higher in individuals with SLE (1.08±0.32) compared to healthy subjects (0.82 0.21; P<0.001). Similarly, the serum level of Asprosin was elevated in patients with SLE (11.91 ± 3.09) compared to the control group (10.28 ± 2.09; P < 0.001). In contrast, the TAC level was lower in subjects with SLE than in healthy controls (0.18±0.23, 0.19 0.51 respectively; p<0.001). Additionally, the serum level of Asprosin was significantly reduced in SLE patients with nephritis (10.17 ± 3.55) compared to those without nephritis (12.4 ± 2.75; p = 0.005).

Conclusion: Elevated levels of Asprosin and CTRP-6 suggest a potential role for these adipokines in SLE pathogenesis. Moreover, the presence of nephritis in SLE patients was associated with reduced plasma levels of Asprosin.

Background: Currently, there is no effective vaccine against feline coronavirus infections. The coronavirus Spike (S) protein plays a critical role in viral binding to cell receptors and contains multiple neutralizing antibody epitopes that trigger the host's immune response to combat infection. Selecting an optimized adjuvant is essential to ensure robust vaccine-induced immunity against pathogenic infections.

Objective: To produce a recombinant S protein for the development of subunit vaccines and evaluate the immune responses elicited by different adjuvants.

Methods: In this study, we developed three subunit vaccines incorporating distinct adjuvants: Alh, ISA201, and CFA FCoV-SP. BALB/c mice were immunized three times via subcutaneous injections with each vaccine formulation. Serum samples were then analyzed to evaluate S protein-specific IgG levels and cytokine concentrations using enzyme-linked immunosorbent assay (ELISA), assessing the magnitude and nature of the vaccine-induced immune responses.

Results: The ISA201 FCoV-SP vaccine induced significantly higher total IgG levels than those in the Alh or CFA groups. All tested protein concentrations resulted in increased serum IgG antibody levels, with the optimal immune dose of recombinant S protein being 15 µg/dose. Additionally, the ISA201 FCoV-SP vaccine led to increased expression of interferon-γ, interleukin-8, and tumor necrosis factor-α.

Conclusion: Collectively, our findings suggest that ISA201 serves as the most effective adjuvant for a recombinant S protein subunit vaccine against FCoV. Additionally, the subunit vaccine developed in this study exhibited acceptable immune responses in mice.

Transforming growth factor-β (TGF-β) signaling plays a complex and dual role in regulating cellular senescence and tumor progression. In normal tissues, TGF-β acts as a tumor suppressor by regulating the cell cycle, inducing apoptosis, and maintaining the integrity of the extracellular matrix (ECM). These functions collectively restrict tumor initiation and support tissue homeostasis. However, in the tumor microenvironment, sustained TGF-β signaling frequently switches to a tumor-promoting role, driving tumor cell proliferation, metastatic dissemination, immune evasion, and therapy resistance. This review aims to clarify the dual role of TGF-β signaling in cellular senescence and tumor progression. It focuses on the molecular mechanisms that drive its transition between tumor suppression and tumor promotion in various biological contexts. We analyze the key determinants governing this functional switch, including tumor type, cellular environment, and signaling crosstalk. Furthermore, we critically evaluate the clinical challenges of therapeutic TGF-β targeting. We highlight emerging strategies to therapeutically modulate TGF-β signaling, focusing on precision medicine approaches that reconcile its tumor-suppressive and oncogenic functions. By providing a comprehensive understanding of TGF-β's dual role, this review offers new insights that may guide personalized cancer therapies and optimize treatment strategies for improved clinical outcomes.

Background: Stonefish (Synanceia spp.) are among the most venomous marine organisms. Their venom contains stonustoxin (SNTX), a heterodimeric toxin that induces severe hemolytic and myotoxic effects primarily mediated by its β-subunit.

Objective: To produce a recombinant SNTX β-subunit and develop neutralizing polyclonal antibodies against SNTX.

Methods: A DNA cassette encoding immunogenic regions of the β-sntx was designed using bioinformatics analysis, and codon-optimized for expression in E. coli. The construct was cloned into pET17b vector, and expressed in E. coli BL21 (DE3). The recombinant protein was purified via Ni-NTA affinity chromatography. For antibody production, rabbits were immunized subcutaneously with the recombinant protein emulsified in Freund's complete adjuvant, followed by booster doses at 2-week intervals. Antiserum was purified using protein G chromatography, and antibody titers were assessed by indirect Enzyme-Linked Immunosorbent Assay (ELISA).

Results: Epitope mapping revealed key immunogenic regions within residues 124-654 of the β-SNTX subunit. Following codon optimization, the codon adaptation index (CAI) increased to 0.93. Recombinant protein production was confirmed by SDS-PAGE and Western blot demonstrating successful purification of a 73 kDa recombinant protein (including TRX/His-tags), with a yield of 40 mg/L. Immunization of rabbits elicited a strong polyclonal IgG response, with antibody titers reaching 1:25,600 following the third booster. Purified IgG (1.8 mg/mL) exhibited high sensitivity in ELISA, detecting the recombinant β-SNTX at concentrations as low as 31.25 ng.

Conclusion: The recombinant β-SNTX subunit demonstrated strong immunogenicity, inducing high-affinity antibodies with specific binding activity against the native toxin. The resulting antiserum demonstrated significant neutralization potential, highlighting its promise for antivenom development.

Background: Psoriasis is a chronic inflammatory skin disorder characterized by elevated levels of proinflammatory cytokines. Mesenchymal stem cells (MSCs) have demonstrated therapeutic potential, yet the specific mechanisms involved are not fully understood.

Objective: To investigated the effectiveness of extracellular vesicles (EVs) derived from MSCs that were genetically modified to overexpress miR-146a, in a mouse model of psoriasis.

Methods: To enhance miR-146a expression, MSCs were transfected, and their EVs were subsequently purified. Thirty mice were randomly assigned to three groups and induced with imiquimod cream to develop psoriasis-like skin lesions. The treatment groups included: (1) a control group administered PBS, (2) a group treated with EVs containing a control miRNA (miR-control EVs), and (3) a group receiving EVs enriched with miR-146a (miR-146a-EVs). EVs were administered intravenously and lesions were evaluated. Following intravenous administration of EVs, the severity of skin lesions was assessed. Concentrations of key cytokines, including IFN-γ, IL-17, TNF-α, IL-23, IL-6, IL-1β, TGF-β, IL-10, and IL-4, were quantified in both spleen and skin tissue lysates using ELISA and qRT-PCR techniques.

Results: The experimental findings demonstrated that the administration of miR-146a-enriched EVs led to a significant improvement in clinical symptoms. There were substantial reductions observed in combined erythema, scaling, and skin thickness measurements compared to untreated controls. Additionally, levels of proinflammatory cytokines IFN-γ, IL-17, TNF-α, IL-23, IL-6, and IL-1β were significantly downregulated in the miR-146a-EV group, while anti-inflammatory TGF-β, IL-10 and IL-4 were upregulated. The same results were obtained in the spleens of mice.

Conclusion: EVs derived from miR-146a-modified MSCs effectively reduced psoriasis-like inflammation by modulating cytokine expression. This novel cell-free therapy holds promise for the treatment of psoriasis.

BLNK deficiency is a subtype of autosomal recessive immune disorders that involves a lack of B cells, agammaglobulinemia, and recurrent infections. We present the case of a 29-year-old Turkish female with BLNK deficiency caused by a novel homozygous CGA > TGA mutation at codon 123 (exon 6) in the BLNK gene. She developed severe liver failure and rickets at the age of 12. Although BLNK mutations are a rare cause of agammaglobulinemia, it is important to consider them in patients with B-cell deficiency and non-immune involvement.

Background: Severe neutropenia significantly increases the risk of bacterial infections. Recent studies have shown that the cytokine interleukin 14 (IL-14) plays an important role in immune cells, but its potential role in neutropenia induced by cytarabine (ara-c) or irradiation is unclear.

Objective: To investigate the role of IL-14 in ara-c or irradiation-induced neutropenia.

Methods: Two neutropenia models were induced by ara-c or irradiation. Neutrophil count was confirmed through flow cytometry and routine blood tests. IL-14 was used to assess the impact on neutropenia. IL-14 expression was analyzed using qPCR, Western blotting and ELISA. A IL-14 receptor (IL-14R) knockout mice model was utilized to confirm the role of IL-14R/STAT3 signaling in vivo.

Results: The results indicated that IL-14 treatment promoted proliferation and increased neutrophil counts in both bone marrow and peripheral blood, while IL-14R knockout suppressed this process. Furthermore, the downstream molecule of IL-14R, STAT3, showed enhanced phosphorylation levels in the presence of IL-14. Finally, we explored the source of IL-14 in the bone marrow, and found that lymphocytes secreted the highest levels of IL-14. Serum levels of IL-14 were significantly reduced in patients after chemotherapy.

Conclusions: These results indicate that IL-14 prevents ara-c or irradiation-induced neutropenia by regulating lymphocytes and activating the IL-14R/STAT3 pathway in neutrophils. This evidence suggests that IL-14 is a potent cytokine for treating ara-c or irradiation-induced neutropenia.

Background: DNA methylation plays a key role in systemic lupus erythematosus (SLE) by regulating gene expression and impacting immune system functions. In SLE, abnormal DNA methylation patterns can lead to the overexpression of pro-inflammatory genes and downregulation of the regulatory genes, contributing to autoimmunity. This dysregulation can increase susceptibility to SLE. Understanding these methylation changes could help discover new therapeutic strategies for managing SLE.

Objective: To evaluate methylation levels of OAS2 and OAS3 in peripheral blood mononuclear cells (PBMCs) in volunteers with SLE were evaluated.

Methods: In this case-control study, we collected 207 peripheral blood samples from 102 SLE patients and 105 healthy subjects. After isolating the PBMCs, methylation analysis was performed using the methylation-quantification of endonuclease-resistant DNA (MethyQESD) method.

Results: The control group had an average OAS2 methylation percentage of 40.02% ± 24.59%, whereas the SLE group had a significantly lower average of 19.46% ± 21.98%. This finding indicates a significant hypomethylation of OAS2 in the SLE cohort (P<0.001). Additionally, a significant difference was observed in the mean methylation levels of OAS3, with SLE patients exhibiting 14.11% ± 19.50% compared to healthy controls at 25.32% ± 20.82% (P<0.001). Patients with renal damage also showed significantly lower OAS2 methylation levels than SLE individuals without renal damage (P<0.001). Furthermore, a negative connection was found between the OAS2 methylation level and creatinine (r= -0.266, P= 0.007).

Conclusion: The pattern of methylation levels observed in OAS2 and OAS3 within PBMCs may provide valuable insights into the mechanisms underlying SLE development.